Müller W A, Becker K, Schäfer A, Koch G, Borkhardt H L
Institut für Medizinische Mikrobiologie der Medizinischen Akademie Magdeburg, DDR.
Angew Parasitol. 1988 Apr;29(1):11-7.
Hybridomas that secrete monoclonal antibody to Toxoplasma gondii have been developed. Two groups of 10 female BALB/c mice each were immunized either over a shorter (71 d) or longer (117 d) period at first with Toxoplasma lysate antigen and afterwards with intact tachyzoites of the RH strain. Higher titres of antibody were obtained with the long-period immunization. The fusion experiments have shown that both schemes of immunization approximately result in the same number (16 and 14% respectively) of hybridoma cell lines producing antigen-specific monoclonal antibodies. Hybridoma cultures secreting antitoxoplasma monoclonal antibodies were screened parallel by indirect immunofluorescence antibody test (IFAT) and enzyme immunoassay (EIA). 16 of the hybridoma cell cultures produced positive results only in the IFAT, 112 reacted only in the EIA and 21 were positive in both tests. The monoclonal antibodies 5B10, 5G6 and 1B2, which are positive in the IFAT form a chemical compound with the major antigens on the surface of RH tachyzoites. The patterns of fluorescence produced by these monoclonal antibodies are in conformity with those produced by using polyclonal sera of Toxoplasma gondii infected hosts (mouse, rabbit, man).
已研制出分泌抗刚地弓形虫单克隆抗体的杂交瘤。将两组各10只雌性BALB/c小鼠,首先分别在较短(71天)或较长(117天)时间内用弓形虫裂解物抗原免疫,然后用RH株完整速殖子免疫。长期免疫获得了更高滴度的抗体。融合实验表明,两种免疫方案产生产生抗原特异性单克隆抗体的杂交瘤细胞系数量大致相同(分别为16%和14%)。通过间接免疫荧光抗体试验(IFAT)和酶免疫测定(EIA)并行筛选分泌抗弓形虫单克隆抗体的杂交瘤培养物。16个杂交瘤细胞培养物仅在IFAT中产生阳性结果,112个仅在EIA中反应,21个在两项试验中均为阳性。在IFAT中呈阳性的单克隆抗体5B10、5G6和1B2与RH速殖子表面的主要抗原形成化合物。这些单克隆抗体产生的荧光模式与使用刚地弓形虫感染宿主(小鼠、兔子、人)的多克隆血清产生的荧光模式一致。