Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University Prague and University Hospital Motol, Prague, Czech Republic.
Bone Marrow Transplant. 2011 Aug;46(8):1089-98. doi: 10.1038/bmt.2010.261. Epub 2010 Nov 8.
Depletion of cellular immunity as a consequence of conditioning before allogeneic hematopoietic SCT (HSCT) frequently results in CMV reactivation, which may in turn lead to life-threatening infections and require timely antiviral treatment. We have investigated the functional signatures of CMV-specific CD4+ and CD8+ T-cells in 191 samples from 118 individuals. We compared healthy donors with both patients with high and undetectable viral loads, and those who controlled and did not control their CMV reactivations. Polychromatic flow-cytometric measurements of CD154 (CD40L), intracellular cytokines (IFNγ, IL2) and a degranulation marker (CD107a) allowed us to assess the functional status of various T-cells simultaneously. We found that dual IFNγ/IL2-producing CD8+ T-cells were present in patients controlling their CMV reactivations but absent from non-controllers. CD8+ T-cells that produce only IFNγ were the most abundant subtype, but they most likely represent non-protective memory cells. Distinct functional signatures were examined by hierarchical clustering, and this revealed that, unlike polyfunctional CD8+ T-cells, CD8+ T-cells that produce IFNγ alone were not functioning in concert with other subsets. In conclusion, our study revealed functional signatures that may be useful for immune monitoring, and it could change the interpretation of previous studies that assessed only IFNγ.
异基因造血干细胞移植（HSCT）前的细胞免疫耗竭常导致 CMV 再激活，继而导致危及生命的感染，并需要及时进行抗病毒治疗。我们研究了 118 名个体的 191 个样本中 CMV 特异性 CD4+和 CD8+T 细胞的功能特征。我们比较了健康供体与高病毒载量和不可检测病毒载量的患者，以及那些控制和未控制 CMV 再激活的患者。通过多色流式细胞术测量 CD154（CD40L）、细胞内细胞因子（IFNγ、IL2）和脱颗粒标志物（CD107a），我们能够同时评估各种 T 细胞的功能状态。我们发现，控制 CMV 再激活的患者中存在产生双重 IFNγ/IL2 的 CD8+T 细胞，但非控制者中不存在。只产生 IFNγ的 CD8+T 细胞是最丰富的亚型，但它们很可能代表非保护性记忆细胞。通过层次聚类分析检查了不同的功能特征，这表明与多效性 CD8+T 细胞不同，单独产生 IFNγ的 CD8+T 细胞不能与其他亚群协同发挥作用。总之，我们的研究揭示了可能有助于免疫监测的功能特征，这可能会改变以前仅评估 IFNγ的研究的解释。
