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在這篇文章里，Roederer和他的團隊提出了一種在BioMark平臺使用多重TaqMan試劑對單個T細胞進行分析的方法。首先，使用BD FACSAria™，根據細胞的標記將其分離以得到單細胞。接下來，Roederer細化其方法學，確保獲得足量的RNA以進行單細胞的表達譜分析。Roederer等展示了低至一個mRNA轉錄本的檢測極限（limit of detection, LOD），結果表明BioMark的高靈敏度和寬的動態范圍，可以檢測單細胞當量的mRNA。
該團隊在單細胞水平監測了T細胞的激活，發現了群體中CD4+細胞的亞群。實際上，CD4+細胞表達類似CSCR5/CCL5和DPPR/TIA1這樣的基因是稀有事件，并非之前使用大量mRNA觀察到的是普遍事件
通過比較CD154+和CD154- T細胞對葡萄球菌腸毒素B（SEB, Staphlyococcal enterotoxin B）的反應，（細胞）的異質性被進一步確立。當被刺激時，分析顯示有一組基因和T細胞的激活強烈相關，但是，只有5-20%的T細胞擁有可以結合SEB的受體Vβ鏈變體。
在這項研究中，細胞異質性被證明有很強的免疫相關性。Roederer和他的團隊正繼續推進系統的、單細胞的方法以更好地理解中樞免疫系統中細胞和分子的相互作用。
參考文獻
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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).
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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). |
