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在這篇文章里，Roederer和他的團(tuán)隊(duì)提出了一種在BioMark平臺(tái)使用多重TaqMan試劑對(duì)單個(gè)T細(xì)胞進(jìn)行分析的方法。首先，使用BD FACSAria™，根據(jù)細(xì)胞的標(biāo)記將其分離以得到單細(xì)胞。接下來(lái)，Roederer細(xì)化其方法學(xué)，確保獲得足量的RNA以進(jìn)行單細(xì)胞的表達(dá)譜分析。Roederer等展示了低至一個(gè)mRNA轉(zhuǎn)錄本的檢測(cè)極限（limit of detection, LOD），結(jié)果表明BioMark的高靈敏度和寬的動(dòng)態(tài)范圍，可以檢測(cè)單細(xì)胞當(dāng)量的mRNA。
該團(tuán)隊(duì)在單細(xì)胞水平監(jiān)測(cè)了T細(xì)胞的激活，發(fā)現(xiàn)了群體中CD4+細(xì)胞的亞群。實(shí)際上，CD4+細(xì)胞表達(dá)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(guò)比較CD154+和CD154- T細(xì)胞對(duì)葡萄球菌腸毒素B（SEB, Staphlyococcal enterotoxin B）的反應(yīng)，（細(xì)胞）的異質(zhì)性被進(jìn)一步確立。當(dāng)被刺激時(shí)，分析顯示有一組基因和T細(xì)胞的激活強(qiáng)烈相關(guān)，但是，只有5-20%的T細(xì)胞擁有可以結(jié)合SEB的受體Vβ鏈變體。
在這項(xiàng)研究中，細(xì)胞異質(zhì)性被證明有很強(qiáng)的免疫相關(guān)性。Roederer和他的團(tuán)隊(duì)正繼續(xù)推進(jìn)系統(tǒng)的、單細(xì)胞的方法以更好地理解中樞免疫系統(tǒng)中細(xì)胞和分子的相互作用。
參考文獻(xiàn)
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