{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:12:14Z","timestamp":1772169134359,"version":"3.50.1"},"reference-count":16,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2023,4,25]],"date-time":"2023-04-25T00:00:00Z","timestamp":1682380800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["108395"],"award-info":[{"award-number":["108395"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["100174"],"award-info":[{"award-number":["100174"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["wellcomeopenresearch.org"],"crossmark-restriction":false},"short-container-title":["Wellcome Open Res"],"abstract":"<ns4:p>\n                    <ns4:bold>Background:<\/ns4:bold>\n                    Interferon-\u03b3 (IFN-\u03b3) secretion by T cells is a key correlate of immune protection against many pathogens including tuberculosis and the neglected tropical disease melioidosis. Clinical studies in tropical regions of immune responses to pathogens and vaccine monitoring studies require the collection of samples in resource-limited rural areas and subsequent shipment to central laboratories for downstream assays and long-term storage.\n                  <\/ns4:p>\n                  <ns4:p>Here, we studied the impact of two different shipping temperatures on the viability, composition and function of peripheral blood mononuclear cells (PBMC) using multi-colour flow cytometry and IFN-g enzyme-linked immunospot assay (IFN-g ELISpot), in order to provide guidance on sample shipment conditions for future clinical studies.<\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Methods:<\/ns4:bold>\n                    Paired peripheral blood mononuclear cell (PBMC) samples from recovered melioidosis patients were stored in liquid nitrogen (-196\u00b0C) and then shipped from Bangkok, Thailand to Oxford, UK at either -80\u00b0C (dry ice) or -196\u00b0C (dry shipper). After thawing, cell viability and composition were assessed by flow cytometry and antigen specific responses to\n                    <ns4:italic>Burkholderia pseudomallei<\/ns4:italic>\n                    (BP) were measured using IFN-g ELISpot.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Results:<\/ns4:bold>\n                    We observed modest lowering of viability in the majority of samples and a reduction in IFN-g responses to BP which correlated to a decrease of monocytes and natural killer cells in samples shipped at -80\u00b0C compared to -196\u00b0C. Despite being lower in magnitude antigen-specific responses remained detectable in the majority of samples.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Conclusions:<\/ns4:bold>\n                    Here we demonstrate that shipment of cryopreserved PBMC at -196\u00b0C has a benefit on cell viability, recovery and T cell responses to bacterial antigens, although useful information can still be obtained from samples shipped at -80\u00b0C, thus providing important guidance for sample management in future clinical trials.\n                  <\/ns4:p>","DOI":"10.12688\/wellcomeopenres.18822.1","type":"journal-article","created":{"date-parts":[[2023,4,26]],"date-time":"2023-04-26T11:55:07Z","timestamp":1682510107000},"page":"188","update-policy":"https:\/\/doi.org\/10.12688\/wellcomeopenres.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Impact of shipping temperature on cell viability and T cell responses to bacterial antigens"],"prefix":"10.12688","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4094-5143","authenticated-orcid":false,"given":"Patpong","family":"Rongkard","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5665-6293","authenticated-orcid":false,"given":"Susanna J.","family":"Dunachie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbara","family":"Kronsteiner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2023,4,25]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"S1-4","DOI":"10.1016\/S0035-9203(08)70002-6","article-title":"The global distribution of Burkholderia pseudomallei and melioidosis: an update.","volume":"102 Suppl 1","author":"B Currie","year":"2008","journal-title":"Trans R Soc Trop Med Hyg."},{"key":"ref-2","doi-asserted-by":"publisher","first-page":"1113-7","DOI":"10.4269\/ajtmh.2010.10-0038","article-title":"Increasing incidence of human melioidosis in Northeast Thailand.","volume":"82","author":"D Limmathurotsakul","year":"2010","journal-title":"Am J Trop Med Hyg."},{"key":"ref-3","doi-asserted-by":"publisher","first-page":"286-9","DOI":"10.1086\/341222","article-title":"Demonstration of a cell-mediated immune response in melioidosis.","volume":"186","author":"N Ketheesan","year":"2002","journal-title":"J Infect Dis."},{"key":"ref-4","doi-asserted-by":"publisher","first-page":"e0004152","DOI":"10.1371\/journal.pntd.0004152","article-title":"T-Cell Responses Are Associated with Survival in Acute Melioidosis Patients.","volume":"9","author":"K Jenjaroen","year":"2015","journal-title":"PLoS Negl Trop Dis."},{"key":"ref-5","doi-asserted-by":"publisher","first-page":"191-204","DOI":"10.1385\/1-59259-903-6:191","article-title":"Interferon-gamma ELISPOT assay for the quantitative measurement of antigen-specific murine CD8+ T-cells.","volume":"302","author":"G Cole","year":"2005","journal-title":"Methods Mol Biol."},{"key":"ref-6","doi-asserted-by":"publisher","first-page":"871-9","DOI":"10.1128\/CDLI.8.5.871-879.2001","article-title":"Development and validation of a gamma interferon ELISPOT assay for quantitation of cellular immune responses to varicella-zoster virus.","volume":"8","author":"J Smith","year":"2001","journal-title":"Clin Diagn Lab Immunol."},{"key":"ref-7","doi-asserted-by":"publisher","first-page":"843-50","DOI":"10.1038\/nm1592","article-title":"Multifunctional TH1 cells define a correlate of vaccine-mediated protection against Leishmania major.","volume":"13","author":"P Darrah","year":"2007","journal-title":"Nat Med."},{"key":"ref-8","doi-asserted-by":"publisher","first-page":"461-9","DOI":"10.1038\/nprot.2009.7","article-title":"The role of IFN-\u03b3 Elispot assay in HIV vaccine research.","volume":"4","author":"H Streeck","year":"2009","journal-title":"Nat Protoc."},{"key":"ref-9","doi-asserted-by":"publisher","first-page":"409-27","DOI":"10.3390\/cells1030409","article-title":"Resting of Cryopreserved PBMC Does Not Generally Benefit the Performance of Antigen-Specific T Cell ELISPOT Assays.","volume":"1","author":"S Kuerten","year":"2012","journal-title":"Cells."},{"key":"ref-10","doi-asserted-by":"publisher","first-page":"e76215","DOI":"10.1371\/journal.pone.0076215","article-title":"Overnight resting of PBMC changes functional signatures of antigen specific T- cell responses: impact for immune monitoring within clinical trials.","volume":"8","author":"S Kutscher","year":"2013","journal-title":"PLoS One."},{"key":"ref-11","doi-asserted-by":"publisher","first-page":"825-33","DOI":"10.1128\/am.20.5.825-833.1970","article-title":"Serological diagnosis of human melioidosis with indirect hemagglutination and complement fixation tests.","volume":"20","author":"A Alexander","year":"1970","journal-title":"Appl Microbiol."},{"key":"ref-12","article-title":"Extended data of impact of shipping temperature on cell viability and T cell responses to bacterial antigens.","author":"P Rongkard","year":"2023"},{"key":"ref-13","article-title":"Underlying data of Impact of shipping temperature on cell viability and T cell responses to bacterial.","author":"P Rongkard","year":"2023"},{"key":"ref-14","doi-asserted-by":"publisher","first-page":"193-200","DOI":"10.1016\/j.cryobiol.2013.06.012","article-title":"Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function.","volume":"67","author":"A Germann","year":"2013","journal-title":"Cryobiology."},{"key":"ref-15","doi-asserted-by":"publisher","first-page":"57-69","DOI":"10.1016\/j.jim.2007.02.003","article-title":"Defining blood processing parameters for optimal detection of cryopreserved antigen-specific responses for HIV vaccine trials.","volume":"322","author":"M Bull","year":"2007","journal-title":"J Immunol Methods."},{"key":"ref-16","doi-asserted-by":"publisher","first-page":"463-471","DOI":"10.3201\/eid2603.190206","article-title":"Human Immune Responses to Melioidosis and Cross-Reactivity to Low-Virulence Burkholderia Species, Thailand1.","volume":"26","author":"P Rongkard","year":"2020","journal-title":"Emerg Infect Dis."}],"container-title":["Wellcome Open Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,19]],"date-time":"2024-06-19T02:50:15Z","timestamp":1718765415000},"score":1,"resource":{"primary":{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,25]]},"references-count":16,"URL":"https:\/\/doi.org\/10.12688\/wellcomeopenres.18822.1","relation":{"has-review":[{"id-type":"doi","id":"10.21956\/wellcomeopenres.20868.r85085","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/wellcomeopenres.20868.r85081","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/wellcomeopenres.20868.r85085","asserted-by":"object"},{"id-type":"doi","id":"10.21956\/wellcomeopenres.20868.r85081","asserted-by":"object"}]},"ISSN":["2398-502X"],"issn-type":[{"value":"2398-502X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,25]]},"assertion":[{"value":"Indexed","URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.21956\/wellcomeopenres.20868.r85085, Munyaradzi Musvosvi, University of Cape Town, Rondebosch, Western Cape, 11 Jun 2024, version 1, indexed","URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1#referee-response-85085","order":0,"name":"referee-response-85085","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.21956\/wellcomeopenres.20868.r85081, Sylvia Chieng, Universiti Kebangsaan Malaysia, Bangi, 19 Jun 2024, version 1, indexed","URL":"https:\/\/wellcomeopenresearch.org\/articles\/8-188\/v1#referee-response-85081","order":1,"name":"referee-response-85081","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was supported by Wellcome [108395, <a href=https:\/\/doi.org\/10.35802\/108395>https:\/\/doi.org\/10.35802\/108395<\/a>; a Master\u2019s Fellowship in Public Health and Tropical Medicine to PR and 100174, <a href=https:\/\/doi.org\/10.35802\/100174>https:\/\/doi.org\/10.35802\/100174<\/a>; an Intermediate Clinical Fellowship award to SD]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":2,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}