Efficacy studies of Treg activation using dsDNA desensitization inhibits immune function without side effect in lupus mice model

The aim of this study was to develop a novel therapeutic method for improving immune system regulation in SLE using escalating dose self-antigen dsDNA immunotherapy. Female Balb/c mice were given a single intraperitoneal injection of 0.5 ml pristane. Starting at 12 weeks after injection, the mice were evaluated for clinical and serological manifestations. Mice with lupus signs (PIL mice) were divided into two groups; a positive control group and PIL (0.1 µg/ml, 1 µg/ml, 10 µg/ml) EDI dsDNA group. EDI dsDNA was administered once every week in consecutively. The doses would increase every week. dsDNA was complexed with the cationic polyethyleneimine (PEI) before injection. Samples were analyzed for autoantibodies levels (dsDNA, ANA) and TGF-β cytokine from serum using ELISA and T-Reg, mature dendritic cells from spleen using flowcytometry. Liver and kidney function was measured by ALT/AST, Ur/Cr, Proteinuria and ALP serum. Efficacy study result showed that immune profile, inflammation marker, proteinuria, liver and renal functions were improved. significantly decreased ANA (p=0.02), anti-dsDNA (p=0.03), dendritic cell mature (p=0.02) compare to positive control, and not significantly decreases Th17 cells (p=0,18) but the result tend to get lower. Desensitization using self-antigen dsDNA was increased T-reg proliferation (p=0.00) and level of TGF-β (p=0.03) significantly compare to positive control. Analysis for side effect revealed that PIL with EDI dsDNA decrease SGOT(p=0.87) SGPT (p=0.22), Ureum (p=0.009), Creatinin (p=0,87), Proteinuria (p=0.009) and ALP (p=0,65) compared to positive control. Desensitization using self-antigen dsDNA coupled to PEI was able to modulate T-Reg as a regulator immune respon and inhibit B and T cell functions and potential to be developed further as new, safe and effective immunotherapy against pristane-induced SLE BALB/c mice.