Effects And Mechanisms Of Anti-p40Inhibiting Experimental Anti-GBM Nephritis

Objective: Both human and experimental glomerulonephritis are characterized by Tcells directed immune reponses and glomerular inflammatory reaction. However, thedifferent T cell subsets, T helper (Th)1and Th17cells are closely linked, andmutually affected. Interleukin(IL)-12and-23share the p40subunit and are importantfor the development of Th1and Th17cells responses in anti-glomerular basementmembrane nephritis. However, It is not well known about the impact of treatmentwith antagonistic p40antibody in inhibiting disease progression of nephrotoxic serumnephritis (NTN).Methods: C57BL/6mice were randomly divided into three groups (n=20), and eachgroup of mice were intraperitoneally injected with both the same proportion of amixture of rabbit IgG and complete Freund’s adjuvant. Since day10, controls wereintraperitonenally injected saline. In nephrotoxic serum nephritis (NTN) groupnephrotoxic sheep serum and incomplete Freund’s adjuvant (ratio between the two4:1)were intraperitoneally injected every other day until the fourth week, according to therenal pathological changes to adjust the dosing frequency. Mice in anti-p40intervention group were injected anti-p40monoclonal antibody for a weekcontinuously, at the same time, given nephrotoxic sheep serum and incompleteFreund’s adjuvant every other day until the fourth week. We impliedimmunofluoresence and periodic acid-Schiff(PAS)-stained to determine the pathologychange of the kidney. And renal function (the blood urea nitrogen levels、creatinineand random urine protein) were observe. FACS was used to assess the T cellsinfiltrated into the kidney and system immune.Results: We can observe the deposition of rabbit IgG and mouse IgG along the GBMby kidney frozen tissue immunofluoresence. Examination of periodic acid-Schiff (PAS)-stained kidney sections and renal function (the blood urea nitrogen levels、creatinine and random urine protein) of mice showed severe focal glomerular damage.Cytokine staining with subsequent FACS analysis showed that anti-p40injectionreduce the Th1and Th17differentiation and infiltration into the kidneys.Conclusion: Anti-p40-injecting mice have reduced numbers of Th1and Th17cells,and developed less severity of nephritis as estimated by renal function, randomproteinuria, and frequency of glomerular crescent formation. These resultsdemonstrated that anti-p40inhibited Th1and Th17cells responses in the experimentalglomerulonephritis.