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CITOKYNES AND IMMUNOGLOBULINS IN ACTINOMYCETOMA PATIENTS

CITOKYNES AND IMMUNOGLOBULINS IN ACTINOMYCETOMA PATIENTS. Luis J. M ÉNDEZ TOVAR Hospital de Especialidades Centro Médico Nacional, IMSS. México, D. F. MYCETOMA. MYCETOMA. Mariat F. Bull Soc Path Exot 1963;56:35-45

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CITOKYNES AND IMMUNOGLOBULINS IN ACTINOMYCETOMA PATIENTS

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  1. CITOKYNES AND IMMUNOGLOBULINS IN ACTINOMYCETOMA PATIENTS Luis J. MÉNDEZ TOVAR Hospital de Especialidades Centro Médico Nacional, IMSS. México, D. F.

  2. MYCETOMA

  3. MYCETOMA • Mariat F. Bull Soc Path Exot 1963;56:35-45 • López-Martínez R, Méndez-Tovar LJ, Lavalle P, Welsh O, Saul A, Macotela Ruíz E. Gac Med Mex 1992;128:477-82

  4. HOST MYCETOMA AGENT ENVIROMENT

  5. Dr. Fernando Latapí Dr. A. González-Ochoa (1902-1989) (1910-1984)

  6. 1960 - 1980 Good prognosis Intradermal reaction (+) Low titres of IgG Intradermal reaction (-) High titres of IgG Bad prognosis González-Ochoa A, Baranda F, Bojalil LF, Bastarrachea F Shibayama H, Félix D

  7. Nocardia asteroides and Nocardiosis • Macrophages and neutrophil cells don’t destroy N. asteroides • Injection of sensibilized T lymphocytes is protective in athymic mice inoculated with N. asteroides. • Superoxido dismutase (SDO) and catalase are present in virulent strains, mutants (SDO-) are not virulent. • Athymic mice inoculated with N. asteroides developed disseminated infections with similar lesions observed in immunosuppresed human patients. Beaman BL, Beaman L, Deem F, Richard L, Doughty A, Gershwin G, Steve M, Bourgeois L, Maslan S, Scates M,

  8. Contributions in 90s • Amikacina treatment • Purified antigens from Nocardia brasiliensis. • Diagnosis by ELISA. • Identification of catalase gene Vera-Cabrera L, Welsh O, Casillas S,, Rodríguez A, Torres-López E, Ramos A, Licón-Trillo A, González-Spencer D.

  9. Interleukines and Immunoglobulines in actinomycetoma murine model • IFN-γ increasing (10 folds) • Increase of IL-4, IL-6, IL-10 • Increase of IgM (7 day) • Increase of IgG (45 day) Salinas-Carmona M, Torres-López E, Ramos AI, et al. Infec Immun 1999;67:2428

  10. OBJECTIVES • By the cytokine profile obtained in proliferation assay of peripheral blood monocyte cell, to know the type of adaptative immune response (Th1 or Th2) exhibited by actinomycetoma patients. • To demonstrate that this response is different to the response elicited in healthy people.

  11. METHODS • N. brasiliensis antigen obtention • Intradermal application of PPD y candidine • Proliferation assay of peripherical blood monocyte cell (PBMC) • Quantification of: IFN-γ, TNF-α, IL-4 and IL-10. using supernatants of proliferation assay • Seric levels of IgG1, IgG2, IgG3, IgG4.

  12. ANTIGENIC FRACTIONSNB1, NB2, …, NB20)

  13. IMMUNOGLOBULIN TITRES ELISA SUPERNATANTS INTERLEUKINS DETERMINATION Proliferationassay

  14. RESULTS

  15. INTRADERMAL RESPONSE(n = 25) Induration diameter (mm) P value in both groups was p = 0.000

  16. Antigenic fractions

  17. Proliferation assay ANTIGENS CONTROL PATIENTS PHA 2588 ± 676 4725 ± 2330 PPD 1685 ± 660 2629 ± 2533 NB 1230 ± 531 2145 ± 2376 NB2 1733 ± 1043 2343 ± 2899 NB4 1924 ± 918 2021 ± 1154 NB6 1053 ± 674 1921 ± 1001 NB8 891 ± 359 1336 ± 776 NB10 1056 ± 551 1075 ± 812

  18. IFN-γ production (n = 25) CONTROL 1400 PATIENTS 1200 P = 0.011 P = 0.000 P = 0.001 1000 800 pg/ml 600 400 200 0 PHA PPD NB NB2 NB4 NB6 NB8 NB10 ANTIGEN

  19. TNF-α production (n = 25) CONTROL 3000 PATIENTS 2500 2000 1500 pg/ml 1000 500 0 PHA PPD NB NB2 NB4 NB6 NB8 NB10 ANTIGEN P = 0.000 P = 0.035 P = 0.001 P = 0.009

  20. IL-4 production(n = 25)

  21. IL-10 production (n = 25) 800 700 CONTROL 600 PATIENTS 500 400 pg/ml 300 200 100 0 PHA PPD NB NB2 NB4 NB6 NB8 NB10 ANTIGEN P = 0.035 P = 0.004 P = 0.035

  22. 0.16 0.14 0.12 0.1 D.O. a 490 nm 0.08 0.06 0.04 0.02 0 PPD PPD NB NB NB2 NB2 NB4 NB4 NB6 NB6 NB8 NB8 NB10 NB10 ANTIGENOS IgG subclass CONTROL PATIENTS IgG1 IgG2 P=0.483 P=0.781 P = 0.861 IgG3 IgG4 P=0.087 P=0.000

  23. Summary

  24. CONCLUSIONS • The patients’ PBMC exhibited a normal antigenic recognizing. • The cytokine profile in patients corresponded to TH2 type response. • The immunoglobulin titres (mainly IgG3 and IgG4) are also compatible with a TH2 response.

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