重组乳酸杆菌Lb.hCGβ-C3d3经鼻腔免疫BALB/c小鼠的抗生育潜能  

The anti-fertility potential of inoculation via nose with recombinant live Lactobacilli of Lb.hCGβ-C3d3

作　　者：王健理[1] 姚晓英[1] 李大金[1] 

机构地区：[1]复旦大学附属妇产科医院暨妇产科研究所生殖免疫研究室,上海200011

出　　处：《中国免疫学杂志》2011年第7期616-619,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金(30700764;31070808)资助

摘　　要：目的：探讨表达hCGβ-C3d3融合蛋白的重组乳酸杆菌避孕疫苗经鼻腔黏膜接种BALB/c小鼠及其抗生育潜能。方法：用野生型乳酸杆菌Lb.及重组乳酸杆菌Lb.hCGβ和Lb.hCGβ-C3d3以两种剂量（10^9个和10^10个）经鼻腔滴注接种68周龄雌性BALB/c小鼠。3周后用相同剂量加强免疫一次。于初次免疫后的第6、7、8周收集血清,分别培养MLTC-1、BeWo细胞;化学发光法检测培养上清中孕酮或hCGβ水平;免疫荧光观察抗血清处理后BeWo细胞的融合情况。结果：用10^10个Lb.hCGβ-C3d3滴鼻免疫BALB/c小鼠产生的抗血清能显著抑制hCG刺激MLTC-1分泌孕酮,明显抑制BeWo细胞分泌hCGβ及细胞融合。结论：重组乳酸杆菌活疫苗Lb.hCGβ-C3d3经鼻腔接种诱导的hCGβ抗体可拮抗hCGβ生物学功能,并干扰滋养细胞功能,因此具有抗生育潜能。Objective：To evaluate the anti-fertility of potential by inoculation via nose with the recombinant live Lactobacilli expressing hCGβ-C3d3 fusion protein.Methods：Female BALB/c mice were immunized with 10^10,10^9 Lb,Lb.hCGβ or Lb.hCGβ-C3d3,respectively,via nose.The anti-serum was collected respectively in the 6-8th week after the primary immunization.MLTC-1 and BeWo cell line were respectively treated with the anti-serum,and then the supernatant was recovered,respectively.Progesterone and hCGβ in the supernatant were determined by Chemoluminescence assay.Fusion of the BeWo cells was investigated by immunofluorescence microscopy.Results：The anti-serum induced by the Lb.hCGβ-C3d3 inoculation with the dose of 10^10 could significantly inhibit the progesterone production in MLTC-1 cells via neutralizing hCG bioactivity,and suppress the hCGβ secretion and fusion in vitro of BeWo cell line.Conclusion：Lb.hCGβ-C3d3 live Lactobacilli are effective in blocking hCG bioactivity,which appears anti-fertility potential.

关 键 词：重组乳酸杆菌 HCGΒ C3d3鼻腔接种 

分 类 号：R392[医药卫生—免疫学]