Group O plasma as a media supplement for CAR-T cells and other adoptive T-cell therapies

Transfusion. 2020 May;60(5):1004-1014. doi: 10.1111/trf.15745. Epub 2020 Mar 13.

Abstract

Background: 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.

Study design and methods: 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.

Results: 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.

Conclusions: 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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Intramural

MeSH terms

  • ABO Blood-Group System / blood*
  • Cells, Cultured
  • Culture Media / chemistry*
  • Flow Cytometry
  • HLA-A Antigens / blood
  • HLA-A Antigens / immunology
  • Humans
  • Immunotherapy, Adoptive* / methods
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / metabolism
  • Leukocytes, Mononuclear / transplantation
  • Lymphocyte Activation
  • Plasma / chemistry
  • Plasma / physiology*
  • Primary Cell Culture / methods*
  • Primary Cell Culture / standards
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / metabolism
  • Receptors, Chimeric Antigen / genetics
  • Receptors, Chimeric Antigen / metabolism
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / metabolism
  • T-Lymphocytes / transplantation
  • Tissue Donors

Substances

  • ABO Blood-Group System
  • Culture Media
  • HLA-A Antigens
  • Receptors, Antigen, T-Cell
  • Receptors, Chimeric Antigen