ABSTRACT
Aim To establish stable and reliable animal models of Blau syndrome (BS) in vivo. Methods C57BL/6J mice were intraperitoneally injected with muramyl dipeptide (MDP) or L18-MDP to induce systemic inflammatory model of BS. Meanwhile, positive drug etanercept (ETN) was set to investigate the response of the model to evaluate effectiveness. SD rats were intravitrealiy injected with MDP to establish BS-associated uveitis model. Serum levels of TNF-a, IL-6 and IL-8 were detected by enzyme linked immunosorbent assay (ELISA). Histopathologic al changes of rat eyeballs were detected by HE staining and the expressions of p65, p-ERK, p-p38, p-JNK, TNF-a, IL-6 and IL-8 in vitreous were determined by immunohistochem-istry (IHC) staining. Results The serum level of TNF-a in mice increased after intraperitoneal injection of MDP (P < 0.05), and increased significantly after L18-MDP injection (P < 0.01). Meanwhile, the levels of IL-6 and IL-8 were also markedly induced by L18-MDP (P < 0. 01, P < 0. 01). ETN treatment evidently inhibited the increased levels of these above cytokines induced by L18-MDP (P < 0. 05, P < 0. 01). After the intravitreous injection of MDP in SD rats, there were numerous inflammatory cells infiltrated in retina and vitreous, and the retina was seriously damaged. The staining levels of p65, p-ERK, p-p38, p-JNK, TNF-a, IL-6 and IL-8 in eyeball tissues were significantly enhanced. Conclusions The systemic animal model of BS can be successfully established by intraperitoneal injection of L18-MDP in C57BL/6J mice, and the good BS-relat-ed uveitis can be induced by intravitreous injection of MDP in SD rats, which provides the simple, convenient, repeatable and i-deal animal models for exploring the pathogenesis of BS and e-valuating the efficacy of drugs.
ABSTRACT
Objective To study the effect of a new immunomodulator composed of muramyl dipeptide(MDP) and anti-CD10monoclonal antibody(MDP-Ab)on the dendritic cells(DC)of children with acute leukemia. Methods DC was adopted to divide the children with acute lymphoblastic leukemia into 6 groups,including the control group,unconjugated anti-CD10alone,unconjugated MDP alone,MDP-Ab alone,lipopolysaccharide(LPS)alone and MDP-Ab + LPS.The immunophenotypes,the endocytosis interleukin-12(IL-12)were detected.The stimulation index of autologous lymphocytes was assayed by adopting 5-(and 6)-carboxyfluorescein diacetate,succinimidyl es-ter(CFSE)-staining method.The supernatants of DC and autologous lymphocytes were used to detect the level of in-terferon-γ(IFN-γ)by using enzyme-linked immunosorbent assay.Results (1)DC immunophenotype:The ex-pressions of human leukocyte antigen-DR(HLA-DR),mature molecule(CD83)and co-stimulatory molecules (CD80and CD86)were increased significantly upon DC triggered with MDP-Ab,compared with the control group,un-conjugated anti-CD10group,and unconjugated MDP group,but lower than those in LPS and combination of MDP-Ab with LPS(F=629.62,P=0.000).(2)The level of IL-12:a significant increase in IL-12 level was detected in MDP-Ab group,LPS group,and combination of MDP-Ab with LPS group,compared with the control group,uncon-jugated anti-CD10group,and unconjugated MDP group(F=857.87,P=0.000). There were significant differences among the first three groups.(3)Endocytosis assay:The uptake of DCs stimulated by unconjugated anti-CD10,un-conjugated MDP,MDP-Ab immunoconjugate,LPS or combination was lower than that of immature DC in the control group which was(81.3 ± 10.1)%.(4)Mixed lymphocyte reaction and IFN-γ level:DC,treated with MDP-Ab, LPS and combination,stimulated more CFSE positive cells and higher level of IFN-γ secretion than the control group and unconjugated anti-CD10group,unconjugated MDP group. The most significance was observed in combination of MDP-Ab with LPS(F=393.36,P=0.000;F=2 497.18,P=0.000).Conclusion It is concluded that MDP-Ab could promote the proliferation and maturation of DC derived from blood of children with acute leukemia.
ABSTRACT
Objective Effects of hepatocellular carcinoma of antisense insulin like growth factor Ⅱ (antisense IGF Ⅱ) combined with muramyl dipeptide (MDP) was studied. Methods A cDNA fragment of human IGF Ⅱ was synthesized and inserted into a eukaryotic expression vector of pcDNA 3 (antisense IGF Ⅱ ). Antisense IGF Ⅱ and MDP encapsulated with liposomes were introduced into HepG 2 human hepatocellular carcinoma cells and were injected into subcutaneous tumors generated in nude mice. The growth of HepG 2 cells transfected with antisense IGF Ⅱ and MDP, and the subcutaneous tumors injected with antisense IGF Ⅱ and MDP were observed. Results The growth of HepG 2 cells was blocked completely, and the subcutaneous tumors disappeared in two of five nude mice. Conclusions The combination therapy of antisense IGF Ⅱ and MDP is effective in the treatment of hepatocellular carcinoma.
ABSTRACT
The in vitro tumoricidal activity of peritoneal macrophages harvested from C57BL mice and activiated by li posomes containing water-soluble muramyl dipeptides (MDP) was measured by ~(125)I.UdR release assay. The results demonstrated that liposomes containing MDP could effectively activiate macrophages to become tumoricidal. The percentage of pu(?)monarymetastasis and the average metastatic pulmonary colonies of mice with experimental tumor metastasis decreased after intravenous injection of MDP encapsulated in liposomes. These results suggested that systemic administration liposomes containing MDP whichinduced macrophage activiation might provide a useful immunot heray for tumor, especial1y for pulmonary mini metastasis.