| Trichinella spiralis is a zoonotic parasitic nematodes and lead to seriously disease in human being and different animal. It is called trichinosis and distribute in worldwide. Eventhough trichinella spiralis infection incur negative effect on public health and animal industry, it still get more focuses for elevating resistance in other disease of host by regulating immunity and especially in tumor. Recently, studies show that that the trichinella spiralis parasite and/or its excretion/secretion antigen can inhibit tumors. The studies support that trichinella spiralis will be used in anti-tumor as a biological agent.In recent years, cancer has been the principal killer to threat life and health of human. According to statistics, liver cancer has ranked top two cancer, after lung cancer ranking in China. Moreover, the poor prognosis of HCC had challenged traditional clinical treatment. So, a new efficient anti-tumor agents to improve the effect of surgery, radiotherapy and chemotherapy is booming.Precursor dendritic cells(DCs) were isolated from the hind legs of the femur and tibia of the C57BL/6 mice and stimulated by cytokines to induce them differentiation and maturation in vitro. The DCs of marrow-derived DC form C57BL/6 mice was identified by microscope, scanning electron microscope, HE staining and flow cytometry. The identified DCs further were stimulated by ES antigen from different developmental stages in lfe cycle of trichinella spiralis. Mouse model of hepatocellular carcinoma H22 were injected the DCs intraperitoneally. IL-4, IL-6, IL-10 and IFN-r of tail vein blood samples collected every 2 days in two weeks were detected. After two weeks, all the mice were sacrificed to evaluated the tumor inhibition rate. It was found that, the inhibition rate in preventive group reached 46.91% with clinical significance. In the trial, the mean body weight of mice in the experimental group were lower than the control group. It indicated that tumor growth in the experimental group mouse were lower than the control group. The average tumor weight was 1.8234 g or 3.4348 g in the experimental group and the control group respectively. Detection of the levels of IL-4 and IL-10 in serum in the experimental group mice were higher than the control mice, while the IL-6 and IFN-r were lower. The result showed that the marrow-derived DC from C57BL/6 mice stimulated by Trichinella ESP has a preventive effect on H22 hepatocellular carcinoma.Next, based on the above studies, the peripheral blood DC isolated healthy adult person and stimulated by the trichinella ESP to test the preventive effect on human H7402 hepatocellular carcinoma(HCC). The healthy adult peripheral blood mononuclear cells(PBMC) isolated by the Ficoll separation liquid were stimulated by adding cytokines to induced them to differentiate and mature in vitro. The DCs were identified by common microscope, scanning electron microscope, HE staining and flow cytometry. The DCs showed perfect characterize of healthy adult peripheral blood DC. Furthermore the DC were stimulated by Trichinella ESP and injected in the nude mice tumor models of H7402 human hepatocellular carcinoma intraperitoneally. The mice were weighed every 3 days.After two weeks, all mices were killed to evaluate tumor inhibitory rate. It was found that, the inhibition rate in preventive group reached 37.04% with clinical significance. The results showed that the mean body weight of in the experimental group mice increase were slower than the control group, but slightly faster than the the positive control group. Tumor weight in the experimental group is lower than the control group of mice and slightly higher than the positive control group. The average tumor weight was 0.9089 g or 1.4437 g in the experimental group and the control group respectively Cytokines level in mice spleen detected by Real-time quantitative PCR of and are consistent with the results of H22 hepatocellular carcinoma.The marrow-derived DC with the perfect characteristics from C57BL/6 mice and the peripheral blood DC with the perfect characteristics from healthy adults were successfully isolated and induced. The effects on mice and H22 and human H7402 hepatocellular carcinoma, of DC stimulated by Trichinella ESP both showed good preventive effects in the study. This study will provide some theoretical and experimental reference. It will positive contribute to application of trichinella spiralis and its production to improve immune function of anti-tumor by stimulating DC from patients and used in clinic. |
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