|
1
|
Yu Q, Dong Y, Wang X, Su C, Zhang R, Xu W, Jiang S, Dang Y, Jiang W. Pharmacological induction of MHC-I expression in tumor cells revitalizes T cell antitumor immunity. JCI Insight 2024; 9:e177788. [PMID: 39106105 PMCID: PMC11385079 DOI: 10.1172/jci.insight.177788] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2023] [Accepted: 07/18/2024] [Indexed: 08/09/2024] Open
Abstract
Antigen presentation by major histocompatibility complex class I (MHC-I) is crucial for T cell-mediated killing, and aberrant surface MHC-I expression is tightly associated with immune evasion. To address MHC-I downregulation, we conducted a high-throughput flow cytometry screen, identifying bleomycin (BLM) as a potent inducer of cell surface MHC-I expression. BLM-induced MHC-I augmentation rendered tumor cells more susceptible to T cells in coculture assays and enhanced antitumor responses in an adoptive cellular transfer mouse model. Mechanistically, BLM remodeled the tumor immune microenvironment, inducing MHC-I expression in a manner dependent on ataxia-telangiectasia mutated/ataxia telangiectasia and Rad3-related-NF-κB. Furthermore, BLM improved T cell-dependent immunotherapeutic approaches, including bispecific antibody therapy, immune checkpoint therapy, and autologous tumor-infiltrating lymphocyte therapy. Importantly, low-dose BLM treatment in mouse models amplified the antitumor effect of immunotherapy without detectable pulmonary toxicity. In summary, our findings repurpose BLM as a potential inducer of MHC-I, enhancing its expression to improve the efficacy of T cell-based immunotherapy.
Collapse
Affiliation(s)
- Qian Yu
- Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, and
| | - Yu Dong
- Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Xiaobo Wang
- Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, and
| | - Chenxuan Su
- Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Runkai Zhang
- Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Wei Xu
- Institute of Immunological Innovation and Translation and
| | - Shuai Jiang
- Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Yongjun Dang
- Basic Medicine Research and Innovation Center for Novel Target and Therapeutic Intervention, Ministry of Education, College of Pharmacy, Chongqing Medical University, Chongqing Medical University, Chongqing, China
| | - Wei Jiang
- Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, and
| |
Collapse
|
|
2
|
Silva RCMC, Lopes MF, Travassos LH. Distinct T helper cell-mediated antitumor immunity: T helper 2 cells in focus. CANCER PATHOGENESIS AND THERAPY 2023; 1:76-86. [PMID: 38328613 PMCID: PMC10846313 DOI: 10.1016/j.cpt.2022.11.001] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/06/2022] [Revised: 10/07/2022] [Accepted: 11/02/2022] [Indexed: 02/09/2024]
Abstract
The adaptive arm of the immune system is crucial for appropriate antitumor immune responses. It is generally accepted that clusters of differentiation 4+ (CD4+) T cells, which mediate T helper (Th) 1 immunity (type 1 immunity), are the primary Th cell subtype associated with tumor elimination. In this review, we discuss evidence showing that antitumor immunity and better prognosis can be associated with distinct Th cell subtypes in experimental mouse models and humans, with a focus on Th2 cells. The aim of this review is to provide an overview and understanding of the mechanisms associated with different tumor outcomes in the face of immune responses by focusing on the (1) site of tumor development, (2) tumor properties (i. e., tumor metabolism and cytokine receptor expression), and (3) type of immune response that the tumor initially escaped. Therefore, we discuss how low-tolerance organs, such as lungs and brains, might benefit from a less tissue-destructive immune response mediated by Th2 cells. In addition, Th2 cells antitumor effects can be independent of CD8+ T cells, which would circumvent some of the immune escape mechanisms that tumor cells possess, like low expression of major histocompatibility-I (MHC-I). Finally, this review aims to stimulate further studies on the role of Th2 cells in antitumor immunity and briefly discusses emerging treatment options.
Collapse
Affiliation(s)
- Rafael Cardoso Maciel Costa Silva
- Laboratory of Immunoreceptors and Signaling, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro 21941-902, Brazil
| | - Marcela Freitas Lopes
- Laboratory of Immunity Biology George DosReis,Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro 21941-902, Brazil
| | - Leonardo Holanda Travassos
- Laboratory of Immunoreceptors and Signaling, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro 21941-902, Brazil
| |
Collapse
|
|
3
|
Suolitiken D, Wang Y, Jin Z, Wang Z. EBV protection- and susceptibility-related HLA alleles and EBV status in the Chinese population: A single-center study. Immun Inflamm Dis 2022; 10:e666. [PMID: 35759244 PMCID: PMC9210551 DOI: 10.1002/iid3.666] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/05/2021] [Revised: 05/29/2022] [Accepted: 05/31/2022] [Indexed: 11/09/2022] Open
Abstract
BACKGROUND Although most adults are infected by Epstein-Barr virus (EBV), some patients develop highly lethal diseases associated with EBV infection, including EBV-hemophagocytic lymphohistiocytosis (EBV-HLH), chronic active EBV infections (CAEBV), and lymphoma, the pathogeneses of which remain to be investigated. The human leukocyte antigen (HLA) complex may be associated with the viral infection pathway, and, therefore, HLA alleles may be associated with EBV-related diseases and subpopulations of infected cells, studies related to EBV-associated diseases, and subpopulations of infected cells that were conducted in China are scarce. METHODS In this study, we analyzed the high-resolution HLA genotypes of 269 patients with EBV-associated diseases and 213 EBV-seronegative hematopoietic stem cell donors using PCR-SBT assay and elucidated the associations of HLA-A, -B, -C, -DRB1, and -DQB1 alleles with EBV-associated diseases in the Chinese population, Benjamini-Hochberg correction to adjust for multiple testing. HLA genotypes were also analyzed in patients with EBV-associated diseases showing EBV-infected lymphocyte subpopulations. RESULTS We found that individuals carrying the following alleles showed the following levels of risks: HLA-DRB1*11 allele, reduced risk of EBV-related disease (OR [odds ratio]: 0.56; 95% confidence interval [95% CI]: 0.32-0.99; p < .05; Adjust p = .71); HLA-DQB1*06:02 allele, reduced risk (OR: 0.5699; 95% CI: 0.3486-0.9317; p < .05; Adjust p = .57); and HLA-B*15:01 allele, increased risk (OR: 1.763; 95% CI: 0.3486-0.9317; p < .05; Adjust p = .57). Patients with EBV-associated diseases showing the B*15:01 genotype had a higher risk of T-cell, NK-cell, and multicell infections than those with other genotype subgroups. CONCLUSIONS These findings highlight the importance of HLA genotype for assessing EBV infectivity.
Collapse
Affiliation(s)
- Dina Suolitiken
- Department of Hematology, Beijing Friendship HospitalCapital Medical UniversityBeijingPeople's Republic of China
| | - Yini Wang
- Department of Hematology, Beijing Friendship HospitalCapital Medical UniversityBeijingPeople's Republic of China
| | - Zhili Jin
- Department of Hematology, Beijing Friendship HospitalCapital Medical UniversityBeijingPeople's Republic of China
| | - Zhao Wang
- Department of Hematology, Beijing Friendship HospitalCapital Medical UniversityBeijingPeople's Republic of China
| |
Collapse
|
|
4
|
Compte M, Sanz L, Álvarez-Vallina L. Applications of trimerbodies in cancer immunotherapy. INTERNATIONAL REVIEW OF CELL AND MOLECULAR BIOLOGY 2022; 369:71-87. [PMID: 35777865 DOI: 10.1016/bs.ircmb.2022.03.008] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/15/2023]
Abstract
Trimerbodies, with their unique structural and functional properties, are the basis of a new generation of therapeutic antibodies, which due to their small size and plasticity are ideal for the generation of novel biological protein drugs with multiple competitive advantages over conventional full-length monoclonal antibodies. Since their emergence, trimerbodies have been used in preclinical cancer diagnosis and therapy. Trimerbodies are highly adaptable molecules, as they allow target-specific modulation of T cell-mediated anti-tumor immunity to enhance preexisting responses or to generate de novo immune responses. In fact, a tumor-specific humanized 4-1BB-agonistic trimerbody has shown a rather impressive safety and efficacy profile in preclinical studies making it a realistic option for clinical development. Moreover, thanks to the avidity effect they are endowed with considerable therapeutic potential as carriers to deliver cytotoxic payloads to tumors. In addition, molecular imaging studies could benefit from some intermediate-sized trivalent trimerbodies as promising candidates for targeted therapy and tumor imaging.
Collapse
Affiliation(s)
- Marta Compte
- Department of Antibody Engineering, Leadartis S.L., Madrid, Spain
| | - Laura Sanz
- Molecular Immunology Unit, Hospital Universitario Puerta de Hierro Majadahonda, Majadahonda, Madrid, Spain
| | - Luis Álvarez-Vallina
- Cancer Immunotherapy Unit (UNICA), Department of Immunology, Hospital Universitario12 de Octubre, Madrid, Spain; Immuno-Oncology and Immunotherapy Group, Instituto de Investigación Sanitaria 12 de Octubre (imas12), Madrid, Spain.
| |
Collapse
|
|
5
|
Hirbod-Mobarakeh A, Shabani M, Keshavarz-Fathi M, Delavari F, Amirzargar AA, Nikbin B, Kutikhin A, Rezaei N. Immunogenetics of Cancer. CANCER IMMUNOLOGY 2020:417-478. [DOI: 10.1007/978-3-030-30845-2_20] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
6
|
Li Q, Cohen JI. Epstein-Barr Virus and the Human Leukocyte Antigen Complex. CURRENT CLINICAL MICROBIOLOGY REPORTS 2019; 6:175-181. [PMID: 33094090 DOI: 10.1007/s40588-019-00120-9] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
Purpose While most adults are infected Epstein-Barr virus (EBV), 3-5% remain uninfected. The human leukocyte antigen (HLA) complex, which controls many pathogens, may influence infection and disease associated with EBV. Recent Findings Numerous EBV proteins and miRNAs down-regulate HLA class I and II expression on the cell surface. HLA class II functions as a receptor for EBV entry into B cells. Specific HLA class II alleles correlate with the susceptibility of B cells to EBV infection in vitro and with EBV seropositivity or seronegativity of humans. HLA class I polymorphisms correlate with development and severity of EBV infectious mononucleosis and with the risk of several virus-associated malignancies including nasopharyngeal carcinoma, Hodgkin lymphoma, and post-transplant lymphoproliferative disease. Significance These findings indicate that while EBV has evolved to use MHC class II as a receptor for virus entry, polymorphisms in MHC class II and class I influence virus infection and disease.
Collapse
Affiliation(s)
- Qingxue Li
- Medical Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD
| | - Jeffrey I Cohen
- Medical Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD
| |
Collapse
|
|
7
|
Garrido F. HLA Class-I Expression and Cancer Immunotherapy. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2019; 1151:79-90. [PMID: 31140107 DOI: 10.1007/978-3-030-17864-2_3] [Citation(s) in RCA: 30] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
The impact of HLA class I loss in cancer immunotherapy is carefully analyzed. Why some metastatic lesions regress and other progress after immunotherapy? Are T lymphocytes responsible for tumour rejection and how these responses can be boosted? These questions are discussed in the context of the molecular mechanisms responsible for MHC/HLA class I alterations. If the metastatic tumour cells harbor "irreversible/hard" HLA lesions, they will escape and kill the host. In contrast, if the molecular lesion is "reversible/soft", tumor cells can potentially recover HLA-class I expression and can finally be destroyed. These important new concepts are integrated together and gain a great importance in the new era of "immune checkpoint antibodies". Finally, the ability to recover HLA-I expression in tumours harboring "structural-irreversible-hard" genetic lesions is seen as a challenge for the future investigation.
Collapse
Affiliation(s)
- Federico Garrido
- Departamento de Analisis Clinicos e Inmunologia, Hospital Universitario Virgen de las Nieves, Facultad de Medicina, Universidad de Granada, Granada, Spain
| |
Collapse
|
|
8
|
Sona P, Hong JH, Lee S, Kim BJ, Hong WY, Jung J, Kim HN, Kim HL, Christopher D, Herviou L, Im YH, Lee KY, Kim TS, Jung J. Integrated genome sizing (IGS) approach for the parallelization of whole genome analysis. BMC Bioinformatics 2018; 19:462. [PMID: 30509173 PMCID: PMC6276166 DOI: 10.1186/s12859-018-2499-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2018] [Accepted: 11/16/2018] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND The use of whole genome sequence has increased recently with rapid progression of next-generation sequencing (NGS) technologies. However, storing raw sequence reads to perform large-scale genome analysis pose hardware challenges. Despite advancement in genome analytic platforms, efficient approaches remain relevant especially as applied to the human genome. In this study, an Integrated Genome Sizing (IGS) approach is adopted to speed up multiple whole genome analysis in high-performance computing (HPC) environment. The approach splits a genome (GRCh37) into 630 chunks (fragments) wherein multiple chunks can simultaneously be parallelized for sequence analyses across cohorts. RESULTS IGS was integrated on Maha-Fs (HPC) system, to provide the parallelization required to analyze 2504 whole genomes. Using a single reference pilot genome, NA12878, we compared the NGS process time between Maha-Fs (NFS SATA hard disk drive) and SGI-UV300 (solid state drive memory). It was observed that SGI-UV300 was faster, having 32.5 mins of process time, while that of the Maha-Fs was 55.2 mins. CONCLUSIONS The implementation of IGS can leverage the ability of HPC systems to analyze multiple genomes simultaneously. We believe this approach will accelerate research advancement in personalized genomic medicine. Our method is comparable to the fastest methods for sequence alignment.
Collapse
Affiliation(s)
- Peter Sona
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Jong Hui Hong
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Sunho Lee
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Byong Joon Kim
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Woon-Young Hong
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Jongcheol Jung
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025
| | - Han-Na Kim
- PGM21 (Personalized Genomic Medicine 21), Ewha Womans University Medical Center, 1071, Anyang Cheon-ro, Yangcheon-gu, Seoul, 158-710, Korea
| | - Hyung-Lae Kim
- PGM21 (Personalized Genomic Medicine 21), Ewha Womans University Medical Center, 1071, Anyang Cheon-ro, Yangcheon-gu, Seoul, 158-710, Korea
| | - David Christopher
- Bioinformatics Solutions, 900 N McCarthy Blvd., Milpitas, CA, 95035, USA
| | - Laurent Herviou
- Bioinformatics Solutions, 900 N McCarthy Blvd., Milpitas, CA, 95035, USA
| | - Young Hwan Im
- Bioinformatics Solutions, 900 N McCarthy Blvd., Milpitas, CA, 95035, USA
| | - Kwee-Yum Lee
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025.,Faculty of Medicine, University of Queensland, QLD, Brisbane, 4072, Australia
| | - Tae Soon Kim
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025.,Department of Clinical Medical Sciences, Seoul National University College of Medicine, 71 Ihwajang-gil, Jongno-gu, Seoul, 03087, South Korea
| | - Jongsun Jung
- Genome Data Integration Center, Syntekabio Incorporated, Techno-2ro B-512, Yuseong-gu, Daejeon, Republic of Korea, 34025.
| |
Collapse
|
|
9
|
Hirbod-Mobarakeh A, Amirzargar AA, Nikbin B, Nicknam MH, Kutikhin A, Rezaei N. Immunogenetics of Cancer. CANCER IMMUNOLOGY 2015:295-341. [DOI: 10.1007/978-3-662-44006-3_17] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
10
|
Tang Q, Zhang J, Qi B, Shen C, Xie W. Downregulation of HLA class I molecules in primary oral squamous cell carcinomas and cell lines. Arch Med Res 2009; 40:256-63. [PMID: 19608014 DOI: 10.1016/j.arcmed.2009.04.004] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2008] [Accepted: 03/30/2009] [Indexed: 10/20/2022]
Abstract
BACKGROUND AND AIMS Loss or downregulation of human leukocyte antigen (HLA) class I expression has been reported in a variety of human tumors including oral squamous cell carcinoma (OSCC). METHODS Expression of HLA class I molecules were evaluated by immunohistochemistry, flow cytometry, semi-quantitative Western blot and RT-PCR in 43 tissue samples of primary oral squamous cell carcinomas (pOSCC) from Chinese patients and two OSCC cell lines. RESULTS HLA class I heavy chain of B/C locus and A locus and beta(2-)microglobulin were obviously lost or downregulated in pOSCC with the percentage of 31, 55 and 35% respectively. The expression of HLA B/C, LMP2, LMP7, LMP10 and PA28beta in OSCC cell lines was also presumably reduced in comparison with normal epithelial cell line. CONCLUSIONS These data suggested that the downregulation of HLA class I molecules in OSCC was closely associated with the low-efficient transcription and abnormality of post-transcription regulation of HLA class I genes and antigen presentation-related genes. These results can add more light to the mechanism by which OSCC escape from immunological surveillance.
Collapse
Affiliation(s)
- Qiusha Tang
- The Key Laboratory of Developmental Genes and Human Disease of Education Ministry, Department of Genetics and Developmental Biology, Southeast University Medical School, Jiangsu Province, China
| | | | | | | | | |
Collapse
|
|
11
|
Ueda Y, Ishikawa K, Shiraishi N, Yokoyama S, Kitano S. Clinical significance of HLA class I heavy chain expression in patients with gastric cancer. J Surg Oncol 2008; 97:451-5. [DOI: 10.1002/jso.20985] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023]
|
|
12
|
Cabrera CM. The Double Role of the Endoplasmic Reticulum Chaperone Tapasin in Peptide Optimization of HLA Class I Molecules. Scand J Immunol 2007; 65:487-93. [PMID: 17523940 DOI: 10.1111/j.1365-3083.2007.01934.x] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
During the assembly of the HLA class I molecules with peptides in the peptide-loading complex, a series of transient interactions are made with ER-resident chaperones. These interactions culminate in the trafficking of the HLA class I molecules to the cell surface and presentation of peptides to CD8(+) T lymphocytes. Within the peptide-loading complex, the glycoprotein tapasin exhibits a relevant function. This immunoglobulin (Ig) superfamily member in the endoplasmic reticulum membrane tethers empty HLA class I molecules to the transporter associated with antigen-processing (TAP) proteins. This review will address the current concepts regarding the double role that tapasin plays in the peptide optimization and surface expression of the HLA class I molecules.
Collapse
Affiliation(s)
- C M Cabrera
- Stem Cell Bank of Andalucia (Spanish Central Node), Hospital Universitario Virgen de las Nieves, Granada, Spain.
| |
Collapse
|
|
13
|
Stühler K, Köper K, Pfeiffer K, Tagariello A, Souquet M, Schwarte-Waldhoff I, Hahn SA, Schmiegel W, Meyer HE. Differential proteome analysis of colon carcinoma cell line SW480 after reconstitution of the tumour suppressor Smad4. Anal Bioanal Chem 2006; 386:1603-12. [PMID: 17043799 DOI: 10.1007/s00216-006-0803-9] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2006] [Revised: 08/23/2006] [Accepted: 08/28/2006] [Indexed: 10/24/2022]
Abstract
The tumour suppressor gene Smad4 is frequently inactivated in gastrointestinal carcinomas. Smad4 plays a pivotal role in transducing signals of the transforming growth factor-beta (TGF-beta) superfamily of proteins. Inactivation of Smad4 seems to occur late during tumour progression when tumours acquire invasive and metastatic properties. Identification of proteins directly or indirectly regulated by Smad4 would, therefore, ease the future design of new diagnostic and therapeutic strategies for gastrointestinal carcinoma. We have used human colon carcinoma cell line SW480 stably transfected with Smad4 as an in-vitro model system to identify Smad4-regulated proteins by applying two-dimensional gel electrophoresis (2DE) then MALDI-PMF/PFF-MS. We identified a total of 47 protein species with a Smad4-dependent expression. From the functions of the candidate proteins we obtained new insights into Smad4's participation in processes, for example apoptosis, differentiation, and proliferation.
Collapse
Affiliation(s)
- Kai Stühler
- Medizinisches Proteom-Center, Ruhr-University Bochum, ZKF E 1.43, Universitätsstrasse 150, 44801, Bochum, Germany.
| | | | | | | | | | | | | | | | | |
Collapse
|
|
14
|
Guest RD, Hawkins RE, Kirillova N, Cheadle EJ, Arnold J, O'Neill A, Irlam J, Chester KA, Kemshead JT, Shaw DM, Embleton MJ, Stern PL, Gilham DE. The role of extracellular spacer regions in the optimal design of chimeric immune receptors: evaluation of four different scFvs and antigens. J Immunother 2005; 28:203-11. [PMID: 15838376 DOI: 10.1097/01.cji.0000161397.96582.59] [Citation(s) in RCA: 230] [Impact Index Per Article: 11.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Human peripheral blood lymphocytes can be transduced to express antigen-dependent CD3zeta chimeric immune receptors (CIRs), which function independently of the T-cell receptor (TCR). Although the exact function of these domains is unclear, previous studies imply that an extracellular spacer region is required for optimal CIR activity. In this study, four scFvs (in the context of CIRs with or without extracellular spacer regions) were used to target the human tumor-associated antigens carcinoembryonic antigen (CEA), neural cell adhesion molecule (NCAM), the oncofetal antigen 5T4, and the B-cell antigen CD19. In all cases human T-cell populations expressing the CIRs were functionally active against their respective targets, but the anti-5T4 and anti-NCAM CIRs showed enhanced specific cytokine release and cytotoxicity only when possessing an extracellular spacer region. In contrast, the anti-CEA and anti-CD19 CIRs displayed optimal cytokine release activity only in the absence of an extracellular spacer. Interestingly, mapping of the scFv epitopes has revealed that the anti-CEA scFv binds close to the amino-terminal of CEA, which is easily accessible to the CIR. In contrast, CIRs enhanced by a spacer domain appear to bind to epitopes residing closer to the cell membrane, suggesting that a more flexible extracellular domain may be required to permit the efficient binding of such epitopes. These results show that a spacer is not necessary for optimal activity of CIRs but that the optimal design varies.
Collapse
Affiliation(s)
- Ryan D Guest
- Cancer Research UK, Department of Medical Oncology, University of Manchester and Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 4BX, UK
| | | | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
15
|
Abstract
AIM: To investigate the expression of several important molecules involved in major histocompatibility complex (MHC) class I presentation pathway in primary hepatocellular carcinoma (HCC), and to determine whether cytotoxic T lymphocyte (CTL) vaccine therapy was suitable for HCC.
METHODS: Labeled streptavidin biotin (LSAB) method of immunohisto-chemistry was used to study 33 HCC tissue specimens.
RESULTS: Most HCC tissues and adjacent histological normal hepatocytes expressed HLA-I antigens,TAP, and B7, expression of B7 was especially strong, and there was no significant difference between them (P>0.05).
CONCLUSION: The MHC class I presentation pathway in primary hepatocellular carcinoma may not be abnormal or dysfunctional, and CTL could kill these tumor cells. Thus, it is suitable and practicable to design and construct CTL vaccine against HCC.
Collapse
Affiliation(s)
- Wei Chen
- Department of Immunology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan Province, China
| | | | | | | |
Collapse
|
|
16
|
Naumova E, Mihaylova A, Stoitchkov K, Ivanova M, Quin L, Toneva M. Genetic polymorphism of NK receptors and their ligands in melanoma patients: prevalence of inhibitory over activating signals. Cancer Immunol Immunother 2005; 54:172-8. [PMID: 15248031 PMCID: PMC11032772 DOI: 10.1007/s00262-004-0575-z] [Citation(s) in RCA: 58] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2004] [Accepted: 05/31/2004] [Indexed: 10/26/2022]
Abstract
Antitumor cytotoxicity of NK cells and T cells expressing NK-associated receptors is regulated by interaction between their cell surface killer immunoglobulin-like receptors (KIRs) and CD94/NKG2 heterodimers with MHC class I ligands on target cells. To test the hypothesis that KIR and/or HLA polymorphisms, and KIR/HLA combinations could contribute to the tumorigenesis, association studies were performed in 50 patients with malignant melanoma (MM) in different stages of disease and 54 controls. Our data showed that the frequency of inhibitory and activating KIR genes and KIR genotypes did not differ significantly between healthy individuals and melanoma patients. HLA haplotype distribution showed statistically significant increased frequencies of A*01-B*35-Cw*04 (0.069 vs 0.000; pc < 0.05; OR = 19.9), A*01-B*08-DRB1*03 (0.079 vs 0.019; pc < 0.05; OR = 4.5), and A*24-B*40-DRB1*11 (0.026 vs 0.000; pc < 0.05; OR = 7.1) in melanoma patients compared with healthy controls. Individuals homozygous for group 2 HLA-C ligands were less frequent in the patient group compared with the control cohort (12% vs 31.5%; p < 0.017). In addition, we observed an increased frequency (88.0% vs 68.5%; p = 0.017; OR = 2.80) of KIR2DL2/2DL3 in combination with their group 1 HLA-C ligands, while the presence of these KIRs in the absence of the putative ligands was decreased (12.0% vs 31.5%; p = 0.017) in the patient group. Furthermore, an increased frequency of activating KIR2DS1 in the absence of the putative HLA-C(Lys80) ligands was found in melanoma patients (16.0% vs 9.2%). In contrast, KIR2DS2 was absent in patients more often (38.0% vs 25.9%) when the presumptive HLA-C(Asn80) ligands were present. A slightly higher incidence of KIR3DL1 in combination with the less effective Bw4(Thr80) ligands was seen in patients with primary (20.8%) compared with metastatic (4.2%) disease. The data obtained in this study imply that there may not be a direct association between KIR gene content in the genome and the presence of malignant melanoma, or melanoma progression. However, some HLA haplotypes could be predisposing to MM in the Bulgarian population. Furthermore, distinct KIR/HLA ligand combinations may be relevant to the development of malignancy whereby inhibition overrides activation of NK cells and T cells expressing NK-associated receptors, which in turn might facilitate tumor escape and progression.
Collapse
Affiliation(s)
- Elissaveta Naumova
- Central Laboratory of Clinical Immunology, University Hospital Alexandrovska, 1, G. Sofiiski St., Sofia, 1431, Bulgaria.
| | | | | | | | | | | |
Collapse
|
|
17
|
Gückel B, Stumm S, Rentzsch C, Marmé A, Mannhardt G, Wallwiener D. A CD80-transfected human breast cancer cell variant induces HER-2/neu-specific T cells in HLA-A*02-matched situations in vitro as well as in vivo. Cancer Immunol Immunother 2005; 54:129-40. [PMID: 15365776 PMCID: PMC11034349 DOI: 10.1007/s00262-004-0583-z] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/22/2004] [Accepted: 06/14/2004] [Indexed: 10/26/2022]
Abstract
Adjuvant treatment is still only working in a small percentage of breast cancer patients. Therefore, new strategies need to be developed. Immunotherapies are a very promising approach because they could successfully attack tumor cells in the stage of dormancy. To assess the feasibility of using an allogeneic approach for vaccination of breast cancer patients, we selected a CD80-transfected breast cancer cell line based on its immunogenic properties. Using CD80+ KS breast cancer cells and human leukocyte antigen (HLA)-A*02-matched peripheral blood mononuclear cells (PBMCs) of breast cancer patients in allogeneic mixed lymphocyte-tumor cell cultures (MLTCs), it was possible to isolate HLA-A*02-restricted cytotoxic T cells (CTLs). Furthermore, a genetically modified KS variant expressing influenza A matrix protein serving as a surrogate tumor-associated antigen (TAA) was able to stimulate flu peptide-specific T cells alongside the induction of alloresponses in MLTCs. KS breast cancer cells were demonstrated to express already known TAAs such as CEA, MUC-1, MAGE-1, MAGE-2, and MAGE-3. To further improve antigenicity, HER-2/neu was added to this panel as a marker antigen known to elicit HLA-A*02-restricted CTLs in patients with breast cancer. Thus, the antigen-processing and antigen-presentation capacity of KS cells was further demonstrated by the stimulation of HER-2/neu-specific CD8+ T cells in PBMCs of breast cancer patients in vitro. These results gave a good rationale for a phase I/II trial, where the CD80+ HER-2/neu-overexpressing KS variant is actually used as a cellular vaccine in patients with metastatic breast cancer. As a proof of principle, we present data from two patients where a significant increase of interferon-gamma (IFN-gamma) release was detected when postvaccination PBMCs were stimulated by allogeneic vaccine cells as well as by HLA-A*02-restricted HER-2/neu epitopes. In whole cell vaccine trials, monitoring is particularly challenging because of strong alloresponses and limited knowledge of TAAs. In this study, a panel of HER-2/neu epitopes, together with the quantitative real time (qRT)-PCR method to analyze vaccine-induced cytokines secreted by T cells, proved to be highly sensitive and feasible to perform an "immunological staging" following vaccination.
Collapse
Affiliation(s)
- Brigitte Gückel
- Department of Gynecology and Obstetrics, University of Tübingen, Calwerstrasse 7, 72076, Tübingen, Germany.
| | | | | | | | | | | |
Collapse
|
|
18
|
Cabrera CM, López-Nevot MA, Jiménez P, Garrido F. Involvement of the chaperone tapasin in HLA-B44 allelic losses in colorectal tumors. Int J Cancer 2005; 113:611-8. [PMID: 15455354 DOI: 10.1002/ijc.20526] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/04/2023]
Abstract
Tumors can exhibit selective allelic losses of HLA class I antigens as part of altered HLA phenotypes. In colorectal tumors, the HLA class I allele most frequently lost is HLA-B44, although the precise mechanism responsible for this loss has not been described to date. From a total of 95 colorectal cryopreserved tumor samples, we selected (by immunohistochemical staining) 13 tumors with HLA-B44-negative expression. Loss of heterozygosity at 6p21.3 was demonstrated to be the cause of the negative expression in 4 cases. In the remaining 9 cases, structural analyses of microdissected tissue samples of the 3 subtypes of HLA-B44 loss in these tumors (B*4402, B*4403 and B*4405) did not reveal any mutations. However, all 3 subtypes of HLA-B44 presented in this study shared a common characteristic: the presence of an aspartic amino acid residue at position 114 in the HLA class I heavy chain. This residue has been described as determining tapasin dependence for the surface expression of these alleles and therefore for antigen presentation. We studied tapasin transcription by RT-PCR in these tumors and found tapasin downregulation in all 9 tumors samples with the HLA-B44-negative phenotype. In contrast, tapasin was normally transcribed in HLA-B44-positive colorectal tumors samples, as well as in 3 HLA-B44-negative laryngeal carcinomas and 1 bladder tumor. Defective tapasin transcription seems to be an alteration responsible for the absence of HLA-B44 expression in colorectal tumors, thus contributing to the generation of tumor immune escape phenotypes.
Collapse
Affiliation(s)
- Carmen M Cabrera
- Departamento de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Avda. Fuerzas Armadas 2, 18014 Granada, Spain
| | | | | | | |
Collapse
|
|
19
|
Kovalcsik E, John J, Turner M, Birchall L, Sage D, Whittle R, Dalgleish A, Pandha H. Differential expression of melanoma-associated antigens and molecules involved in antigen processing and presentation in three cell lines established from a single patient. Melanoma Res 2004; 14:463-71. [PMID: 15577316 DOI: 10.1097/00008390-200412000-00005] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
Tumour cells are able to evade the immune system by using several 'escape mechanisms'. Downregulation of molecules involved in the processing and presentation of self-antigens has been reported. However, these adaptations have not been compared in metastases in different anatomical locations but derived from a single patient. We investigated three melanoma cell lines--MJT1 from the parietal lobe of the brain, MJT3 from the cerebellum and MJT5 from the left side of the neck--established from biopsies excised from a 45 year old female patient. Although human leukocyte antigen (HLA) class I was detected in all three cell lines by flow cytometry using an anti-HLA monomorphic antibody, further serological analysis demonstrated HLA B38 loss in all three cell lines, HLA B7 downregulation in MJT5 (skin metastases) and B7 loss in MJT3 and MJT1 (brain metastases) compared with the HLA type of the patient's normal autologous lymphocytes. Interferon-gamma (IFNgamma) treatment increased the expression of HLA class I and transporters associated with antigen processing 1 (TAP1) in all three cell lines. De novo HLA class II molecule expression was observed after IFNgamma treatment in MJT3 and MJT5. Western blot and reverse transcription-polymerase chain reaction results revealed heterogeneity of melanoma-associated antigen (MAA) expression in the cell lines: MJT3 cells expressed higher levels of MAAs than the other two cell lines. In conclusion, this study has demonstrated that three metastatic lesions from a single patient can have differential expression of molecules involved in antigen processing (TAP1) and presentation (HLA I and II), but that expression of these molecules is modulated by IFNgamma to a similar degree in all cell lines. In contrast, the downregulation of expression of specific MAAs between the three cell lines was unaffected by the addition of IFNgamma.
Collapse
Affiliation(s)
- Edit Kovalcsik
- Department of Oncology, St. George's Hospital Medical School, London, UK
| | | | | | | | | | | | | | | |
Collapse
|
|
20
|
Monti P, Marchesi F, Reni M, Mercalli A, Sordi V, Zerbi A, Balzano G, Di Carlo V, Allavena P, Piemonti L. A comprehensive in vitro characterization of pancreatic ductal carcinoma cell line biological behavior and its correlation with the structural and genetic profile. Virchows Arch 2004; 445:236-47. [PMID: 15258755 DOI: 10.1007/s00428-004-1053-x] [Citation(s) in RCA: 46] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2004] [Accepted: 05/15/2004] [Indexed: 01/07/2023]
Abstract
There are a large number of stable pancreatic ductal carcinoma cell lines (PDCL) that are used by researchers worldwide. Detailed data about their differentiation status and genetic alterations are present in the literature, but a systematic correlation with cell biological behavior is often lacking. PDCL ( n=12) were clustered by source of tumor cell (ascites, primary tumor, metastasis), and the data of functional cell biology were correlated with the reported structural and genetic profiles. Major histocompatibility complex expression, chemosensitivity and aneuploidia appeared to be related to the source of PDCL, and proliferative capacity appeared to be related to the grade of differentiation. No correlation between genetic/structural features of PDCL and biological behavior was found. All the cell lines appeared generally insensitive to in vitro treatment with 5-fluorouracil and showed variable degrees of susceptibility to gemcitabine, raltitrexed and oxaliplatin. All the PDCL showed resistance to Fas-mediated apoptosis but were significantly sensitive to the pro-apoptotic effect of inflammatory cytokines [interleukin (IL)-1beta, tumor necrosis factor (TNF)alpha and interferon gamma]. PDCL were characterized for the secretion of several factors relevant to the tumor-immune cross talk. Vascular endothelial growth factor, CCL2, CCL5 and transforming growth factor beta were the factors most frequently released; less frequent was the secretion of CXCL8, CCL22, IL-6 and sporadically CXCL12, IL-10 and hepatocyte growth factor. The cytokines IL-1beta and TNFalpha were always undetectable. In conclusion, a clear correlation between structural/genetic features and function could not be detected, suggesting the weakness of a "morphological" classification for the in vitro studies of pancreatic cancer.
Collapse
Affiliation(s)
- Paolo Monti
- Laboratory of Experimental Surgery, Department of Diabetes and Transplant Immunology, S. Raffaele Scientific Institute, Via Olgettina 60, 20132 Milan, Italy
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
21
|
Li Y, Tredget EE, Ghahary A. Cell surface expression of MHC class I antigen is suppressed in indoleamine 2,3-dioxygenase genetically modified keratinocytes: implications in allogeneic skin substitute engraftment. Hum Immunol 2004; 65:114-23. [PMID: 14969766 DOI: 10.1016/j.humimm.2003.11.004] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/11/2003] [Revised: 11/17/2003] [Accepted: 11/18/2003] [Indexed: 10/26/2022]
Abstract
Indoleamine 2,3-dioxygenase (IDO) has been indicated to prevent the fetus from maternal T-cell rejection. A longer survival of IDO genetically modified islets transplanted into NOD mouse kidney capsules has also been demonstrated. As IDO mediated mechanism of graft protection has not been elucidated, in our study we hypothesize that the expression of IDO may prevent immune rejection by suppressing the major histocompatibility complex (MHC) class I antigen. To test this hypothesis, an IDO adenoviral vector was constructed and the effect of IDO on MHC class I expression was evaluated on recombinant adenoviral transfected keratinocytes. Following a successful construction of IDO expressing adenoviral vector, the catabolic activity of IDO enzyme was evaluated by measuring the levels of its product, kynurenine in keratinocyte conditioned medium. The results indicated a higher level of kynurenine in IDO expressing cells relative to those of control cells. The results of MHC class I experiments revealed a significant downregulation of cell membrane associated MHC class I antigen in IDO genetically modified keratinocytes relative to that of either nontransfected or empty vector transfected cells. Further experiments demonstrated that an addition of tryptophan or IDO inhibitor markedly restored the expression of MHC class I on IDO transfected keratinocytes. The findings of this study suggest that downregulation of MHC class I expression by IDO might be one of the mechanisms through which IDO mediates local immunosuppression.
Collapse
Affiliation(s)
- Yunyuan Li
- Department of Surgery, University of Alberta, Edmonton, Alberta, Canada
| | | | | |
Collapse
|
|
22
|
Wiendl H, Mitsdoerffer M, Weller M. Hide-and-seek in the brain: a role for HLA-G mediating immune privilege for glioma cells. Semin Cancer Biol 2003; 13:343-51. [PMID: 14708714 DOI: 10.1016/s1044-579x(03)00025-7] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/27/2022]
Abstract
This review summarizes the current knowledge on the expression and functional role of HLA-G in normal CNS cells and brain tumor cells in vitro and in vivo. The CNS has classically been viewed as an immune-privileged organ. Here we discuss some of the particularities of anti-tumoral responses within this compartment. Special emphasis is dedicated to the possible role of the non-classical MHC molecule HLA-G as an alternative mechanism of immune escape. We review the mechanisms how glioma cell-derived HLA-G may paralyze the immune system and which cellular subsets of the immune system are affected. Possible therapeutic implications derived from these observations include the targeting of HLA-G expression within the framework of inducing glioma-specific immunity.
Collapse
Affiliation(s)
- Heinz Wiendl
- Department of Neurology, University of Tübingen, Hoppe-Seyler-Strasse 3, D-72076 Tübingen, Germany.
| | | | | |
Collapse
|
|
23
|
Kayser S, Watermann I, Rentzsch C, Weinschenk T, Wallwiener D, Gückel B. Tumor-associated antigen profiling in breast and ovarian cancer: mRNA, protein or T cell recognition? J Cancer Res Clin Oncol 2003; 129:397-409. [PMID: 12836015 DOI: 10.1007/s00432-003-0445-7] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2003] [Accepted: 04/04/2003] [Indexed: 02/04/2023]
Abstract
PURPOSE The absence of tumor-associated antigens (TAA) which might elicit an immune response is one reason for the disappointing results of therapeutical vaccines in cancer patients. Moreover, impaired expression of MHC class-I and components involved in antigen processing, such as TAP-1, -2, LMP-2, -7, and MECL-1, may lead to tumor escape from immune recognition. Expression profiling of TAA is one approach towards the design of well-defined and individualized anti-tumor vaccines. METHODS Quantitative polymerase chain reaction (qRT-PCR) is the method of choice to characterize immunologically relevant properties of individual tumors. However, the application of qRT-PCR as a surrogate parameter for the expression of TAAs depends upon the assumption that the level of an mRNA species correlates with the cellular level of the protein it encodes. Therefore, we additionally analyzed TAA expression by immunofluorescence and T cell recognition. RESULTS In the present study we were unable to confirm that impaired TAP-1 or -2 (transporter associated with antigen processing) expression characterized at the mRNA level is an appropriate surrogate parameter for down-regulated MHC class-I expression in breast cancer. In addition, we analyzed the expression pattern of TAAs in breast and ovarian cancer cell lines. Besides the well-known over-expression of HER-2/neu, CEA, and MUC-1, multiple antigens of the MAGE-family were frequently co-expressed. We investigated whether detection of TAAs by qRT-PCR correlates with monoclonal antibody staining, and which method could predict T cell recognition. We demonstrated a correlation between tumor cell lysis by HLA-A*0201-restricted, MUC-1-specific CTL and threshold levels of MUC-1-specific mRNA. CONCLUSION MUC-1 is an example that TAA profiling by RT-PCR and flow cytometry can fail to correlate with each other and are of limited value in the prediction of T cell recognition.
Collapse
Affiliation(s)
- Simone Kayser
- Department of Gynecology and Obstetrics, University of Tübingen, Calwerstrasse 7, 72076, Tübingen, Germany
| | | | | | | | | | | |
Collapse
|
|
24
|
Garcia-Lora A, Algarra I, Garrido F. MHC class I antigens, immune surveillance, and tumor immune escape. J Cell Physiol 2003; 195:346-55. [PMID: 12704644 DOI: 10.1002/jcp.10290] [Citation(s) in RCA: 367] [Impact Index Per Article: 16.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Oncogenic transformation in human and experimental animals is not necessarily followed by the appearance of a tumor mass. The immune system of the host can recognize tumor antigens by the presentation of small antigenic peptides to the receptor of cytotoxic T-lymphocytes (CTLs) and reject the nascent tumor. However, cancer cells can sometimes escape these specific T-cell immune responses in the course of somatic (genetic and phenotypic) clonal evolution. Among the tumor immune escape mechanisms described to date, the alterations in the expression of major histocompatibility complex (MHC) molecules play a crucial step in tumor development due to the role of MHC antigens in antigen presentation to T-lymphocytes and the regulation of natural killer cell (NK) cell function. In this work, we have (1) updated information on the mechanisms that allow CTLs to recognize tumor antigens after antigen processing by transformed cells, (2) described the altered MHC class I phenotypes that are commonly found in human tumors, (3) summarized the molecular mechanisms responsible for MHC class I alteration in human tumors, (4) provided evidence that these altered human leukocyte antigens (HLA) class I phenotypes are detectable as result of a T-cell immunoselection of HLA class I-deficient variants by an immunecompetent host, and (5) presented data indicating the MHC class I phenotype and the immunogenicity of experimental metastatic tumors change drastically when tumors develop in immunodeficient mice.
Collapse
Affiliation(s)
- Angel Garcia-Lora
- Servicio de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Spain
| | | | | |
Collapse
|
|
25
|
Abstract
AIM: To investigate whether CTL vaccine therapy is suitable for primary hepatocellular carcinoma (HCC) from the viewpoint of HLA class I antigens expression.
METHODS: The immunocytochemistry, image analysis, flow cytometry, and labeled streptavidin biotin (LSAB) method of immunohistochemistry were applied respectively to study 4 HCC cell lines (e.g. Alexander, HepG2, SMMC-7721, and QGY-7703) cultured in vitro and 6 frozen tissue specimens of HCC.
RESULTS: The positive control cell line Raji had very strong positive staining. Most mitotic and nonmitotic cells of the 4 HCC cell lines had various intensity of HLA class I antigens expression. The negative control cell K562 and the control slides of all the cell lines had no positive staining. In the 6 HCC specimens immunohistochemically studied, histological normal hepatocytes had no or very weak positive staining and the liver sinus had very strong positive staining. Most HCC cells in the sections from the 6 HCC specimens had strong positive HLA class I antigens staining. The positive staining was located in the cytoplasm, the perinuclear area, and at the cell membrane of the liver cancer cells. Flow cytometry also revealed that Raji and those 4 HCC cell lines had strong HLA class I antigens expression, which was confirmed quantitatively by the image analysis. It showed that the objective grayscale values of Raji and those 4 HCC cell lines were significantly different from that of K562 (Raji 114.04 ± 10.94, Alexander 165.97 ± 5.35, HepG2167.02 ± 12.60, QGY-7703161.46 ± 7.13, SMMC-7721165.93 ± 5.21, K562244.89 ± 4.60, P < 0.01). Significant differences were also found between Raji and the 4 HCC cell lines.
CONCLUSION: HCC cells express HLA class I antigens strongly. From this point of view, the active specific immunotherapy of CTL vaccine is suitable and practicable for HCC.
Collapse
Affiliation(s)
- Jian Huang
- Bioinfo Tech Incorporated Company, 10F Zhuangsen No.8 Dongsheng Street, Chengdu 610015, Sichuan Province,China.
| | | | | |
Collapse
|
|
26
|
Affiliation(s)
- F Garrido
- Departamento de Analisis Clinicos, Hospital Universitario Virgen de las Nieves, Granada, Spain
| | | |
Collapse
|
|
27
|
Brouwer RE, van der Heiden P, Schreuder GMT, Mulder A, Datema G, Anholts JDH, Willemze R, Claas FHJ, Falkenburg JHF. Loss or downregulation of HLA class I expression at the allelic level in acute leukemia is infrequent but functionally relevant, and can be restored by interferon. Hum Immunol 2002; 63:200-10. [PMID: 11872238 DOI: 10.1016/s0198-8859(01)00381-0] [Citation(s) in RCA: 68] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
Human leukocyte antigen (HLA) class I expression at the allelic level was analyzed in 397 acute myeloid leukemia (AML) and 186 acute lymphoid leukemia (ALL) using a complement-dependent cytotoxicity assay. Impaired recognition possibly due to HLA downregulation was observed in 2% of the patients with AML and ALL in complete remission, and in 8%-15% in the groups with blasts. In 15 instances of diminished cytotoxicity, leukemic cells and control PHA blasts from the same patients were further analyzed using flow cytometry. In 4/6 ALL and 4/9 AML patients HLA downregulation or complete loss (2 patients) of cell surface expression could be confirmed. No genomic abnormalities were observed. In addition, 12 AML and 13 ALL patients were tested during relapse using flow cytometry. In 1/12 AML patients and 1/13 ALL patients allelic downregulation of cell surface expression was found. In two patients tested, downregulation or loss of cell surface expression of HLA class I antigens corresponded with impaired T cell mediated lysis by HLA restricted cytotoxic T lymphocyte.Treatment of the cells with alpha- or gamma-interferon could restore HLA class I expression and T-cell recognition. In conclusion, downregulation of cell surface expression of HLA class I expression at the allelic level in AML and ALL is infrequent but functionally relevant. HLA downregulation was reversible and T-cell recognition could be restored by alpha- or gamma-interferon.
Collapse
Affiliation(s)
- Rolf E Brouwer
- Laboratory of Experimental Hematology, Department of Hematology, Leiden, The Netherlands
| | | | | | | | | | | | | | | | | |
Collapse
|
|
28
|
Abstract
Cancers in mouse and man express multiple tumor-specific as well as tumor-associated antigens. Immunodominance in the host response to these antigens can result in successive selection of heritable antigen loss variants. Immunodominance may also prevent the development of responses to new tumor-specific antigens that may arise during tumor progression. Some tumor-specific antigens are retained during tumor progression possibly because they are essential for survival of the malignant phenotype. Immunodominance may allow cancer cells to escape even after loss of a single MHC Class I allele because cross-presentation of the retained antigen by this allele that must be expressed on the surrounding antigen presenting cells sustains the immunodominant response. This prevents effective responses to secondary antigens that may remain as potential targets. Immunization with in vitro selected cancer cell variants that lack the immunodominant antigen can break the immunodominance and prevent escape of cancers from host immunity.
Collapse
Affiliation(s)
- H Schreiber
- Department of Pathology, The University of Chicago, IL 60637, USA.
| | | | | | | |
Collapse
|
|
29
|
Pangault C, Le Friec G, Caulet-Maugendre S, Léna H, Amiot L, Guilloux V, Onno M, Fauchet R. Lung macrophages and dendritic cells express HLA-G molecules in pulmonary diseases. Hum Immunol 2002; 63:83-90. [PMID: 11821155 DOI: 10.1016/s0198-8859(01)00373-1] [Citation(s) in RCA: 63] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
HLA-G is selectively expressed in extravillous trophoblast of human placenta, which does not express classical HLA-A and -B molecules. Several studies report the role of HLA-G as a molecule involved in immune tolerance. By interacting with NK and T cells inhibitory receptors, HLA-G may downregulate their cytotoxicity functions. To appreciate the biologic and clinical relevance of HLA-G expression in lung diseases, HLA class I and HLA-G expression were analyzed in a panel of 36 ex vivo neoplastic tissues and 8 non-neoplastic lung tissues. Immunohistochemical analysis was performed using a pan-HLA class I antibody (W6/32) and three different specific anti-HLA-G antibodies (87G, MEMG/9 and 4H84). These findings demonstrated that HLA-G products were not expressed in pulmonary structural cells. However, HLA-G molecules were detected in activated macrophages and dendritic cells infiltrating lung carcinomas (33%) and nontumoral pulmonary diseases (25%). HLA-G expression was not correlated with classical HLA alterations. No statistical correlation was found between HLA-G expression and clinical or biologic parameters except high tumor size. The expression of HLA-G in myelo-monocytic cells infiltrating lung pathologic tissues could alter antigenic presentation and contribute to decrease immune response efficiency, subsequently favoring the progression of tumoral or inflammatory processes.
Collapse
Affiliation(s)
- Celine Pangault
- Laboratoire Universitaire d'Hématologie et de la Biologie des Cellules Sanguines, Faculté de Médecine, Université de Rennes I, Rennes Cedex, France.
| | | | | | | | | | | | | | | |
Collapse
|
|
30
|
Falk CS, Schendel DJ. Allogeneic MHC class I ligands and their role in positive and negative regulation of human cytotoxic effector cells. Hum Immunol 2002; 63:8-19. [PMID: 11916173 DOI: 10.1016/s0198-8859(01)00360-3] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
The allogeneic mixed lymphocyte culture (MLC) has served as an important experimental system for elucidating the cellular and molecular basis of human lymphocyte responses. Complex mixtures of lymphocytes are stimulated by disparate alloantigens, inducing cellular activation and generating a cytokine milieu that is an excellent breeding ground for the proliferation and differentiation of many distinct lymphocyte subsets. Cloning of individual lymphocytes following alloactivation has allowed various cytotoxic lymphocytes to be isolated and characterized with respect to phenotype and specificity. These analyses have revealed that all types of cytotoxic effector cells are regulated by interactions with MHC-peptide ligands, however, the consequences of these interactions can result in opposite functional outcomes. In this review we summarize how allogeneic MHC class I-peptide ligands positively or negatively regulate the activities of four distinct groups of cytotoxic lymphocytes and how this information might be transferred into clinical use.
Collapse
Affiliation(s)
- Christine S Falk
- Institute of Molecular Immunology, GSF National Research Center for the Environment and Health, Munich, Germany.
| | | |
Collapse
|
|
31
|
Abstract
Attempts to generate an anticancer immune response in vivo in patients with cancer have taken several forms. Although to date there have been relatively few published studies describing the effects of the approach in hematologic malignancy, that circumstance is expected to change rapidly during the next few years. In solid tumors, it is not known which, if any, of the approaches being explored will be able to produce responses of sufficient effectiveness and duration to be of general clinical value. Despite the documented increase in survival of patients developing an immune response to tumor immunization, no randomized clinical trial has been entirely convincing. As knowledge of the molecular basis of the immune response and of the immune defenses used by cancer cells improves, it is reasonable to expect to see increasing benefits from tumor vaccines, which are likely to complement, long before they replace, conventional therapies.
Collapse
Affiliation(s)
- Peter J. DeMaria
- Genitourinary Malignancies Branch, Center for Cancer Research, National
Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
| | - Marijo Bilusic
- Genitourinary Malignancies Branch, Center for Cancer Research, National
Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
| |
Collapse
|
|
32
|
So T, Takenoyama M, Sugaya M, Yasuda M, Eifuku R, Yoshimatsu T, Osaki T, Yasumoto K. Unfavorable prognosis of patients with non-small cell lung carcinoma associated with HLA-A2. Lung Cancer 2001; 32:39-46. [PMID: 11282427 DOI: 10.1016/s0169-5002(00)00204-x] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
BACKGROUND HLA class I molecules present antigenic peptides to cytotoxic T lymphocytes and, thus, play an important role in immune surveillance. Since 1970s there have been many reports of an increased frequency of one or more HLA haplotype in association with autoimmune disease, and malignancy. We studied types of HLA class I antigens in 204 resected non-small cell lung carcinoma (NSCLC) patients and also examined its correlation with clinicopathologic features and prognosis. METHOD Serological typing for HLA class I antigens was performed using a microcytotoxicity test. The disease-free survival curves were calculated by the Kaplan-Meier method and then compared using the Logrank test. Multivariate analysis was carried out by Cox's proportional hazard method. RESULTS The difference in disease-free survival time between the HLA-A2 present group and A2 absent group was significant (P = 0.040). The 3-year disease-free survival rate of all patients was 44% in HLA-A2 present group and 66% in A2 absent group. When a comparison was made within the group with stage I, expression of HLA-A2 was the only independent factor that affected survival time by multivariate analysis (P = 0.0457). CONCLUSIONS Expression of HLA-A2 was considered as one of the unfavorable prognostic factors in NSCLC patients. Our results suggested expression of HLA-A2 in NSCLC patients was one of the mechanisms of escape from immune surveillance.
Collapse
Affiliation(s)
- T So
- Department of Surgery II, School of Medicine, University of Occupational and Environmental Health, Iseigaoka 1-1, Yahatanishi-ku, Kitakyushu 807-8555, Japan.
| | | | | | | | | | | | | | | |
Collapse
|
|
33
|
Fernández MA, Ruiz-Cabello F, Oliva MR, Cabrera T, Jimenez P, López Nevot MA, Garrido F. Beta2-microglobulin gene mutation is not a common mechanism of HLA class I total loss in human tumors. INTERNATIONAL JOURNAL OF CLINICAL & LABORATORY RESEARCH 2001; 30:87-92. [PMID: 11043502 DOI: 10.1007/bf02874164] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
Abstract
One hundred and sixty-two tumor samples were analyzed for HLA class I expression using immunohistological techniques. HLA class I total loss (phenotype no. I) was detected in 31 cases (19%), comprising 20 colorectal, 3 laryngeal, and 2 bladder carcinomas and 6 melanomas. Twenty-one cases were selected for molecular analysis due to a higher proportion of tumor cells versus stroma cells (75%). We investigated whether beta2-microglobulin mutation was responsible for HLA downregulation. Single-strand conformation polymorphism and sequencing analysis of DNA samples was performed. Alterations were detected only in melanomas M78 (a point mutation in the initiation ATG sequence), M79 (a mutation in codon 31 producing a stop codon), and M34 (a TTCT deletion introducing a termination codon signal). We found no beta2-microglobulin gene mutation in the other 18 samples. Loss of heterozygosity in 15q close to the beta2-microglobulin gene was found in 5 cases. We conclude that HLA class I total loss can frequently occur without beta2-microglobulin gene mutations.
Collapse
Affiliation(s)
- M A Fernández
- Servicio de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Spain
| | | | | | | | | | | | | |
Collapse
|
|
34
|
Melief CJ, Toes RE, Medema JP, van der Burg SH, Ossendorp F, Offringa R. Strategies for immunotherapy of cancer. Adv Immunol 2001; 75:235-82. [PMID: 10879286 DOI: 10.1016/s0065-2776(00)75006-1] [Citation(s) in RCA: 119] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
MESH Headings
- Adjuvants, Immunologic
- Animals
- Antibodies, Monoclonal/therapeutic use
- Antibodies, Neoplasm/therapeutic use
- Antigen Presentation
- Antigens, CD/physiology
- Antigens, Neoplasm/immunology
- Apoptosis
- Cancer Vaccines/therapeutic use
- Cytokines/genetics
- Cytokines/physiology
- Disease Susceptibility
- Genetic Therapy
- Humans
- Immune Tolerance
- Immunity, Innate
- Immunoglobulin Idiotypes/immunology
- Immunologic Deficiency Syndromes/complications
- Immunologic Deficiency Syndromes/immunology
- Immunotherapy/methods
- Immunotherapy, Active
- Immunotherapy, Adoptive
- Lymphocyte Cooperation
- Lymphocytes, Tumor-Infiltrating/immunology
- Mice
- Neoplasm Proteins/immunology
- Neoplasms/etiology
- Neoplasms/immunology
- Neoplasms/prevention & control
- Neoplasms/therapy
- Neoplasms, Experimental/immunology
- Neoplasms, Experimental/therapy
- Oncogenic Viruses/immunology
- Receptors, Tumor Necrosis Factor/physiology
- T-Lymphocyte Subsets/immunology
- Tumor Virus Infections/immunology
Collapse
Affiliation(s)
- C J Melief
- Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, The Netherlands
| | | | | | | | | | | |
Collapse
|
|
35
|
Palmisano GL, Pistillo MP, Capanni P, Pera C, Nicolò G, Salvi S, Perdelli L, Pasciucco G, Ferrara GB. Investigation of HLA class I downregulation in breast cancer by RT-PCR. Hum Immunol 2001; 62:133-9. [PMID: 11182222 DOI: 10.1016/s0198-8859(00)00241-x] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
Downregulation of HLA class I antigen expression has been reported in a significant proportion of primary breast carcinomas suggesting an escape mechanism from CTL mediated lysis leading to tumor dissemination and metastasis. We have previously reported the biochemical and immunohistochemical analysis of HLA total class I (W6/32 mAb), alpha-chain (Q1/28,TP25.99 mAbs) and beta(2)-microglobulin (Namb-1 mAb) subunits expression in 25 primary breast carcinomas. This study at protein level resulted in the observation of three different HLA class I expression patterns by both techniques: high, low, and absent downregulation patterns. To better characterize the HLA class I antigens downregulation we extended such analysis also at RNA level by RT-PCR using HLA-A, HLA-B, HLA-C, and beta(2)-microglobulin specific primers either in breast cancer or normal tissues derived from the same patient. None (100%) of the alpha-chain genes analyzed in patient tumor tissues showed significant reduction of expression. In 10 patients out of 25 (40%) the beta(2)-microglobulin gene showed complete loss of expression compared with the corresponding normal tissue counterpart, which showed a constitutive expression, whereas in 2 patients (12.5%) its expression was comparable with the normal counterpart. Sequence analysis at genomic level revealed no defects affecting beta(2)-microglobulin gene in those patients showing lack of expression. Also TAP1 and TAP2 genes expression were investigated in order to confirm or exclude involvement of the MHC class I molecules assembling machinery. The RT-PCR approach mainly confirmed our beta(2)-microglobulin biochemical analysis indicating that in breast cancer specimens it is possible to address the HLA class I gene downregulation as a phenomenon occurring at post-transcriptional level mainly affecting the beta(2)-microglobulin gene expression.
Collapse
Affiliation(s)
- G L Palmisano
- Immunogenetics Laboratory, National Cancer Institute and Department of Oncology, Biology and Genetics c/o Advanced Biotechnology Center (CBA), University of Genova, Italy
| | | | | | | | | | | | | | | | | |
Collapse
|
|
36
|
Garcia-Lora A, Algarra I, Gaforio JJ, Ruiz-Cabello F, Garrido F. Immunoselection by T lymphocytes generates repeated MHC class I-deficient metastatic tumor variants. Int J Cancer 2001; 91:109-19. [PMID: 11149409 DOI: 10.1002/1097-0215(20010101)91:1<109::aid-ijc1017>3.0.co;2-e] [Citation(s) in RCA: 64] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Alteration of MHC class I molecule expression is a widespread mechanism used by tumor cells to evade T cell responses. It has long been proposed that the origin of these MHC class I-negative or -deficient tumor variants is T cell immune selection. However, there are no experimental or clinical data to substantiate this hypothesis, and this issue is currently the subject of debate. Here we report that an H-2 class I-negative fibrosarcoma tumor clone generated MHC class I-negative spontaneous lung metastases in immunocompetent syngeneic BALB/c mice. Interestingly, the same B9 clone generated MHC class I-positive metastatic nodes, under basal conditions, in athymic nu/nu BALB/c mice. This phenomenon was observed in the metastatic nodules generated after a period of in vivo growth but not in the primary tumors growing locally in the footpad. These findings support the hypothesis that the H-2 phenotype of metastatic nodes is influenced by the T cell repertoire of the host, since in the absence of this T cell pressure (i.e., in nude mice) the metastatic nodes 'recovered' H-2 class I expression. In addition, 2 different phenotypes were found when the metastatic nodules obtained from immunocompetent mice were treated with IFN-gamma. One phenotype, present in 83% of the colonies, was characterized by resistance of the Ld molecule to IFN-gamma induction, due to a deletion involving the Ld gene. The second phenotype (17% of the colonies) was similar to the original B9 clone and was characterized by the response of K, D and L class I genes to IFN-gamma. These data provide evidence that the changes in MHC class I expression during tumor development might not be random but could be predictable.
Collapse
Affiliation(s)
- A Garcia-Lora
- Servicio de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Spain
| | | | | | | | | |
Collapse
|
|
37
|
Seliger B, Schreiber K, Delp K, Meissner M, Hammers S, Reichert T, Pawlischko K, Tampé R, Huber C. Downregulation of the constitutive tapasin expression in human tumor cells of distinct origin and its transcriptional upregulation by cytokines. TISSUE ANTIGENS 2001; 57:39-45. [PMID: 11169257 DOI: 10.1034/j.1399-0039.2001.057001039.x] [Citation(s) in RCA: 50] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
Human tumor cells frequently exhibit abnormalities in the major histocompatibility complex (MHC) class I surface expression which can be due to structural alterations and/or dysregulation of various components of the MHC class I antigen processing machinery, such as HLA class I heavy and light chains, the peptide transporter and the proteasome subunits. Although several cofactors critical for proper MHC class I assembly have been identified, their contribution to the immune escape phenotype of tumor cells has not been analyzed. In order to determine whether tapasin deficits are an integral part of immune escape mechanisms of human tumors, we studied the constitutive and cytokine-regulated expression pattern of tapasin in malignant cells of distinct histology. Heterogeneous and reduced expression levels of tapasin were found in small-cell lung carcinoma, pancreatic carcinoma, colon carcinoma, head an neck squamous cell carcinoma and renal cell carcinoma cell lines. Tapasin downregulation was also prominent in surgically removed tumor lesions when compared to normal controls. The impaired tapasin expression is often associated with low MHC class I cell surface expression. In addition, various cytokines, including interferon (IFN)-alpha, IFN-gamma, tumor necrosis factor (TNF)-alpha and interleukin (IL)-4, but not granulocyte-macrophage colony stimulating factor (GM-CSF), transcriptionally upregulate to a distinct extent and in a time-dependent manner tapasin expression in tumor cells. Thus, deficient tapasin expression appears to be a frequent event in human tumor cells. Its restoration by cytokines further suggests that impaired tapasin expression in tumors is rather due to dysregulation than to structural alterations.
Collapse
Affiliation(s)
- B Seliger
- The Johannes Gutenberg-University, IIIrd Department of Internal Medicine, Mainz, Germany.
| | | | | | | | | | | | | | | | | |
Collapse
|
|
38
|
Extensive genetic alterations of the HLA region, including homozygous deletions of HLA class II genes in B-cell lymphomas arising in immune-privileged sites. Blood 2000. [DOI: 10.1182/blood.v96.10.3569.h8003569_3569_3577] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
In B-cell lymphomas, loss of human leukocyte antigen (HLA) class I and II molecules might contribute to immune escape from CD8+ and CD4+ cytotoxic T cells, especially because B cells can present their own idiotype. Loss of HLA expression and the possible underlying genomic alterations were studied in 28 testicular, 11 central nervous system, and 21 nodal diffuse large B-cell lymphomas (DLCLs), the first two sites are considered as immune-privileged sites. The analysis included immunohistochemistry, loss of heterozygosity analysis, and fluorescent in situ hybridization (FISH) on interphase cells and isolated DNA fibers. Total loss of HLA-A expression was found in 60% of the extranodal cases and in 10% of the nodal cases (P < .01), whereas loss of HLA-DR expression was found in 56% and 5%, respectively (P < .01). This was accompanied by extensive loss of heterozygosity within the HLA region in the extranodal DLCLs. In 3 cases, retention of heterozygosity for D6S1666 in the class II region suggested a homozygous deletion. This finding was confirmed by interphase FISH that showed homozygous deletions in the class II genes in 11 of the 18 extranodal lymphomas but in none of the 7 nodal DLCLs (P < .001). Mapping by fiber FISH showed variable deletions that always included HLA-DQ and HLA-DR genes. Hemizygous deletions and mitotic recombinations often involving all HLA genes were found in 13 of 18 extranodal and 2 of 7 nodal lymphomas. In conclusion, a structural loss of HLA class I and II expression might help the B-cell lymphoma cells to escape from immune attack.
Collapse
|
|
39
|
Extensive genetic alterations of the HLA region, including homozygous deletions of HLA class II genes in B-cell lymphomas arising in immune-privileged sites. Blood 2000. [DOI: 10.1182/blood.v96.10.3569] [Citation(s) in RCA: 146] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Abstract
In B-cell lymphomas, loss of human leukocyte antigen (HLA) class I and II molecules might contribute to immune escape from CD8+ and CD4+ cytotoxic T cells, especially because B cells can present their own idiotype. Loss of HLA expression and the possible underlying genomic alterations were studied in 28 testicular, 11 central nervous system, and 21 nodal diffuse large B-cell lymphomas (DLCLs), the first two sites are considered as immune-privileged sites. The analysis included immunohistochemistry, loss of heterozygosity analysis, and fluorescent in situ hybridization (FISH) on interphase cells and isolated DNA fibers. Total loss of HLA-A expression was found in 60% of the extranodal cases and in 10% of the nodal cases (P < .01), whereas loss of HLA-DR expression was found in 56% and 5%, respectively (P < .01). This was accompanied by extensive loss of heterozygosity within the HLA region in the extranodal DLCLs. In 3 cases, retention of heterozygosity for D6S1666 in the class II region suggested a homozygous deletion. This finding was confirmed by interphase FISH that showed homozygous deletions in the class II genes in 11 of the 18 extranodal lymphomas but in none of the 7 nodal DLCLs (P < .001). Mapping by fiber FISH showed variable deletions that always included HLA-DQ and HLA-DR genes. Hemizygous deletions and mitotic recombinations often involving all HLA genes were found in 13 of 18 extranodal and 2 of 7 nodal lymphomas. In conclusion, a structural loss of HLA class I and II expression might help the B-cell lymphoma cells to escape from immune attack.
Collapse
|
|
40
|
Friedl J, Stift A, Paolini P, Roth E, Steger GG, Mader R, Jakesz R, Gnant MF. Tumor antigen pulsed dendritic cells enhance the cytolytic activity of tumor infiltrating lymphocytes in human hepatocellular cancer. Cancer Biother Radiopharm 2000; 15:477-86. [PMID: 11155819 DOI: 10.1089/cbr.2000.15.477] [Citation(s) in RCA: 21] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023] Open
Abstract
OBJECTIVE Tumor infiltrating lymphocytes (TILs) stimulated with interleukin-2 (IL-2) ex vivo have been successfully used therapeutically in some cancer patients, but their potency in eliciting an effective anti-tumor response is variable. We have tried to augment killing activity of tumor infiltrating lymphocytes derived from hepato-cellular carcinoma (HCC) using autologous monocytes derived dendritic cells. METHODS Tumor infiltrating lymphocytes (TILs) from 6 patient with hepatocellular carcinoma were isolated and the phenotype were further characterized. From the same patients, autologous dendritic cells were generated from CD14+ monocytes that were cultured for 6 days in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4). Those professional antigen presenting cells were pulsed with whole autologous hepatoma tumor lysates (pDC). TILs were cocultured with pDC or unpulsed DC. To assess the cytotoxic potency of TILs, the ability to lyse the tumor cell targets K652, Daudi and an allogeneic HCC celline was determined in a standard cytotoxic assay. RESULTS Tumor cells targets in vitro are poorly lysed by tumor infiltrating lymphocytes indicating T-cell hyporesponsiveness. In contrast, the killing activity of HCC derived TILs against Daudi (9.15% +/- 7.5) and allogeneic HCC tumor target (18.2% +/- 9.2) could be significantly augmented when stimulated with pDC (Daudi: 38% +/- 6.8 and allogeneic HCC: 55% +/- 10). The killing activity of TILs against K562 was unaffected by pDC. CONCLUSION The low cytotoxic activity profile of HCC derived TILs in vitro can be increased by tumor lysate pulsed dendritic cells and may therefore be more effective in vivo when used for adoptive immunotherapy.
Collapse
Affiliation(s)
- J Friedl
- Department of Surgery, University of Vienna, General Hospital, Waehringer Guertel 18-20, A-1090 Wien, Austria
| | | | | | | | | | | | | | | |
Collapse
|
|
41
|
Cabrera T, Salinero J, Fernandez MA, Garrido A, Esquivias J, Garrido F. High frequency of altered HLA class I phenotypes in laryngeal carcinomas. Hum Immunol 2000; 61:499-506. [PMID: 10773352 DOI: 10.1016/s0198-8859(00)00097-5] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/23/2022]
Abstract
The exact frequency of HLA class I losses in human tumors is unknown. We have previously shown that primary breast and colorectal carcinomas frequently lose HLA class I molecules (88% and 73%, respectively). Now we report that this phenomenon is also a frequent event in laryngeal carcinomas. Of a total of 76 laryngeal carcinomas analyzed, 66% of the tumors showed an alteration in HLA class I phenotype. These altered HLA phenotypes were classified as total HLA loss (10.52%) (phenotype I); HLA-A locus-specific loss (13.15%) (phenotype IIIa); HLA-B locus-specific loss (10.52%) (phenotype IIIb); HLA allelic loss (27.63%) (phenotype IV); and HLA-A and B locus loss (3.9%). Comparison of histopathological parameters with HLA expression showed that poorly differentiated tumors had the lowest levels of HLA class I expression (p < 0.05).
Collapse
Affiliation(s)
- T Cabrera
- Departamento de Análisis Clínicos, Universidad de Granada, Spain
| | | | | | | | | | | |
Collapse
|
|
42
|
Tortorella D, Gewurz B, Schust D, Furman M, Ploegh H. Down-regulation of MHC class I antigen presentation by HCMV; lessons for tumor immunology. Immunol Invest 2000; 29:97-100. [PMID: 10854174 DOI: 10.3109/08820130009062289] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Affiliation(s)
- D Tortorella
- Harvard Medical School, Department of Pathology, Boston, Massachusetts 02115, USA
| | | | | | | | | |
Collapse
|
|
43
|
Abstract
It is generally accepted that human and experimental tumor cells can lose major histocompatibility complex (MHC) class I molecules. These human leukocyte antigen (HLA) losses are detected when the primary tumor breaks the basal membrane, invades the surrounding tissues, and starts to metastasize. These altered HLA class I phenotypes probably constitute the major tumor escape mechanism facing anti-tumor T-cell mediated responses. Thus, it is important to characterize these phenotypes in clinical tumor samples, analyze the mechanism(s) responsible for them, and counsel patients before and during peptide anti-cancer immunotherapy. The present paper summarizes the most relevant altered HLA class I phenotypes found in human tumor samples, indicates their frequency, and outlines the mechanisms implicated. This review also points out that the natural killer (NK) escape mechanism of HLA class I deficient cancer cells is yet to be defined. Knowledge accumulated to date reveals that HLA class I molecules are an important crossroad in tumor immunology.
Collapse
Affiliation(s)
- I Algarra
- Departamento de Ciencias de la Salud, Facultad de Ciencias Experimentales, Universidad de Jaen, Spain
| | | | | |
Collapse
|
|
44
|
Jiménez P, Cantón J, Collado A, Cabrera T, Serrano A, Real LM, García A, Ruiz-Cabello F, Garrido F. Chromosome loss is the most frequent mechanism contributing to HLA haplotype loss in human tumors. Int J Cancer 1999; 83:91-7. [PMID: 10449614 DOI: 10.1002/(sici)1097-0215(19990924)83:1<91::aid-ijc17>3.0.co;2-4] [Citation(s) in RCA: 91] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
Loss of heterozygosity (LOH) in the short arm of chromosome 6 (6p) was detected in samples obtained from colon (13.8%), larynx (17.6%) and melanoma (15.3%) tumors. The parallel study of HLA-antigen expression in tumor tissues using locus- and polymorphic-specific antibodies in combination with LOH microsatellite analysis on 6p allowed us to establish that LOH in chromosome 6 is a representative phenomenon in most tumor cells present in a given tumor tissue. In most cases, specific HLA alleles had been lost in a predominant population of tumor cells, indicating that LOH is a non-irrelevant mutation that probably confers a selective advantage for survival of the mutant cell. We also demonstrate that LOH frequently occurred through chromosome loss rather than somatic recombination. LOH at all loci studied on the p and q arms of chromosome 6 was observed in at least 56.2% (9/17) cases. This HLA-associated microsatellite analysis was a useful tool for classifying tumors as LOH-positive or -negative, and therefore to consider a patient as a potential non-responder or responder in a vaccination trial.
Collapse
Affiliation(s)
- P Jiménez
- Servicio de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Granada, Spain
| | | | | | | | | | | | | | | | | |
Collapse
|
|
45
|
Suzuki K, Sahara H, Okada Y, Yasoshima T, Hirohashi Y, Nabeta Y, Hirai I, Torigoe T, Takahashi S, Matsuura A, Takahashi N, Sasaki A, Suzuki M, Hamuro J, Ikeda H, Wada Y, Hirata K, Kikuchi K, Sato N. Identification of Natural Antigenic Peptides of a Human Gastric Signet Ring Cell Carcinoma Recognized by HLA-A31-Restricted Cytotoxic T Lymphocytes. THE JOURNAL OF IMMUNOLOGY 1999. [DOI: 10.4049/jimmunol.163.5.2783] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/02/2023]
Abstract
Abstract
Peptides of human melanomas recognized by CD8+ CTLs have been identified, but the nature of those of nonmelanoma tumors remains to be elucidated. Previously, we established a gastric signet ring cell carcinoma HST-2 and HLA-A31 (A*31012)-restricted autologous CTL clone, TcHST-2. In the present study, we determined the natural antigenic peptides of HST-2 cells. The purified preparation of acid-extracted Ags was submitted to the peptide sequencer, and one peptide, designated F4.2 (Tyr-Ser-Trp-Met-Asp-Ile-Ser-Cys-Trp-Ile), appeared to be immunogenic. To confirm the antigenicity of F4.2 further, we constructed an expression minigene vector (pF4.2ss) coding adenovirus E3, a 19-kDa protein signal sequence plus F4.2. An introduction of pF4.2ss minigene to HST-2 and HLA-A31(+) allogeneic tumor cells clearly enhanced and induced the TcHST-2 reactivity, respectively. Furthermore, when synthetic peptides of F4.2 C-terminal-deleted peptides were pulsed to HST-2 cells, F4.2-9 (nonamers), but not F4.2-8 or F4.2-7 (octamer or heptamer, respectively), enhanced the reactivity of TcHST-2, suggesting that the N-terminal ninth Trp might be a T cell epitope. This was confirmed by lack of antigenicity when using synthetic substituted peptides as well as minigenes coding F4.2 variant peptides with Ala or Arg at the ninth position of F4.2. Meanwhile, it was indicated that the sixth position Ile was critically important for the binding to HLA-A31 molecules. Thus, our data indicate that F4.2 may work as an HLA-A31-restricted natural antigenic peptide recognized by CTLs.
Collapse
Affiliation(s)
- Kazuhiro Suzuki
- *Pathology and
- †Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan
| | - Hiroeki Sahara
- *Pathology and
- ‡Marine Biomedical Institute, Sapporo Medical University School of Medicine, Rishirifuji-cho, Hokkaido, Japan
| | - Yohjiro Okada
- *Pathology and
- †Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan
| | | | - Yoshihiko Hirohashi
- *Pathology and
- §Department of Otolaryngology, Wakayama Medical College, Wakayama, Japan
| | | | | | | | | | | | - Nobuaki Takahashi
- *Pathology and
- ‡Marine Biomedical Institute, Sapporo Medical University School of Medicine, Rishirifuji-cho, Hokkaido, Japan
| | | | - Manabu Suzuki
- ¶Central Research Laboratories, Ajinomoto Co. Inc., Kawasaki, Japan
| | - Junji Hamuro
- ¶Central Research Laboratories, Ajinomoto Co. Inc., Kawasaki, Japan
| | - Hideyuki Ikeda
- ∥Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan; and
| | - Yoshimasa Wada
- †Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan
| | - Koichi Hirata
- †Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan
| | - Kokichi Kikuchi
- *Pathology and
- #Sapporo Immunodiagnostic Laboratory, Sapporo, Japan
| | | |
Collapse
|