resting of cryopreserved pbmc does not generally benefit the performance of antigen-specific t cell elispot？assays休息的低温贮藏pbmc一般不会受益有关酶联免疫斑点抗原t细胞的性能分析.pdfVIP

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resting of cryopreserved pbmc does not generally benefit the performance of antigen-specific t cell elispot？assays休息的低温贮藏pbmc一般不会受益有关酶联免疫斑点抗原t细胞的性能分析.pdf

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resting of cryopreserved pbmc does not generally benefit the performance of antigen-specific t cell elispot？assays休息的低温贮藏pbmc一般不会受益有关酶联免疫斑点抗原t细胞的性能分析

Cells 2012, 1, 409-427; doi:10.3390/cells1030409 OPEN ACCESS cells ISSN 2073-4409 /journal/cells Article Resting of Cryopreserved PBMC Does Not Generally Benefit the Performance of Antigen-Specific T Cell ELISPOT Assays Stefanie Kuerten 1,*, Helena Batoulis 1, Mascha S. Recks 1, Edith Karacsony 2, Wenji Zhang 2, Ramu A. Subbramanian 2 and Paul V. Lehmann 2,* 1 Department of Anatomy I, University of Cologne, Joseph-Stelzmann-Str. 9, 50931 Cologne, Germany; E-Mails: helena.batoulis@uk-koeln.de (H.B.); mascha.recks@uk-koeln.de (M.S.R.) 2 Cellular Technology Ltd. (C.T.L), Shaker Heights, OH 44122, USA; E-Mails: edith.karacsony@ (E.K.); wenji.zhang@ (W.Z.); ramu.subbramanian@ (R.A.S.) * Authors to whom correspondence should be addressed; E-Mails: stefanie.kuerten@uk-koeln.de (S.K.); paul.lehmann@uk-koeln.de (P.V.L.); Tel.: +49-221-478-5414 (S.K.); Fax: +49-221-478-6711 (S.K.). Received: 13 June 2012; in revised form: 12 July 2012 / Accepted: 14 July 2012 / Published: 30 July 2012 Abstract: T cell monitoring is increasingly performed using cryopreserved PBMC. It has been suggested that resting of PBMC after thawing, that is, culturing them overnight in test medium, produces higher antigen-induced spot counts in ELISPOT assays. To evaluate the importance of overnight resting, we systematically tested cryopreserved PBMC from 25 healthy donors. CEF peptides (comprising CMV, EBV and flu antigens) were used to stimulate CD8 cells and mumps antigen to stimulate CD4 cells. The da