研究HIV治疗过程的单细胞免疫反应

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用于疫苗开发的免疫保护相关性

研究提前注射疫苗以保护CD4记忆细胞不被破坏，以及淋巴细胞预防病毒扩增的功能。

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促进和发展细胞研究技术

使用先进的仪器，包括FACS系统和Fluidigm的仪器、试剂、分析和方法。

图：HIV感染的H9 T细胞；

Roederer博士和他的团队有一篇方法学论文，题目为：Highly multiplexed quantitation of gene expression on single cells (Journal of Immunological Methods, 2013, 391(1-2):133-145)。 

“在免疫学领域，极大的细胞异质性已被广泛认识，但相应的工作还进行的很少。”- Mario Roederer，Ph.D. 在这篇文章里，Roederer和他的团队提出了一种在BioMark平台使用多重TaqMan试剂对单个T细胞进行分析的方法。首先，使用BD FACSAria™，根据细胞的标记将其分离以得到单细胞。接下来，Roederer细化其方法学，确保获得足量的RNA以进行单细胞的表达谱分析。Roederer等展示了低至一个mRNA转录本的检测极限（limit of detection, LOD），结果表明BioMark的高灵敏度和宽的动态范围，可以检测单细胞当量的mRNA。 该团队在单细胞水平监测了T细胞的激活，发现了群体中CD4+细胞的亚群。实际上，CD4+细胞表达类似CSCR5/CCL5和DPPR/TIA1这样的基因是稀有事件，并非之前使用大量mRNA观察到的是普遍事件通过比较CD154+和CD154- T细胞对葡萄球菌肠毒素B（SEB, Staphlyococcal enterotoxin B）的反应，（细胞）的异质性被进一步确立。当被刺激时，分析显示有一组基因和T细胞的激活强烈相关，但是，只有5-20%的T细胞拥有可以结合SEB的受体Vβ链变体。 在这项研究中，细胞异质性被证明有很强的免疫相关性。Roederer和他的团队正继续推进系统的、单细胞的方法以更好地理解中枢免疫系统中细胞和分子的相互作用。 参考文献1、Dominguez, M.H., Chattopadhyay, P.K., Ma, S., Lamoreaux, L., McDavid, A., Finak, G., Gottardo, R., Koup, R., &Roederer, M. Highly multiplexed quantitation of gene expression on single cells. Journal of Immunological Methods in press 2013 (13 pages).2、Kern, F., LiPira, G., Gratama, J., Manca, F. &Roederer, M. Measuring antigen specific immune responses: Understanding immunopathogenesis and improving diagnostics in infectious disease, autoimmunity and cancer. Trends in Immunology 26, 477-484 (2005).3、Chattopadhyay, P. K., Yu, J. &Roederer, M. A live-cell assay to detect antigen-specific CD4 T cells with diverse cytokine profiles. Nature Medicine 11, 1113-1117 (2005).4、Mattapallil, J. M., Douek, D. C., Hill, B., Nishimura, Y., Martin, M. &Roederer, M. Massive infection and loss of memory CD4 T cells in multiple tissues during acute SIV infection. Nature 434, 1093-1097 (2005).5、Chattopadhyay, P. K., Perfetto, S. P. &Roederer, M. The colorful future of cell analysis by flow cytometry. Discovery Medicine 4, 255-262 (2004).6、De Rosa, S. C., Lu, F. X., Perfetto, S. P., Yu, J., Moser, S., Miller, C. J., Evans, T. G. &Roederer, M. Vaccination in humans generates broad T cell cytokine responses. J. Immunol. 173, 5372-5380 (2004).7、Perfetto, S. P., Chattopadhyay, P. K. &Roederer, M. 17-Color Flow Cytometry: Unraveling the Immune System. Nature Reviews Immunology 4, 648-655 (2004).8、De Rosa, S. C., Andrus, J. P., Perfetto, S. P., Mantovani, J. J., Herzenberg, L. A., Herzenberg, L. A. &Roederer, M. Ontogeny of gamma-delta T cells in man. J. Immunol. 172, 1637-45 (2004).9、Berhanu, D., Mortari, F. &Roederer, M. Optimized lymphocyte isolation methods for analysis of chemokine receptor expression. J. Immunol. Methods 279, 199-207 (2003).De Rosa, S. C., Brenchley, J. M. &Roederer, M. Beyond six colors: A new era in flow cytometry. Nature Medicine 9, 112-117 (2003).10、De Rosa, S. C., Herzenberg, L. A. &Roederer, M. 11-color, 13-parameter flow cytometry: identification of human naive T cells by phenotype, function, and T-cell receptor diversity. Nature Medicine 7, 245-8. (2001).11、Wille-Reece, U., Flynn, B. J., Lore, K., Koup, R. A., Kedl, R. M., Mattapallil, J. M., Weiss, W. R., Roederer, M. & Seder, R. A. HIV Gag protein conjugated to a TLR7/8 agonist influences the magnitude and quality of Th1 and CD8+ T cell responses in nonhuman primates. ProcNatlAcadSci (USA), in press (2005).12、Nishimura, Y., Brown, C. R., Mattapallil, J. M., Igarashi, T., Buckler-White, A., Lafont, B. A. P., Hirsch, V. M., Roederer, M. & Martin, M. Non-activated naive CD4 T cells are the principal source of progeny virus during acute infections of monkeys with highly pathogenic X4-tropic SHIVs. ProcNatlAcadSci (USA) 102, 8000-8005 (2005).13、Song, K., Rabin, R. L., Hill, B. J., De Rosa, S., Perfetto, S. P., Zhang, H. H., Foley, J. F., Reiner, J. S., Douek, D. C., Roederer, M. & Farber, J. M. CXCR3 and CCR4 Reveal Polarized Early Central Memory CD4+ T Cells in Humans. ProcNatlAcadSci USA 102, 7916-7921 (2005).14、Betts, M. R., Exley, B., Price, D. A., Bansal, A., Camacho, Z. T., Teaberry, V., West, S. M., Ambrozak, D. R., Tomaras, G., Roederer, M., Kilby, J. M., Tartaglia, J., Belshe, R., Gao, F., Douek, D. C., Weinhold, K. J., Koup, R. A., Goepfert, P. & Ferrari, G. Characterization of functional and phenotypic changes in anti-Gag vaccine-induced T cell responses and their role in protection after HIV-1 infection. ProcNatlAcadSci (USA) 102, 4512-7 (2005).15、Stuge, T., Holmes, S. P., Saharan, S., Rubio, V., Roederer, M., Weber, J. & Lee, P. P. Complexity of endogenous versus vaccine-elicited T cell responses to cancer.PLoS Medicine 1, e28 (2004).16、Betts, M. R., Price, D. A., Brenchley, J. M., Lore, K., Guenaga, F., Smed-Sorensen, A., Ambrozak, D. R., Migueles, S. A., Connors, M., Roederer, M., Douek, D. C. &Koup, R. The functional profile of of primary human antiviral CD8(+) T cell effector activity is dictated by cognate peptide concentration. J. Immunol. 10, 6407-6417 (2004).17、Sullivan, N. J., Geisbert, T. W., Geisbert, J. B., Xu, L., Yang, Z.-y., Roederer, M., Koup, R. A., Jahrling, P. B. &Nabel, G. Accelerated vaccination for Ebola virus hemorrhagic fever in nonhuman primates. Nature 424, 681-684 (2004).18、Rubio, V., Stuge, T., Betts, M. R., Weber, J., Roederer, M. & Lee, P. P. Ex vivo identification, isolation, and analysis of tumor-cytolytic T cells. Nature Medicine 9, 1377-1382 (2003).19、Brenchley, J. M., Karandikar, N. J., Betts, M. R., Ambrozak, D. R., Hill, B. J., Crotty, L. E., Casazza, J. P., Kuruppu, J., Migueles, S. A., Connors, M., Roederer, M., Douek, D. C. &Koup, R. A. Expression of CD57 defines replicative senescence and antigen-induced apoptotic death of CD8+ T-cells. Blood 101, 2711-2720 (2003).20、Yang Shih, T. A., Meffre, E., Roederer, M. &Nussenzweig, M. C. Role of BCR affinity in T cell dependent antibody responses in vivo. Nature Immunology 3, 570-5. (2002).21、Shih, T. A., Roederer, M. &Nussenzweig, M. C. Role of antigen receptor affinity in T cell-independent antibody responses in vivo. Nature Immunology 3, 399-406. (2002).