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.
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The authors have no conflicts of interest to disclose.
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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