ABSTRACT
Objective To investigate the role of IFN-γ and FOXP3 expression in subpopulation distribution and functions of tumor-infiltrating lymphocytes (TILs) in the microenvironment of renal cell cancer.Methods 30 renal cell cancer tissue samples were freshly collected from the laparoscopic radical nephrectomy in the first hospital of Jiaxing.After frozen sectioning,immunofluorescent staining was conducted to detect the infiltrating CD4 positive and CDs positive cells,and the expression of FOXP3 and IFN-γ as well.In addition,TILs were isolated from the tumor tissues by density-gradient centrifugation.TILs from tumor center or tumor invasive edge were purified independently and measured for the mRNA levels of FOXP3 and IFN-γ by qRT (quantitative reverse transcription)-PCR.Results Tumor-infiltrating CD4+ and CD8+ T cells were concentrated in the invasive edge of renal cell cancer tissues.The expression of FOXP3 was found to be inversely related to that of IFN-γ from the immunofluorescent staining.The relative FOXP3 mRNA levels for the TILs from tumor center and invasive edge were 64.6 ± 9.4 and 36.2 ± 1.8,respectively,with significant difference(P ＜0.05).The relative IFN-γ mRNA levels were 631.8 ± 151.4 and 1 726.0 ± 344.1 (P ＜ 0.05).The trend of relative expression of FOXP3 was reversed in terms of IFN-γ.Conclusions The study on the renal cell cancer tissue samples suggested that the tumor-specific cytotoxic immune cells relatively concentrated in the tumor invasive edge.
ABSTRACT
Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.
ABSTRACT
Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.
ABSTRACT
Objective To study the inhibitory effects of angelica A_3 active fraction (A_3) inflammation and up-regulation of cyclooxygenase-2 (Cox-2) expression of rat uterus induced by lipopolysaccharides (LPS). Methods The anti-inflammatory effects of A_3 were investigated in rats using the carrageenin-induced paw swelling model and in mice using dimethylbenzene-induced ear edema model; RT-PCR and Western blotting were used to analyze Cox-2 mRNA and the protein expression levels. Results A_3 (1, 5, and 10 mg/kg) dose-dependently inhibited dimethylbenzene-induced ear edema in mice and paw swelling in rats by ig administration. LPS 1 ?g/mL could significantly increase the level of Cox-2 mRNA and protein expression. A_3(10—320 mg/L) could concentration-dependently inhibit Cox-2 mRNA and protein over-expression stimulated by LPS. Conclusion A_3 possesses better anti-inflammatory effects than angelica oil, which maybe relates to its inhibitory effects on Cox-2 overexpression.