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HLA-DRB1 allelic combinations differentially shape dendritic cell antigen presentation enhanced by tumor cell line lysate-pulsing

ABSTRACT: The study explores the anti-tumor immune response through the analysis of the HLA-II immunopeptidome of dendritic cells (DCs) from HLA-heterozygous donors pulsed with a protein extract from the MCF-7 tumor cell line. The objective was to characterize how different HLA-DRB1 allele combinations influence peptide presentation and how DC pulsing affects the immunopeptidome profile. Our findings demonstrate that HLA-DR heterozygosity significantly shapes the peptide repertoire in an allele-specific manner, with allele dominance modulated by specific allelic combinations. Notably, tumor-derived peptides such as annexin A2, galectin-1, and elongation factor 1-alpha 2 were identified in association with particular HLA-DRB1 alleles. These insights provide valuable information for the design of personalized immunotherapies based on peptide-pulsed DCs aimed at enhancing CD4+ tumor-infiltrating lymphocyte responses.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Dendritic Cell

SUBMITTER: Gonzalo Lázaro

LAB HEAD: Mercè Martí

PROVIDER: PXD064999 | Pride | 2026-02-09

REPOSITORIES: Pride

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HLA-DRB1 Allelic Combinations Differentially Shape Dendritic Cell Antigen Presentation Enhanced by Tumour Cell Line Lysate-Pulsing.

Lázaro Gonzalo G Cedano Juan A JA Faus Maitane M Roura-Mir Carme C Garrigós Laia L Pérez-García José J Román-García Javier J Cortés Javier J Aran Andrea A Martí Mercè M

HLA 20260201 2

The anti-tumour immune response plays a pivotal role in eliminating tumour cells, with the presence of tumour-infiltrating lymphocytes (TILs) often correlated with improved patient outcomes. Among these, CD4+ T lymphocytes act as key orchestrators of the immune response, functioning as effector and regulatory cells, and are essential for establishing immunological memory. To better understand the role of CD4+ T cells in anti-tumour immunity, we analysed the HLA-II immunopeptidome of dendritic ce ...[more]

PMID: 41607090

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Project description:Tumor-Infiltrating Lymphocytes (TILs) are composed by several immune subpopulations including NK, NKT, Tab, Tgd and B cells. Although some of these cells can have antitumoral activities, there is also a large number of cells which are not tumor-specific,known as bystander cells. These cells may not have a direct role in the antitumoral response but could lead to a modulation of the immune response after time. Recent studies have described fatal encephalitis and myocarditis in cancer patients after immune checkpoint therapies, in which EBV-specific T cells may be involved. Since EBV is able to infect B cells and epithelial cells, both present in the primary tumor site, the first events of these fatal responses could be a consequence of a dysregulation of the immune system in the tumor microenvironment, although this have not been deeply studied. Thus, the main objectives of this study are (i) to use EBV-transformed B cells from the tumor site as a model to study the tumor-infiltrating EBV+ B cells and (ii) to use these EBV-derived B cells to study the relationship with autologous T cells isolated from the tumor site. T cells from TILs have been isolated from a triple negative breast cancer patient before neoadjuvancy and were expanded directly from the biopsy. A Patient-Derived Xenograft (PDX) generated from the breast tumor, progressed into a lymphocytic neoplasm. EBVtransformed B cells were obtained from this PDX (PDX-derived B cells). T Cell Receptor high-throughput sequencing has been used to monitor the clonotypes present in the different samples. We have found 43 clonotypes infiltrating the PDX-562 tissue and 18 of them were also present in the human biopsy or in the biopsy-derived T cell cultures. 5 of these sequences were found in one of the biopsy-derived T cells analyzed. Thus, PDXderived B cells have been used as antigen presenting cells to expand these cells. Few clonotypes were obtained after expansions, although the analysis of the middle CDR3 have revealed a similar pattern with the 18 sequences from the PDX sample. Furthermore, peptides presented by HLA have been eluted from PDX-derived B cells, followed by MS analysis. Peptides obtained derived from 31 different proteins and >40% are cancerrelated in terms of expression.

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