摘要
制备抗PbPH单克隆抗体(mAb),检测伯氏疟原虫(Plasmodium berghei)PbPH的表达特点和传播阻断能力。主要通过蛋白免疫印迹(Western blot,WB)和间接免疫荧光法(Indirect immunofluoscent assay,IFA),确定PbPH的表达阶段以及抗PbPH单克隆抗体的特异性。1×10^(6)个P.berghei感染雌性BALB/c小鼠3 d后,尾静脉收集感染小鼠的血液,与鼠源的抗PbPH单克隆抗体/PBS对照,按照不同的稀释倍数(15、110、150)进行混合培养,观察15 min后,伯氏疟原虫配子体出丝中心数及24 h后动合子形成数的变化。WB和IFA检测抗PbPH单克隆抗体可以识别雌雄配子体、雄配子、雌配子/合子、retort和动合子的表面抗原。体外传播阻断实验中,与PBS对照组相比,在加入不同浓度(15、110)的抗PbPH单克隆抗体培养后,配子体出丝中心数分别减少了41.7%、32.7%,差异具有统计学意义(P<0.05);动合子形成数分别减少了45.1%、14.8%,差异具有统计学意义(P<0.05)。抗PbPH单克隆抗体可以有效阻断体外配子体出丝中心和动合子的形成,从而影响伯氏疟原虫在蚊体内的进一步发育和继续传播。
Characteristics and transmission blocking ability of PbPH were studied by preparing monoclonal antibodies(mAb) against PbPH. The expression phase and the specificity of mAb of PbPH were mainly detected by Western blot(WB) and indirect immunofluoscent assay(IFA). In vitro assays, 3 days after Plasmodium berghei infected female BALB/c mice, the blood of the infected mice was obtained from the tail vein, and mixed culture with the anti-PbPH mAb/PBS at different dilution concentrations(1:5,1:10,1:50), and the number of exflagellation centers of male gametocytes and the number of ookinete were observed and counted 15 min after. Anti-PbPH mAb can identify the surface antigens of male and female gametocytes, male gametes, female gametes/zygotes, retort and ookinetes by WB and IFA. In vitro experiment of transmission blocking, compared with the PBS control group, after adding different concentrations(1:5,1:10) of anti-PbPH mAb, the exflagellation centers of male gametocytes were significantly decreased by 41.7% and 32.7% respectively with difference possessing statistic significance(P<0.05). The number of ookinete was significantly reduced by 45.1% and 14.8%, respectively with difference possessing statistic significance(P<0.05). Anti-PbPH mAb can effectively block the exflagellation centers and ookinetes In vitro, thereby affecting further development and continuous transmission of plasmodium in mosquito′s bodies.
作者
张耀文
刘鹏波
寇旭
郑文琪
刘飞
王美莲
ZHANG Yao-wen;LIU Peng-bo;KOU Xu;ZHENG Wen-qi;LIU Fei;WANG Mei-lian(Teach.&Res.Div.of Pathogen Biol.,Coll.of Basic Med.Sci.,China Med.Uni.,Shenyang 110021;Teach&Res.Div.of Immunol.,Coll.of Basic Med.Sci.,China Med.Uni.,Shenyang 110021)
出处
《微生物学杂志》
CAS
CSCD
2021年第2期67-72,共6页
Journal of Microbiology
基金
辽宁省科技厅项目(QN2019023)。
关键词
伯氏疟原虫
PbPH
动合子
配子体
单克隆抗体
传播阻断疫苗
Plasmodium berghei
PbPH
Ookinete
Gametocytes
Monoclonal antibodies(mAb)
Transmission-blocking vaccines
