弓形虫单克隆抗体的制备及其保护作用观察

STUDY OF PROTECTION MCAB AGAINST TOXOPLASMA GONDII AND IT'S PROTECTIVE TEST

应用杂交瘤技术制备弓形虫单克隆抗体（ＭｃＡｂ）。经克隆化获得２Ｂ１０、２Ｂ１１、２Ｇ８、２Ｄ１２；和１Ｄ１ＭｃＡｂ。用ＩＦＡ鉴定筛选，２Ｂ１０、２Ｂ１１为抗弓形虫整虫的ＭｃＡｂ、２Ｇ８、２Ｄ１２和１Ｄ１为抗弓形虫膜抗原的ＭｃＡｂ。用以上五株ＭｃＡｂ进行小鼠保护性试验：１．感染前２４小时、４８小时分别将５株ＭｃＡｂ稀释液注入小鼠腹腔，以１０４弓形虫速殖子进行攻击感染，结果２Ｂ１０、２Ｂ１１，和２Ｇ８对高毒株弓形虫感染鼠有明显的保护作用；２．将弓形虫速殖子与２Ｂ１０、２Ｂ１１和２Ｇ８孵育后再攻击感染能明显延长感染鼠的平均死亡时间（ＭＴＤ）；３．５株ＭｃＡｂ混合液于感染前２４小时注入小鼠体内及与速殖子孵育后也能明显延长感染鼠的ＭＴＤ。

McAb against T. gondii FR strain was produced by hybridoma technology. 5 strain were selected and performed by timiting ditution method in 96 cells culturs plates subcloning 3 times to get the strong positive McAb.They are 2B_(10).2B_(11), 2G_8, 2D_(12) and 1D_1.Two of them(2B_(10),2B_(11) were strong reacted against somatic antigen of T gondii(FR strain)identified by IFA method, Three of them (2G_8,2D_(12),1D_1) were reacted against only with the surface antigen.protective tests were examined with the five monoclonal antibodies.1,The experimental mice were injected 5 McAb 24 or 48hr before for each group,then they were challenged by 10￣4 tachyzoites for each one, three group, which were injected by 2B_(10), 2B_(11), 2G_8 McAb, Were conferred significant protection to the contro1.2 Miee were challenged with highly virulent tachyzoites, which had McAb,were gained a significant delay the mean-time to death (MTD). 3, 24h after an intraperitoneal injection of the mixture of the five monoclonal antibodies were get the significantly prolonged MTD of the mice.