{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,15]],"date-time":"2023-10-15T05:40:19Z","timestamp":1697348419622},"reference-count":24,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2007,6,6]],"date-time":"2007-06-06T00:00:00Z","timestamp":1181088000000},"content-version":"vor","delay-in-days":5,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion Medicine"],"published-print":{"date-parts":[[2007,6]]},"abstract":"<jats:p><jats:bold><jats:sc>summary<\/jats:sc><\/jats:bold> Information is lacking regarding efficiency of removal of circulating dendritic cells (DCs) by leucoreduction (LR) of blood. This is important since DCs may play a role in transporting abnormal prion, the likely infectious agent of variant Creutzfeldt\u2013Jakob disease. In this study, we report development of a real\u2010time polymerase chain reaction (RT\u2010PCR) assay to quantify residual DCs in LR whole blood via measurement of selected messenger RNA (mRNA) markers.<\/jats:p><jats:p>Taqman\u2010based RT\u2010PCR assays were set up for CD83 as a marker of mature DCs, and CD1c, CD11c, CD303 and CD304 as markers for plasmacytoid and myeloid DCs along with the pan\u2010leucocyte marker CD45. We then assayed 46 paired pre\u2010\/post\u2010LR whole blood samples and determined the log<jats:sub>10<\/jats:sub> reduction of their CD83 and CD45 mRNA.<\/jats:p><jats:p>Our data indicate that RT\u2010PCR can be used to detect suitably low CD83 mRNA levels. We measured a median log<jats:sub>10<\/jats:sub> reduction for CD83 mRNA of 4\u00b75 [standard error of the mean (SEM) 0\u00b707] and 4\u00b71 (SEM 0\u00b710) for CD45 mRNA. These reductions are comparable to cell removal, where flow cytometry indicated a reduction in total white cell counts of 4\u00b73 log<jats:sub>10<\/jats:sub> (SEM 0\u00b709). Our other group of markers were reduced to their detection limits, CD1c (3\u00b79 log<jats:sub>10<\/jats:sub>, SEM 0\u00b73), CD11c (5\u00b70 log<jats:sub>10<\/jats:sub>, SEM 0), CD303 (3 log<jats:sub>10<\/jats:sub>, SEM 0\u00b71), CD304 (4\u00b70 log<jats:sub>10<\/jats:sub>, SEM 0) which are all higher than the minimum specifications for LR products.<\/jats:p><jats:p>In conclusion, we successfully developed an RT\u2010PCR assay to quantify suitably low numbers of DC cells. We show for the first time that DCs are effectively removed using a standard whole blood filter.<\/jats:p>","DOI":"10.1111\/j.1365-3148.2007.00735.x","type":"journal-article","created":{"date-parts":[[2007,6,7]],"date-time":"2007-06-07T06:58:44Z","timestamp":1181199524000},"page":"175-181","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Measurement of CD83 mRNA by real\u2010time polymerase chain reaction to determine removal of dendritic cells by leucoreduction of whole blood"],"prefix":"10.1111","volume":"17","author":[{"given":"Alex","family":"Morrison","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ian","family":"MacGregor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2007,6,6]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI200113155"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-3148.2000.00230.x"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2257.2001.00356.x"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1999.39111169.x"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/39057"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V98.13.3733"},{"key":"e_1_2_6_8_1","unstructured":"CJD Surveilance unit Edinburgh CJD Statistics[WWW document]. 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