An Intracameral Injection of Antigen Induces In Situ Chemokines and Cytokines Required for the Generation of Circulating Immunoregulatory Monocytes
Figure 4
Regulation of the increase in TNF-α and CCL2 in the anterior chamber after an intracameral injection.
Aqueous humor was collected at at 3 and 6 hour intervals following the intracameral injection of OVA. CCL2 and TNF-α levels in the aqueous humor were measured by ELISA. (A) Neutralization of TNF-α prevents the early rise of CCL2. Mice received an intracameral injection of OVA +/– anti-TNF-α antibody or isotype control. Neutralization of TNF-α via anti-TNF-α antibody at the time of AC injection significantly (P<0.05) reduced the early (3 hr) rise of (CCL2) MCP-1 but not at 6 hours. The data were pooled from 2 independent experiments and a minimum of 6 replicates were used in each group for each experiment. (B) Neutralization of TGF-β via a neutralizing antibody or TGF-β receptor kinase inhibitor (SB-431542) results in a significant reduction(P<0.05) in the rise of TNF-α induced by an intracameral injection of OVA. The data were pooled from 2 independent experiments. A minimum of 6 replicates were used for each experiment. (C) Neutralization of TGF-β via anti-TGFβ neutralizing antibody, Soluble TGF-β receptor or by TGF-β receptor kinase inhibitor (SB-431542) does not influence the CCL2 levels in the aqueous humor.(D) Neutralization of TGF-β by intracameral injection of anti-TGF-β with OVA does not block the infiltration of monocytes into the anterior chamber. Irides were removed 16 hr after intracameral injection and cell suspensions stained for Gr1 and CÎ11b. Experiments were repeated 3X.