|
1
|
Xu C, Khin LW, Tam HZ, Goh LL, Koh ET, Dalan R, Leong KP. Haptoglobin 2-2 genotype is associated with increased risk of cardiovascular disease in patients with rheumatoid arthritis: a matched case-control study. Front Med (Lausanne) 2024; 11:1442858. [PMID: 39741512 PMCID: PMC11685008 DOI: 10.3389/fmed.2024.1442858] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2024] [Accepted: 11/18/2024] [Indexed: 01/03/2025] Open
Abstract
Introduction Traditional risk factors do not fully explain the increased risk of cardiovascular disease (CVD) in patients with rheumatoid arthritis (RA). The Haptoglobin (Hp) 2-2 genotype confers a lower anti-oxidant and higher inflammatory effect on the vasculature compared to the non-Hp 2-2 genotype. This study investigates the association of the Hp genotype with CVD in patients with RA. Methods Data from 69 RA patients with CVD and 207 sex- and ethnicity-matched RA patients without CVD, collected from 1 January 2000 to 31 December 2020, were retrieved from the Tan Tock Seng Hospital RA Registry. CVD was examined against demographics, clinical and laboratory variables in univariate models. Associations between the Hp genotypes and CVD were analyzed using conditional logistic regression. Results We studied 276 patients (65.2% female, 82.6% Chinese, median age 60.9 years). Most participants were in low disease activity or remission (79.3%). The Hp 2-2 genotype was present in 49.6% (137/276). In the group with CVD, the prevalence of the Hp 2-2 genotype was 50.9% (29/57) in the Chinese, 100% (5/5) in the Indians, and 28.6% (2/7) in the Malays. In the non-CVD group, the respective prevalence was 46.8% (80/171), 66.7% (10/15), and 52.4% (11/21). In univariate analysis, the matched odds ratio (OR) of the Hp 2-2 genotype for CVD in RA was 1.34 [95% confidence interval (CI): 1.22-1.47; p < 0.001]. The Hp 2-2 genotype was significantly associated with CVD (adjusted matched OR: 1.13; 95% CI: 1.01-1.27; p = 0.033) in the multivariate logistic regression model after adjusting the confounding factors, including age, smoking, diabetes, hypertension, hyperlipidemia, anti-CCP autoantibodies, and disease activity. Conclusion The Hp 2-2 genotype is associated with an increased risk of CVD in patients with RA in this multi-ethnic cohort.
Collapse
Affiliation(s)
- Chuanhui Xu
- Department of Rheumatology, Allergy and Immunology, Tan Tock Seng Hospital, Singapore, Singapore
- Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore
| | - Lay Wai Khin
- Clinical Research & Innovation Office, Tan Tock Seng Hospital, Singapore, Singapore
| | - Hui Zhen Tam
- Clinical Research & Innovation Office, Tan Tock Seng Hospital, Singapore, Singapore
| | - Liuh Ling Goh
- Molecular Diagnostic Laboratory, Tan Tock Seng Hospital, Singapore, Singapore
| | - Ee Tzun Koh
- Department of Rheumatology, Allergy and Immunology, Tan Tock Seng Hospital, Singapore, Singapore
| | - Rinkoo Dalan
- Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore
- Department of Endocrinology, Tan Tock Seng Hospital, Singapore, Singapore
| | - Khai Pang Leong
- Department of Rheumatology, Allergy and Immunology, Tan Tock Seng Hospital, Singapore, Singapore
| |
Collapse
|
|
2
|
Allegra A, Cicero N, Mirabile G, Giorgianni CM, Gangemi S. Novel Biomarkers for Diagnosis and Monitoring of Immune Thrombocytopenia. Int J Mol Sci 2023; 24:ijms24054438. [PMID: 36901864 PMCID: PMC10003036 DOI: 10.3390/ijms24054438] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2023] [Revised: 02/12/2023] [Accepted: 02/21/2023] [Indexed: 03/12/2023] Open
Abstract
Lower-than-normal platelet counts are a hallmark of the acquired autoimmune illness known as immune thrombocytopenia, which can affect both adults and children. Immune thrombocytopenia patients' care has evolved significantly in recent years, but the disease's diagnosis has not, and it is still only clinically achievable with the elimination of other causes of thrombocytopenia. The lack of a valid biomarker or gold-standard diagnostic test, despite ongoing efforts to find one, adds to the high rate of disease misdiagnosis. However, in recent years, several studies have helped to elucidate a number of features of the disease's etiology, highlighting how the platelet loss is not only caused by an increase in peripheral platelet destruction but also involves a number of humoral and cellular immune system effectors. This made it possible to identify the role of immune-activating substances such cytokines and chemokines, complement, non-coding genetic material, the microbiome, and gene mutations. Furthermore, platelet and megakaryocyte immaturity indices have been emphasized as new disease markers, and prognostic signs and responses to particular types of therapy have been suggested. Our review's goal was to compile information from the literature on novel immune thrombocytopenia biomarkers, markers that will help us improve the management of these patients.
Collapse
Affiliation(s)
- Alessandro Allegra
- Division of Hematology, Department of Human Pathology in Adulthood and Childhood “Gaetano Barresi”, University of Messina, 98100 Messina, Italy
- Correspondence:
| | - Nicola Cicero
- Department of Biomedical, Dental, Morphological and Functional Imaging Sciences (BIOMORF), University of Messina, 98100 Messina, Italy
| | - Giuseppe Mirabile
- Division of Hematology, Department of Human Pathology in Adulthood and Childhood “Gaetano Barresi”, University of Messina, 98100 Messina, Italy
| | - Concetto Mario Giorgianni
- Department of Biomedical, Dental, Morphological and Functional Imaging Sciences (BIOMORF), University of Messina, 98100 Messina, Italy
| | - Sebastiano Gangemi
- Allergy and Clinical Immunology Unit, Department of Clinical and Experimental Medicine, University of Messina, 98100 Messina, Italy
| |
Collapse
|
|
3
|
Serek P, Oleksy-Wawrzyniak M. The Effect of Bacterial Infections, Probiotics and Zonulin on Intestinal Barrier Integrity. Int J Mol Sci 2021; 22:11359. [PMID: 34768787 PMCID: PMC8583036 DOI: 10.3390/ijms222111359] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2021] [Revised: 10/15/2021] [Accepted: 10/19/2021] [Indexed: 12/12/2022] Open
Abstract
The intestinal barrier plays an extremely important role in maintaining the immune homeostasis of the gut and the entire body. It is made up of an intricate system of cells, mucus and intestinal microbiota. A complex system of proteins allows the selective permeability of elements that are safe and necessary for the proper nutrition of the body. Disturbances in the tightness of this barrier result in the penetration of toxins and other harmful antigens into the system. Such events lead to various digestive tract dysfunctions, systemic infections, food intolerances and autoimmune diseases. Pathogenic and probiotic bacteria, and the compounds they secrete, undoubtedly affect the properties of the intestinal barrier. The discovery of zonulin, a protein with tight junction regulatory activity in the epithelia, sheds new light on the understanding of the role of the gut barrier in promoting health, as well as the formation of diseases. Coincidentally, there is an increasing number of reports on treatment methods that target gut microbiota, which suggests that the prevention of gut-barrier defects may be a viable approach for improving the condition of COVID-19 patients. Various bacteria-intestinal barrier interactions are the subject of this review, aiming to show the current state of knowledge on this topic and its potential therapeutic applications.
Collapse
Affiliation(s)
- Paweł Serek
- Department of Biochemistry and Immunochemistry, Wroclaw Medical University, 50-368 Wroclaw, Poland
| | - Monika Oleksy-Wawrzyniak
- Department of Pharmaceutical Microbiology and Parasitology, Wroclaw Medical University, 50-556 Wroclaw, Poland;
| |
Collapse
|
|
4
|
Assessment of haptoglobin alleles in autism spectrum disorders. Sci Rep 2020; 10:7758. [PMID: 32385356 PMCID: PMC7210291 DOI: 10.1038/s41598-020-64679-w] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/11/2019] [Accepted: 04/01/2020] [Indexed: 02/07/2023] Open
Abstract
Gene-environment interactions, by means of abnormal macromolecular intestinal adsorption, is one of the possible causes of autism spectrum disorders (ASD) predominantly in patients with gastrointestinal disorders. Pre-haptoglobin-2 (zonulin), encoded by the Haptoglobin (HP) allele-2 gene, enhances the intestinal permeability by modulation of intercellular tight junctions. The two alleles of HP, HP1 and HP2, differ for 2 extra exons in HP2 that result in exon duplication undetectable by classic genome-wide association studies. To evaluate the role of HP2 in ASD pathogenesis and to set up a method to discriminate HP alleles, Italian subjects with ASD (n = 398) and healthy controls (n = 379) were genotyped by PCR analysis; subsequently, the PCR results were integrated with microarray genotypes (Illumina Human Omni 1S-8), obtained using a subset from the same subjects, and then we developed a computational method to predict HP alleles. On the contrary to our expectations, there was no association between HP2 and ASD (P > 0.05), and there was no significant allele association in subjects with ASD with or without gastrointestinal disorders (P > 0.05). With the aid of bioinformatics analysis, from a window frame of ~2 Mb containing 314 SNPs, we obtain imputation accuracy (r2) between 0.4 and 0.9 (median 0.7) and correct predictions were between 70% and 100% (median 90%). The conclusions endorse that enhanced intestinal permeability in subjects with ASD should not be imputed to HP2 but to other members of the zonulin family and/or to environmental factors.
Collapse
|
|
5
|
Leonard MM, Camhi S, Kenyon V, Betensky RA, Sturgeon C, Yan S, Fasano A. Targeted genotyping for the prediction of celiac disease autoimmunity development in patients with type 1 diabetes and their family members. World J Diabetes 2019; 10:189-199. [PMID: 30891154 PMCID: PMC6422857 DOI: 10.4239/wjd.v10.i3.189] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/04/2019] [Revised: 03/04/2019] [Accepted: 03/09/2019] [Indexed: 02/05/2023] Open
Abstract
BACKGROUND Patients with type 1 diabetes (T1D) and their first-degree relatives (FDRs) have an increased risk of developing celiac disease (CD) compared to the general population. This is largely explained by the shared association with major histocompatibility class II human leukocyte antigen (HLA) DQ2 and/or DQ8 between the two disease states.
AIM To describe the frequency of CD autoimmunity (CDA) and the distribution of HLA and haptoglobin genotypes in patients with T1D and their FDRs. Additionally, we aimed at identifying predictors associated with an increased risk of developing CDA in patients with T1D and their family members.
METHODS We obtained clinical information and blood samples from 1027 participants (302 with T1D and 725 FDRs) over a five-year period. Samples were tested for autoantibodies associated with CD, HLA-DQ alleles, and haptoglobin genotype. We fit univariate and multiple logistic regression models for CDA separately for subjects with T1D and for FDRs of subjects with T1D.
RESULTS Implementation of a screening program increased the frequency of CDA by 2-fold in participants with T1D and 2.8-fold in their FDRs. Multivariate analysis found that, in participants with T1D, having both DR7-DQ2 and DR4-DQ8 was associated with an increased frequency of CDA. In FDRs of T1D patients, reported CD in the family was associated with an increased frequency of CDA during screening. Haptoglobin 2 genotype was not associated with developing CDA in the multivariate analysis.
CONCLUSION Patients with T1D and their FDRs have a high frequency of CDA. Carrying both DR7-DQ2 and DR4-DQ8 was associated with development of CDA in patients with T1D.
Collapse
Affiliation(s)
- Maureen M Leonard
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| | - Stephanie Camhi
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| | - Victoria Kenyon
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| | - Rebecca A Betensky
- Department of Biostatistics, Harvard T.H. Chan School of Public Health, Boston, MA 02115, United States
| | - Craig Sturgeon
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| | - Shu Yan
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| | - Alessio Fasano
- Mucosal Immunology and Biology Research Center, Mass General Hospital for Children, Boston, MA 02115, United States
- Center for Celiac Research and Treatment, Mass General Hospital for Children, Boston, MA 02115, United States
- Department of Pediatric Gastroenterology and Nutrition, Mass General Hospital for Children, Boston, MA 02114, United States
| |
Collapse
|
|
6
|
Assadsangabi A, Evans CA, Corfe BM, Lobo A. Application of Proteomics to Inflammatory Bowel Disease Research: Current Status and Future Perspectives. Gastroenterol Res Pract 2019; 2019:1426954. [PMID: 30774653 PMCID: PMC6350533 DOI: 10.1155/2019/1426954] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/17/2018] [Accepted: 10/08/2018] [Indexed: 12/11/2022] Open
Abstract
Inflammatory bowel disease (IBD) is a chronic relapsing/remitting inflammatory illness of the gastrointestinal tract of unknown aetiology. Despite recent advances in decoding the pathophysiology of IBD, many questions regarding disease pathogenesis remain. Genome-wide association studies (GWAS) and knockout mouse models have significantly advanced our understanding of genetic susceptibility loci and inflammatory pathways involved in IBD pathogenesis. Despite their important contribution to a better delineation of the disease process in IBD, these genetic findings have had little clinical impact to date. This is because the presence of a given gene mutation does not automatically correspond to changes in its expression or final metabolic or structural effect(s). Furthermore, the existence of these gene susceptibility loci in the normal population suggests other driving prerequisites for the disease manifestation. Proteins can be considered the main functional units as almost all intracellular physiological functions as well as intercellular interactions are dependent on them. Proteomics provides methods for the large-scale study of the proteins encoded by the genome of an organism or a cell, to directly investigate the proteins and pathways involved. Understanding the proteome composition and alterations yields insights into IBD pathogenesis as well as identifying potential biomarkers of disease activity, mucosal healing, and cancer progression. This review describes the state of the art in the field with respect to the study of IBD and the potential for translation from biomarker discovery to clinical application.
Collapse
Affiliation(s)
- Arash Assadsangabi
- Gastroenterology Unit, Salford Royal Hospital, Salford, UK
- Molecular Gastroenterology Research Group, Academic Unit of Surgical Oncology, Department of Oncology and Insigneo Institute, University of Sheffield, Sheffield, UK
| | - Caroline A. Evans
- Department of Chemical and Biological Engineering, University of Sheffield, Sheffield, UK
| | - Bernard M. Corfe
- Molecular Gastroenterology Research Group, Academic Unit of Surgical Oncology, Department of Oncology and Insigneo Institute, University of Sheffield, Sheffield, UK
| | - Alan Lobo
- Gastroenterology Unit, Salford Royal Hospital, Salford, UK
| |
Collapse
|
|
7
|
Pedroza-Díaz J, Chavarria TPL, Vahos CHM, Hernández Ramírez DF, Olivares-Martínez E, Vásquez G, Llorente L, Fragoso-Loyo H, Röthlisberger S, Ortiz Reyes BL. Proteomic Analysis of Cerebrospinal Fluid: A Search for Biomarkers of Neuropsychiatric Systemic Lupus Erythematosus. CURR PROTEOMICS 2019. [DOI: 10.2174/1570164615666180911125252] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Abstract
Background:
Neuropsychiatric systemic lupus erythematosus or NPSLE, as its name suggests, refers to the neurological and psychiatric manifestations of Systemic Lupus Erythematosus (SLE). In clinical practice, it is often difficult to reach an accurate diagnosis, as this disease presents differently in different patients, and the available diagnostic tests are often not specific enough.
Objectives:
The aim of this study was to search for proteomic biomarkers in cerebrospinal fluid that could be proposed as diagnostic aids for this disease.
Methods:
The proteomic profile of cerebrospinal fluid samples of 19 patients with NPSLE, 12 patients with SLE and no neuropsychiatric manifestation (SLEnoNP), 6 patients with neuropsychiatric symptoms but no SLE (NPnoSLE), 5 with Other Autoimmune Disorders without neuropsychiatric manifestations (OADs), and 4 Healthy Controls (HC), were obtained by two-dimensional gel electrophoresis and compared using ImageMaster Platinum 7.0 software.
Results:
The comparative analysis of the different study groups revealed three proteins of interest that were consistently over-expressed in NPSLE patients. These were identified by mass spectrometry as albumin (spot 16), haptoglobin (spot 160), and beta-2 microglobulin (spot 161).
Conclusion:
This work is one of the few proteomic studies of NPSLE that uses cerebrospinal fluid as the biological sample. Albumin has previously been proposed as a potential biomarker of rheumatoid arthritis and SLE, which is coherent with these results; but this is the first report of haptoglobin and beta-2 microglobulin in NPSLE, although haptoglobin has been associated with increased antibody production and beta-2 microglobulin with lupus nephritis.
Collapse
Affiliation(s)
- Johanna Pedroza-Díaz
- Instituto Tecnologico Metropolitano-ITM-, Facultad de Ciencias Exactas y Aplicadas, Grupo de Investigacion e Innovacion Biomedica GI2B, Medellín, Colombia
| | - Tania Paola Luján Chavarria
- Universidad de Antioquia, Facultad de Medicina, Grupo de Inmunologia Celular e Inmunogenetica-GICIG-, Medellin, Colombia
| | - Carlos Horacio Muñoz Vahos
- Universidad de Antioquia, Facultad de Medicina, Grupo de Reumatologia Universidad de Antioquia - GRUA-, Medellín, Colombia
| | | | - Elizabeth Olivares-Martínez
- Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Department of Immunology & Rheumatology, Mexico DF, Mexico
| | - Gloria Vásquez
- Universidad de Antioquia, Facultad de Medicina, Grupo de Inmunologia Celular e Inmunogenetica-GICIG-, Medellin, Colombia
| | - Luis Llorente
- Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Department of Immunology & Rheumatology, Colombia
| | - Hilda Fragoso-Loyo
- Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Department of Immunology & Rheumatology, Mexico DF, Mexico
| | - Sarah Röthlisberger
- Instituto Tecnologico Metropolitano-ITM-, Facultad de Ciencias Exactas y Aplicadas, Grupo de Investigacion e Innovacion Biomedica GI2B, Medellín, Colombia
| | - Blanca Lucía Ortiz Reyes
- Universidad de Antioquia, Facultad de Medicina, Grupo de Inmunologia Celular e Inmunogenetica-GICIG-, Medellin, Colombia
| |
Collapse
|
|
8
|
Madda R, Lin SC, Sun WH, Huang SL. Differential expressions of plasma proteins in systemic lupus erythematosus patients identified by proteomic analysis. JOURNAL OF MICROBIOLOGY, IMMUNOLOGY, AND INFECTION = WEI MIAN YU GAN RAN ZA ZHI 2018; 52:816-826. [PMID: 30170966 DOI: 10.1016/j.jmii.2018.02.004] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/27/2017] [Revised: 01/18/2018] [Accepted: 02/21/2018] [Indexed: 10/17/2022]
Abstract
INTRODUCTION Systemic lupus erythematosus (SLE) is a chronic and complex autoimmune disease with a wide range of clinical manifestations that affects multiple organs and tissues. Therefore the differential expression of proteins in the serum/plasma have potential clinical applications when treating SLE. METHODS We have compared the plasma/serum protein expression patterns of nineteen active SLE patients with those of twelve age-matched and gender-matched healthy controls by proteomic analysis. To investigate the differentially expressed proteins among SLE and controls, a 2-dimensional gel electrophoresis coupled with high-resolution liquid chromatography tandem mass spectrometry was performed. To further understand the molecular and biological functions of the identified proteins, PANTHER and Gene Ontology (GO) analyses were employed. RESULTS A total of 14 significantly expressed (p < 0.05, p < 0.01) proteins were identified, and of these nine were up-regulated and five down-regulated in the SLE patients. The functional enrichment analysis assigned the majority of the identified proteins including alpha 2 macroglobulin, complement C4, complement factor H, fibrinogen beta chain, and alpha-1-antitrypsin were part of the complement/coagulation cascade, which is an important pathway that plays a crucial role in SLE pathogenesis. In addition to these proteins the differential expressions of ceruloplasmin, transthyretin, and haptoglobin play a potential role in the renal system abnormalities of SLE. CONCLUSION Therefore, the identified differentially expressed proteins are relevant to SLE patient's cohort. Most importantly the up-regulated proteins might be the potential candidates for renal system involvement in SLE disease pathogenesis. In order to confirm the diagnostic/therapeutic potential of the identified proteins, future validation studies are required.
Collapse
Affiliation(s)
- Rashmi Madda
- Department of Life Sciences, National Central University, Taiwan
| | - Shih-Chang Lin
- Department of Life Sciences, National Central University, Taiwan; Department of Medicine, College of Medicine, Fu-Jen Catholic University, Taiwan; Division of Rheumatology and Immunology, Cathay General Hospital, Taiwan
| | - Wei-Hsin Sun
- Department of Life Sciences, National Central University, Taiwan.
| | - Shir-Ly Huang
- Institute of Microbiology and Immunology, National Yang-Ming University, Taiwan.
| |
Collapse
|
|
9
|
Madda R, Lin SC, Sun WH, Huang SL. Plasma proteomic analysis of systemic lupus erythematosus patients using liquid chromatography/tandem mass spectrometry with label-free quantification. PeerJ 2018; 6:e4730. [PMID: 29761050 PMCID: PMC5947061 DOI: 10.7717/peerj.4730] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2018] [Accepted: 04/18/2018] [Indexed: 12/13/2022] Open
Abstract
CONTEXT Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease with unknown etiology. OBJECTIVE Human plasma is comprised of over 10 orders of magnitude concentration of proteins and tissue leakages. The changes in the abundance of these proteins have played an important role in various human diseases. Therefore, the research objective of this study is to identify the significantly altered expression levels of plasma proteins from SLE patients compared with healthy controls using proteomic analysis. The plasma proteome profiles of both SLE patients and controls were compared. METHODS A total of 19 active SLE patients and 12 healthy controls plasma samples were analyzed using high-resolution electrospray ionization liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) followed by label-free quantification. RESULTS A total of 19 proteins showed a significant level of expression in the comparative LC-ESI-MS/MS triplicate analysis; among these, 14 proteins had >1.5- to three-fold up-regulation and five had <0.2- to 0.6-fold down-regulation. Gene ontology and DAVID (Database Annotation Visualization, and Integrated Discovery) functional enrichment analysis revealed that these proteins are involved in several important biological processes including acute phase inflammatory responses, complement activation, hemostasis, and immune system regulation. CONCLUSION Our study identified a group of differentially expressed proteins in the plasma of SLE patients that are involved in the imbalance of the immune system and inflammatory responses. Therefore, these findings may have the potential to be used as prognostic/diagnostic markers for SLE disease assessment or disease monitoring.
Collapse
Affiliation(s)
- Rashmi Madda
- Department of Life Sciences, National Central University, Zhongli, Taiwan
| | - Shih-Chang Lin
- Division of Medicine, College of Medicine, Fu Jen Catholic University, Taipei, Taiwan
- Department of Rheumatology and Immunology, Cathay General Hospital, Taipei, Taiwan
| | - Wei-Hsin Sun
- Department of Life Sciences, National Central University, Zhongli, Taiwan
| | - Shir-Ly Huang
- Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan
| |
Collapse
|
|
10
|
Pinna S, Pasella S, Deiana M, Baralla A, Mannu A, Masala AGE, Pileri PV, Deiana N, Scognamillo F, Pala C, Zinellu A, Carru C, Deiana L. Proteomic analysis of human plasma and peripheral blood mononuclear cells in Systemic Lupus Erythematosus patients. J Immunol Methods 2017; 446:37-46. [DOI: 10.1016/j.jim.2017.03.019] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2016] [Revised: 03/09/2017] [Accepted: 03/28/2017] [Indexed: 01/22/2023]
|
|
11
|
Wongtrakul J, Thongtan T, Roytrakul S, Praparattanapan J, Wipasa J, Kumrapich B, Supparatpinyo K. Identification of novel biomarkers for adult-onset-immunodeficiency (AOID) syndrome using serum proteomics. ASIAN PAC J TROP MED 2017. [PMID: 28647184 DOI: 10.1016/j.apjtm.2017.05.014] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/19/2022] Open
Abstract
OBJECTIVE To identify the candidate protein biomarkers of adult-onset-immunodeficiency (AOID) syndrome using serum proteomics. METHODS Screening and verification phases were performed in the study. A total of 97 serum samples were classified into three groups: AOID patients with opportunistic infections (active AOID), AOID patients without opportunistic infections (inactive AOID), and healthy control. In the screening phase, pooled sera collected from patients and healthy control in each group were separated by 2D-gel electrophoresis, analyzed for differentially expressed proteins and identified for biomarkers using LC/MS. In the verification phase, the protein candidates were selected for confirmation by western blotting. RESULTS The analysis revealed 35 differentially expressed proteins. Three proteins including haptoglobin, gelsolin, and transthyretin, were selected for verification. The results showed that the levels of haptoglobin in both active and inactive AOID groups were significantly higher than that in the control group, while the levels of gelsolin in the active AOID group were significantly lower than that in the inactive AOID group. The level of transthyretin in the active AOID group was also significantly lower than that in the control group. CONCLUSIONS The comparison of serum proteins between the three groups revealed three candidates which are related to chronic inflammatory diseases. Haptoglobin and transthyretin biomarkers could be applied in clinical assessment for monitor of disease outcome, including for the study of AOID pathogenesis.
Collapse
Affiliation(s)
- Jeerang Wongtrakul
- Research Institute for Health Sciences, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand.
| | - Thananya Thongtan
- Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, 1873 Rama 4 Road, Pathumwan, Bangkok 10330, Thailand
| | - Sittiruk Roytrakul
- Proteomics Research Laboratory, Genome Institute, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathumthani 12120, Thailand
| | - Jutarat Praparattanapan
- Department of Medicine, Faculty of Medicine, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand
| | - Jiraprapa Wipasa
- Research Institute for Health Sciences, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand
| | - Benjawan Kumrapich
- Research Institute for Health Sciences, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand
| | - Khuanchai Supparatpinyo
- Department of Medicine, Faculty of Medicine, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand
| |
Collapse
|
|
12
|
Nicolaou O, Kousios A, Hadjisavvas A, Lauwerys B, Sokratous K, Kyriacou K. Biomarkers of systemic lupus erythematosus identified using mass spectrometry-based proteomics: a systematic review. J Cell Mol Med 2017; 21:993-1012. [PMID: 27878954 PMCID: PMC5387176 DOI: 10.1111/jcmm.13031] [Citation(s) in RCA: 30] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/24/2016] [Accepted: 09/29/2016] [Indexed: 12/21/2022] Open
Abstract
Advances in mass spectrometry technologies have created new opportunities for discovering novel protein biomarkers in systemic lupus erythematosus (SLE). We performed a systematic review of published reports on proteomic biomarkers identified in SLE patients using mass spectrometry-based proteomics and highlight their potential disease association and clinical utility. Two electronic databases, MEDLINE and EMBASE, were systematically searched up to July 2015. The methodological quality of studies included in the review was performed according to Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines. Twenty-five studies were included in the review, identifying 241 SLE candidate proteomic biomarkers related to various aspects of the disease including disease diagnosis and activity or pinpointing specific organ involvement. Furthermore, 13 of the 25 studies validated their results for a selected number of biomarkers in an independent cohort, resulting in the validation of 28 candidate biomarkers. It is noteworthy that 11 candidate biomarkers were identified in more than one study. A significant number of potential proteomic biomarkers that are related to a number of aspects of SLE have been identified using mass spectrometry proteomic approaches. However, further studies are required to assess the utility of these biomarkers in routine clinical practice.
Collapse
Affiliation(s)
- Orthodoxia Nicolaou
- Department of Electron Microscopy/Molecular PathologyThe Cyprus Institute of Neurology and GeneticsNicosiaCyprus
- Department of Electron Microscopy/Molecular PathologyCyprus School of Molecular MedicineNicosiaCyprus
| | - Andreas Kousios
- Department of Electron Microscopy/Molecular PathologyCyprus School of Molecular MedicineNicosiaCyprus
| | - Andreas Hadjisavvas
- Department of Electron Microscopy/Molecular PathologyThe Cyprus Institute of Neurology and GeneticsNicosiaCyprus
- Department of Electron Microscopy/Molecular PathologyCyprus School of Molecular MedicineNicosiaCyprus
| | - Bernard Lauwerys
- Department of RheumatologyUniversité catholique de LouvainBruxellesBelgium
| | - Kleitos Sokratous
- Department of Electron Microscopy/Molecular PathologyThe Cyprus Institute of Neurology and GeneticsNicosiaCyprus
| | - Kyriacos Kyriacou
- Department of Electron Microscopy/Molecular PathologyThe Cyprus Institute of Neurology and GeneticsNicosiaCyprus
- Department of Electron Microscopy/Molecular PathologyCyprus School of Molecular MedicineNicosiaCyprus
| |
Collapse
|
|
13
|
Aggarwal A, Gupta R, Negi VS, Rajasekhar L, Misra R, Singh P, Chaturvedi V, Sinha S. Urinary haptoglobin, alpha-1 anti-chymotrypsin and retinol binding protein identified by proteomics as potential biomarkers for lupus nephritis. Clin Exp Immunol 2017; 188:254-262. [PMID: 28120479 DOI: 10.1111/cei.12930] [Citation(s) in RCA: 30] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/12/2017] [Indexed: 12/15/2022] Open
Abstract
The study was aimed at identification by proteomics and validation by enzyme-linked immunosorbent assay (ELISA) of potential urinary biomarkers for lupus nephritis. Study subjects comprised 88 systemic lupus erythematosus (SLE) patients and 60 controls (rheumatoid arthritis, diabetes mellitus and healthy individuals). Based on the SLE disease activity index (SLEDAI), patients were classified as active renal (AR), active non-renal (ANR) or inactive disease (ID). Urinary proteins from a group of patients with AR or ID were resolved by two-dimensional gel electrophoresis and identified by matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS/MS). The selected biomarkers were validated by ELISA using samples from all patients and controls. AR patients were followed-up for 12 months after start of therapy. Three urinary proteins, alpha-1 anti-chymotrypsin (ACT), haptoglobin (HAP) and retinol binding protein (RBP), were detected in patients with AR and not ID. Upon validation, ACT levels were higher in AR patients than the other groups (P < 0·001) and showed good correlation with renal SLEDAI (r = 0·577, P < 0·001) as well as SLEDAI (r = 0·461, P < 0·001). Similarly, HAP levels were > 10-fold higher in AR than other groups (P < 0·001) and correlated well with renal SLEDAI (r = 0·594, P < 0·001) and SLEDAI (r = 0·371, P < 0·01). RBP levels were also higher in AR patients than in other groups (P < 0·05), except diabetes, and showed moderate correlation with renal SLEDAI (r = 0·284, P < 0·008) and SLEDAI (r = 0·316, P < 0·003). Upon follow-up with treatment, levels of all three proteins declined at 6 and 12 months (P < 0·01). Multiple logistic regression identified ACT as the best marker to differentiate AR from ANR. Urinary HAP, ACT and RBP are potential biomarkers for lupus nephritis activity.
Collapse
Affiliation(s)
- A Aggarwal
- Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - R Gupta
- Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - V S Negi
- Department of Clinical Immunology, JIPMER, Puducherry, India
| | - L Rajasekhar
- Department of Rheumatology, Nizam Institute of Medical Sciences, Hyderabad, India
| | - R Misra
- Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - P Singh
- Biochemistry Division, Central Drug Research Institute, Lucknow, India
| | - V Chaturvedi
- Biochemistry Division, Central Drug Research Institute, Lucknow, India
| | - S Sinha
- Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India.,Biochemistry Division, Central Drug Research Institute, Lucknow, India
| |
Collapse
|
|
14
|
Timlin H, Machireddy K, Petri M. Low Level of Haptoglobin in Lupus. J Investig Med High Impact Case Rep 2017; 5:2324709616689106. [PMID: 28203576 PMCID: PMC5298417 DOI: 10.1177/2324709616689106] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/03/2016] [Revised: 11/23/2016] [Accepted: 12/11/2016] [Indexed: 11/15/2022] Open
Abstract
Haptoglobin levels are measured in systematic lupus erythematosus patients as part of the workup for anemia, with low levels indicating hemolysis. Haptoglobin is an acute phase protein. We present 2 lupus patients who were found to have low haptoglobin levels in the absence of other evidence of hemolysis.
Collapse
Affiliation(s)
- Homa Timlin
- Johns Hopkins University, Baltimore, MD, USA
| | | | | |
Collapse
|
|
15
|
Proteomic analysis reveals aberrant expression of CALR and HSPA5 in thyroid tissues of Graves' disease. Clin Biochem 2017; 50:40-45. [DOI: 10.1016/j.clinbiochem.2016.08.014] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/01/2016] [Revised: 08/18/2016] [Accepted: 08/19/2016] [Indexed: 02/06/2023]
|
|
16
|
Identification of multiple transferrin species in the spleen and serum from mice with collagen-induced arthritis which may reflect changes in transferrin glycosylation associated with disease activity: The role of CD38. J Proteomics 2016; 134:127-137. [DOI: 10.1016/j.jprot.2015.11.023] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2015] [Revised: 11/11/2015] [Accepted: 11/26/2015] [Indexed: 12/12/2022]
|
|
17
|
Supporting data for the MS identification of distinct transferrin glycopeptide glycoforms and citrullinated peptides associated with inflammation or autoimmunity. Data Brief 2016; 6:587-602. [PMID: 26909372 PMCID: PMC4731419 DOI: 10.1016/j.dib.2015.12.045] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2015] [Revised: 12/17/2015] [Accepted: 12/23/2015] [Indexed: 01/27/2023] Open
Abstract
This data article presents the results of all the statistical analyses applied to the relative intensities of the detected 2D-DiGE protein spots for each of the 3 performed DiGE experiments. The data reveals specific subsets of protein spots with significant differences between WT and CD38-deficient mice with either Collagen-induced arthritis (CIA), or with chronic inflammation induced by CFA, or under steady-state conditions. This article also shows the MS data analyses that allowed the identification of the protein species which serve to discriminate the different experimental groups used in this study. Moreover, the article presents MS data on the citrullinated peptides linked to specific protein species that were generated in CIA(+) or CFA-treated mice. Lastly, this data article provides MS data on the efficiency of the analyses of the transferrin (Tf) glycopeptide glycosylation pattern in spleen and serum from CIA(+) mice and normal controls. The data supplied in this work is related to the research article entitled "identification of multiple transferrin species in spleen and serum from mice with collagen-induced arthritis which may reflect changes in transferrin glycosylation associated with disease activity: the role of CD38" [1]. All mass spectrometry data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with identifiers PRIDE: PXD002644, PRIDE: PXD002643, PRIDE: PXD003183 and PRIDE: PXD003163.
Collapse
|
|
18
|
Rosal-Vela A, García-Rodríguez S, Postigo J, Iglesias M, Longobardo V, Lario A, Merino J, Merino R, Zubiaur M, Sancho J. Distinct serum proteome profiles associated with collagen-induced arthritis and complete Freund's adjuvant-induced inflammation in CD38−/−
mice: The discriminative power of protein species or proteoforms. Proteomics 2015; 15:3382-93. [DOI: 10.1002/pmic.201400536] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/14/2015] [Revised: 04/25/2015] [Accepted: 07/10/2015] [Indexed: 11/06/2022]
Affiliation(s)
- Antonio Rosal-Vela
- Departamento de Biología Celular e Inmunología; Instituto de Parasitología y Biomedicina López Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| | - Sonia García-Rodríguez
- Departamento de Biología Celular e Inmunología; Instituto de Parasitología y Biomedicina López Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| | - Jorge Postigo
- Departamento de Biología Molecular; Instituto de Formación e Investigación Marqués de Valdecilla, Universidad de Cantabria; Cantabria Spain
| | - Marcos Iglesias
- Departamento de Biología Molecular; Instituto de Formación e Investigación Marqués de Valdecilla, Universidad de Cantabria; Cantabria Spain
| | - Victoria Longobardo
- Unidad de Proteómica; Instituto de Parasitología y Biomedicina López-Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| | - Antonio Lario
- Unidad de Proteómica; Instituto de Parasitología y Biomedicina López-Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| | - Jesús Merino
- Departamento de Biología Molecular; Instituto de Formación e Investigación Marqués de Valdecilla, Universidad de Cantabria; Cantabria Spain
| | - Ramón Merino
- Instituto de Biomedicina y Biotecnología de Cantabria/CSIC-Universidad de Cantabria-SODERCAN; Cantabria Spain
| | - Mercedes Zubiaur
- Departamento de Biología Celular e Inmunología; Instituto de Parasitología y Biomedicina López Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| | - Jaime Sancho
- Departamento de Biología Celular e Inmunología; Instituto de Parasitología y Biomedicina López Neyra (IPBLN), Consejo Superior de Investigaciones Científicas (CSIC), PTS Granada; Granada Spain
| |
Collapse
|
|
19
|
Behavioral Deficits Are Accompanied by Immunological and Neurochemical Changes in a Mouse Model for Neuropsychiatric Lupus (NP-SLE). Int J Mol Sci 2015; 16:15150-71. [PMID: 26151848 PMCID: PMC4519892 DOI: 10.3390/ijms160715150] [Citation(s) in RCA: 40] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/28/2015] [Revised: 06/11/2015] [Accepted: 06/24/2015] [Indexed: 12/27/2022] Open
Abstract
Neuropsychiatric symptoms of systemic lupus erythematosus (NP-SLE) have been understudied compared to end-organ failure and peripheral pathology. Neuropsychiatric symptoms, particularly affective and cognitive indications, may be among the earliest manifestations of SLE. Among the potential pathophysiological mechanisms responsible for NP-SLE are increased peripheral pro-inflammatory cytokines, subsequent induction of indoleamine-2,3-dioxygenase (IDO) and activation of the kynurenine pathway. In the MRL/MpJ-Faslpr (MRL/lpr) murine model of lupus, depression-like behavior and cognitive dysfunction is evident before significant levels of autoantibody titers and nephritis are present. We examined the behavioral profile of MRL/lpr mice and their congenic controls, a comprehensive plasma cytokine and chemokine profile, and brain levels of serotonin and kynurenine pathway metabolites. Consistent with previous studies, MRL/lpr mice had increased depression-like behavior and visuospatial memory impairment. Plasma levels of different inflammatory molecules (Haptoglobin, interleukin 10 (IL-10), interferon γ-inducible protein 10 (IP-10/CXCL10), lymphotactin, macrophage inhibitory protein 3β (MIP-3β/CCL19), monocyte chemotactic protein 1, 3 and 5 (MCP-1/CCL2, MCP-3/CCL7, MCP-5/CCL12), vascular cell adhesion molecule 1 (VCAM-1), lymphotactin and interferon γ (IFN-γ)) were increased in MRL/lpr mice. In cortex and hippocampus, MRL/lpr mice had increased levels of kynurenine pathway metabolites (kynurenine, 3-hydroxykynurenine, 3-hydroxynthranilic acid and quinolinic acid). Therefore, our study suggests that increased cytokine expression may be critical in the regulation subtle aspects of brain function in NP-SLE via induction of IDO and tryptophan/kynurenine metabolism.
Collapse
|
|
20
|
Kazemipour N, Qazizadeh H, Sepehrimanesh M, Salimi S. Biomarkers identified from serum proteomic analysis for the differential diagnosis of systemic lupus erythematosus. Lupus 2014; 24:582-7. [PMID: 25391542 DOI: 10.1177/0961203314558860] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2014] [Accepted: 10/03/2014] [Indexed: 01/22/2023]
Abstract
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that involves different organs. Its most important feature is the production of specific autoantibodies against nuclear or cytoplasmic antigens. Proteomic analysis of serum, as one of the most readily available body fluids, can be used as a method for clarifying the pathogenesis of SLE. In this study the serum proteome of 13 patients with SLE was evaluated and compared with seven healthy control participants. A specific kit was used to remove high-abundance proteins. After depletion, the protein expression patterns created by two-dimensional gel electrophoresis (2-DE) and MALDI-TOF/TOF-MS were used to identify disease-associated proteins. We found differential expression of 15 protein spots, including seven up-regulated and eight down-regulated proteins in SLE samples, in comparison with healthy participants. These spots were identified by MALDI-TOF/TOF-MS and classified into three groups include keratins, apolipoproteins and albumin, and individual proteins such as transthyretin, haptoglobin and prothrombin. These findings can help to clarify the pathophysiology and mechanism of SLE.
Collapse
Affiliation(s)
- N Kazemipour
- Department of Biochemistry, School of Veterinary Medicine, Shiraz University, Shiraz, Iran Department of Biology, College of Science, University of Sistan and Baluchestan, Zahedan, Iran
| | - H Qazizadeh
- Department of Biology, College of Science, University of Sistan and Baluchestan, Zahedan, Iran
| | - M Sepehrimanesh
- Gastroenterohepatology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - S Salimi
- Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan, Iran
| |
Collapse
|
|
21
|
IraL is an RssB anti-adaptor that stabilizes RpoS during logarithmic phase growth in Escherichia coli and Shigella. mBio 2014; 5:e01043-14. [PMID: 24865554 PMCID: PMC4045071 DOI: 10.1128/mbio.01043-14] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
UNLABELLED RpoS (σ(S)), the general stress response sigma factor, directs the expression of genes under a variety of stressful conditions. Control of the cellular σ(S) concentration is critical for appropriately scaled σ(S)-dependent gene expression. One way to maintain appropriate levels of σ(S) is to regulate its stability. Indeed, σ(S) degradation is catalyzed by the ClpXP protease and the recognition of σ(S) by ClpXP depends on the adaptor protein RssB. Three anti-adaptors (IraD, IraM, and IraP) exist in Escherichia coli K-12; each interacts with RssB and inhibits RssB activity under different stress conditions, thereby stabilizing σ(S). Unlike K-12, some E. coli isolates, including uropathogenic E. coli strain CFT073, show comparable cellular levels of σ(S) during the logarithmic and stationary growth phases, suggesting that there are differences in the regulation of σ(S) levels among E. coli strains. Here, we describe IraL, an RssB anti-adaptor that stabilizes σ(S) during logarithmic phase growth in CFT073 and other E. coli and Shigella strains. By immunoblot analyses, we show that IraL affects the levels and stability of σ(S) during logarithmic phase growth. By computational and PCR-based analyses, we reveal that iraL is found in many E. coli pathotypes but not in laboratory-adapted strains. Finally, by bacterial two-hybrid and copurification analyses, we demonstrate that IraL interacts with RssB by a mechanism distinct from that used by other characterized anti-adaptors. We introduce a fourth RssB anti-adaptor found in E. coli species and suggest that differences in the regulation of σ(S) levels may contribute to host and niche specificity in pathogenic and nonpathogenic E. coli strains. IMPORTANCE Bacteria must cope with a variety of environmental conditions in order to survive. RpoS (σ(S)), the general stress response sigma factor, directs the expression of many genes under stressful conditions in both pathogenic and nonpathogenic Escherichia coli strains. The regulation of σ(S) levels and activity allows appropriately scaled σ(S)-dependent gene expression. Here, we describe IraL, an RssB anti-adaptor that, unlike previously described anti-adaptors, stabilizes σ(S) during the logarithmic growth phase in the absence of additional stress. We also demonstrate that iraL is found in a large number of E. coli and Shigella isolates. These data suggest that strains containing iraL are able to initiate σ(S)-dependent gene expression under conditions under which strains without iraL cannot. Therefore, IraL-mediated σ(S) stabilization may contribute to host and niche specificity in E. coli.
Collapse
|
|
22
|
Tan W, Wang F, Guo D, Ke Y, Shen Y, Lv C, Zhang M. High serum level of haptoglobin is associated with the response of 12 weeks methotrexate therapy in recent-onset rheumatoid arthritis patients. Int J Rheum Dis 2014; 19:482-9. [PMID: 24863583 DOI: 10.1111/1756-185x.12380] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
BACKGROUND We previously found, using microarray, haptoglobin (HP) expression signal was 5.1-fold increased in peripheral blood mononuclear cells (PBMCs) from methotrexate (MTX)-resistant rheumatoid arthritis (RA) patients. OBJECTIVES To investigate whether serum levels of HP are associated with the response of 12 weeks MTX therapy in recent-onset RA patients. METHODS Sixty-nine active RA patients with recent onset (< 24 months) were treated with MTX. Clinical variables, levels of HP messenger RNA (mRNA) in PBMCs and HP serum levels were tested at week 0 and week 12. RESULTS After 12 weeks of MTX treatment, 34.7% of RA patients were categorized as responders according to European League Against Rheumatism (EULAR) response criteria (Week 12 Disease Activity Score of 28 joints [DAS-28] ≤ 3.2 and decrease > 1.2) and all others (65.2%) were defined as non-responders. The baseline HP mRNA in PBMCs from non-responders is significantly higher than those in responders (P < 0.05). Similar to mRNA expression, non-responders showed significantly elevated serum HP levels at baseline (369.9 ± 159.8 mg/dL) compared to those in responders (255.3 ± 143.9 mg/dL) (P = 0.01). Serum HP levels were decreased significantly from 255.3 ± 143.9 mg/dL at baseline to 186.4 ± 108.5 mg/dL at week 12 (P = 0.04) in responders, but remained at high levels in non-responders. CONCLUSIONS High serum levels of HP at baseline are associated with inadequate response of 12 weeks MTX treatment in recent-onset RA patients. Further replication studies in larger samples are needed to validate HP as a potential predictive biomarker for response to MTX therapy in RA.
Collapse
Affiliation(s)
- Wenfeng Tan
- Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Fang Wang
- Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Dunming Guo
- Department of Orthopedics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Yao Ke
- Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Youxuan Shen
- Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Chengyin Lv
- Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Miaojia Zhang
- Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| |
Collapse
|
|
23
|
A plasma proteomic approach in Rett syndrome: classical versus preserved speech variant. Mediators Inflamm 2013; 2013:438653. [PMID: 24453418 PMCID: PMC3884802 DOI: 10.1155/2013/438653] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2013] [Revised: 10/16/2013] [Accepted: 10/17/2013] [Indexed: 12/31/2022] Open
Abstract
Rett syndrome (RTT) is a progressive neurodevelopmental disorder mainly caused by mutations in the gene encoding the methyl-CpG-binding protein 2 (MeCP2). Although over 200 mutations types have been identified so far, nine of which the most frequent ones. A wide phenotypical heterogeneity is a well-known feature of the disease, with different clinical presentations, including the classical form and the preserved speech variant (PSV). Aim of the study was to unveil possible relationships between plasma proteome and phenotypic expression in two cases of familial RTT represented by two pairs of sisters, harbor the same MECP2 gene mutation while being dramatically discrepant in phenotype, that is, classical RTT versus PSV. Plasma proteome was analysed by 2-DE/MALDI-TOF MS. A significant overexpression of six proteins in the classical sisters was detected as compared to the PSV siblings. A total of five out of six (i.e., 83.3%) of the overexpressed proteins were well-known acute phase response (APR) proteins, including alpha-1-microglobulin, haptoglobin, fibrinogen beta chain, alpha-1-antitrypsin, and complement C3. Therefore, the examined RTT siblings pairs proved to be an important benchmark model to test the molecular basis of phenotypical expression variability and to identify potential therapeutic targets of the disease.
Collapse
|
|
24
|
Vanuytsel T, Vermeire S, Cleynen I. The role of Haptoglobin and its related protein, Zonulin, in inflammatory bowel disease. Tissue Barriers 2013; 1:e27321. [PMID: 24868498 DOI: 10.4161/tisb.27321] [Citation(s) in RCA: 100] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/21/2013] [Revised: 11/22/2013] [Accepted: 11/22/2013] [Indexed: 02/08/2023] Open
Abstract
Crohn's disease (CD) and ulcerative colitis (UC), collectively called inflammatory bowel disease (IBD), are immune-mediated conditions characterized by a chronic inflammation of the gut. Their precise etiology is unknown, although an increased intestinal permeability has been shown to play a central role in the pathogenesis of IBD. The intestinal epithelium provides the largest interface between the external environment and the host, and is thus a crucial regulation site of innate and adaptive immunity. Zonulin is one of the few known physiological mediators of paracellular intestinal permeability. It was found upregulated in different immune diseases like Celiac disease and Type 1 Diabetes (T1D). Recently, human zonulin was identified as prehaptoglobin-2 (pre-HP2) which before only had been regarded as the inactive precursor for HP2. Haptoglobin (HP) is a hemoglobin-binding protein with immunomodulatory properties. Its gene harbors a common polymorphism with 2 different alleles: HP1 and HP2. Allele HP2 and genotype HP22 has been shown to be overrepresented in different immune diseases like Rheumatoid Arthritis (RA), Systemic Lupus Erythematosus (SLE) and T1D, and has also been found to be more frequent in patients with IBD (UC and CD) than in healthy controls. In order to get some clues about the mechanism of action of HP(2) in IBD pathogenesis, we here review the current state of knowledge about zonulin and haptoglobin structure and function, and their plausible role in immune mediated diseases with an emphasis on IBD.
Collapse
Affiliation(s)
- Tim Vanuytsel
- Department of Clinical and Experimental Medicine; TARGID; KU Leuven
| | | | - Isabelle Cleynen
- Department of Clinical and Experimental Medicine; TARGID; KU Leuven
| |
Collapse
|
|
25
|
Garcia-Rodriguez S, Arias-Santiago S, Perandrés-López R, Castellote L, Zumaquero E, Navarro P, Buendía-Eisman A, Ruiz JC, Orgaz-Molina J, Sancho J, M Zubiaur. Increased gene expression of Toll-like receptor 4 on peripheral blood mononuclear cells in patients with psoriasis. J Eur Acad Dermatol Venereol 2013; 27:242-50. [PMID: 23457721 DOI: 10.1111/j.1468-3083.2011.04372.x] [Citation(s) in RCA: 45] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
BACKGROUND A role for the innate immune system in driving the autoimmune T cell cascade in psoriasis has been proposed. Toll-like receptors-(TLR)-2 and -4 play a role in inflammation, atherosclerosis, and their specific role in psoriasis remains unclear. OBJECTIVE To evaluate TLR2 and TLR4 gene expression levels in peripheral blood mononuclear cells from psoriatic patients. METHODS Changes in TLR2 ⁄ 4 gene expressions were evaluated using quantitative real-time reverse transcription polymerase chain reaction in peripheral blood mononuclear cells, from twenty-one patients with severe psoriasis, and analysed whether there was any correlation with cytokine plasma levels (T-helper 0-, T-helper 1-, T-helper 2- or regulatory T cells-type), or Calprotectin and with S100A8 and S100A9 gene expression levels. Eleven non-psoriatic healthy controls were analysed. RESULTS A clear increase in TLR4 gene expression was observed (3.84 ± 0.93, n = 21) together with a moderate increase in TLR2 expression (1.522 ± 0.31, n = 21). Both TLR4 and TLR2 gene expressions were significantly augmented in psoriatic patients compared with controls (all P < 0.001). Correlations between TLR2 and S100A9 gene expressions (r = 0.5145, P = 0.0170, n = 21); and between TLR2 expression and plasma interleukin-2 (r = 0.5667, P = 0.0074); interleukin-4 (r = 0.4766, P = 0.0289), interleukin-10 (r = 0.4355, P = 0.0484) and interleukin-13 (r = 0.4603, P = 0.0358), were found. When patients with atheroma plaque were considered (n = 7), both TLR4 (3.47 ± 0.99, P = 0.0156) and TLR2 (1.63 ± 0.31, P = 0.0156) expressions were significantly increased vs. controls and correlated with plasma TNF-a (r = 0.8929, P = 0.0123, in both cases). CONCLUSION Differential TLR4 ⁄ 2 gene expressions on psoriatic peripheral blood mononuclear cells and correlations with regulatory and ⁄ or proinflammatory cytokines and ⁄ or damage-associated molecular pattern molecule S100A9 emphasize innate immune response role in psoriasis.
Collapse
Affiliation(s)
- S Garcia-Rodriguez
- Department of Cellular Biology and Immunology, Instituto de Parasitología y Biomedicina López-Neyra, IPBLN-CSIC, PT Ciencias de Salud, Armilla, Granada, Spain
| | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
26
|
Sokolowska I, Gawinowicz MA, Ngounou Wetie AG, Darie CC. Disulfide proteomics for identification of extracellular or secreted proteins. Electrophoresis 2012; 33:2527-36. [PMID: 22899260 DOI: 10.1002/elps.201200182] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
The combination of SDS-PAGE and MS is one of the most powerful and perhaps most frequently used gel-based proteomics approaches in protein identification. However, one drawback of this method is that separation takes place under denaturing and reducing (R) conditions and as a consequence, all proteins with identical apparent molecular mass (Mr) will run together. Therefore, low-abundant proteins may not be easily identified. Another way of investigating proteins by proteomics is by analyzing subproteomes from a total proteome such as phosphoproteomics, glycoproteomics, or disulfide proteomics. Here, we took advantage of the property of secreted proteins to form disulfide bridges and investigated disulfide-linked proteins, using SDS-PAGE under nonreducing (NR) conditions. We separated sera from normal subjects and from patients with various diseases by SDS-PAGE (NR) and (R) conditions, followed by LC-MS/MS analysis. Although we did not see any detectable difference between the sera separated by SDS-PAGE(R), we could easily identify the disulfide-linked proteins separated by SDS-PAGE (NR). LC-MS/MS analysis of the disulfide-linked proteins correctly identified haptoglobin (Hp), a disulfide-linked protein usually found as a heterotetramer or as a disulfide-linked heteropolymer. Western blotting under NR and R conditions using anti-Hp antibodies confirmed the LC-MS/MS experiments and further confirmed that upon reduction, the disulfide-linked Hp heterotetramers and polymers were no longer disulfide-linked polymers. These data suggest that simply by separating samples on SDS-PAGEunder NR conditions, a different, new proteomics subset can be revealed and then identified.
Collapse
Affiliation(s)
- Izabela Sokolowska
- Biochemistry & Proteomics Group, Department of Chemistry & Biomolecular Science, Clarkson University, Potsdam, NY 13699-5810, USA
| | | | | | | |
Collapse
|
|
27
|
Zheng CX, Ji ZQ, Zhang LJ, Wen Q, Chen LH, Yu JF, Zheng D. Proteomics-based identification of haptoglobin as a favourable serum biomarker for predicting long-term response to splenectomy in patients with primary immune thrombocytopenia. J Transl Med 2012; 10:208. [PMID: 23039040 PMCID: PMC3506455 DOI: 10.1186/1479-5876-10-208] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/07/2012] [Accepted: 10/02/2012] [Indexed: 12/19/2022] Open
Abstract
BACKGROUND Splenectomy is the most effective treatment for patients with primary immune thrombocytopenia (ITP) who fail to respond to steroid therapy. Thus far, there is no effective means to predict the long-term haematological response of the procedure. The purpose of this study was to identify serum biomarkers as predictors of long-term response based on a proteomics approach. METHODS The serum samples of ITP patients were collected before splenectomy and seven days after surgery. After depletion of the abundant serum proteins, pooled preoperative serum samples from four responders to splenectomy, four nonresponders and four healthy controls were subjected to two-dimensional gel electrophoresis (2-DE). Nine protein spots with at least a five-fold alteration in expression between responders and nonresponders were all identified as haptoglobin (Hp) by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometer (MS) analysis. The validation of serum Hp expression was performed using enzyme-linked immunosorbent assays (ELISA) in thirty-seven responders, thirteen nonresponders and twenty-one healthy controls. RESULTS The preoperative serum levels of Hp in the nonresponders (925.9 ± 293.5 μg/ml) were significantly lower than those in the responders (1417.4 ± 315.0 μg/ml, p <0.001) and the healthy controls (1409.1 ± 354.2 μg/ml, p <0.001), while there was no significant difference between the latter two groups. The postoperative serum levels of Hp in responders and nonresponders were (1414.1 ± 225.0 μg/ml) and (952.9 ± 202.4 μg/ml), respectively. There were no significant differences between the serum Hp levels before and after surgery in both responders and nonresponders (p>0.05). The preoperative serum levels of Hp did not significantly correlate with preoperative platelet count of the same blood samples (r = 0.244, p = 0.087), while it positively correlated with postoperative peak platelet count (r = 0.622, p < 0.001). The optimal cutoff value of preoperative serum Hp levels (1173.80 μg/ml) derived from the receiver operating characteristic (ROC) curve led to 78.4% sensitivity and 84.6% specificity. CONCLUSIONS These results suggest that serum Hp levels may serve as a favourable predictor for the long-term response to splenectomy in ITP and may help to understand the pathophysiological differences between responders and nonresponders.
Collapse
Affiliation(s)
- Chao-Xu Zheng
- Department of Haematology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China
| | | | | | | | | | | | | |
Collapse
|
|
28
|
Monte AA, Vasiliou V, Heard KJ. Omics Screening for Pharmaceutical Efficacy and Safety in Clinical Practice. JOURNAL OF PHARMACOGENOMICS & PHARMACOPROTEOMICS 2012; S5:001. [PMID: 23264882 PMCID: PMC3526192 DOI: 10.4172/2153-0645.s5-001] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
As molecular techniques have improved, investigators have attempted to improve pharmaceutical efficacy and safety by making trait associations with genomic, epigenomic, transcriptomic, proteomic, and metabolomic polymorphisms. The 'omics era has seen screening assays for pharmaceutical efficacy and safety translated into clinical practice. This manuscript will discuss each 'omic field and the screening assays available to the clinician. While success has been demonstrated in each 'omic field, many challenges remain. Assays need wider availability, predictive values remain low, and costs remain high. In order for clinicians to realize improved efficacy and safety due 'omic screens, development of improved techniques, combining of 'omic assays, and increased clinical utilization is necessary. This is an exciting time for investigators and clinicians that desire improved pharmaceutical therapy.
Collapse
Affiliation(s)
- Andrew A. Monte
- Rocky Mountain Poison and Drug Center, Denver, CO, USA
- University of Colorado School of Medicine, Department of Emergency Medicine Aurora, CO, USA
| | - Vasilis Vasiliou
- Molecular Toxicology & Environmental Health Sciences Program, Department of Pharmaceutical Sciences, University of Colorado Denver, Aurora, CO, USA
| | - Kennon J. Heard
- Rocky Mountain Poison and Drug Center, Denver, CO, USA
- University of Colorado School of Medicine, Department of Emergency Medicine Aurora, CO, USA
| |
Collapse
|
|
29
|
Korte EA, Gaffney PM, Powell DW. Contributions of mass spectrometry-based proteomics to defining cellular mechanisms and diagnostic markers for systemic lupus erythematosus. Arthritis Res Ther 2012; 14:204. [PMID: 22364570 PMCID: PMC3392812 DOI: 10.1186/ar3701] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022] Open
Abstract
Systematic lupus erythematosus (SLE) is a complex disease for which molecular diagnostics are limited and pathogenesis is not clearly understood. Important information is provided in this regard by identification and characterization of more specific molecular and cellular targets in SLE immune cells and target tissue and markers of early-onset and effective response to treatment of SLE complications. In recent years, advances in proteomic technologies and applications have facilitated such discoveries. Here we provide a review of insights into SLE pathogenesis, diagnosis and treatment that have been provided by mass spectrometry-based proteomic approaches.
Collapse
Affiliation(s)
- Erik A Korte
- Department of Biochemistry and Molecular Biology, University of Louisville School of Medicine, 570 South Preston St, Baxter Research Building I, Room 204E, Louisville, KY 40202, USA
| | | | | |
Collapse
|
|
30
|
Pavón EJ, García-Rodríguez S, Zumaquero E, Perandrés-López R, Rosal-Vela A, Lario A, Longobardo V, Carrascal M, Abián J, Callejas-Rubio JL, Ortego-Centeno N, Zubiaur M, Sancho J. Increased expression and phosphorylation of the two S100A9 isoforms in mononuclear cells from patients with systemic lupus erythematosus: a proteomic signature for circulating low-density granulocytes. J Proteomics 2011; 75:1778-91. [PMID: 22230807 DOI: 10.1016/j.jprot.2011.12.020] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2011] [Revised: 11/03/2011] [Accepted: 12/14/2011] [Indexed: 10/14/2022]
Abstract
Proteins differentially expressed in peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients versus Normal controls were identified by 2-DE and MALDI-MS. Thus, S100A9 expression was significantly increased in SLE PBMCs relative to Normal PBMCs at both mRNA and protein levels. Increased S100A9 levels in SLE PBMCs correlated positively with the abnormal presence of low-density granulocytes (LDGs) detected by flow-cytometry in the mononuclear cell fractions. Another set of proteins that were differentially expressed in SLE PBMCs formed S100A9-independent clusters, suggesting that these differences in protein expression are in fact reflecting changes in the abundance of specific cell types. In SLE PBMCs spots of the two S100A9 isoforms, S100A9-l and S100A9-s, and their phosphorylated counterparts were identified and confirmed to be phosphorylated at Thr(113) by MS/MS analyses. In addition, the phorbol ester PMA alone or in combination with ionomycin induced a stronger increase in threonine phosphorylation of S100A9 in SLE than in Normal PBMCs, while the same stimuli caused the opposite effect on phosphorylation and activation of Erk1/2, suggesting the existence of an abnormal S100A9 signaling in SLE PBMCs. Therefore, the expansion and activation of LDGs in SLE seems to underlie this prominent S100A9 signature.
Collapse
Affiliation(s)
- Esther J Pavón
- Departamento de Biología Celular e Inmunología, Instituto de Parasitología y Biomedicina "López-Neyra", IPBLN-CSIC, Armilla, Spain
| | | | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
31
|
Speeckaert R, Van Vlierberghe H, Troisi R, De Bacquer D, Speeckaert MM, De Buyzere ML, Claeys L, de Hemptinne B, Delanghe JR. Donor haptoglobin phenotype determines outcome following liver transplantation. Transpl Int 2011; 24:619-26. [PMID: 21401731 DOI: 10.1111/j.1432-2277.2011.01246.x] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
Abstract
Haptoglobin (Hp) is a polymorphic plasma protein with multiple functions defined by three major phenotypes (Hp 1-1, Hp 2-1, and Hp 2-2). In this article, the effects of the donor Hp phenotype (determined by starch gel electrophoresis) on the outcome and the iron status after liver transplantation were investigated. A total of 450 liver transplant patients were enrolled in this study with a median follow-up of 37 months. Kaplan-Meier and Cox regression survival analyses showed a significantly worse graft survival for liver transplantation cases with an Hp 2-2 donor phenotype, which was associated with an increased mortality rate in this group. In male patients, the Hp 2-2 phenotype was associated with higher serum ferritin concentrations, which may be linked to the significantly increased likelihood of infectious complications in this phenotype. Liver transplant patients with Hp 1-1 and Hp 2-1 grafts had a better outcome probability than recipients of an Hp 2-2 graft, which may be explained by differences in iron metabolism induced by the Hp genotype of the graft.
Collapse
Affiliation(s)
- Reinhart Speeckaert
- Department of Clinical Chemistry, Ghent University Hospital, De Pintelaan 85, Gent, Belgium
| | | | | | | | | | | | | | | | | |
Collapse
|
|
32
|
De Franceschi L, Bosello S, Scambi C, Biasi D, De Santis M, Caramaschi P, Peluso G, La Verde V, Bambara LM, Ferraccioli G. Proteome analysis of biological fluids from autoimmune-rheumatological disorders. Proteomics Clin Appl 2011; 5:78-89. [PMID: 21246742 DOI: 10.1002/prca.201000069] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2010] [Revised: 10/27/2010] [Accepted: 11/01/2010] [Indexed: 11/10/2022]
Abstract
Autoimmune-rheumatological diseases are worldwide distributed disorders and represent a complex array of illnesses characterized by autoreactivity (reactivity against self-antigens) of T-B lymphocytes and by the synthesis of autoantibodies crucial for diagnosis (biomarkers). Yet, the effects of the autoimmune chronic inflammation on the infiltrated tissues and organs generally lead to profound tissue and organ damage with loss of function (i.e., lung, kidney, joints, exocrine glands). Although progresses have been made on the knowledge of these disorders, much still remains to be investigated on their pathogenesis and identification of new biomarkers useful in clinical practice. The rationale of using proteomics in autoimmune-rheumatological diseases has been the unmet need to collect, from biological fluids that are easily obtainable, a summary of the final biochemical events that represent the effects of the interplay between immune cells, mesenchymal cells and endothelial cells. Proteomic analysis of these fluids shows encouraging results and in this review, we addressed four major autoimmune-rheumatological diseases investigated through proteomic techniques and provide evidence-based data on the highlights obtained in systemic sclerosis, primary and secondary Sjogren's syndrome, systemic lupus erythematosus and rheumatoid arthritis.
Collapse
Affiliation(s)
- Lucia De Franceschi
- Department of Medicine, Section of Internal Medicine, University of Verona, Verona, Italy
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
33
|
Zumaquero E, Muñoz P, Cobo M, Lucena G, Pavón EJ, Martín A, Navarro P, García-Pérez A, Ariza-Veguillas A, Malavasi F, Sancho J, Zubiaur M. Exosomes from human lymphoblastoid B cells express enzymatically active CD38 that is associated with signaling complexes containing CD81, Hsc-70 and Lyn. Exp Cell Res 2010; 316:2692-706. [DOI: 10.1016/j.yexcr.2010.05.032] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/13/2010] [Revised: 05/28/2010] [Accepted: 05/28/2010] [Indexed: 12/14/2022]
|
|
34
|
Accelerated atherosclerosis in systemic lupus erythematosus: role of proinflammatory cytokines and therapeutic approaches. J Biomed Biotechnol 2010; 2010. [PMID: 20936125 PMCID: PMC2948929 DOI: 10.1155/2010/607084] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/15/2010] [Revised: 06/21/2010] [Accepted: 07/26/2010] [Indexed: 12/02/2022] Open
Abstract
Systemic lupus erythematosus (SLE), a chronic multisystem autoimmune disease with a broad range of clinical manifestations, is associated with accelerated atherosclerosis (AT) and increased risk of cardiovascular complications. Relevant factors directly influencing the development of AT comprise immune complex generation, complement activation, and changes in the production and activity of a complex network of cytokines, including type I and II interferons, B lymphocyte stimulator (BLyS), TNFα, IL-6, IL-17 and migration macrophage inhibitor (MIF). Autoantibodies, also responsible for cytokine expression and activation, play a supplementary key role in the development of AT. Genomic and proteomic studies have contributed to the discovery of genes and proteins involved in AT, including some that may be suitable to be used as biomarkers. All that data has allowed the development of new drugs, most of them evaluated in clinical trials: inhibitors of IFN and TNFα, B cell directed therapies, synthetic oligodeoxynucleotides, intravenous immunoglobulin, or statins. The focus of the present paper is to summarize recent evidence showing the role of cytokines in the development of AT in SLE and the rationale, and safety concerns, in the use of combined therapy to prevent AT and cardiovascular disease.
Collapse
|
|
35
|
|
|
36
|
Guerranti R, Bertocci E, Fioravanti A, Papakostas P, Montella A, Guidelli G, Cortelazzo A, Nuti R, Giordano N. Serum Proteome of Patients with Systemic Sclerosis: Molecular Analysis of Expression and Prevalence of Haptoglobin Alpha Chain Isoforms. Int J Immunopathol Pharmacol 2010; 23:901-9. [DOI: 10.1177/039463201002300326] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
Abstract
Haptoglobin (Hpt) is an acute phase protein characterized by three major phenotypes (Hpt 1-1, Hpt 2-1 and Hpt 2-2). The Hpt 2-2 phenotype is associated with increased prevalence of various systemic diseases, including autoimmune disorders. Moreover, the Hpt 2-2 phenotype induces a shift from Th1 to Th2 response and increases fibrotic processes. On this basis, we performed serum proteomic analysis of with Systemic Sclerosis (SSc), a connective tissue disorder associated with Th2-type immune response and characterized by interstitial and perivascular fibrosis due to different factors (including genetic, environmental, immunological and microchimeric factors). Serum of 23 SSc outpatients (4 males, 19 females, mean age 54±5.3 years) diagnosed according to the American Rheumatism Association (ARA) criteria, were considered for the proteomic analysis and compared to serum of 21 control subjects. Serum depleted of HAP was analyzed by 2-DE, and Hpt chain spots were identified by WB. The expression frequency of each Hpt a chain in SSc patients and controls was compared and quantitative analysis of spot expression (% Vol) was performed. Above all, our study amplifies the limited data in the literature on proteomic analysis in SSc, also confirming previous data that revealed a significant increase of haptoglobin type 2-2 and a concomitant decrease of the 1-1 phenotype in SSc patients. Moreover, our results demonstrate that c spots are more prevalent in SSc patients than in controls (91.3% vs 55.5%, p<0.05), while the expression frequency of a and b spots does not change. In patients Hpt 2-1 or Hpt 1-1 e spot is less abundant. According to our results, the c and e spots can be considered markers for SSc and thus be of use for the early diagnosis of connective tissue disorders and in establishing appropriate treatment.
Collapse
Affiliation(s)
| | | | - A. Fioravanti
- Department of Clinical Medicine and Immunology, Rheumatology Unit, University of Siena, Siena, Italy
| | | | | | - G.M. Guidelli
- Department of Clinical Medicine and Immunology, Rheumatology Unit, University of Siena, Siena, Italy
| | | | | | | |
Collapse
|
|
37
|
Identification of human zonulin, a physiological modulator of tight junctions, as prehaptoglobin-2. Proc Natl Acad Sci U S A 2009; 106:16799-804. [PMID: 19805376 DOI: 10.1073/pnas.0906773106] [Citation(s) in RCA: 312] [Impact Index Per Article: 19.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
Increased intestinal permeability (IP) has emerged recently as a common underlying mechanism in the pathogenesis of allergic, inflammatory, and autoimmune diseases. The characterization of zonulin, the only physiological mediator known to regulate IP reversibly, has remained elusive. Through proteomic analysis of human sera, we have now identified human zonulin as the precursor for haptoglobin-2 (pre-HP2). Although mature HP is known to scavenge free hemoglobin (Hb) to inhibit its oxidative activity, no function has ever been ascribed to its uncleaved precursor form. We found that the single-chain zonulin contains an EGF-like motif that leads to transactivation of EGF receptor (EGFR) via proteinase-activated receptor 2 (PAR(2)) activation. Activation of these 2 receptors was coupled to increased IP. The siRNA-induced silencing of PAR(2) or the use of PAR(2)(-/-) mice prevented loss of barrier integrity. Proteolytic cleavage of zonulin into its alpha(2)- and beta-subunits neutralized its ability to both activate EGFR and increase IP. Quantitative gene expression revealed that zonulin is overexpressed in the intestinal mucosa of subjects with celiac disease. To our knowledge, this is the initial example of a molecule that exerts a biological activity in its precursor form that is distinct from the function of its mature form. Our results therefore characterize zonulin as a previously undescribed ligand that engages a key signalosome involved in the pathogenesis of human immune-mediated diseases that can be targeted for therapeutic interventions.
Collapse
|
|
38
|
López-Pedrera C, Barbarroja N, Villalba JM. Novel biomarkers of atherosclerosis and cardiovascular risk in autoimmune diseases: Genomics and proteomics approaches. Proteomics Clin Appl 2009; 3:213-25. [DOI: 10.1002/prca.200800147] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2008] [Indexed: 01/15/2023]
|
|
39
|
Camafeita E, Lamas JR, Calvo E, López JA, Fernández-Gutiérrez B. Proteomics: New insights into rheumatic diseases. Proteomics Clin Appl 2009; 3:226-241. [DOI: 10.1002/prca.200800146] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 08/30/2023]
|
|
40
|
Dai Y, Hu C, Huang Y, Huang H, Liu J, Lv T. A proteomic study of peripheral blood mononuclear cells in systemic lupus erythematosus. Lupus 2009; 17:799-804. [PMID: 18755861 DOI: 10.1177/0961203308089444] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
Our objective was to analyze the changes in the protein expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE). Peripheral blood was obtained from patients with SLE and healthy controls. 2-D gel electrophoresis was performed, and gels were silver-stained. Differentially expressed protein spots were detected, some of which were identified by MALDI-TOF spectrometry. Match rates of 71% +/- 4% and 72% +/- 4% were gotten for control and patient gels, respectively. 791 +/- 17 spots were detected for control gels and 781 +/- 17 for patient gels. Eleven protein spots were up-regulated, and 9 protein spots were down-regulated in patients with SLE. Five differentially expressed proteins were identified as immunoglobulin J chain, apolipoprotein A-IV precursor, calprotectin L1H and zinc finger protein subfamily 1A (all up-regulated) and glutathione S-transferase (down-regulated), some of which had previously been shown to play a potential role in the pathogenesis of SLE. We conclude there are significant changes in the 2-D maps of PBMCs in patients with SLE and applying this proteomic approach may be a useful way to gain novel insights into SLE.
Collapse
Affiliation(s)
- Y Dai
- The Second Clinical Medical College, Jinan University, Shenzhen People's Hospital, Guangdong Province, China.
| | | | | | | | | | | |
Collapse
|
|
41
|
The association of Haptoglobin polymorphism with Crohn's disease in Israel. J Crohns Colitis 2008; 2:214-8. [PMID: 21172213 DOI: 10.1016/j.crohns.2008.03.005] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/04/2008] [Accepted: 03/18/2008] [Indexed: 02/08/2023]
Abstract
OBJECTIVES Haptoglobin is a α(2)-sialoglycoprotein with hemoglobin binding capacity. Functional differences between the Hp phenotypes with the Hp 1-1 protein being a superior anti-inflammatory to the Hp 2-2 protein have been identified. The aim of our study was to investigate the possible role of Hp polymorphism in the susceptibility to Crohn's disease and its clinical course. METHODS Hp phenotypes were determined for 382 Israeli CD patients and 3243 healthy controls. Phenotypic data for all Crohn's disease patients were carefully characterized. Analysis was preformed to evaluate the association between Hp polymorphism and Crohn's disease. RESULTS The frequency of Haptoglobin 1-1 was lower in Crohn's disease patients than in healthy individuals (6.28% vs. 9.28%, P=0.057). There was no association between Haptoglobin phenotypes and disease location, behavior or extra-intestinal manifestations. No association was found between the Haptoglobin polymorphism and the frequency of the three Crohn's disease associated NOD2 mutations examined. CONCLUSIONS We found a borderline significant decrease of the Haptoglobin 1-1 phenotype in Israeli Crohn's disease patients compared to healthy controls. Our findings may support the importance of inflammation in Crohn's disease pathogenesis and the protective function of Haptoglobin 1-1 in the susceptibility for Crohn's disease.
Collapse
|
|
42
|
Mao L, Dong H, Yang P, Zhou H, Huang X, Lin X, Kijlstra A. MALDI-TOF/TOF-MS reveals elevated serum haptoglobin and amyloid A in Behcet's disease. J Proteome Res 2008; 7:4500-7. [PMID: 18754684 DOI: 10.1021/pr800279m] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
Behcet's disease (BD) is a multisystemic autoimmune disease with unclear etiology and pathogenesis. To screen aberrant serum proteins in BD, serum samples were obtained from eight male BD patients with active uveitis and eight male healthy volunteers with informed consent. The serum samples from active BD patients and normal controls were pooled. Highly abundant serum proteins (albumin and IgG) were depleted from these two samples using an affinity capture based kit. The obtained samples were subjected to two-dimensional gel electrophoresis (2-DE). Protein spots were visualized with the "blue silver" staining. Differently expressed proteins were subsequently identified by matrix-assisted laser desorption /ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). Western blot and enzyme-linked immunosorbent assay (ELISA) were performed using the serum samples from 18 patients with active BD, 6 patients with inactive BD, 22 patients with Vogt-Koyanagi-Harada (VKH) syndrome, and 20 healthy volunteers to validate the results of 2-DE and MS. Proteomic profiles of the pooled samples were compared, and approximately 800 protein spots were observed in each of the gels. Expression levels of four of the protein spots in active BD were significantly higher than those in the normal controls. Mass spectrometric protein identification revealed that the four protein spots corresponded to two proteins: haptoglobin (Hp) and serum amyloid A (SAA). Western blot and ELISA showed that Hp was only overexpressed in active BD but not in inactive BD, VKH syndrome, or healthy controls. An obvious band of SAA was detected in 72.2% of the serum samples from BD patients, whereas a vague band of this protein was found in 10.0% of the tested normal samples and 9.1% of VKH samples. Our results revealed a significantly increased expression of Hp and SAA in serum of active BD patients. These two proteins may be involved in the development of BD.
Collapse
Affiliation(s)
- Liming Mao
- Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology of Sun Yat-sen University, Guangzhou, P.R. China
| | | | | | | | | | | | | |
Collapse
|
|
43
|
Fagoonee S, Caorsi C, Giovarelli M, Stoltenberg M, Silengo L, Altruda F, Camussi G, Tolosano E, Bussolati B. Lack of plasma protein hemopexin dampens mercury-induced autoimmune response in mice. JOURNAL OF IMMUNOLOGY (BALTIMORE, MD. : 1950) 2008; 181:1937-1947. [PMID: 18641331 DOI: 10.4049/jimmunol.181.3.1937] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 03/10/2025]
Abstract
Several factors affect the autoimmune response, including iron-dependent modulation of T cells. Hemopexin is the plasma protein with the highest binding affinity to heme. It mediates heme-iron recovery in the liver, thus controlling heme-iron availability in peripheral cells. The aim of the present study was to investigate the role of hemopexin in the progress of an autoimmune response. To this end, we chose a mouse model of mercury-induced autoimmunity and evaluated the susceptibility of hemopexin-null mice to mercury treatment compared with wild-type controls. In this study we show that lack of hemopexin dampens mercury-induced autoimmune responses in mice. Hemopexin-null mice produced fewer antinuclear autoantibodies and had reduced deposits of immune complexes in the kidney after mercuric chloride treatment compared with wild-type mice. These features were associated with a reduction in activated T cells and lower absolute B cell number in spleen and impaired IgG1 and IgG2a production. In contrast, in hemopexin-null mice the response to OVA/CFA immunization was maintained. In addition, hemopexin-null mice had reduced transferrin receptor 1 expression in T cells, possibly due to the increase in heme-derived iron. Interestingly, CD4(+)T cells isolated from mercury-treated hemopexin-null mice show reduced IFN-gamma-dependent STAT1 phosphorylation compared with that of wild-type mice. Our data suggest that hemopexin, by controlling heme-iron availability in lymphocytes, modulates responsiveness to IFN-gamma and, hence, autoimmune responses.
Collapse
Affiliation(s)
- Sharmila Fagoonee
- Department of Genetics, Biology and Biochemistry, and Molecular Biotechnology Center, University of Turin, Turin, Italy.
| | | | | | | | | | | | | | | | | |
Collapse
|