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Biotechnical Methods Section (BTS)

A novel approach to identify antigens recognized by CD4 T cells using complement-opsonized bacteria expressing a cDNA library

Abstract

In patients with hematological malignancies receiving HLA-matched stem cell transplantation, T cells specific for minor histocompatibility antigens play a major role in graft rejection, induction of graft-versus-host disease and beneficial graft-versus-leukemia reactivity. Several human minor histocompatibility antigens recognized by T cells have been identified, but only two are presented by HLA class II molecules. In search of an efficient approach to identify antigenic peptides processed through the HLA class II pathway, we constructed a cDNA library in bacteria that were induced to express proteins. Bacteria were opsonized with complement to enforce receptor-mediated uptake by Epstein–Barr virus immortalized B cells that were subsequently used as antigen-presenting cells. This approach was validated with an HLA class II-restricted antigen encoded by gene DBY. We were able to identify bacteria expressing DBY diluted into a 300-fold excess of bacteria expressing a nonrelevant gene. Screening of a bacterial library using a DBY-specific CD4 T cell clone resulted in the isolation of several DBY cDNAs. We propose this strategy for a rapid identification of HLA class II-restricted antigenic peptides recognized by CD4 T cells.

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Acknowledgements

We would like to thank CA Davis (Department of Developmental Molecular Genetics, Ben Gurion University of the Negev, Beer Sheva, Israel) for providing us with the pKE-1 vector, and Dr Edus H Warren (Fred Hutchinson Cancer Research Center, Seattle, WA, USA) for interesting discussion, careful reading and English editing of the manuscript. We also thank Mrs C Wildmann for her precious assistance, and Mrs N Krack for editorial assistance.

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Correspondence to P van der Bruggen.

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van de Corput, L., Chaux, P., van der Meijden, E. et al. A novel approach to identify antigens recognized by CD4 T cells using complement-opsonized bacteria expressing a cDNA library. Leukemia 19, 279–285 (2005). https://doi.org/10.1038/sj.leu.2403583

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