Immunoprofiling of Breast Cancer Antigens Using Antibodies Derived from Local Lymph Nodes

Abstract

Tumor antigens are responsible for initiating an immune response in cancer patients, and their identification may provide new biomarkers for cancer diagnosis and targets for immunotherapy. The general use of serum antibodies to identify tumor antigens has several drawbacks, including dilution, complex formation, and background reactivity. In this study, antibodies were generated from antibody-secreting cells (ASC) present in tumor-draining lymph nodes of 20 breast cancer patients (ASC-probes) and were used to screen breast cancer cell lines and protein microarrays. Half of the ASC-probes reacted strongly against extracts of the MCF-7 breast cancer cell line, but each with a distinct antigen recognition profile. Three of the positive ASC-probes reacted differentially with recombinant antigens on a microarray containing cancer-related proteins. The results of this study show that lymph node-derived ASC-probes provide a highly specific source of tumor-specific antibodies. Each breast cancer patient reacts with a different antibody profile which indicates that targeted immunotherapies may need to be personalized for individual patients. Focused microarrays in combination with ASC-probes may be useful in providing immune profiles and identifying tumor antigens of individual cancer patients.

Identification of Tumor Antigens in Ovarian Cancers Using Local and Circulating Tumor-Specific Antibodies

“…In this study, most ASC probes isolated from a small cohort of ovarian cancer patients (n = 9/11) reacted to extracts of a high-grade serous ovarian cancer cell line (OVCAR3) with varying intensity. While antigen reactivity was lower than previously detected in breast cancer, both shared and unique patterns were also seen [19]. Interestingly, higher antigen reactivities were seen in soluble cell fractions containing cytoplasmic antigens, when compared to insoluble fractions mostly including membranous antigens.…”

“…Antibody generation requires the induction of antibody-secreting B cells (ASCs) in tissue-draining lymph nodes, with the subsequent release of antibodies into the blood. Previous studies have shown that ASCs recovered from tissue-draining lymph nodes are highly enriched for disease-related antigens compared to those in the blood [17,18] and can be used to identify novel tumor antigens [19,20]. To identify the cognate antigens recognized by tumor antigen-induced antibodies, high-density protein microarrays have emerged as useful tools, although challenges with cross-reactivity and high background are common [21].…”

High Numbers and Densities of PD1+ T-Follicular Helper Cells in Triple-Negative Breast Cancer Draining Lymph Nodes Are Associated with Lower Survival

“…Hence, shaping a TDLN towards a humoral immune response would lead to a lower survival rate in TN breast cancer patients. An approach to identify a possible source of this immune response is to directly isolate plasma cells from tumor draining LNs and check whether they secrete antibodies binding to tumor antigens, e.g., by indirect immunofluorescence of tumor sections incubated with supernatant from cultured plasma cells [ 21 ]. However, the role of antibodies against tumor antigens needs further clarification and could be complemented by techniques to identify foreign peptides [ 22 , 23 ] as antibodies against tumor antigens are insufficient to carry an antitumoral immune response [ 24 ].…”

Tumor antigens heterogeneity and immune response-targeting neoantigens in breast cancer

“…Ding et al [127] analyzed 47 luminal breast cancer samples and identified an average of 31 nsSNVs in each breast cancer genome investigated, demonstrating that nsSNVs are expressed and recognized by the immune system in many cases. Young et al [131] compared the individual antibody response in breast cancer patients in which antibody-secreting cells (ASCs) were generated from the tumor-draining lymph nodes of 20 subjects. Reflecting the large molecular heterogeneity of breast cancer, a unique antibody profile was defined for each patient through a one-dimensional western blot screening of MCF7 extracts, for which only a few bands were found in common with additional subjects.…”