• Humans
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/pathology
• Carcinoma, Hepatocellular/metabolism
• Liver Neoplasms/genetics
• Liver Neoplasms/pathology
• Liver Neoplasms/metabolism
• Proto-Oncogene Proteins c-akt/metabolism
• Proto-Oncogene Proteins c-akt/genetics
• TOR Serine-Threonine Kinases/metabolism
• TOR Serine-Threonine Kinases/genetics
• Phosphatidylinositol 3-Kinases/metabolism
• Phosphatidylinositol 3-Kinases/genetics
• Signal Transduction
• Disease Progression
• Gene Expression Regulation, Neoplastic
• Cell Proliferation/genetics
• Cell Line, Tumor
• Tumor Microenvironment/genetics
• Single-Cell Analysis
• Sequence Analysis, RNA
• Prognosis
• Cell Movement/genetics
• Male
• Female
• Adaptor Proteins, Signal Transducing/genetics
• Adaptor Proteins, Signal Transducing/metabolism
• Biomarkers, Tumor/genetics

• National Natural Science Foundation of China
• Key Project with Chinese Medicine Characteristics and Advantages, Ruikang Hospital, 2023 (Funding for Chinese Medicine Service System and Capacity Building)
• Nature Science Foundation of Guangxi

• Colorectal Cancer
• Immune Checkpoint Inhibitor
• Macrophage

• Colorectal Neoplasms/drug therapy
• Colorectal Neoplasms/pathology
• Colorectal Neoplasms/metabolism
• Humans
• Animals
• Tumor-Associated Macrophages/metabolism
• Tumor-Associated Macrophages/immunology
• Mice
• Drug Resistance, Neoplasm
• Signal Transduction
• Macrophage Colony-Stimulating Factor/metabolism
• Immune Checkpoint Inhibitors/pharmacology
• Immune Checkpoint Inhibitors/therapeutic use
• Tumor Microenvironment
• Cell Line, Tumor
• Mice, Inbred C57BL
• Female

• BAMBI
• HCC
• cell signaling pathways
• macrophage polarization
• prognosis

• Liver Neoplasms/metabolism
• Liver Neoplasms/pathology
• Liver Neoplasms/genetics
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/pathology
• Carcinoma, Hepatocellular/genetics
• Humans
• Glycolysis/genetics
• Lipid Metabolism/genetics
• Prognosis
• Biomarkers, Tumor/metabolism
• Biomarkers, Tumor/genetics
• Membrane Proteins/metabolism
• Membrane Proteins/genetics
• Macrophages/metabolism
• Animals
• Epithelial-Mesenchymal Transition/genetics
• Gene Expression Regulation, Neoplastic
• Male
• Mice
• Cell Line, Tumor
• Female
• Cell Proliferation

• immune cell
• oncolytic virus
• treatment
• tumor
• tumor microenvironment

• Humans
• Tumor Microenvironment/immunology
• Oncolytic Viruses/immunology
• Oncolytic Viruses/genetics
• Oncolytic Virotherapy/methods
• Neoplasms/therapy
• Neoplasms/immunology
• Animals

• National Natural Science Foundation of China

The relevant mechanism of tumor-associated macrophages (TAMs) in the treatment of colorectal cancer patients with immune checkpoint inhibitors (ICIs) is discussed, and the application prospects of TAMs in reversing the treatment tolerance of ICIs are discussed to provide a reference for related studies. As a class of drugs widely used in clinical tumor immunotherapy, ICIs can act on regulatory molecules on cells that play an inhibitory role-immune checkpoints-and kill tumors in the form of an immune response by activating a variety of immune cells in the immune system. The sensitivity of patients with different types of colorectal cancer to ICI treatment varies greatly. The phenotype and function of TAMs in the colorectal cancer microenvironment are closely related to the efficacy of ICIs. ICIs can regulate the phenotypic function of TAMs, and TAMs can also affect the tolerance of colorectal cancer to ICI therapy. TAMs play an important role in ICI resistance, and making full use of this target as a therapeutic strategy is expected to improve the immunotherapy efficacy and prognosis of patients with colorectal cancer.

• Colorectal cancer
• Immune checkpoint inhibitor resistance
• Tumor microenvironment
• Tumor-associated macrophages
• Review

• CCA
• TAMs
• immune evasion
• macrophage reprogramming
• tumor microenvironment

• Humans
• Cholangiocarcinoma/immunology
• Cholangiocarcinoma/pathology
• Cholangiocarcinoma/etiology
• Cholangiocarcinoma/therapy
• Tumor-Associated Macrophages/immunology
• Tumor-Associated Macrophages/metabolism
• Bile Duct Neoplasms/immunology
• Bile Duct Neoplasms/pathology
• Bile Duct Neoplasms/therapy
• Bile Duct Neoplasms/etiology
• Tumor Microenvironment/immunology
• Disease Progression
• Animals
• Tumor Escape

• Intestinal flora metabolites
• Macrophage
• Polarization

• Colorectal cancer
• M1/M2
• Macrophages
• Phytochemicals

• Colorectal Neoplasms/drug therapy
• Phytochemicals/pharmacology
• Phytochemicals/therapeutic use
• Macrophages/drug effects
• Humans
• Animals
• Disease Progression
• Antineoplastic Agents, Phytogenic/pharmacology
• Tumor Microenvironment/drug effects
• Reactive Oxygen Species/metabolism

• cancer
• drug resistance
• immunotherapy
• tumor microenvironment
• tumor-associated macrophages

• Humans
• Drug Resistance, Neoplasm
• Tumor-Associated Macrophages/drug effects
• Tumor-Associated Macrophages/immunology
• Tumor-Associated Macrophages/metabolism
• Tumor Microenvironment/drug effects
• Tumor Microenvironment/immunology
• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use
• Neoplasms/drug therapy
• Neoplasms/immunology
• Animals

• CCL2
• CCL5
• CXCL8 (IL8)
• CXCR2
• chemokine receptor
• chemokines
• endometrial cancer
• single nucleotide polymorphisms (SNPs)

• Inflammasome
• Interferon-γ
• Macrophage
• NLRP3
• Placental mesenchymal stem cells
• Premature ovarian insufficiency
• Pyroptosis

• No. 81901439 the National Natural Science Foundation of China
• 2020BEG03042 The Key R&D Program of Ninxia, Yinchuan, China

• biomarkers
• clinical implementation
• colorectal cancer
• genetic susceptibility factors
• immune system
• immunotherapy
• mCRC
• personalized medicine
• pharmacogenetics
• precision medicine

• hepatocellular carcinoma
• immunotherapy
• intrahepatic cholangiocarcinoma
• tumor-associated macrophages

Skin malignant melanoma is a highly aggressive skin tumor, which is also a major cause of skin cancer-related mortality. It can spread from a relatively small primary tumor and metastasize to multiple locations, including lymph nodes, lungs, liver, bone, and brain. What’s more metastatic melanoma is the main cause of its high mortality. Among all organs, the lung is one of the most common distant metastatic sites of melanoma, and the mortality rate of melanoma lung metastasis is also very high. Elucidating the mechanisms involved in the pulmonary metastasis of cutaneous melanoma will not only help to provide possible explanations for its etiology and progression but may also help to provide potential new therapeutic targets for its treatment. Increasing evidence suggests that tumor-associated macrophages (TAMs) play an important regulatory role in the migration and metastasis of various malignant tumors. Tumor-targeted therapy, targeting tumor-associated macrophages is thus attracting attention, particularly for advanced tumors and metastatic tumors. However, the relevant role of tumor-associated macrophages in cutaneous melanoma lung metastasis is still unclear. This review will present an overview of the origin, classification, polarization, recruitment, regulation and targeting treatment of tumor-associated macrophages, as well as the soluble mediators involved in these processes and a summary of their possible role in lung metastasis from cutaneous malignant melanoma. This review particularly aims to provide insight into mechanisms and potential therapeutic targets to readers, interested in pulmonary metastasis melanoma.

• Humans
• Lung/pathology
• Lung Neoplasms/pathology
• Melanoma/pathology
• Neoplasms, Second Primary
• Skin Neoplasms/pathology
• Tumor-Associated Macrophages

Onco-immunogenic molecule CD155 is overexpressed in various tumor microenvironments (TME) including in colorectal cancer (CRC). Tumor-associated macrophages (TAMs) are the most abundant immune cells in CRC TME and play a vital role in CRC progression and metastasis. Most studies have focused on investigating the role of CRC cell-specific CD155 on CRC progression, while the contribution of TAMs-specific CD155 is still unknown. Here, we sought to investigate the expression pattern of CD155 in CRC TAMs and its role in tumor immunity and progression.

CD155 expression patterns in CRC TAMs and macrophages in paratumor or adjacent normal tissue were analyzed in 50 patients with CRC using flow cytometry and in 141 patients with CRC using immunohistochemistry. The correlation of CD155 expression level in TAMs with M1 and M2 phenotypic transition was analyzed. The role of macrophage-specific CD155 in CRC progression and tumor immune response was investigated in vitro and in vivo. We further analyzed the effect of CRC cells on the regulation of CD155 expression in macrophages.

CRC TAMs from clinical samples showed robustly higher expression of CD155 than macrophages from paratumor and adjacent normal tissues. The CD155 expression level was higher in TAMs of CRC at III/IV stages compared with the I/II stages and was negatively associated with the survival of patients with CRC. CD155⁺ TAMs showed an M2 phenotype and higher expression of interleukin (IL)-10 and transforming growth factor (TGF)-β. CD155⁺ macrophages promoted CRC cell migration, invasion, and tumor growth supporting the findings from the clinical tissue analysis. This effect was mainly regulated by TGF-β-induced STAT3 activation-mediated release of matrix metalloproteinases (MMP)2 and MMP9 in CRC cells. CD155^–⁄– bone marrow transplantation in wild-type mice, as well as CD155– macrophages treatment, promoted the antitumor immune response in the mice ectopic CRC model. Additionally, CRC cells released IL-4 to trigger CD155 expression in macrophages indicating the regulatory role of CRC cells in the development of CD155⁺ TAMs.

These findings indicated that CD155⁺ TAMs are responsible for the M2-phenotype transition, immunosuppression, and tumor progression in CRC. The specific localization of CD155⁺ TAMs in CRC tissue could turn into a potential therapeutic target for CRC treatment.

• immunity
• tumor escape
• tumor microenvironment

• ADME
• biomarkers
• colorectal cancer
• germline variants
• immunity
• immunogenetics
• inflammation
• microenviroment
• pharmacogenetics

• cancer
• innate immunity
• macrophage
• macrophage activation
• macrophage biology
• mitochondrial DNA

• 82071984; BK20191223; K2019024; SJCX21_1723 National Natural Science Foundation of China; the Social Development Foundation of Jiangsu Province; the Key Programme of Jiangsu Commission of Health; the Postgraduate Innovation Project of Jiangsu Province

Colorectal cancer accounts for approximately 10% of all annually diagnosed cancers worldwide being liver metastasis, the most common cause of death in patients with colorectal cancer. The interplay between tumor and stromal cells in the primary tumor microenvironment and at distant metastases are rising in importance as potential mechanisms of the tumor progression. In this review we discuss the new biomarkers derived from tumor microenvironment and liquid biopsy as emerging prognostic and treatments response markers for metastatic colorectal cancer. We also review the developing new clinical strategies based on tumor microenvironmental cells to tackle metastatic disease in metastatic colorectal cancer patients.

Colorectal cancer (CRC) is one of the most common cancers in western countries. Its mortality rate varies greatly, depending on the stage of the disease. The main cause of CRC mortality is metastasis, which most commonly affects the liver. The role of tumor microenvironment in tumor initiation, progression and metastasis development has been widely studied. In this review we summarize the role of the tumor microenvironment in the liver pre-metastatic niche formation, paying attention to the distant cellular crosstalk mediated by exosomes. Moreover, and based on the prognostic and predictive capacity of alterations in the stromal compartment of tumors, we describe the role of tumor microenvironment cells and related liquid biopsy biomarkers in the delivery of precise medication for metastatic CRC. Finally, we evaluate the different clinical strategies to prevent and treat liver metastatic disease, based on the targeting of the tumor microenvironment. Specifically, targeting angiogenesis pathways and regulating immune response are two important research pipelines that are being widely developed and promise great benefits.

• biomarkers
• colorectal cancer
• liquid biopsy
• liver metastases
• tumor microenvironment

Angiogenesis pathway genes show substantial genetic variability causing inter-individual differences in responses to anti-angiogenic drugs. We examined 20 single nucleotide polymorphisms (SNPs) in 13 of these genes to predict tumour response and clinical outcome measured as progression free survival (PFS) and overall survival (OS) in 57 patients with metastatic colorectal cancer (mCRC) given bevacizumab plus chemotherapy. SNPs were detected (iPLEX® Assay) in genomic DNA extracted from formalin-fixed paraffin-embedded tumour specimens. The variant allele CD39 rs11188513 was associated with a good tumour response (p = 0.024). Patients homozygous for the wild-type allele FGF2 rs1960669 showed a median PFS of 10.95 months versus 5.44 months for those with at least one variant allele-A (HR 3.30; 95% CI: 1.52–7.14; p = 0.001). Patients homozygous for wild-type MMP9 rs2236416 and rs2274755 showed a median PFS of 9.48 months versus 6 and 6.62 months, respectively, for those with at least one variant allele (p = 0.022, p = 0.043, respectively). OS was also lengthened to 30.92 months (p = 0.034) in carriers of wild-type ANGPT1 rs2445365 versus 22.07 months for those carrying at least one variant allele-A. These gene variants were able to predict clinical outcome and tumour response in mCRC patients given bevacizumab-based therapy.

• angiogenesis
• bevacizumab
• colorectal cancer
• polymorphisms

• Adult
• Aged
• Aged, 80 and over
• Angiopoietin-1/genetics
• Antineoplastic Combined Chemotherapy Protocols/pharmacology
• Antineoplastic Combined Chemotherapy Protocols/therapeutic use
• Apyrase/genetics
• Bevacizumab/pharmacology
• Bevacizumab/therapeutic use
• Biomarkers, Tumor/genetics
• Biopsy
• Colorectal Neoplasms/drug therapy
• Colorectal Neoplasms/genetics
• Colorectal Neoplasms/mortality
• Colorectal Neoplasms/pathology
• Drug Resistance, Neoplasm/genetics
• Female
• Fibroblast Growth Factor 2/genetics
• Humans
• Male
• Matrix Metalloproteinase 9/genetics
• Middle Aged
• Neovascularization, Pathologic/drug therapy
• Neovascularization, Pathologic/genetics
• Neovascularization, Pathologic/mortality
• Neovascularization, Pathologic/pathology
• Polymorphism, Single Nucleotide
• Progression-Free Survival
• Retrospective Studies

• biomarker
• chemotherapy
• hematogenous metastasis
• immunotherapy
• lymphatic metastasis
• monocyte, CD68
• tumor-associated macrophage

• cancer epidemiology
• colorectal cancer

• Adaptor Proteins, Signal Transducing/genetics
• Aged
• Angiogenesis Inhibitors/therapeutic use
• Apoptosis Regulatory Proteins/genetics
• Bevacizumab/therapeutic use
• Biomarkers, Tumor/genetics
• Cohort Studies
• Colorectal Neoplasms/drug therapy
• Colorectal Neoplasms/genetics
• Colorectal Neoplasms/therapy
• Combined Modality Therapy
• Endoplasmic Reticulum/genetics
• Endoplasmic Reticulum/metabolism
• Female
• Genetic Association Studies
• Humans
• Inflammation/genetics
• Machine Learning
• Male
• Membrane Proteins/genetics
• Middle Aged
• NLR Proteins
• Outcome and Process Assessment, Health Care/methods
• Polymorphism, Single Nucleotide
• Progression-Free Survival
• Proto-Oncogene Proteins p21(ras)/genetics
• Signal Transduction

• EC | EC Seventh Framework Programm | FP7 Health (FP7-HEALTH - Specific Programme "Cooperation": Health)
• Science Foundation Ireland (SFI)

• 12-chemkine signature
• TLS
• colorectal cancer
• recurrence
• tumor infiltrating immune cells

• Biomarkers, Tumor/genetics
• Chemokines/genetics
• Colorectal Neoplasms/genetics
• Colorectal Neoplasms/pathology
• Colorectal Neoplasms/surgery
• Databases, Genetic
• Female
• France
• Gene Expression Regulation, Neoplastic
• Humans
• Japan
• Male
• Neoplasm Staging
• Prognosis
• Survival Analysis
• Tertiary Lymphoid Structures/genetics
• United States

• P30 CA014089 NCI NIH HHS
• P30CA014089 NCI NIH HHS

• Hypertrophic scar
• Keloid
• Macrophages
• Predisposition
• Wound healing

• Youth Doctor Collaborative Innovation Team
• Shanghai Ninth People’s Hospital
• Shanghai Jiaotong University School of Medicine
• Shanghai Youth Top-Notch Talent Program
• National Natural Science Foundation of China
• Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant
• Shanghai “Rising Stars of Medical Talent” Youth Development Program
• Shanghai Jiaotong University “Chenxing” Youth Development Program

• angiogenesis
• pro-metastasis risk
• tumor metastasis
• vessel targeting drugs

• Antineoplastic Agents/therapeutic use
• Humans
• Neoplasm Metastasis/drug therapy
• Neoplasms/drug therapy
• Neovascularization, Pathologic/drug therapy

• Colorectal cancer
• NF-κB signaling
• Pharmacological inhibitors
• Tumorigenesis

• M2-polarized
• Tumor-associated macrophages
• breast cancer
• colitis-associated colon cancer
• triptolide
• tumor growth

• cancer immunology
• innate immunity
• monocytes
• myeloid cell heterogeneity
• neutrophils
• tumor microenvironment

• Animals
• Cell Differentiation/immunology
• Dendritic Cells/immunology
• Disease Progression
• Endothelial Cells/immunology
• Humans
• Immunity, Innate/immunology
• Macrophages/immunology
• Mice
• Monocytes/cytology
• Monocytes/immunology
• Neoplasms/immunology
• Neutrophils/immunology
• Tumor Microenvironment/immunology

Significant progress in the development of new immunotherapies has led to successful clinical trials for malignant melanoma and non-small cell lung cancer; however, for the majority of solid tumours of the gastrointestinal tract, little or no progress has been seen. The efficacy of immunotherapies is limited by the complexities of a diverse set of immune cells, and interactions between the tumour cells and all other cells in the local microenvironment of solid tumours. A large fraction of immune cells present in and around solid tumours derive from the innate arm of the immune system and using these cells against tumours offers an alternative immunotherapeutic option, especially as current strategies largely harness the adaptive arm of the immune system. This option is currently being investigated and attempts at using the innate immune system for gastrointestinal cancers are showing initial results. Several important factors, including cytokines, chemotherapeutics and the microbiome, influence the plasticity and functionality of innate (myeloid) cells in the microenvironment, and this complexity of regulation has limited translation into successful trials so far. In this review, current concepts of the immunobiology of the innate arm in the tumour microenvironment are presented in the context of clinical translation.

Macrophages play an important role in a wide variety of physiologic and pathologic processes. Plasticity and functional polarization are hallmarks of macrophages. Macrophages commonly exist in two distinct subsets: classically activated macrophages (M1) and alternatively activated macrophages (M2). M2b, a subtype of M2 macrophages, has attracted increasing attention over the past decade due to its strong immune‐regulated and anti‐inflammatory effects. A wide variety of stimuli and multiple factors modulate M2b macrophage polarization in vitro and in vivo. M2b macrophages possess both protective and pathogenic roles in various diseases. Understanding the mechanisms of M2b macrophage activation and the modulation of their polarization might provide a great perspective for the design of novel therapeutic strategies. The purpose of this review is to discuss current knowledge of M2b macrophage polarization, the roles of M2b macrophages in a variety of diseases and the stimuli to modulate M2b macrophage polarization.

• Autoimmunity
• Chemokines
• Disease pathogenesis
• Inflammation
• Manipulation of immune response
• Monocyte/Macrophage

Experimental models have often been at the origin of immunological paradigms such as the M1/M2 dichotomy following macrophage polarization. However, this clear dichotomy in animal models is not as obvious in humans, and the separating line between M1-like and M2-like macrophages is rather represented by a continuum, where boundaries are still unclear. Indeed, human infectious diseases, are characterized by either a back and forth or often a mixed profile between the pro-inflammatory microenvironment (dominated by interleukin (IL)-1β, IL-6, IL-12, IL-23 and Tumor Necrosis Factor (TNF)-α cytokines) and tissue injury driven by classically activated macrophages (M1-like) and wound healing driven by alternatively activated macrophages (M2-like) in an anti-inflammatory environment (dominated by IL-10, Transforming growth factor (TGF)-β, chemokine ligand (CCL)1, CCL2, CCL17, CCL18, and CCL22). This review brews the complexity of the situation during infectious diseases by stressing on this continuum between M1-like and M2-like extremes. We first discuss the basic biology of macrophage polarization, function, and role in the inflammatory process and its resolution. Secondly, we discuss the relevance of the macrophage polarization continuum during infectious and neglected diseases, and the possibility to interfere with such activation states as a promising therapeutic strategy in the treatment of such diseases.

• M1-like/M2-like
• human macrophage
• infectious diseases
• inflammation
• leishmaniasis
• polarization

Macrophages play a crucial role in the interaction between tumor and immune system, and iNOS is known as a surrogate marker of M1 macrophages activation. The goal of the study was to investigate the role of iNOS polymorphisms as prognostic marker in mCRC patients.

Functional significant polymorphisms in the promoter of INOS gene were analyzed by PCR-based and direct DNA sequencing in 4 cohorts of patients receiving bevacizumab based first-line chemotherapy: two evaluation cohorts (TRIBE ARM A and ARM B) and two validation cohorts (FIRE 3 arm A and MOMA). The relation of the SNPs with PFS and OS was evaluated through Kaplan-Meier method and log-rank test. Subgroup analyses according to RAS status were preplanned.

In the exploratory cohort 1 (TRIBE A), patients with CCTTT any>13repeats (N = 57) showed improved median PFS compared with patients carrying the ≤13/≤13 repeats variant (N = 152) (HR, 0.64; 95%CI 0.44–0.92, p = 0.010). Similar results were shown adopting the >26repeats/≤26 repeats (HR, 0.56; 95%CI 0.36–0.87, p = 0.005). In RAS mutant, patient with any>13 repeats (N = 24) had improved PFS results compared with those carrying the ≤13/≤13 repeats variant (N = 81) (HR, 0.51; 95%CI 0.30–0.87, p = 30.009). Similar results were found adopting the >26 repeats/≤26 repeats cut off: (HR, 0.52; 95%CI 0.27–0.98, p = 0.035). These data were partially confirmed in the exploratory cohort 2 (TRIBE B): a better median PFS was observed in patients with >26 repeats vs ≤26 repeats (N = 205) patients. However, these data were not confirmed in the two validation cohorts.

We failed to replicate the exploratory findings in both validation sets. The CCTTT polymorphic region of the INOS gene does not predict outcome in mCRC receiving bevacizumab based first line chemotherapy. Further investigations are needed to reveal mechanisms between tumor, immune system and chemotherapy response.

• Aged
• Bevacizumab/pharmacology
• Bevacizumab/therapeutic use
• Clinical Trials as Topic
• Cohort Studies
• Colorectal Neoplasms/diagnosis
• Colorectal Neoplasms/drug therapy
• Colorectal Neoplasms/genetics
• Colorectal Neoplasms/pathology
• Female
• Genotype
• Humans
• Male
• Middle Aged
• Neoplasm Metastasis
• Nitric Oxide Synthase Type II/genetics
• Polymorphism, Single Nucleotide
• Prognosis
• Treatment Outcome

• P30 CA014089 NCI NIH HHS
• P30CA014089 NIH HHS
• National Cancer Institute

• bevacizumab
• inflammation
• metastatic colorectal cancer
• predictive biomarkers
• vasculature

The possibility of detecting and classifying living cells in a label-free and non-invasive manner holds significant theranostic potential. In this work, Hyperspectral Imaging (HSI) has been successfully applied to the analysis of macrophagic polarization, given its central role in several pathological settings, including the regulation of tumour microenvironment. Human monocyte derived macrophages have been investigated using hyperspectral reflectance confocal microscopy, and hyperspectral datasets have been analysed in terms of M1 vs. M2 polarization by Principal Components Analysis (PCA). Following PCA, Linear Discriminant Analysis has been implemented for semi-automatic classification of macrophagic polarization from HSI data. Our results confirm the possibility to perform single-cell-level in vitro classification of M1 vs. M2 macrophages in a non-invasive and label-free manner with a high accuracy (above 98% for cells deriving from the same donor), supporting the idea of applying the technique to the study of complex interacting cellular systems, such in the case of tumour-immunity in vitro models.