Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Solna, Sweden..
Immunology. 2010 Sep;131(1):128-40. doi: 10.1111/j.1365-2567.2010.03284.x. Epub 2010 May 10.
A better understanding of similarities and differences in the composition of the cellular immune system in non-human primates (NHPs) compared with human subjects will improve the interpretation of preclinical studies. It will also aid in addressing the usefulness of NHPs as subjects for studying chronic diseases, vaccine development and immune reconstitution. We employed high content colour flow cytometry and analysed simultaneously the expression of CD3, CD4, CD8alpha, CD8beta, CD16/CD56, CD45RA, CCR7, CD27, CD28, CD107a and the interleukin-7 receptor alpha-chain (IL-7Ralpha) in peripheral blood mononuclear cells (PBMCs) of 27 rhesus macaques and 16 healthy human subjects. Regulatory T cells (Tregs) were identified using anti-CD3, -CD4, -CD25, -FoxP3, and -IL-7Ralpha monoclonal antibodies. Responsiveness to IL-7 was gauged in a signal transducer and activation of transcription 5 (STAT-5) phosphorylation assay. Human and NHP PBMCs showed a similar T-cell composition pattern with some remarkable differences. Similarities: human and NHP CD4(+) and CD8(+) cells showed a similar STAT-5 phosphorylation pattern in response to IL-7. Multicolour flow cytometric analysis identified a CD4(+) CD8alphaalpha(+) CD8alphabeta(+) T-cell population in NHPs as well as in human subjects that expressed the degranulation marker CD107a and may represent a unique CD4(+) T-cell subset endowed with cytotoxic capacity. Differences: we identified in PBMCs from NHPs a higher proportion (5.16% in CD3(+) T cells) of CD8alphaalpha(+) T cells when compared with human donors (1.22% in CD3(+) T cells). NHP CD8alphaalpha(+) T cells produced tumour necrosis factor-alpha / interferon-gamma (TNF-alpha/IFN-gamma) or TNF-alpha, whereas human CD8alphaalpha(+) T cells produced simultaneously TNF-alpha/IFN-gamma and IL-2. A minor percentage of human CD8(+) T cells expressed CD25(bright) and FoxP3 (0.01%). In contrast, 0.07% of NHP CD8(+) T cells exhibited the CD25(bright) FoxP3(+) phenotype. PBMCs from NHPs showed less IL-7Ralpha-positive events in all T-cell subsets including CD4(+) Tregs (median 5%) as compared with human (median 12%). The data visualize commonalities and differences in immune cell subsets in humans and NHPs, most of them in long-lived memory cells and cells with suppressive functions. This provides a matrix to assess future efforts to study diseases and vaccines in NHPs.
为了更好地理解非人类灵长类动物（NHPs）与人类之间细胞免疫组成的相似性和差异性，我们需要提高对临床前研究的解读能力。此外，这还有助于确定 NHPs 作为研究慢性疾病、疫苗开发和免疫重建的研究对象的有效性。我们采用高内涵彩色流式细胞术，同时分析了外周血单个核细胞（PBMCs）中 CD3、CD4、CD8alpha、CD8beta、CD16/CD56、CD45RA、CCR7、CD27、CD28、CD107a 和白细胞介素-7 受体 alpha 链（IL-7Ralpha）的表达情况，这些细胞取自 27 只恒河猴和 16 名健康人类志愿者。使用抗 CD3、CD4、CD25、FoxP3 和 IL-7Ralpha 单克隆抗体鉴定调节性 T 细胞（Tregs）。通过信号转导和转录激活因子 5（STAT-5）磷酸化检测评估 IL-7 的反应性。人类和 NHP 的 PBMCs 表现出相似的 T 细胞组成模式，同时也存在一些显著差异。相似性：人类和 NHP 的 CD4(+) 和 CD8(+) 细胞对 IL-7 的反应呈现出相似的 STAT-5 磷酸化模式。多色流式细胞术分析在 NHPs 中以及人类志愿者中鉴定出 CD4(+) CD8alphaalpha(+) CD8alphabeta(+) T 细胞群，该细胞群表达脱颗粒标记物 CD107a，可能代表具有细胞毒性能力的独特 CD4(+) T 细胞亚群。差异：与人类供体（CD3(+) T 细胞中的 1.22%）相比，我们在 NHPs 的 PBMCs 中发现 CD8alphaalpha(+) T 细胞的比例更高（CD3(+) T 细胞中的 5.16%）。NHP CD8alphaalpha(+) T 细胞产生肿瘤坏死因子-α/干扰素-γ（TNF-alpha/IFN-gamma）或 TNF-alpha，而人类 CD8alphaalpha(+) T 细胞同时产生 TNF-alpha/IFN-gamma 和 IL-2。人类 CD8(+) T 细胞中仅有一小部分（0.01%）表达 CD25(bright)和 FoxP3。相比之下，0.07%的 NHP CD8(+) T 细胞表现出 CD25(bright) FoxP3(+)表型。与人类（中位数 12%）相比，NHPs 的 PBMCs 中所有 T 细胞亚群（包括 CD4(+) Tregs）的 IL-7Ralpha 阳性事件较少（中位数 5%）。该数据直观地展示了人类和 NHPs 中免疫细胞亚群的相似性和差异性，其中大多数是在长寿命记忆细胞和具有抑制功能的细胞中观察到的。这为评估未来在 NHPs 中研究疾病和疫苗的努力提供了基础。
