| PartⅠThe Antitumor Effect of BCG in Treatment of Transplanted Human promyelocytic leukemia HL-60 cells in Nude MiceObjective It has been a long time to explore a new type of tumor treatment,which only selectively act on tumor cells with limited toxicity for normal tissues.Adjuvant therapy is appropriate treatment.Now BCG as the adjuvant treatment in Bacillus Calmette-Guerin treatment of bladder cancer and melanoma have achieved significant efficacy.But the efficacy of BCG used for the treatment of acute leukemia in patients were very few studied and reported in the literature.In this paper,through the establishment of human promyelocytic leukemia cell mutant xenograft models,and to explore the anti-leukemia effect of the BCG(Bacillus Calmette-Guerin vaccine,BCG).Methods An animal model was established by inoculating the Human promyelocytic leukemia HL-60 cells into the BALB/C(8～10 weeks of age) nude mice.The mice were randomly divided into two groups and received BCG therapy,The two groups include control group and experimental group,control group with normal saline in the tumor inoculation,and the experimental group was divided into T1 BCG group,T2 inactivated BCG group.Observed in each group the survival of nude mice with tumors,as well as through a number of tumor tissues and paraffin sections line HE staining,light microscope observation of morphological changes within the tumor.And it is to observe the immune response in infiltration of leukemia after the organization vaccinated with the immunohistochemical method.Results After HL-60 cells into nude mice,nude mice in each group were detected an swelling in the body surface against injection site,and gradually there is partial transfer, and transfer of liver and spleen.In pathology BCG group,inactivated BCG group showed different degrees of tumor necrosis phenomenon.It is to see the CD20-positive cells in the internal and marginal zone of the tumor tissue in the control group and experimental group,in particular afer using the BCG in the tumor tissue and necrotic area it can been observated the aggregation of a large number of CD20 positive cells of,and especially in the alive BCG group. Conclusion 1 Human promyelocytic leukemia HL-60 leukemia cells transplanted in nude mice can establish the Animal model of leukemia,and there is the phenomenon of systemic metastasis.2 Comparative study by the groups it is found that there is obvious inflammatory cell infiltration in Leukemia organization,and BCG on tumor tissue has its own significant role in damage.3 BCG can delay the growth of leukemia cells and extend the survival period of nude mouse with leukemia,which suggested that BCG has a clear anti-leukemia effect. PartⅡThe study of the mechanism of anti leukemia about the lymphocytes activated by BCG in vitroObjective Cancer of minimal residual disease(minimal residual disease,MRD) and multidrug resistance(multiple drug resistance,MDR),respectively,are the main cause of refractory and become difficult to capture the blood tumor.Study shows that after leukemia achieved complete remission immunotherapy can improve the immune efficacy, and is expected to reach the purpose to cure leukemia.In this paper,through the establishment of human promyelocytic leukemia cell mutant xenograft models,and Discussion BCG(BCG) in nude mice leukemia inhibit the growth of leukemia cells and its mechanism.It will provide a theoretical basis for the clinical application of BCG.Methods An animal model was established by inoculating the Human promyelocytic leukemia HL-60 cells into the BALB/C(8～10 weeks of age) nude mice.First of all,the application of Ficoll-Hypaque separation collect the peripheral blood mononuclear cells (PBMC) from Normal volunteer,and then injected the actival immune cells cultured in vitro into the tumor tissue of the nude mice;According to DC-activated lymphocytes the nude mouse are divided into different.Groupl:vaccine cultured DC induced by rhGM-CSF+rhIL-4+rhTNF-α(GIT);Group2:GIT United with BCG in different ways (live BCG group;Group3:inactivated BCG group;Group4:United BCG and HL-60 cell antigen preparation of cytotoxic cells group).It is to observe in each group the survival of nude mice with tumors,as well as the tumor through a number of organizations and dirty Organize line browser HE stained paraffin sections,light microscopic observation of morphological changes within the tumor,DC to further explore the role of tumor cell killing effects of the different.And it is to observe the immune response in infiltration of leukemia after the organization vaccinated with the immunohistochemical method.Results After HL-60 cells into nude mice,nude mice in each group were detected an swelling in the body surface against injection site,and gradually there is partial transfer, and transfer of liver and spleen.After the injection of the activation of immune cells,the groups showed different levels of tumor necrosis.The group using the actival cells cultured by antigen-presenting cells in vitro the nude mouse prolonged survival of about 1～2 days than the other applications group.Immunohistochemical results suggest that in each group there are a large number of inflammatory cells in the tumor tissue,around necrotic tissue the large number of their B lymphocytes can be seen in mice,Group4 Group more visible tumor necrosis broader region.The results also showed that CD35, CD45RO and CD8-positive cells cultured in vitro in nude mice after injection of short-term survival,has played the role of anti-tumor,and also prompted that the DC played a vital role in anti-tumor mechanisms,CTL is more significant in the destruction of tumor cellsConclusion 1 Human promyelocytic leukemia HL-60 leukemia cells transplanted in nude mice can establish the Animal model of leukemia.2 BCG can induced DC maturation and enhance their antigen-presenting function.3 It didn't find that activation of live BCG and the fired BCG in inducing the role of DC is a significant difference.4 By antigen-presenting dendritic cells,the immune activity of lymphocytes can be a significant anti-tumor,and extended of the survival time of mice.5 Leukemia tumor cells has its own nude CD20 lymphocytic infiltration,as well as CD35,CD45RO positive cells,Group4 Group in CDS-positive cells still exist. The anti-tumor effects of BCG was prompted by the promotion of DC cells,B lymphocytes,T lymphocyte activity and a variety of factors. |
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