Martin V, Arcavi M, Santillan G, Amendoeira M R, De Souza Neves E, Griemberg G, Guarnera E, Garberi J C, Angel S O
Departamento de Parasitología, ANLIS Dr. Carlos G. Malbran, Buenos Aires, Argentina.
Clin Diagn Lab Immunol. 1998 Sep;5(5):627-31. doi: 10.1128/CDLI.5.5.627-631.1998.
The Toxoplasma gondii rhoptry protein Rop2 was expressed in Escherichia coli as a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196-561). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196-561 as the antigen substrate. The analyzed sera were divided according to T. gondii-specific serological tests (IgG, IgA, or IgM indirect immunofluorescence and IgA or IgM immunosorbent agglutination assay) as group A (IgG+ IgA- IgM-; n = 35), group B (IgG+ IgA+ IgM+; n = 21), group C (IgG+ IgA+ IgM-; n = 5), and group D (IgG+ IgA- IgM+; n = 16). Twenty-six T. gondii-seronegative sera from individuals with other infections were also included (group E). Anti-Rop2 IgG antibodies were detected in 82.8% of group A sera and in 97.6% of the sera with acute-phase marker immunoglobulins (groups B, C, and D). The percentage of IgA antibody reactivity against Rop2196-561 was 17.1% in group A, 50% in group D, and 80.8% in groups B and C. The percentage of IgM antibody reactivity was 0% in groups A and C and 62% in groups B and D. Sera from group E failed to show IgA, IgM, or IgG antibody reactivity. Since T. gondii Rop2 elicits a strong humoral response from an early stage of infection, it is suggested that recombinant Rop2196-561 would be suitable for use in diagnostic systems, in combination with other T. gondii antigens, to detect specific IgG, IgA, and IgM antibodies.
弓形虫棒状体蛋白Rop2在大肠杆菌中表达为一种融合蛋白,包含55 kDa成熟Rop2的44 kDa片段,在N端带有六个组氨酸残基(Rop2196 - 561)。以Rop2196 - 561作为抗原底物,通过免疫球蛋白G(IgG)、IgA和IgM酶联免疫吸附测定法分析人类弓形虫感染期间的体液免疫反应。根据弓形虫特异性血清学检测(IgG、IgA或IgM间接免疫荧光以及IgA或IgM免疫吸附凝集试验)将分析的血清分为A组(IgG + IgA - IgM - ;n = 35)、B组(IgG + IgA + IgM + ;n = 21)、C组(IgG + IgA + IgM - ;n = 5)和D组(IgG + IgA - IgM + ;n = 16)。还纳入了来自患有其他感染的个体的26份弓形虫血清阴性血清(E组)。在A组血清的82.8%以及具有急性期标志物免疫球蛋白的血清(B组、C组和D组)的97.6%中检测到抗Rop2 IgG抗体。针对Rop2196 - 561的IgA抗体反应性百分比在A组中为17.1%,在D组中为50%,在B组和C组中为80.8%。IgM抗体反应性百分比在A组和C组中为0%,在B组和D组中为62%。E组血清未显示出IgA、IgM或IgG抗体反应性。由于弓形虫Rop2在感染早期引发强烈的体液免疫反应,因此建议重组Rop2196 - 561与其他弓形虫抗原联合用于诊断系统,以检测特异性IgG、IgA和IgM抗体。