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在這篇文章里，Roederer和他的團隊提出了一種在平臺使用多重TaqMan試劑對單個T細胞進行分析的方法。首先，使用BD FACSAria™，根據(jù)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(zhuǎn)錄本的檢測極限（limit of detection, LOD），結果表明的高靈敏度和寬的動態(tài)范圍，可以檢測單細胞當量的mRNA。
該團隊在單細胞水平監(jiān)測了T細胞的激活，發(fā)現(xiàn)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(zhì)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(zhì)性被證明有很強的免疫相關性。Roederer和他的團隊正繼續(xù)推進系統(tǒng)的、單細胞的方法以更好地理解中樞免疫系統(tǒng)中細胞和分子的相互作用。
參考文獻
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