The Role Of Borrelia Burgdorfei Recombinant Membrane Proteins RBmpA In The Research Of Mouse Macrophages And Lymphocytes In Vitro

Objective:Use recombinant Borrelia burgdorfei membrance protein A (rBmpA) to stimulate the cell culture in vitro, including Kunming (KM) mice peritoneal macrophage, spleen lymphocyte and mouse monocyte macrophage cell line (RAW264.7), developing the cell proliferation experiment (MTT) on these cells, after stimulating cell culture in vitro with different concentration rBmpA, collect the cell cultural supernatant for detecting contents of IL-1β, IL-6, IL-17, through this way to research roles of various cells in pathogenic mechanism of Lyme arthritis.Methods:1. To explore extraction technique of Kunming mice spleen lymphocyte, optimize the conditions of extraction and culture to obtain more spleen lymphocyte.2. To explore extraction technique of Kunming mice peritoneal macrophage,optimize the conditions of extraction and culture to obtain more peritoneal macrophages.3. To explore the culture condition and culture technique of RAW264.7macrophage cell line.4. To develop MTT test on Kunming mouse spleen lymphocyte and RAW264.7macrophage cell line.5. To detect inflammatory cytokines concentration of IL-6, IL-17and IL-1β by ELISA and explore the roles of various cells in the pathogenesis of Lyme arthritis.Results:1. Through researching and exploring, develop extraction and culture technology of Kunming mouse spleen lymphocyte and peritoneal macrophage successfully.2. Develop the culture technique of RAW264.7macrophage cell line successfully.3. MTT test on spleen lymphocyte showed increase in cell proliferation (P<0.05), compared with the normal control group.4. Inflammatory cytokine, IL-6and IL-17, in the culture supernatant of spleen lymphocyte increased significantly (P<0.05), compared with the normal control group.5. Inflammatory cytokine, IL-1β, in the culture supernatant of peritoneal macrophage demonstrated no signifant difference (P>0.05), compared with the normal control group, through analyzing and exploring the relative reasons to lay the foundation for further research later.6. MTT test on RAW264.7macrophage cell line demonstrated increase in cell proliferation (P<0.05), compared with the normal control group.Conclusion:1. rBmpA plays a role in stimulating spleen lymphocyte by secreting inflammary cytokines (IL-6, IL-17).2. rBmpA caused an increase in cell proliferation in both spleen lymphocyte and RAW264.7macrophage cell line.3. IL-1β in peritoneal macrophages was not affected by rBmpA.4. rBmpA is closely related with Lyme arthritis.