IL 22 and Paclitaxel human endothelial cells IL 17A ELISA kits were purchased from R D Systems, Inc. and were performed based on kit protocols. Apoptosis Assay Apoptotic Inhibitors,Modulators,Libraries cells were detected by staining cells with both the annexin V FITC and TUNEL tech nology according to the manufacturers instructions. Phospho Bad expression was detected by western blot using anti Phospho Bad antibody. SDS PAGE and Western blotting A total of 5 million T cells were lysed in 100 ul lysis buffer. Complete cell lysis was achieved by immedi ately vortexing the cells and then boiling in an equal amount of 2 SDS protein loading buffer at 95 C for 5 minutes. Cell debris was removed by centrifugation at 12, 000 rpm for 3 min. Twenty microliter of each sam ple was loaded into a 12% SDS polyacrylamide gel con taining a 4% stacking gel.

Immunoblotting was Inhibitors,Modulators,Libraries carried out. Primary antibodies of anti Phospho Bad, anti Bad were purchased from Cell Signaling Technology. Anti b actin antibody was from Santa Cruz Biotechnology, Inc. SNP Genotyping Genomic DNA was extracted from the peripheral blood of each individual using Promega Wizard Genomic DNA Purification kit. The samples were analyzed by TaqMan genotyping assay using the Real time PCR system 7500. The primers and probes for C2CFB rs9332739 and C3 rs2230199 were from the inventory SNP assay while CFH rs1061170 were custom designed from Applied Biosystems. Geno types were determined based on the fluorescence intensi ties of FAM and VIC. The call rates of 3 assays were 98. 5% and the call accuracies were 100%. Statistical Analysis Non parametric methods were used since the expression of IL17 and IL22 does not follow a parametric distribution.

To evaluate if the expression of these 2 cytokines follows a normal distri bution, we visually checked the histograms as well as used the Kolmogorov Smirnov method. For the associa tion study between IL 22IL 17 and Inhibitors,Modulators,Libraries some characteristics of patients, Wilcoxons nonparametric two sample rank sum test was used. Age was analyzed using Pearson cor relation. The software used for all the analyses was The SAS System, version 9. 2. Results We listed the demographic, clinical information for both controls and AMD patients in Table 1 and Table 2. Ocular therapies, co morbidities and complement related genetic variance were also included to AMD patients for later data analysis. These information will be used to evaluate con founding factors.

All the subjects in this study are Cauca sians. There are 45 controls and the age range was from 59 to 87. Fifty three percent are females and 47% are males. There are 40 AMD patients in this study and the age range was from 57 to 97. Fifty percent are females and 50% are males. C5a promotes the expression of IL 22 and IL 17 from human T cells in Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries vitro To study the role of C5a on human CD4 T cells, we used ELISA selleck and intracellular staining to detect cytokine expression. PBMCs from AMD patients and controls were treated with or without C5a and a C5aR antagonist for 3 days.