Abstract

Detection of Human Toxoplasma -Specific Immunoglobulins A, M, and G with a Recombinant Toxoplasma gondii Rop2 Protein Valentina Martin 1 , Miriam Arcavi 2 , Graciela Santillan 1 , Maria Regina R. Amendoeira 3 , Elizabeth De Souza Neves 4 , Gloria Griemberg 2 , Eduardo Guarnera 1 , Juan C. Garberi 1 , and Sergio O. Angel 1 , * Departamento de Parasitologı́a, ANLIS Dr. Carlos G. Malbran, 1 and Immunologı́a Clı́nica, Departamento de Bioquı́mica Clı́nica, Facultad de Farmacia y Bioquı́mica, Hospital de Clı́nicas José de San Martin-UBA, 2 Buenos Aires, Argentina, and Instituto Oswaldo Cruz-Fundaçao Oswaldo Cruz, 3 and Hospital Evandro Chagas-FIOCRUZ, 4 Rio de Janeiro, Brazil ABSTRACT 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 (Rop2 196–561 ). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2 196–561 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 Rop2 196–561 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 Rop2 196–561 would be suitable for use in diagnostic systems, in combination with other T. gondii antigens, to detect specific IgG, IgA, and IgM antibodies.