Figure 7

Altered Mixed Lymphocyte Reaction in post-OVA KO mice. 0.1 × 105 CD4+ T cells isolated by magnetic activated cell separation (MACS) by positive selection from spleen of the mice were co-cultured with γ-irradiated 0.1x103 adhering alveolar macrophages from BALf of the same experimental animal. Both cell types were from the experimental animals themselves viz. C57Bl/6 WT and KO mice. This is Syngeneic MLR where the APCs (alveolar macrophages) were γ-irradiated (3300rads). Allogeneic MLR reaction involves co-culturing 0.1x105 CD4+ T cells from the spleen of BALB/c mice with 0.1x103 γ-irradiated APCs from BALf of the experimental mice. Both control (saline-treated) and OVA-treated of each group were tested. Each culture was incubated with 1μg/ml OVA.The responders here are the T cells and the stimulators are the APCs, viz. alveolar macrophages which are γ-irradiated to inhibit their own proliferation. Since acute allergic asthma is a Th2 mediated phenomenon, interaction between T cells and macrophages will elucidate functional cross-talk between the two cell types when responders are autologous as well as when they are from a different species. 2-fold increase in post-OVA NOX vs. post-OVA WT and 2.16-fold increase in post OVA-DKO vs. post-OVA shows that T cell:APC interaction is actually more efficient in the absence of the gp91phox and MMP-12 as well as gp91phox.