The Role Of Borrelia Burgdorferi Recombinant Membrane Protein Rbmpa In The Research Of Human Macrophage Cell Line THP-1Cell In Vitro

Objective:To use recombinant Borrelia burgdorferi membrance protein A (rBmpA) to stimulate the human macrophage cell line(THP-1) in vitro to collect the cell cultural supernatant for detecting concentrations of HumanIL-1β、IL-6、IL-8、IL-12p70、TNF-α and MIF, and researching the role of Borrelia burgdorferi membrance protein A (BmpA) in pathogenic mechanism of Lyme arthritis.Methods:1. Cultured condition of THP-1cells:We cultured THP-1cell in37℃,5%incubator—co2with1640medium with10%FBS.2. THP-1cells were differentiated into adherent cells:THP-1cells (5X105/mL) were cultured in96-well plates (100ul/well) with100ng/mL PMA medium for24h.3. To use rBmpA to stimulate THP-1cells:THP-1cells(5X105/mL) were cultured in96-well plates(100ul/well) with20ug/mL,40ug/ml rBmpA for24h,48h and72h.4. To detect inflammatory cytokines concentration of Human IL-1β、IL-6、IL-8、 IL-12p70、TNF-α and MIF by ELISA in the THP-1cells cultural supernatant and to explore the role of BmpA in the pathogenesis of Lyme arthritis.Results:1. The culture technique of THP-1cells was successfully developed.2. We successfully induced the THP-1cells into adherent cell.3. Compared with the control group, inflammatory cytokine, including Human IL-1β、 IL-6、IL、IL-12p70and TNF-α, in the culture supernatant of THP-1cells increased significantly (P<0.05).4. Compared with the control group, inflammatory cytokine, Human-MIF, in the culture supernatant of THP-1cells was no significant difference (P>0.05).Conclusion:1. rBmpA plays an important role in stimulating THP-1cell to secrete inflammatory cytokines (Human IL-1β、IL-6、IL-8、IL-12p70and TNF-α).2. Human-MIF in THP-1cells were not affected by rBmpA at the experimental concentration.3. BmpA may be closely related with the development of Lyme arthritis by stimulating the human macrophage cell.