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Cancer-testis antigen ACRBP: Cytotoxic response to its HLA-A2 restricted peptide and immune features in ovarian cancer.

While our prior study identified the HLA-A *0201-restricted ACRBP epitope peptide and demonstrated its capacity to generate cytotoxic T lymphocytes (CTLs) in vitro, the clinical relevance of the peptide-induced T cell reactivity in ovarian cancer (OC) patients and the in vivo anti-tumor efficacy of these CTLs remain unexplored. In this study, dendritic cells were sensitized with ACRBP peptide (ALLVLCYSI) and co-cultured with autologous CD8+T cells to induce the production of specific cytotoxic T lymphocytes (Pep-CTLs). The anti-tumor effects of Pep-CTLs were evaluated in SCID mice bearing human ovarian cancer (OC) OVCAR-3 cells. Concurrently, we co-cultured ALLVLCYSI peptide with peripheral blood mononuclear cells (PBMCs) from OC patients (HLA-A2+, ACRBP+) and assessed the number of specific T cells using ELISPOT assays. The immunological impact of the ACRBP peptide against human OC was validated through both in vitro and in vivo experiments. These findings establish a preclinical foundationfor developing ACRBP peptide-based vaccines in OC immunotherapy. To further elucidate ACRBP's role in OC treatment, the study analyzed single-cell RNA sequencing data from 8 OC patients and bulk RNA sequencing data from the Cancer Genome Atlas Project (TCGA) comprising 308 ovarian cancer cases. This analysis aimed to explore the heterogeneity among ACRBP-expressing tumor cell populations and to investigate the correlation between ACRBP expression and immune molecule expression (including MHC and chemokines) alongside chemotherapy response. These insights furnish a theoretical framework supporting the future application of ACRBP in tumor immunotherapy and strategies to prevent immune escape.

Humans↗

Detergent solubilization, purification, and separation of specificities of HLA antigens from a cultured human lymphoblastoid line, RPMI 4265.

HLA antigens have been purified to homogeneity after detergent solubilization from RPMI 4265, a human lymphoblastoid line. The inhibition of cytotoxicity assay for HLA antigen was modified, using preincubation with bovine serum albumin of antigen samples containing detergent to prevent lysis of target cells by detergent. Solubilization was tested with many types of detergents. A polyethyleneglycol oleyl ether nonionic detergent mixture, Brij 99:Brij 97 (2:1) was selected for solubilization, since it selectively solubilized HLA antigens, had a low absorbance at 280 nm and was uncharded. HLA antigens were then purified by Lens culinaris lectin affinity chromatography and Bio-Gel A-5m filtration. The antigen specifity HLA-A2 was separated from specificities HLA-B7,12 by isoelectric focusing. Purified HLA antigens contained a subunit of Mr=44,000 with NH2-terminal glycine, and a subunit of Mr=12,000, beta2-microglobulin, with NH2-terminal isoleucine.

Amino Acids↗

Primate HLA antibodies are specifically mitogenic for human lymphocytes and act synergistically with beta2m antibodies.

A monkey antiserum against HLA-A28 was raised by immunization with papain-solubilized antigens. The titre of this serum was 1/370 for beta2-microglobulin (beta2m) 1/750 for A28 and 1/200 for the cross-reacting HLA-A2 determinant. This serum was highly mitogenic for human blood lymphocytes. Mitogenicity was reduced or abolished after absorption of the serum on a beta2m-spharose column. Column-eluted anti-beta2m antibodies were not mitogenic. However, mitogenicity was restored by reconstitution with unabsorbed and absorbed antibodies. Mitogenicity was target cell specific. Tuhs, cells from all individuals were activated by the unabsorbed serum. The absorbed anti-A28 serum was mitogenic only for target cells carrying A28 and/or the cross-reacting determinant A2, but not for cells lacking either of these antigens. A definite gene-dose effect was recorded, since target cells homozygous for A2 responded better to polyclonal activation by the primate serum than cells which were heterozygous.

Animals↗

Strong association between B-lymphocyte group-2 specificity and asthma.

30 families in which at least one member has asthma were tested for five new specificities of B lymphocytes as well as for twenty-five HLA antigens. 41 other asthmatic patients were also tested. HLA-B8 was slightly less common than normal in the 71 patients with asthma and there was a trend towards an increased frequency of HLA-A2 in these patients. However, 88% of the 30 asthma patients had B lymphocyte group 2 compared with 24% of the 109 controls. This strong association between asthma and B lymphocyte group 2 was not completely paralleled by linkage with a postulated susceptibility gene of the HLA complex in 9 families whose members were investigated in detail.

Adult↗

B- and T-cell-specific alloantigens in man.

Screening for B-cell-specific antibodies in unabsorbed pregnancy sera preselected for weak reactivity in regular HLA-A, -B, and -C screening yielded a relatively high proportion (35/81) of B-cell-specific antibodies. Most B-cell-specific antibodies react broadly, showing inclusion phenomena suggesting 'cross-reactivity' analogous to that observed in HLA-A and -B serology. In control experiments with T-cell-enriched suspensions three antisera reacted with T cells and not with B cells. These antisera are highly associated with HLA-A2 in the unrelated population and segregate with HLA haplotypes in families and with HLA-A in a family with HLA-A, B recombination, Thus it appears that the human equivalents of Ia antigens may include--in analogy to the murine Ia antigens--B cell- as well as T-cell-specific alloantigens.

Antibody Specificity↗