Efficient Culture of Human Naive and Memory B Cells for Use as APCs.
The ability to culture and expand B cells in vitro has become a useful tool for studying human immunity. A limitation of current methods for human B cell culture is the capacity to support mature B cell proliferation. We developed a culture method to support the efficient activation and proliferation of naive and memory human B cells. This culture supports extensive B cell proliferation, with ∼103-fold increases following 8 d in culture and 106-fold increases when cultures are split and cultured for 8 more days. In culture, a significant fraction of naive B cells undergo isotype switching and differentiate into plasmacytes. Culture-derived (CD) B cells are readily cryopreserved and, when recovered, retain their ability to proliferate and differentiate. Significantly, proliferating CD B cells express high levels of MHC class II, CD80, and CD86. CD B cells act as APCs and present alloantigens and microbial Ags to T cells. We are able to activate and expand Ag-specific memory B cells; these cultured cells are highly effective in presenting Ag to T cells. We characterized the TCR repertoire of rare Ag-specific CD4+T cells that proliferated in response to tetanus toxoid (TT) presented by autologous CD B cells. TCR Vβ usage by TT-activated CD4+T cells differs from resting and unspecifically activated CD4+T cells. Moreover, we found that TT-specific TCR Vβ usage by CD4+T cells was substantially different between donors. This culture method provides a platform for studying the BCR and TCR repertoires within a single individual.
Department of Immunology, Duke University, Durham, NC 27710.|Tzu Chi Medical Center, Hualien 970, Taiwan; and.
Kuei-Ying
0000-0002-7927-2310
KY
Su
Department of Immunology, Duke University, Durham, NC 27710.
Akiko
0000-0003-4741-4705
A
Watanabe
Department of Immunology, Duke University, Durham, NC 27710.
Chen-Hao
0000-0001-5801-2918
CH
Yeh
Department of Immunology, Duke University, Durham, NC 27710; ghkelsoe@duke.edu.|Human Vaccine Institute, Duke University, Durham, NC 27710.
Garnett
G
Kelsoe
Department of Immunology, Duke University, Durham, NC 27710.
Masayuki
M
Kuraoka
D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000502:physiology; D001402:B-Lymphocytes / Q000276:immunology* / Q000502:physiology; D015704:CD4 Antigens; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D007156:Immunologic Memory; D008213:Lymphocyte Activation; D011948:Receptors, Antigen, T-Cell / Q000276:immunology / Q000378:metabolism; D013745:Tetanus Toxoid / Q000276:immunology
D015704:CD4 Antigens; D011948:Receptors, Antigen, T-Cell; D013745:Tetanus Toxoid