1887

Abstract

Despite the development of serological and molecular methods in recent years, the diagnosis of cerebral toxoplasmosis in human immunodeficiency virus (HIV)-infected patients still presents difficulties. In the present study, we investigated whether cerebral toxoplasmosis induced changes in the reactivity of serum toward excreted–secreted antigens (ESA) in order to develop an assay for evaluating HIV-infected patients with cerebral toxoplasmosis. The antigen selection was based on those produced by tachyzoites, since it is the form of the organism responsible for disseminating the infection, as well as stimulation of the humoral and cellular immune responses. By using an ELISA containing pooled ESA recovered from infected culture supernatants with tachyzoites-RH strain (ESA-ELISA), we found that ESA had a high specificity for sera from patients with cerebral toxoplasmosis. The reactions were compared with an ELISA using crude tachyzoites antigen, widely used in traditional serology. The assays were performed on 293 serum samples separated as follows: 100 sera from patients with cerebral toxoplasmosis and AIDS (symptomatic), 99 sera from individuals with chronic toxoplasmosis (asymptomatic) and 94 sera from healthy individuals without toxoplasmosis (control). The crude tachyzoite antigen in ELISA was able to distinguish both groups of sera with toxoplasmosis, as similar reactivity were observed in sera from patients with cerebral toxoplasmosis and those from chronic individuals. In contrast, ESA-ELISA distinguished sera from symptomatic and asymptomatic individuals (three times more reactive in the former group, 12.6 versus 4.2). The assays were reproducible based on immunoblotting and statistical analysis. These data suggest the utility of ESA-ELISA in the diagnosis of cerebral toxoplasmosis in HIV-infected patients, since it provided clear evidence that anti-ESA antibodies are present principally in patients with active infection. The absence of a significant amount of antibodies distinguished the patients without clinical symptoms of infection.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47687-0
2008-07-01
2019-10-14
Loading full text...

Full text loading...

/deliver/fulltext/jmm/57/7/845.html?itemId=/content/journal/jmm/10.1099/jmm.0.47687-0&mimeType=html&fmt=ahah

References

  1. Antinori, A., Larussa, D., Cingolani, A., Lorenzini, P., Bossolasco, S., Finazzi, M.G., Bongiovanni, M., Guaraldi, G., Grisetti, S. & other authors ( 2004; ). Prevalence, associated factors, and prognostic determinants of AIDS-related toxoplasmic encephalitis in the era of advanced highly active antiretroviral therapy. Clin Infect Dis 39, 1681–1691.[CrossRef]
    [Google Scholar]
  2. Bastien, P. ( 2002; ). Molecular diagnosis of toxoplasmosis. Trans R Soc Trop Med Hyg 96 (Suppl. 1), S205–S215.[CrossRef]
    [Google Scholar]
  3. Bretagne, S. ( 2003; ). Molecular diagnostics in clinical parasitology and mycology: limits of the current polymerase chain reaction (PCR) assays and interest of the real-time PCR assays. Clin Microbiol Infect 9, 505–511.[CrossRef]
    [Google Scholar]
  4. Carruthers, V. B. ( 2002; ). Host cell invasion by the opportunistic pathogen Toxoplasma gondii. Acta Trop 81, 111–122.[CrossRef]
    [Google Scholar]
  5. Cesbron-Delauw, M. F. & Capron, A. ( 1993; ). Excreted-secreted antigens of Toxoplasma gondii: their origin and role in the host-parasite interaction. Res Immunol 144, 41–44.[CrossRef]
    [Google Scholar]
  6. Cohen, B. A. ( 1999; ). Neurological manifestations of toxoplasmosis in AIDS. Semin Neurol 19, 201–211.[CrossRef]
    [Google Scholar]
  7. Colombo, F. A., Vidal, J. E., Penalva de Oliveira, A. C., Hernández, A. V., Bonasser-Filho, F., Nogueira, R. S., Focaccia, R. & Pereira-Chioccola, V. L. ( 2005; ). Diagnosis of cerebral toxoplasmosis in AIDS patients in Brazil: importance of molecular and immunological methods using peripheral blood samples. J Clin Microbiol 43, 5044–5047.[CrossRef]
    [Google Scholar]
  8. Derouin, F., Leport, C., Pueyo, S., Morlat, P., Letrillart, B., Chene, G., Ecobichon, J. L., Luft, B., Aubertin, J. & other authors ( 1996; ). Predictive value of Toxoplasma gondii antibody titres on the occurrence of toxoplasmic encephalitis in HIV-infected patients. AIDS 10, 1521–1527.[CrossRef]
    [Google Scholar]
  9. Dubey, J. P. ( 1996; ). Toxoplasma gondii. In Medical Microbiology, 5th edn. Edited by S. Baron, R. C. Peake, D. A. James, M. Susman, C. A. Kennedy, M. J. D. Singleton & S. Schuenke. Galveston, TX: University of Texas Medical Branch.
  10. Dubey, J. P. ( 1998; ). Advances in the life cycle of Toxoplasma gondii. Int J Parasitol 28, 1019–1024.[CrossRef]
    [Google Scholar]
  11. Duquesne, V., Auriault, C., Darcy, F., Decavel, J. P. & Capron, A. ( 1990; ). Protection of nude rats against Toxoplasma infection by excreted–secreted antigen-specific helper T-cells. Infect Immun 58, 2120–2126.
    [Google Scholar]
  12. Ferreira, I. M. R., Vidal, J. E., Costa-Silva, T. A., Meira, C. S., Hiramoto, R. M., Penalva de Oliveira, A. C. & Pereira-Chioccola, V. L. ( 2008; ). Toxoplasma gondii: genotyping of strains from Brazilian AIDS patients with cerebral toxoplasmosis by multilocus PCR-RFLP markers. Exp Parasitol 118, 221–227.[CrossRef]
    [Google Scholar]
  13. Hellerbrand, C., Goebel, F. D. & Disko, R. ( 1996; ). High predictive value of Toxoplasma gondii IgG antibody levels in HIV-infected patients for diagnosis of cerebral toxoplasmosis. Eur J Clin Microbiol Infect Dis 15, 869–872.[CrossRef]
    [Google Scholar]
  14. Hill, D. E., Chirukandoth, S. & Dubey, J. P. ( 2005; ). Biology and epidemiology of Toxoplasma gondii in man and animals. Anim Health Res Rev 6, 41–61.[CrossRef]
    [Google Scholar]
  15. Hoffmann, C. ( 2005; ). Opportunistic infections. In HIV Medicine pp. 365–344. Edited by C. Hoffmann, J. K. Rockstroh & B. S. Kamps. Paris: Flying Publisher. www.HIVMedicine.com.
  16. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–681.[CrossRef]
    [Google Scholar]
  17. Luft, B. J. & Remington, J. S. ( 1992; ). Toxoplasmic encephalitis. Clin Infect Dis 15, 211–222.[CrossRef]
    [Google Scholar]
  18. Montoya, J. G. & Liesenfeld, O. ( 2004; ). Toxoplasmosis. Lancet 363, 1965–1976.[CrossRef]
    [Google Scholar]
  19. Pereira-Chioccola, V. L., Fragata-Filho, A. A., Levy, A. M., Rodrigues, M. M. & Schenkman, S. ( 2003; ). Enzyme-linked immunoassay using recombinant trans-sialidase of Trypanosoma cruzi can be employed for monitoring of patients with Chagas' disease after drug treatment. Clin Diagn Lab Immunol 10, 826–830.
    [Google Scholar]
  20. Portegies, P., Solod, L., Cinque, P., Chaudhuri, A., Begovac, J., Everall, I., Weber, T., Bojar, M., Martinez-Martin, P. & Kennedy, G. E. ( 2004; ). Guidelines for the diagnosis and management of neurological complications of HIV infection. Eur J Neurol 11, 297–304.[CrossRef]
    [Google Scholar]
  21. Prigione, I., Facchetti, P., Lecordier, L., Deslee, D., Chiesa, S., Cesbron-Delauw, M. F. & Pistoia, V. ( 2000; ). T cell clones raised from chronically infected healthy humans by stimulation with Toxoplasma gondii excretory–secretory antigens cross-react with live tachyzoites: characterization of the fine antigenic specificity of the clones and implications for vaccine development. J Immunol 164, 3741–3748.[CrossRef]
    [Google Scholar]
  22. Skiest, D. J. ( 2002; ). Focal neurological disease in patients with acquired immunodeficiency syndrome. Clin Infect Dis 34, 103–115.[CrossRef]
    [Google Scholar]
  23. Tilley, M., Fichera, M. E., Jerome, M. E., Roos, D. S. & White, M. W. ( 1997; ). Toxoplasma gondii sporozoites from a transient parasitophorus vacuole that is impermeable and contains only a subset of dense granule proteins. Infect Immun 65, 4598–4605.
    [Google Scholar]
  24. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76, 4350–4357.[CrossRef]
    [Google Scholar]
  25. Vidal, J. E., Colombo, F. A., Penalva de Oliveira, A. C., Focaccia, R. & Pereira-Chioccola, V. L. ( 2004; ). PCR assay using cerebrospinal fluid for diagnosis of cerebral toxoplasmosis in Brazilian AIDS patients. J Clin Microbiol 42, 4765–4768.[CrossRef]
    [Google Scholar]
  26. Vidal, J. E., Hernandez, A. V., Penalva de Oliveira, A. C., Dauar, R. F. & Barboza, S. P. Jr Focaccia, R. ( 2005; ). Cerebral toxoplasmosis in HIV-positive patients in Brazil: clinical features and predictors of treatment response in the HAART era. AIDS Patient Care STDS 19, 626–634.[CrossRef]
    [Google Scholar]
  27. Zenner, L., Estaquier, J., Darcy, F., Maes, P., Capron, A. & Caesbron-Delauw, M. F. ( 1999; ). Protective immunity in the rat model of congenital toxoplasmosis and the potential vaccinal role of excreted-secreted antigens (ESA). Parasite Immunol 21, 261–272.[CrossRef]
    [Google Scholar]
  28. Zhou, X. W., Kafsack, B. F. C., Cole, R. N., Beckett, P., Shen, R. F. & Carruthers, V. B. ( 2005; ). The opportunistic pathogen Toxoplasma gondii deploys a diverse legion of invasion and survival proteins. J Biol Chem 280, 34233–34244.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47687-0
Loading
/content/journal/jmm/10.1099/jmm.0.47687-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error