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Reported incidence of TA\u2010GVHD is lower when leukoreduced RBCs and older blood products are transfused; therefore, the impact of leukoreduction and storage was evaluated as an alternative prevention strategy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design and Methods<\/jats:title><jats:p>Effectiveness of leukoreduction filters on white blood cell (WBC) proliferation was evaluated by filtering buffy coat (BC) products and isolating residual WBCs. Additionally, leukoreduced RBCs were spiked with 5\u2009\u00d7\u200910<jats:sup>6<\/jats:sup> WBCs on Day 21 of hypothermic storage, then stored and processed on Days 7, 14, and 21 to obtain residual WBCs to investigate the impact of hypothermic storage on their viability and proliferative ability. Viability of residual WBCs was assessed by staining with annexin V and an antibody cocktail for flow cytometry analysis. Proliferative ability was assessed by placing carboxyfluorescein diacetate succinimidyl ester\u2013labeled residual WBCs into culture for 6\u2009days with phytohemagglutinin before flow cytometry assessment.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Filtration of BC units depleted WBCs, particularly T lymphocytes, to 0.001%\u2009\u00b1\u20090.003% cells\/unit, although proliferative activity remained consistent with prefiltration levels of WBCs. WBCs in stored RBCs remained viable even on Day 21 of storage; however, the proliferative activity decreased to 0.24%\u2009\u00b1\u20090.41%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Hypothermic storage of RBCs for 21\u2009days or more is sufficient to inactivate T lymphocytes, which may help prevent TA\u2010GVHD when irradiated RBCs are not available.<\/jats:p><\/jats:sec>","DOI":"10.1111\/trf.16273","type":"journal-article","created":{"date-parts":[[2021,1,23]],"date-time":"2021-01-23T04:03:25Z","timestamp":1611374605000},"page":"1247-1257","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Hypothermic storage of leukoreduced red blood cells for greater than 21\u2009days is a safe alternative to irradiation"],"prefix":"10.1111","volume":"61","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8854-2872","authenticated-orcid":false,"given":"Olga","family":"Mykhailova","sequence":"first","affiliation":[{"name":"Centre for Innovation Canadian Blood Services  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tracey R.","family":"Turner","sequence":"additional","affiliation":[{"name":"Centre for Innovation Canadian Blood Services  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carly","family":"Olafson","sequence":"additional","affiliation":[{"name":"Centre for Innovation Canadian Blood Services  Edmonton Alberta Canada"},{"name":"Department of Laboratory Medicine and Pathology University of Alberta  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anita","family":"Howell","sequence":"additional","affiliation":[{"name":"Centre for Innovation Canadian Blood Services  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susan N.","family":"Nahirniak","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta  Edmonton Alberta Canada"},{"name":"Alberta Precision Laboratories  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juanita","family":"Wizniak","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology University of Alberta  Edmonton Alberta Canada"},{"name":"Alberta Precision Laboratories  Edmonton Alberta Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanan Y. 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