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- Title
Group O plasma as a media supplement for CAR-T cells and other adoptive T-cell therapies.
- Authors
Nelson, Randin C.; Fellowes, Vicki; Jin, Ping; Liu, Hui; Highfill, Steven L.; Ren, Jiaqiang; Szymanski, James; Flegel, Willy A.; Stroncek, David F.
- Abstract
<bold>Background: </bold>Most chimeric antigen receptor T (CAR-T) cells and other adoptive T-cell therapies (ACTs) are currently manufactured by ex vivo expansion of patient lymphocytes in culture media supplemented with human plasma from group AB donors. As lymphocytes do not express A or B antigens, the isoagglutinins of non-AB plasmas are unlikely to cause deleterious effects on lymphocytes in culture.<bold>Study Design and Methods: </bold>Seeding cultures with peripheral blood mononuclear cell (PBMNC) concentrates from group A1 donors and using a CAR-T culture protocol, parallel cultures were performed, each with unique donor plasmas as media supplements (including group O plasmas with high-titer anti-A and group AB plasmas as control). An additional variable, a 3% group A1 red blood cell (RBC) spike, was added to simulate a RBC-contaminated PBMNC collection. Cultures were monitored by cell count, viability, flow cytometric phenotype, gene expression analysis, and supernatant chemokine analysis.<bold>Results: </bold>There was no difference in lymphocyte expansion or phenotype when cultured with AB plasma or O plasma with high-titer anti-A. Compared to controls, the presence of contaminating RBCs in lymphocyte culture led to poor lymphocyte expansion and a less desirable phenotype-irrespective of the isoagglutinin titer of the plasma supplement used.<bold>Conclusions: </bold>This study suggests that ABO incompatible plasma may be used as a media supplement when culturing cell types that do not express ABO antigens-such as lymphocytes for CAR-T or other ACT. The presence of contaminating RBCs in culture was disadvantageous independent of isoagglutinin titer.
- Subjects
ERYTHROCYTES; MASS media; CHIMERIC antigen receptors; BLOOD cells; PLASMA confinement; ABO blood group system; FLOW cytometry; CELL culture; IMMUNIZATION; MONONUCLEAR leukocytes; HLA-B27 antigen; CULTURE media (Biology); BLOOD plasma; CELL receptors; IMMUNOLOGY technique; RESEARCH funding; T cells; BLOOD
- Publication
Transfusion, 2020, Vol 60, Issue 5, p1004
- ISSN
0041-1132
- Publication type
journal article
- DOI
10.1111/trf.15745