HLA Class Ia and Ib Polyreactive Anti-HLA-E IgG2a Monoclonal Antibodies (TFL-006 and TFL-007) Suppress Anti-HLA IgG Production by CD19+ B Cells and Proliferation of CD4+ T Cells While Upregulating Tregs

Figure 5

Phytohemagglutin (PHA) mediated activation of CD3+/CD4+ T lymphocytes. PHA induces blastogenesis and proliferation of the T cells. Experiments were done in triplicate (see [47]). Blastogenesis of was determined by counting the lymphoblasts, after culturing purified lymphocytes with or (as control) without PHA for 72 h. Proliferation was monitored by labelling the purified lymphocytes with the intracellular fluorescent dye carboxyfluorescein succinimidyl ester (CFSE). After 72 h, the labelling of the cells was measured: PHA-treated T cells undergo four to six divisions. Using flow cytometry, the mitotic activity is measured by the successive twofold reductions in fluorescent intensity of the T cells placed in culture for 72 h. Experiments were done in triplicate [48]. In the no PHA box, the number of cells in the M4 column is highly negligible. The box “with PHA” shows increase in the cell numbers in M4 column (shown with ).