Low oxygen tension and autologous plasma enhance T-cell proliferation and CD49d expression density in serum-free media.

BACKGROUND: As cellular immunotherapy with ex vivo expanded cells becomes more widely used to treat a variety of illnesses, optimization of culture parameters, to maximize cell production and function, is essential for continued success. The effects of reduced oxygen tension and autologous plasma on T-cell expansion, receptor expression, apoptosis, and cytolytic activity in serum-free media were investigated.
METHODS: PBMCs derived from whole blood samples were activated with anti-CD3 and anti-CD28 MAb in serum-free (AIM V) medium containing IL-2, and maintained at 5% and 20% oxygen tension. In some cases cultures were supplemented with 2% autologous plasma.
RESULTS: Low oxygen enhanced T-cell expansion 13- and 4.8-fold in serum-free and plasma-supplemented media, respectively. Autologous plasma also had a beneficial effect on T-cell cultures. Plasma-supplemented cultures expanded 74-fold more than serum-free cultures at low oxygen tension, and 43-fold more at high oxygen tension. Several samples expanded very poorly under serum-free conditions, and reasonable cell numbers were obtained only from plasma-supplemented cultures. CD49d expression density increased 3-fold to 4-fold in cultures supplemented with plasma. In contrast to our previous findings in serum-containing media, IL-2 receptor expression kinetics were unaffected by oxygen tension. No effects caused by oxygen tension or autologous plasma on expression of other surface antigens (CD4, CD8, CD44, CD95) were observed. DISCUSSION: Low oxygen tension and autologous plasma greatly increase expansion of T cells, thereby decreasing the time needed for production of cells for prophylaxis. Increased CD49d expression density may translate into improved migration and cytotoxicity.