CTL酶聯(lián)免疫斑點分析儀應(yīng)用新發(fā)表關(guān)于“COVID”研究的文獻
我們的一份關(guān)于COVID的論文已經(jīng)提交-預(yù)印本已經(jīng)可以在線獲取并免費分發(fā)(下面的“ biorxiv”鏈接):
亞歷山大·雷曼(Alexander Lehmann),格雷格(Greg A)解卷積T細胞對SARS-CoV-2的反應(yīng):特異性與偶然交叉和同源交叉反應(yīng)。 2020年。提交給免疫學(xué)前沿。 bioRxiv:
主要消息是:
通過使用IFN-g ELISPOT(可檢測效應(yīng)細胞),可以區(qū)分SARS-CoV-2感染者與未感染人類。根據(jù)文獻,其他基于流式細胞術(shù)的技術(shù),如AID,多聚體,CFSE稀釋在未感染的個體中提供了假陽性結(jié)果,據(jù)稱是由于檢測到與季節(jié)性冠狀病毒交叉反應(yīng)的中央記憶細胞而引起的普通感冒(或者只是沒有被足夠敏感,可以在頻率相當(dāng)?shù)蜁r提供干凈的數(shù)據(jù))
SARS-CoV-2上的幾種蛋白質(zhì)抗原具有免疫原性,需要進行監(jiān)測以檢測所有感染的個體(祝您好運的多聚體僅觀察一種/幾種肽)。
我們在這里表明,T細胞親和力測量(肽庫的串行稀釋)對于明確區(qū)分SARS-CoV2感染/未感染也很重要。由于(a)有效的電池利用率,以及(b)高吞吐能力且?guī)缀醪簧婕叭斯?,因此可以使用ELISPOT輕松實現(xiàn)。
無關(guān)的大型肽池作為特異性控制:隨著越來越多的科學(xué)家開始理解T細胞免疫監(jiān)測不能依賴于幾種肽(多聚體),越來越多的人(包括SARS-CoV-2 T細胞診斷中的大多數(shù))使用巨型肽池。但是,到目前為止,尚未確定這是一種有效的方法–我們在這里做到了!Our first paper on COVID has just been submitted – the pre-print is already available online and free for distribution (the “biorxiv” link below):
Alexander Lehmann, Greg A Kirchenbaum, Ting Zhang, Pedro A Reche, Paul V Lehmann. Deconvoluting the T cell response to SARS-CoV-2: specificity versus chance- and cognate cross-reactivity. 2020. Submitted to Frontiers in Immunology. bioRxiv:
The main messages are:
By using IFN-g ELISPOT (that detects effector memory cells) one can distinguish between SARS-CoV-2 infected vs non-infected humans. Based on the literature, other flowcytometry-based techniques like AID, multimers, CFSE dilution provide false positive results in non-infected individuals allegedly due to detecting central memory cells that cross-react with seasonal coronaviruses causing common cold (or, just by not being sensitive enough to provide clean data when the frequencies are rather low)
Several protein antigens on SARS-CoV-2 are immunogenic and need to be monitored to detect all infected individuals (good luck multimers looking at one/few peptide(s) only.)
We show here that T cell affinity measurements (serial dilution of peptide pools)are also important for a clear SARS-CoV2 infected/non-infected distinction. Such can be easily done with ELISPOT due to (a) efficient cell utilization, and (b) high throughput capability with little labor involved.
Irrelevant mega peptide pools as specificity controls: as more and more scientists start to understand that T cell immune monitoring cannot rely on few peptides (multimers), more and more (including most in SARS-CoV-2 T cell diagnostic) use mega peptide pools. However, it has not been so far established that this is a valid approach – we did it here!
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