Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.
Research Institute for Oncology, Hematology and Cell Therapy, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran.
BMC Immunol. 2022 Jun 10;23(1):31. doi: 10.1186/s12865-022-00497-1.
BKPyV associated hemorrhagic cystitis (BKPyV-HC) is a major and prevalent outcome of hematopoietic stem cell transplantation (HSCT) with no standard treatment option. Adoptive T cell therapy (ACT) against transplant-associated viruses has shown promising potential. We sought to produce virus-specific T cells (VSTs) against BKPyV with the aim of treating refractory HSCT-associated HC.
Peripheral blood mononuclear cells (PBMC) from healthy donors were isolated by Ficoll-Hypaque density gradient centrifugation. BKPyV-pulsed, monocyte-derived dendritic cells (mo-DCs) and T cells were co-cultured and expanded over 2-3 weeks with the addition of IL-2. The T cells were examined for various functional assays.
Comparison analysis of Carboxyfluorescein diacetate succinimidyl ester (CFSE) indicated that the percentage of proliferated cells were significantly higher in donors (49.62 ± 7.09%) than controls (7.96 ± 4.55%). Furthermore, expanded T cells exhibited specificity to BKPyV antigens by IFN-γ ELISPOT assay. The expanded cells showed cytotoxic function versus human lymphoblastoid cell line (LCL). Final VST products mainly comprised of CD8/CD69 double-positive T cells, which were significantly higher in donors (46.8 ± 7.1%) than controls (16.91 ± 3.40%).
In this study we demonstrated the feasibility of producing functional BKPyV-specific T cells in healthy donors using BKPyV PepMixes. These functional cells were able to proliferate and produce IFN-γ cytokine in response to BKPyV PepMixes. In addition, these T cells had cytotoxic ability against BKPyV antigen-expressing target cells.
BK 多瘤病毒相关出血性膀胱炎（BKPyV-HC）是造血干细胞移植（HSCT）的主要且普遍的结果，目前尚无标准的治疗方法。针对移植相关病毒的过继性 T 细胞疗法（ACT）显示出了很有前景的潜力。我们试图针对 BKPyV 产生病毒特异性 T 细胞（VST），以治疗难治性 HSCT 相关 HC。
通过 Ficoll-Hypaque 密度梯度离心法从健康供者中分离外周血单个核细胞（PBMC）。将 BKPyV 脉冲的单核细胞衍生的树突状细胞（mo-DC）和 T 细胞共培养并在 2-3 周内进行扩增，同时添加 IL-2。对 T 细胞进行各种功能测定。
Carboxyfluorescein diacetate succinimidyl ester（CFSE）的比较分析表明，供者中增殖细胞的百分比明显高于对照（49.62 ± 7.09%）。此外，通过 IFN-γ ELISPOT 测定，扩增的 T 细胞显示出对 BKPyV 抗原的特异性。扩增的细胞对人淋巴母细胞系（LCL）表现出细胞毒性作用。最终的 VST 产物主要由 CD8/CD69 双阳性 T 细胞组成，供者中（46.8 ± 7.1%）明显高于对照（16.91 ± 3.40%）。
在这项研究中，我们使用 BKPyV PepMixes 证明了在健康供者中产生功能性 BKPyV 特异性 T 细胞的可行性。这些功能性细胞能够增殖并对 BKPyV PepMixes 产生 IFN-γ 细胞因子。此外，这些 T 细胞对表达 BKPyV 抗原的靶细胞具有细胞毒性作用。
