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Efficient Culture of Human Naive and Memory B Cells for Use as APCs

Kuei-Ying Su, Akiko Watanabe, Chen-Hao Yeh, Garnett Kelsoe and Masayuki Kuraoka
J Immunol November 15, 2016, 197 (10) 4163-4176; DOI: https://doi.org/10.4049/jimmunol.1502193
Kuei-Ying Su
*Department of Immunology, Duke University, Durham, NC 27710;
†Tzu Chi Medical Center, Hualien 970, Taiwan; and
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Akiko Watanabe
*Department of Immunology, Duke University, Durham, NC 27710;
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Chen-Hao Yeh
*Department of Immunology, Duke University, Durham, NC 27710;
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Garnett Kelsoe
*Department of Immunology, Duke University, Durham, NC 27710;
‡Human Vaccine Institute, Duke University, Durham, NC 27710
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Masayuki Kuraoka
*Department of Immunology, Duke University, Durham, NC 27710;
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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 ∼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.

Footnotes

  • This work was supported in part by Duke Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery Grant AI100645-02 and Autoimmunity Center of Excellence Grant AI56363.

  • The TCRβ sequence data sets presented in this article have been submitted to the immunoSEQ Analyzer database (https://clients.adaptivebiotech.com/pub/9816555b-5673-4316-85e2-244acb293f0b) under accession numbers 14_d0_CD4naive-2016881253, 14_d7_activatedCD4-2016881253, 14_d7_TTspecificCD4-2016881253, 6_d0_CD4naive-2016881253, 6_d7_activatedCD4-2016881253, and 6_d7_TTspecificCD4-2016881253.

  • The online version of this article contains supplemental material.

  • Abbreviations used in this article:

    7-AAD
    7-aminoactinomycin D
    CD
    culture-derived
    DC
    dendritic cell
    FSC
    forward scatter
    HA
    recombinant influenza hemagglutinin
    MFI
    mean fluorescent intensity
    MHCII
    MHC class II
    rPA
    recombinant Bacillus anthracis protective Ag
    Td
    tetanus–diphtheria toxoid
    TT
    tetanus toxoid
    TT-PE
    TT conjugated with PE.

  • Received October 19, 2015.
  • Accepted August 30, 2016.
  • Copyright © 2016 by The American Association of Immunologists, Inc.
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The Journal of Immunology: 197 (10)
The Journal of Immunology
Vol. 197, Issue 10
15 Nov 2016
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Efficient Culture of Human Naive and Memory B Cells for Use as APCs
Kuei-Ying Su, Akiko Watanabe, Chen-Hao Yeh, Garnett Kelsoe, Masayuki Kuraoka
The Journal of Immunology November 15, 2016, 197 (10) 4163-4176; DOI: 10.4049/jimmunol.1502193

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Efficient Culture of Human Naive and Memory B Cells for Use as APCs
Kuei-Ying Su, Akiko Watanabe, Chen-Hao Yeh, Garnett Kelsoe, Masayuki Kuraoka
The Journal of Immunology November 15, 2016, 197 (10) 4163-4176; DOI: 10.4049/jimmunol.1502193
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