Analysis of Neisseria gonorrhoeae outer membrane protein Porin B regulates the immune response by in-hibiting the proliferation of CD4+T-cells
Objective To investigate whether the outer membrane protein Porin B of Neisseria gonorrhoeae can regulate the immune response by inhibiting the proliferation of CD4+T cells.Method The DNA sequence of the Porin B gene of NG strains was obtained from GenBank,and the DNA of NG strain FA1090 was used as a template to amplify the Porin B gene,and the Porin B gene was de-tected by 1.5%agarose gel electrophoresis.The PCR product was double digested with Bam HⅠ and Eco RⅠ and cloned into the di-gested vector pET30a to obtain the recombinant plasmid pET30a-Porin B.The pET30a-Porin B was detected by 1.5%agarose gel elec-trophoresis.CD4+T cells were isolated from spleens and lymph nodes of C57BL/6 mice and divided into control,LPB,MPB and HPB groups.The control group was not treated in any way,and the LPB,MPB and HPB groups were co-cultured with low,medium and high doses of pET30a-Porin B,respectively.Flow cytometry was used to detect the proliferation and apoptosis of CD4+T cells in each group and comparatively analysed.Results The PCR amplification products were subjected to 1.5%agarose gel electrophoresis,and a clear band at 1.0 kb was obtained,indicating that the correct Porin B gene was amplified.pET30a-Porin B was subjected to 1.5%agarose gel electrophoresis,and 1.0 kb of the target fragment and 5.4 kb of vector fragment appeared,which indicated that the recom-binant plasmid pET30a-Porin B was successfully constructed.Porin B was successfully constructed.The proliferation index of CD4+T cells in LPB,MPB and HPB groups were significantly decreased compared with the control group,and with the increase of the dose of recombinant plasmid pET30a-Porin B,the proliferation index of CD4+T cells was decreased about significantly.The apoptotic proportion of CD4+T cells in LPB,MPB and HPB groups were significantly increased compared with the control group,and with the increase of the dose of recombinant plasmid pET30a-Porin B dose increased,the increase in the proportion of CD4+T cell apoptosis was more sig-nificant.Conclusion Porin B can inhibit the proliferation of CD4+T cells and promote their apoptosis in a dose-dependent manner.This indicates that Porin B can regulate the body's immune response by inhibiting CD4+T cell proliferation.A large amount of research and clinical trials are needed in the later stage to demonstrate the clinical efficacy of Porin B and provide basic clinical data for the study of NG vaccines in clinical practice.
Neisseria gonorrhoeaeOuter membrane proteinPorin B proteinCD4+T cellImmunity
万方数据
