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To define protocols facilitating sample shipment and longitudinal analysis, this study evaluated the susceptibility of different functional assays for <jats:italic>A. fumigatus<\/jats:italic>\u2010specific T\u2010cell quantification and characterisation to pre\u2010analytic delays. <jats:styled-content style=\"fixed-case\">PBMC<\/jats:styled-content>s from 6 healthy donors were analysed after immediate isolation, after 6\u00a0hours whole blood storage or after cryopreservation using 3 different common media. Functional responses to <jats:italic>A. fumigatus<\/jats:italic> lysate stimulation were comparatively assessed by flow cytometry, <jats:styled-content style=\"fixed-case\">ELISPOT<\/jats:styled-content> and 14\u2010plex cytokine assay. After 6\u00a0hours pre\u2010analytic storage, all functional assays showed reduced detection rates, higher coefficients of variation (<jats:styled-content style=\"fixed-case\">CV<\/jats:styled-content>) and widely varying individual recovery indices of specific T\u2010cell response. While cryopreservation resulted in sufficient yields and largely unaltered cellular composition, outcomes of functional readouts significantly differed from freshly processed samples. For <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>154\u2010based flow cytometry, only cryopreservation in <jats:styled-content style=\"fixed-case\">RPMI<\/jats:styled-content> supplemented with autologous serum resulted in satisfactory detection rates and <jats:styled-content style=\"fixed-case\">CV<\/jats:styled-content>s. For <jats:styled-content style=\"fixed-case\">ELISPOT<\/jats:styled-content> and cytokine secretion assays, none of the cryopreservation protocols provided sufficient concordance with immediately processed samples. Even using the same readout platform, individual analytes widely varied in their susceptibility to cryopreservation, highlighting that distinct optimisation is required depending on the downstream assay.<\/jats:p>","DOI":"10.1111\/myc.12780","type":"journal-article","created":{"date-parts":[[2018,4,3]],"date-time":"2018-04-03T08:32:52Z","timestamp":1522744372000},"page":"549-560","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Susceptibility of <i>A. fumigatus<\/i>\u2010specific T\u2010cell assays to pre\u2010analytic blood storage and <scp>PBMC<\/scp> cryopreservation greatly depends on readout platform and analytes"],"prefix":"10.1111","volume":"61","author":[{"given":"Chris D.","family":"Lauruschkat","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7784-7256","authenticated-orcid":false,"given":"Sebastian","family":"Wurster","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lukas","family":"Page","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria","family":"Lazariotou","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariola","family":"Dragan","sequence":"additional","affiliation":[{"name":"Department of Surgery I University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philipp","family":"Weis","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew J.","family":"Ullmann","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hermann","family":"Einsele","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u00fcrgen","family":"L\u00f6ffler","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine II University Hospital of Wuerzburg  Wuerzburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2018,6,11]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2013.00421"},{"key":"e_1_2_8_3_1","first-page":"253","article-title":"Using ELISpot technology to improve the diagnosis of tuberculosis infection: from the bench to the T\u2010SPOT.TB assay. 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