Cyclophosphamide effect on coccidioidomycosis in the rat

Remesar, Mirta C.; Blejer, Jorgelina L.; Negroni, Ricardo; Nejamkis, Marta R.

doi:10.1590/S0036-46651989000600009

Abstracts

Coccidioidomycosis is a systemic mycosis, endemic in arid areas of the American continent. The rat was employed as an experimental host, since it had been shown to reproduce human lesions and present a chronic course of disease with granulomas mainly restricted to lungs. Given the influence of immunosuppressive therapy on the clinical course of human coccidioidomycosis, we studied the effect of cyclophosphamide (CY) in the experimental rat model. Accordingly, animals were inoculated with 400 Coccidioides immitis arthroconidia of the Acosta strain, by intracardiacal route. As single CY doses failed to alter the course of disease, three schedules were used: A) 4 daily doses of 20 mg/kg each, prior to C. immitis inoculation; B) 4 similar daily doses after infection; and C); 6 doses of 20 mg/kg each, given from day +1 to +4, then on days +8 and +9, post infection (pi), taking day 0 as the time of fungal inoculation. The first two schedules inhibited antibody formation up to day 28 pi, without modifying cellular response to coccidioidin as measured by foodpad swelling. Initially, there was greater fungal spread than in controls receiving C. immitis alone, which proved self-limiting in the latter. In contrast, schedule C led to 559r mortality, with both humoral and cellular response abrogation, accompanied by extensive C. immitis dissemination. Histology disclosed significant alterations, such as the persistence of primary infection sporangia, corresponding to the acute stage of coccidioidomycosis in the absence of granuloma development. Therefore, the observed depression in cellular immunity seems responsible for the lack of inflammatory reaction capable of restricting sporangia proliferation in tissues which, in turn, enhances pathogen spread and mortality rate.

Cyclophosphamide; Coccidioidomycosis; Immunosuppression

El propósito de este trabajo fue estudiar el efecto de la inmunosupresión causada por la droga ciclofosfamida (CY) sobre la infección de la rata con Coccidioides immitis por vía intracardíaca. Este huésped fue empleado como modelo experimental, ya que presenta una evolución de la enfermedad semejante a la del hombre, alcanzando una etapa crónica con granulomas principalmente restringidos a los pulmones. Se utilizaron tres esquemas de CY: A) 4 dosis de 20 mg/kg cada una, antes de la inoculación de Ci; B) 4 dosis de igual cantidad de CY, luego de la infección; y C) 6 dosis de 20 mg/kg cada una, administradas desde el día +1 hasta +4 y continuando los días + 8 y +9 post-infección (pi). Los dos primeros esquemas inhibieron la formación de anticuerpos hasta el día 28 pi, sin modificar la respuesta celular a la coccidioidina, medida como hinchazón de la almohadilla plantar. Se observó una mayor diseminación fúngica inicial, autolimi-tándose más tarde. Por el contrario, el esquema C provocó un 55% de mortalidad, disminución de la respuesta humoral y celular, acompañada de una extensa diseminación del Ci. La histología mostró alteraciones significativas, tales como persistencia de esporangios de primoinfección, correspondientes al estadio agudo de la coccidioidomicosis, con ausencia de desarrollo de granulomas. Por lo tanto, la depresión observada en la respuesta celular debido al tratamiento con CY sería la responsable de la ausencia de la reacción inflamatoria capaz de restringir la proliferación de esporangios en los tejidos, lo cual a su vez favorece la diseminación del microorganismo patógeno y el aumento de morta lidad.

THERAPEUTIC ASSAYS

Cyclophosphamide effect on coccidioidomycosis in the rat

Efecto de la ciclofosfamida en la infección por Coccidioides immitis en la rata

Mirta C. Remesar; Jorgelina L. Blejer; Ricardo Negroni; Marta R. Nejamkis

Departamento de Microbiología, Facultad de Medicina. Buenos Aires, Argentina

^{Address for correspondence} Address for correspondence: Dr. M. C. Remesar. Departamento de Microbiología. Facultad de Medicina - UBA. Paraguay 2155, Piso 12, 1121 Buenos Aires, Argentina.

SUMMARY

Coccidioidomycosis is a systemic mycosis, endemic in arid areas of the American continent.

The rat was employed as an experimental host, since it had been shown to reproduce human lesions and present a chronic course of disease with granulomas mainly restricted to lungs.

Given the influence of immunosuppressive therapy on the clinical course of human coccidioidomycosis, we studied the effect of cyclophosphamide (CY) in the experimental rat model. Accordingly, animals were inoculated with 400 Coccidioides immitis arthroconidia of the Acosta strain, by intracardiacal route. As single CY doses failed to alter the course of disease, three schedules were used: A) 4 daily doses of 20 mg/kg each, prior to C. immitis inoculation; B) 4 similar daily doses after infection; and C); 6 doses of 20 mg/kg each, given from day +1 to +4, then on days +8 and +9, post infection (pi), taking day 0 as the time of fungal inoculation. The first two schedules inhibited antibody formation up to day 28 pi, without modifying cellular response to coccidioidin as measured by foodpad swelling. Initially, there was greater fungal spread than in controls receiving C. immitis alone, which proved self-limiting in the latter. In contrast, schedule C led to 559r mortality, with both humoral and cellular response abrogation, accompanied by extensive C. immitis dissemination. Histology disclosed significant alterations, such as the persistence of primary infection sporangia, corresponding to the acute stage of coccidioidomycosis in the absence of granuloma development.

Therefore, the observed depression in cellular immunity seems responsible for the lack of inflammatory reaction capable of restricting sporangia proliferation in tissues which, in turn, enhances pathogen spread and mortality rate.

Key words: Cyclophosphamide; Coccidioidomycosis; Immunosuppression.

RESUMEN

El propósito de este trabajo fue estudiar el efecto de la inmunosupresión causada por la droga ciclofosfamida (CY) sobre la infección de la rata con Coccidioides immitis por vía intracardíaca. Este huésped fue empleado como modelo experimental, ya que presenta una evolución de la enfermedad semejante a la del hombre, alcanzando una etapa crónica con granulomas principalmente restringidos a los pulmones. Se utilizaron tres esquemas de CY: A) 4 dosis de 20 mg/kg cada una, antes de la inoculación de Ci; B) 4 dosis de igual cantidad de CY, luego de la infección; y C) 6 dosis de 20 mg/kg cada una, administradas desde el día +1 hasta +4 y continuando los días + 8 y +9 post-infección (pi). Los dos primeros esquemas inhibieron la formación de anticuerpos hasta el día 28 pi, sin modificar la respuesta celular a la coccidioidina, medida como hinchazón de la almohadilla plantar. Se observó una mayor diseminación fúngica inicial, autolimi-tándose más tarde. Por el contrario, el esquema C provocó un 55% de mortalidad, disminución de la respuesta humoral y celular, acompañada de una extensa diseminación del Ci. La histología mostró alteraciones significativas, tales como persistencia de esporangios de primoinfección, correspondientes al estadio agudo de la coccidioidomicosis, con ausencia de desarrollo de granulomas. Por lo tanto, la depresión observada en la respuesta celular debido al tratamiento con CY sería la responsable de la ausencia de la reacción inflamatoria capaz de restringir la proliferación de esporangios en los tejidos, lo cual a su vez favorece la diseminación del microorganismo patógeno y el aumento de morta lidad.

Full text available only in PDF format.

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ACKNOWLEDGMENTS

This work was supported by grants provided by the National Research Council (CONICET, Argentina) and by the University of Buenos Aires.

Recebido para publicação em 25 1 1989.

1. ADLER, B. & FAINE, S. - Susceptibility of mice treated with cyclophosphamide to lethal infection with Leptospira interrogans Serovar pomona Infect. Immun., 14.703-708, 1976.
2. AJELLO, L. - Coccidiodomycosis. In: PROCEEDINGS OF SECOND COCCIDIODOMYCOSIS SYMPOSIUM. Tuc son, Arizona, The University of Arizona Press, 1967.
3. BEAMAN. B. L. & MASLAN, S. - Effect of cyclophosphamide on experimental Nocardia asteroides infection in mice. Infect. Immun., 16: 995- 1004, 1977.
4. BEAMAN, B. L.: PAPPAGIANIS. D. & BENJAMINI, E. - Significance of T cells in resistance to experimental murine coccidioidomycosis. Infect. Immun., 17: 580-585. 1977.
5. BISTONI, F.: BACCARINI. M.; BLASI, E.: MARCONI. P.; PUCCETTI, P. & GARACI. E. - Correlation between in vivo and in vitro studies of modulation of resistance to experimental Candida albicans infection by cyclophos phamide in mice. Infect. Immun., 40: 46-55, 1983.
6. BORELLI, D. - Prevalence of systemic mycosis in Latin America. In: INTERNATIONAL SYMPOSIUM ON MY COSIS. Washington, 1970. PROCEEDINGS. Washington, Pan American Health Organization, 1970. p. 28 - 38 (Seien tific Publication N°. 205).
7. CALHOUN, D. L.; GALGIANI, J. N.; ZUKOSKI, Ch. & COPELAND, J. G. - Coccidiodomycosis in recent renal or cardiac transplant recipients. In: EINSTEIN, H. E. & CANTANZARO, A. - Coccidioidomycosis. International Conference on Coccidioidomycosis, 4°, Washington, 1985. PROCEEDINGS, p. 312-318.
8. CONAT, N. F.; SMITH, D. T.: BAKER, R. D. & CALLA WAY, J. L - MANUAL OF CLINICAL MYCOLOGY. 3rd. ed. Philadelphia, W. B. Saunders Company, 1971. p. 134-169.
9. COZAD, G. C. & LINDSEY, T. S. - Effect of cyclophos phamide on Histoplasma capsulatum infections in mice. Infect. Immun., 9: 261-265, 1974.
10. EMMONS, C. W.; BINFORD, C. H. & UTZ, J. P. - Cocci dioidomycosis. In: EMMONS. C. W.: BINFORD, C. H. & UTZ, J. P. - Medical mycology. 2nd. ed. Philadelphia. Lea & Febiger, 1970. p. 207-229.
11. GRAYBILL, J. R. & MITCHELL, L. - Cyclophosphamide effects on murine cryptococcosis. Infect. Immun., 21: 674-677, 1978.
12. HENDEL, E. E.: HUI, A. N.: DIAZ, J. & BOYLEN, C. T.- Pneumocystis carinii pneumonia and malignant lym phoma in a homosexual male with disseminated coccidioi domycosis. In: EINSTEIN, H. E. & CATANZARO, A. - Coccidioidomycosis. International Conference on Cocci dioidomycosis, 4°., Washington, 1985. Proceedings, p. 305-311.
13. LASCANO, E. F.; BLEJER, J. L.; GALASSI, N. V. & NE JAMKIS, M. R. - Brain inflammatory exudate in Junin Virus infected rats: its characterization by the immunope roxidase (PAP) technique. J. Neuroimmunol, 11: 105-116, 1986.
14. MOSER. S. A. & DOMER, J. E. - Effects of cyclophos phamide on murine candidiasis. Infect. Immun., 27: 376-386, 1980.
15. NEGRONI, P. - Coccidiodomicosis. In: NEGRONI, P. - Las blastomicosis y coccidiodomicosis. Micosis profundas y viscerales. Buenos Aires, Comisión de Investi gación Clínica, 1966. v. 3. p. 235-354.
16. NEGRONI, R.; FINQUELIEVICH, J. L. & ELIAS COSTA. M. R. I. - Estúdio de la coccidioidomicosis experimental en ratas Wistar. Rev. argent. Micol., 8: 7-11, 1985.
17. RUBINSTEIN, P. & NEGRONI, R. - Coccidioidomicosis. In: RUBINSTEIN, P. & NEGRONI. R. - Micosis bronco-pulmonares del adulto y del nino. 2. ed Buenos Aires. Ed. Beta. 1981. p. 291-323.
18. RUBINSTEIN, H. R.; MASIH, D. T.: MARTICORENA. B. & RIERA, C. M. - Experimental coccidioidomycosis: effects of cyclophosphamide in immunologic responses. Mycopathologia (Den Haag), 94: 91-95, 1986
19. SHARBAUGH, R. J. & GROGAN, J. B. - Suppression of reticuloendothelial function in the rat with cyclophos phamide. J. Bact. 100: 117-122, 1969.
20. STANDARD, P. G. & KAUFMAN. L. - Immunological procedure for the rapid and specific identification of Coccidioides immitis cultures. J. clin. Microbiol., 5: 149-153. 1977.
21. STEVENS, D. A.: PAPPAGIANIS, D.; MARINCOVICH. V. & WADDER, T. F. - Immunotherapy in recurrent coc cidioidomycosis. Cell. Immunol., 12: 37-48, 1974.
22. STEVENS, D. A. - Coccidioidomycosis. New York, Pie num Plublishing Corporations. 1980.
23. STEVENS, D. A. - Clinical manifestations and manage ment of coccidioidomycosis in the compromised patient. In: WARNOCK, D. W. & RICHARDSON. M. D. - Fungal infection in the compromised patient. New York, J. Wiley & Sons, 1982. p. 199-205.
24. ZAROR, L.; ROBLES, A. M. & NEGRONI. R. - Pruebas de inmunodifusión en médios gelosados con agregado de polietilenglicol 6000 para el serodiagnóstico de las micosis. Rev. argent. Microbiol., 10: 61-64, 1978.

Address for correspondence:

Dr. M. C. Remesar.

Departamento de Microbiología.

Facultad de Medicina - UBA.

Paraguay 2155, Piso 12, 1121 Buenos Aires, Argentina.

Publication Dates

Publication in this collection
01 Dec 2010
Date of issue
Dec 1989

History

Received
25 Jan 1989

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ADLER, B. & FAINE, S. - Susceptibility of mice treated with cyclophosphamide to lethal infection with Leptospira interrogans Serovar pomona Infect. Immun., 14.703-708, 1976.

[2] 2. AJELLO, L. - Coccidiodomycosis. In: PROCEEDINGS OF SECOND COCCIDIODOMYCOSIS SYMPOSIUM. Tuc son, Arizona, The University of Arizona Press, 1967.

[3] 3. BEAMAN. B. L. & MASLAN, S. - Effect of cyclophosphamide on experimental Nocardia asteroides infection in mice. Infect. Immun., 16: 995- 1004, 1977.

[4] 4. BEAMAN, B. L.: PAPPAGIANIS. D. & BENJAMINI, E. - Significance of T cells in resistance to experimental murine coccidioidomycosis. Infect. Immun., 17: 580-585. 1977.

[5] 5. BISTONI, F.: BACCARINI. M.; BLASI, E.: MARCONI. P.; PUCCETTI, P. & GARACI. E. - Correlation between in vivo and in vitro studies of modulation of resistance to experimental Candida albicans infection by cyclophos phamide in mice. Infect. Immun., 40: 46-55, 1983.

[6] 6. BORELLI, D. - Prevalence of systemic mycosis in Latin America. In: INTERNATIONAL SYMPOSIUM ON MY COSIS. Washington, 1970. PROCEEDINGS. Washington, Pan American Health Organization, 1970. p. 28 - 38 (Seien tific Publication N°. 205).

[7] 7. CALHOUN, D. L.; GALGIANI, J. N.; ZUKOSKI, Ch. & COPELAND, J. G. - Coccidiodomycosis in recent renal or cardiac transplant recipients. In: EINSTEIN, H. E. & CANTANZARO, A. - Coccidioidomycosis. International Conference on Coccidioidomycosis, 4°, Washington, 1985. PROCEEDINGS, p. 312-318.

[8] 8. CONAT, N. F.; SMITH, D. T.: BAKER, R. D. & CALLA WAY, J. L - MANUAL OF CLINICAL MYCOLOGY. 3rd. ed. Philadelphia, W. B. Saunders Company, 1971. p. 134-169.

[9] 9. COZAD, G. C. & LINDSEY, T. S. - Effect of cyclophos phamide on Histoplasma capsulatum infections in mice. Infect. Immun., 9: 261-265, 1974.

[10] 10. EMMONS, C. W.; BINFORD, C. H. & UTZ, J. P. - Cocci dioidomycosis. In: EMMONS. C. W.: BINFORD, C. H. & UTZ, J. P. - Medical mycology. 2nd. ed. Philadelphia. Lea & Febiger, 1970. p. 207-229.

[11] 11. GRAYBILL, J. R. & MITCHELL, L. - Cyclophosphamide effects on murine cryptococcosis. Infect. Immun., 21: 674-677, 1978.

[12] 12. HENDEL, E. E.: HUI, A. N.: DIAZ, J. & BOYLEN, C. T.- Pneumocystis carinii pneumonia and malignant lym phoma in a homosexual male with disseminated coccidioi domycosis. In: EINSTEIN, H. E. & CATANZARO, A. - Coccidioidomycosis. International Conference on Cocci dioidomycosis, 4°., Washington, 1985. Proceedings, p. 305-311.

[13] 13. LASCANO, E. F.; BLEJER, J. L.; GALASSI, N. V. & NE JAMKIS, M. R. - Brain inflammatory exudate in Junin Virus infected rats: its characterization by the immunope roxidase (PAP) technique. J. Neuroimmunol, 11: 105-116, 1986.

[14] 14. MOSER. S. A. & DOMER, J. E. - Effects of cyclophos phamide on murine candidiasis. Infect. Immun., 27: 376-386, 1980.

[15] 15. NEGRONI, P. - Coccidiodomicosis. In: NEGRONI, P. - Las blastomicosis y coccidiodomicosis. Micosis profundas y viscerales. Buenos Aires, Comisión de Investi gación Clínica, 1966. v. 3. p. 235-354.

[16] 16. NEGRONI, R.; FINQUELIEVICH, J. L. & ELIAS COSTA. M. R. I. - Estúdio de la coccidioidomicosis experimental en ratas Wistar. Rev. argent. Micol., 8: 7-11, 1985.

[17] 17. RUBINSTEIN, P. & NEGRONI, R. - Coccidioidomicosis. In: RUBINSTEIN, P. & NEGRONI. R. - Micosis bronco-pulmonares del adulto y del nino. 2. ed Buenos Aires. Ed. Beta. 1981. p. 291-323.

[18] 18. RUBINSTEIN, H. R.; MASIH, D. T.: MARTICORENA. B. & RIERA, C. M. - Experimental coccidioidomycosis: effects of cyclophosphamide in immunologic responses. Mycopathologia (Den Haag), 94: 91-95, 1986

[19] 19. SHARBAUGH, R. J. & GROGAN, J. B. - Suppression of reticuloendothelial function in the rat with cyclophos phamide. J. Bact. 100: 117-122, 1969.

[20] 20. STANDARD, P. G. & KAUFMAN. L. - Immunological procedure for the rapid and specific identification of Coccidioides immitis cultures. J. clin. Microbiol., 5: 149-153. 1977.

[21] 21. STEVENS, D. A.: PAPPAGIANIS, D.; MARINCOVICH. V. & WADDER, T. F. - Immunotherapy in recurrent coc cidioidomycosis. Cell. Immunol., 12: 37-48, 1974.

[22] 22. STEVENS, D. A. - Coccidioidomycosis. New York, Pie num Plublishing Corporations. 1980.

[23] 23. STEVENS, D. A. - Clinical manifestations and manage ment of coccidioidomycosis in the compromised patient. In: WARNOCK, D. W. & RICHARDSON. M. D. - Fungal infection in the compromised patient. New York, J. Wiley & Sons, 1982. p. 199-205.

[24] 24. ZAROR, L.; ROBLES, A. M. & NEGRONI. R. - Pruebas de inmunodifusión en médios gelosados con agregado de polietilenglicol 6000 para el serodiagnóstico de las micosis. Rev. argent. Microbiol., 10: 61-64, 1978.

Brasil

Brasil

Cyclophosphamide effect on coccidioidomycosis in the rat

Efecto de la ciclofosfamida en la infección por Coccidioides immitis en la rata

Abstracts

Publication Dates

History