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The role of CD4+ T cells in BKV-specific T cell immunity

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Abstract

Reactivation of polyomavirus BK (BKV) infection represents a severe complication in kidney transplant (KTX) patients. We previously reported an association between a declining BK viral load and the reconstitution of CD4+ T cell BKV-specific immunity in patients following kidney transplantation. However, the specific contribution of CD4+ T cells in the regulation of BKV-replication is unknown. Nevertheless, in vitro enrichment of BKV-specific T cells and subsequent adoptive T cell transfer may improve the restoration of immune competence in KTX patients with BKV infection. To date, strategies to capture human BKV-specific T cells with the ensuing expansion to clinically useful numbers are lacking. Here, we demonstrated a comprehensive flow cytometric analysis of the BKV-specific T cell response that permits access to the majority of T cells specific for immunodominant BKV antigens. A full-spectrum evaluation of the BKV-specific T cell response was performed by stimulating peripheral blood mononuclear cells (PBMC) with a mixture of BKV immunodominant peptide pools at varying concentrations and measuring activation marker expression and cytokine secretion. We also examined the effects of co-stimulation and PBMC resting time prior to activation. We defined the narrow range of stimulation conditions that permit the capture and expansion of functional BKV-specific T cell lines. The generated BKV-specific T cell lines showed the highest specificity and functionality when the T cells were captured according to IFNγ-secretion. This study highlights the multifunctional and cytolytic BKV-specific CD4+ T cells as a dominant population within the generated T cell product. This method offers a novel approach for the generation of BKV-specific T cell lines for adoptive immunotherapy and underscores the critical role of CD4+ T cells in the clearance of BKV.

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Abbreviations

BKV:

Polyomavirus BK

BKV-PPM:

BKV overlapping peptide pool mix

SOT:

Solid organ transplantation

BKVAN:

BKV-associated nephropathy

PBMC:

Peripheral blood mononuclear cells

TCL:

T cell line

SEB:

Staphylococcus enterotoxin B

HAdV:

Human adenovirus

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Acknowledgments

We thank Dr. Jason Millward for a critical reading and editing of the manuscript. The study was supported by Profit (EFRE grant) and Novartis grant to NB.

Conflict of interest

The authors have no conflicts of interest to disclose.

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Authors and Affiliations

  1. Department of Nephrology, Charité University Medicine, Berlin, Germany

    B. J. D. Weist, H. Fuehrer, A. Sattler, P. Reinke & N. Babel

  2. Berlin Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany

    B. J. D. Weist, M. Schmueck, H. Fuehrer, A. Sattler, P. Reinke & N. Babel

  3. Institute of Medical Immunology, Charité University Medicine, Berlin, Germany

    M. Schmueck

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  1. B. J. D. Weist
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  2. M. Schmueck
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Correspondence to N. Babel.

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Weist, B.J.D., Schmueck, M., Fuehrer, H. et al. The role of CD4+ T cells in BKV-specific T cell immunity. Med Microbiol Immunol 203, 395–408 (2014). https://doi.org/10.1007/s00430-014-0348-z

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  • DOI: https://doi.org/10.1007/s00430-014-0348-z

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