Efficient culture of human naive and memory B cells for use as APCs

KY Su, A Watanabe, CH Yeh, G Kelsoe… - The Journal of …, 2016 - journals.aai.org
KY Su, A Watanabe, CH Yeh, G Kelsoe, M Kuraoka
The Journal of Immunology, 2016journals.aai.org
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∼ 10 3-fold increases following 8 d in culture and 10 6-fold
increases when cultures are split and cultured for 8 more days. In culture, a significant …
Abstract
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∼ 10 3-fold increases following 8 d in culture and 10 6-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.
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