Immunomodulatory Effects of Hemagglutinin- (HA-) Modified A20 B-Cell Lymphoma Expanded as a Brain Tumor on Adoptively Transferred HA-Specific CD4+ T Cells
Figure 3
HA-specific CD4+Thy1.1+ T cells from cervical lymph nodes of A20HA brain tumor-bearing mice show increased response to restimulation in vitro with MHC class II synthetic HA peptide. ((a), (b), (c)) Flow cytometry of CD4+Thy1.1+CD44+ T cells from cervical lymph nodes on day 2 after adoptive transfer. HA-specific CD4+Thy1.1+ T cells were injected i.v. into A20HA brain tumor-bearing mice 5 days after i.c. tumor inoculation (A20HA + T cells; dot plots in the upper row) and a tumor-free control group (T cells; dot plots in the middle row). A20HA brain tumor-bearing mice in the absence of HA-specific CD4+Thy1.1+ T cells represented a negative control (A20HA; dot plots in the lower row). Cervical lymph nodes were pooled and analyzed for CD4+Thy1.1+CD44+ T cells (a) before incubation (in vivo column; right upper quadrants) and ((b), (c)) after 72 h incubation in vitro in triplicates (b) without (in vitro column; right upper quadrants) and (c) with (in vitro column; right upper quadrants) HA peptide. Histograms under the dot plots show the level of CD44 expression in the gated populations of CD4+Thy-1.1+ T cells (shown in the right upper quadrants). Numbers above histograms indicate percent of gated CD4+Thy-1.1+ T cells in the designed dot plots (left numbers) and intensity of CD44 expression by these cells (right numbers). One representative experiment of equivalent two is shown.
