Abstract:
The functional competence of leukemia inhibitory factor (LIF), as immunocontraceptive vaccine in mice, was investigated. Balb/c mice were divided into two groups of vaccinated and controls. The recombinant human LIF (rhLIF) protein and phosphate buffer saline was emulsified with Freund\'s adjuvant and injected into vaccinated and control groups, respectively. Theinhibition of implantation was evaluated in mice uterine. The concentration of secreted interferon-γ (IFN-γ) and interleukin (IL)-4 were measured in cultured splenocyte of mice stimulated by rhLIF. The expressions of immune responsive gene 1 (IRG-1), cochlin (COCH), amphiregulin(Ar), and heparin-binding EGF-like growth factor (HB-EGF) genes were determined. Mice were assessed for inhibition of fertility after delivery, reversibility of immune response against rhLIF, and survival rate. Active immunization of mice with rhLIF resulted in reduction of the implantation and fertility rate up to 80.49% and 75%, respectively. All mice produced a high titer of anti-rhLIF antibodies in serums and vaginal fluids washes after 16 weeks; however, these antibodies were cleared from vaginal fluid washes after six months. A significant down-regulation in mRNA levels of IRG-1, Ar and HB-EGF was observed in vaccinated group compared to controls; however, no significant change in the expression profile of cochlin gene was detected. The results showed that rhLIF prevented pregnancy in a high percentage of female mice. Although the immunization of female Balb/c mice with rhLIF inhibited fertility and expression of genes associated with this molecule, further studies are needed to support this protein as a suitable candidate for contraceptive vaccine in mammals.
摘要:
白血病抑制因子(LIF)的功能,作为小鼠的免疫避孕疫苗,被调查。将Balb/c小鼠分为两组接种和对照。重组人LIF(rhLIF)蛋白和磷酸盐缓冲盐水与弗氏佐剂乳化,并注射到接种组和对照组,分别。在小鼠子宫中评估植入的抑制作用。在rhLIF刺激的小鼠的培养脾细胞中测量分泌的干扰素-γ(IFN-γ)和白介素(IL)-4的浓度。免疫应答基因1(IRG-1)的表达,cochlin(COCH),双调蛋白(Ar),并测定肝素结合EGF样生长因子(HB-EGF)基因。评估小鼠分娩后对生育力的抑制作用,针对rhLIF的免疫应答的可逆性,和存活率。用rhLIF主动免疫小鼠导致植入和生育率降低高达80.49%和75%,分别。16周后,所有小鼠在血清和阴道洗液中产生高滴度的抗rhLIF抗体;然而,这些抗体在6个月后从阴道洗液中清除.与对照组相比,在接种组观察到IRG-1、Ar和HB-EGF的mRNA水平显著下调;然而,未检测到cochlin基因表达谱的显著改变。结果表明,rhLIF在高比例的雌性小鼠中阻止了怀孕。尽管用rhLIF免疫雌性Balb/c小鼠抑制了生育力和与该分子相关的基因表达,需要进一步的研究来支持这种蛋白质作为哺乳动物避孕疫苗的合适候选物。