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Ex-vivo clonally expanded B lymphocytes infiltrating colorectal carcinoma are of mature immunophenotype and produce functional IgG

PLoS ONE, ISSN: 1932-6203, Vol: 7, Issue: 2, Page: e32639
2012
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Article Description

Background: Tumor infiltrating B cells (TiBc) have not yet been investigated in detail. This may at least in part be due to technical difficulties. Here we describe a straightforward and reproducible method to isolate and culture TiBc from primary colorectal carcinomas (CRC). Methods/Results: TiBc cultures were generated by Epstein-Barr virus (EBV) immortalization. With this method, monoclonal TiBc cultures were obtained for 14/19 CRCs. As assessed by flow cytometry and ELISA, TiBc showed an activated immunophenotype (CD23 , CD80 ) and produced immunoglobulin (Ig; IgG secretion in 55% of the cultures). In functional in vitro analysis, most of the IgGs specifically bound to allogeneic CRC target cells. These data suggest that TiBc are antigen-experienced and thus may exhibit functionality in situ. Additionally, mini-cultures generated from 12 further CRCs revealed TiBc outgrowth exclusively in the presence of EBV. Conclusion: In summary, this simple method provides a cellular tool and our data set the stage for analysing the bivalent role of TiBc; being antigen-presenting cells on the one hand and tumor-specific antibody producers on the other. Additionally, the generation of long-term TiBc cultures and their monoclonal Ig may serve to identify novel tumor-specific antigens. © 2012 Maletzki et al.

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http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84857681294&origin=inward; http://dx.doi.org/10.1371/journal.pone.0032639; http://www.ncbi.nlm.nih.gov/pubmed/22393427; https://dx.plos.org/10.1371/journal.pone.0032639.g002; http://dx.doi.org/10.1371/journal.pone.0032639.g002; https://dx.plos.org/10.1371/journal.pone.0032639.t003; http://dx.doi.org/10.1371/journal.pone.0032639.t003; https://dx.plos.org/10.1371/journal.pone.0032639.g003; http://dx.doi.org/10.1371/journal.pone.0032639.g003; https://dx.plos.org/10.1371/journal.pone.0032639.g001; http://dx.doi.org/10.1371/journal.pone.0032639.g001; https://dx.plos.org/10.1371/journal.pone.0032639.t002; http://dx.doi.org/10.1371/journal.pone.0032639.t002; https://dx.plos.org/10.1371/journal.pone.0032639.t001; http://dx.doi.org/10.1371/journal.pone.0032639.t001; https://dx.plos.org/10.1371/journal.pone.0032639; https://dx.doi.org/10.1371/journal.pone.0032639.t001; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.t001; https://dx.doi.org/10.1371/journal.pone.0032639.t002; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.t002; https://dx.doi.org/10.1371/journal.pone.0032639.g001; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.g001; https://dx.doi.org/10.1371/journal.pone.0032639; https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0032639; https://dx.doi.org/10.1371/journal.pone.0032639.g003; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.g003; https://dx.doi.org/10.1371/journal.pone.0032639.t003; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.t003; https://dx.doi.org/10.1371/journal.pone.0032639.g002; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0032639.g002; http://dx.plos.org/10.1371/journal.pone.0032639.g003; http://dx.plos.org/10.1371/journal.pone.0032639.t001; https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032639&type=printable; http://dx.plos.org/10.1371/journal.pone.0032639.g001; http://dx.plos.org/10.1371/journal.pone.0032639.t003; http://dx.plos.org/10.1371/journal.pone.0032639.t002; http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0032639; http://dx.plos.org/10.1371/journal.pone.0032639; http://dx.plos.org/10.1371/journal.pone.0032639.g002; http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0032639; http://www.plosone.org/article/metrics/info:doi/10.1371/journal.pone.0032639; http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032639&type=printable

Claudia Maletzki; Annika Jahnke; Christiane Ostwald; Ernst Klar; Friedrich Prall; Michael Linnebacher; Ann M. Moormann

Public Library of Science (PLoS)

Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences; Multidisciplinary

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