Liver morphology with emphasis on bile ducts changes and survival analysis in mice submitted to multiple Schistosoma mansoni infections and chemotherapy

Silva, L. C. da; Vianna, M. Regina; Abrantes, C. P.; Lima, D. M. C.; Falavigna, A. L.; Antonelli-Cardoso, R. H.; Gallucci, S. D. Deperon; Brito, T. de

doi:10.1590/S0036-46651990000500004

Abstracts

In an attempt to be as close as possible to the infected and treated patients of the endemic areas of schistosomiasis (S. mansoni) and in order to achieve a long period of follow-up, mice were repeatedly infected with a low number of cercariae. Survival data and histological variables such as schistosomal granuloma, portal changes, hepatocellular necrosis, hepatocellular regeneration, schistosomotic pigment, periductal fibrosis and chiefly bile ducts changes were analysed in the infected treated and non treated mice. Oxamniquine chemotherapy in repeatedly infected mice prolonged survival significantly when compared to non-treated animals (chi-square 9.24, p = 0.0024), thus confirming previous results with a similar experimental model but with a shorter term follow-up. Furthermore, mortality decreased rapidly after treatment suggesting an abrupt reduction in the severity of hepatic lesions. A morphological and immunohistochemical study of the liver was carried out. Portal fibrosis, with a pattern resembling human Symmers fibrosis was present at a late phase in the infected animals. Bile duct lesions were quite close to those described in human Mansonian schistosomiasis. Schistosomal antigen was observed in one isolated altered bile duct cell. The pathogenesis of the bile duct changes and its relation to the parasite infection and/or their antigens are discussed.

Experimental mansonian schistosomiasis; Oxamniquine therapy; Bile duct changes; Survival analysis

Numa tentativa de estar o mais próximo possível a pacientes infectados e tratados nas áreas endêmicas de esquistosomose (S. mansoni) e também para obter um período mais longo de seguimento, camundongos foram repetidamente infectados com um número baixo de cercarias. Dados de sobrevivência e variáveis histológicas tais como granuloma esquistosomótico, alterações portais, necrose hepatocelular, regeneração hepática, pigmento esquistosomótico, fi-brose periductal e principalmente, alterações dos ductos biliares foram analisados nos animais infectados tratados e não tratados. A terapêutica por oxamniquina nos animais repetidamente infectados prolonga a sobrevivência de maneira significante (Chi-quadrado 9,24, p = 0,0024), portanto confirmando resultados anteriores com um modelo semelhante mas com um período mais curto de seguimento. Ainda, a mortalidade decresce rapidamente depois do tratamento, sugerindo uma abrupta redução na gravidade das lesões hepáticas. O fígado foi ainda estudado sob o ponto de vista morfológico e imunohistoquímico. Fibrose portal, com um quadro que lembra a fibrose humana do tipo Symmers está presente na fase tardia da infecção. As alterações de ductos biliares são muito próximas daquelas descritas na esquistosomose mansônica humana. Antígeno esquistosomótico foi observado em uma célula isolada do revestimento alterado de duetos biliares. A patogênese das alterações ductais e sua possível relação com a infecção parasitária e/ou seus antígenos foi discutida.

ORIGINAL ARTICLES

Liver morphology with emphasis on bile ducts changes and survival analysis in mice submitted to multiple Schistosoma mansoni infections and chemotherapy

Alterações morfológicas hepáticas, com especial ênfase nas alterações dos ductos biliares e análise de sobrevivência em camundongos submetidos a infecções múltiplas por S. mansoni e a quimioterapia

L. C. da Silva^I; M. Regina Vianna^II,III; C. P. Abrantes^I; D. M. C. Lima^I; A. L. Falavigna^I; R. H. Antonelli-Cardoso^I; S. D. Deperon Gallucci^I; T. de Brito^I,II,III

^IInstitute of Tropical Medicine of S. Paulo. São Paulo, SP, Brazil (LIM-47)

^IIDepartment of Pathology, S. Paulo Medical School, University of S. Paulo

^III"Instituto Adolfo Lutz", S. Paulo Health Service, S. Paulo, S. Paulo, SP, Brazil

^{Address for correspondence} Address for correspondence: Thales F. de Brito, M. D. Faculdade de Medicina da Universidade de São Paulo, Departamento de Patologia Av. Dr. Arnaldo, 455 CEP 01246 São Paulo, SP, Brasil

SUMMARY

In an attempt to be as close as possible to the infected and treated patients of the endemic areas of schistosomiasis (S. mansoni) and in order to achieve a long period of follow-up, mice were repeatedly infected with a low number of cercariae.

Survival data and histological variables such as schistosomal granuloma, portal changes, hepatocellular necrosis, hepatocellular regeneration, schistosomotic pigment, periductal fibrosis and chiefly bile ducts changes were analysed in the infected treated and non treated mice.

Oxamniquine chemotherapy in repeatedly infected mice prolonged survival significantly when compared to non-treated animals (chi-square 9.24, p = 0.0024), thus confirming previous results with a similar experimental model but with a shorter term follow-up. Furthermore, mortality decreased rapidly after treatment suggesting an abrupt reduction in the severity of hepatic lesions.

A morphological and immunohistochemical study of the liver was carried out. Portal fibrosis, with a pattern resembling human Symmers fibrosis was present at a late phase in the infected animals. Bile duct lesions were quite close to those described in human Mansonian schistosomiasis. Schistosomal antigen was observed in one isolated altered bile duct cell. The pathogenesis of the bile duct changes and its relation to the parasite infection and/or their antigens are discussed.

Key words: Experimental mansonian schistosomiasis; Oxamniquine therapy; Bile duct changes; Survival analysis.

RESUMO

Numa tentativa de estar o mais próximo possível a pacientes infectados e tratados nas áreas endêmicas de esquistosomose (S. mansoni) e também para obter um período mais longo de seguimento, camundongos foram repetidamente infectados com um número baixo de cercarias.

Dados de sobrevivência e variáveis histológicas tais como granuloma esquistosomótico, alterações portais, necrose hepatocelular, regeneração hepática, pigmento esquistosomótico, fi-brose periductal e principalmente, alterações dos ductos biliares foram analisados nos animais infectados tratados e não tratados.

A terapêutica por oxamniquina nos animais repetidamente infectados prolonga a sobrevivência de maneira significante (Chi-quadrado 9,24, p = 0,0024), portanto confirmando resultados anteriores com um modelo semelhante mas com um período mais curto de seguimento. Ainda, a mortalidade decresce rapidamente depois do tratamento, sugerindo uma abrupta redução na gravidade das lesões hepáticas.

O fígado foi ainda estudado sob o ponto de vista morfológico e imunohistoquímico. Fibrose portal, com um quadro que lembra a fibrose humana do tipo Symmers está presente na fase tardia da infecção. As alterações de ductos biliares são muito próximas daquelas descritas na esquistosomose mansônica humana. Antígeno esquistosomótico foi observado em uma célula isolada do revestimento alterado de duetos biliares. A patogênese das alterações ductais e sua possível relação com a infecção parasitária e/ou seus antígenos foi discutida.

Full text available only in PDF format.

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ACKNOWLEDGEMENTS

The authors would like to thank Edna A. Leick and Dr. R. Telma M. Santos for technical help and Maria Elí P. de Castro for secretarial assistance in the preparation of this manuscript.

Recebido para publicação em 10/8/1990

1. ABDUL-ALL, G. & ATTALLAH, A.M. Immunopathology of experimental Schistosoma mansoni infection: immunohistochemical localization of parasite antigens in the host tissue. Int. Arch. Allergy, 82: 89-94, 1987.
2. ACOSTA-FERREIRA, W.; VERCELLI-RELTA, J. & FALCONI, L.M. Fasciola hepatica human infection. Histopathological study of sixteen cases. Virchows Arch. A Path. Anat. Histol., 383: 319-327, 1979.
3. ANDRADE, Z.A. & GRIMAUD, J.A. Evolution of the schistosomal hepatic lesions in mice after curative chemotherapy. Amer. J. Path., 124: 59-65, 1986.
4. ANDREWS, P.; DYCKA, J. & FRANK, G. Effects of praziquantel on clinical-chemical parameters in healthy and schistosome infected mice. Ann. trop. Med. Parasit., 74: 167-177, 1980.
5. CHOU, S.T. & GIBSON, J.B. The histochemistry of biliary mucins and the changes caused by infestation with Clonorchis sinensis. J. Path., 101: 185-197, 1970.
6. DA SILVA, L.C.; ALVES, V.A.F.; ABRANTES, C.P.; LIMA, D.M.; CHRISTO, C.H. & DE BRITO, T. Effects of chemotherapy on mice submitted to multiple Schistosoma mansoni infection. A controlled randomized prospective study. Tropenmed. Parasit., 36: 150-154, 1985.
7. GÖNNERT, R.V. Schistosomiasis studien IV Zur Pathologie der Schistosomiasis der Maus. Z. Tropenmed. Parasit., 6: 279-336, 1955.
8. HOU, P.C. The pathology of Clonorchis sinensis infestation of the liver. J. Path. Bact., 70: 53-64, 1955.
9. HOU, P.C. The relationship between primary carcinoma of the liver and infestation with Clonorchis sinensis. J. Path. Bact., 72: 239-246, 1956.
10. HOU, P.C. Primary carcinoma of bile duct of the liver of the cat (Felis catus) infested with Clonorchis sinensis. J. Path. Bact., 87: 239-244, 1964.
11. KAPLAN, E.L. & MEIER, P. Nonparametric estimation from incomplete observations. J. Amer. Statist. Ass., 53: 457-481, 1958.
12. KOZUKA, S.; KURASHINA, M.; TSUBONE, M.; HACHISUKA, K. & YASUI, A. Significance of intestinal metaplasia for the evolution of cancer in the biliary tract. Cancer, 54: 2277-2285, 1984.
13. KURASHINA, M.; KOZUKA, S.; NAKASIMA, N.; HIRABAYASI, N. & MASAFUMI, I. Relationship of intrahepatic bile duct hyperplasia to cholangiocellular carcinoma. Cancer, 61: 2469-2474, 1988.
14. PETO, R. & PETO, J. Asymptotically efficient rank invariant procedures. J. roy. Statist. Soc., Series A, 135: 185-207, 1972.
15. STERNBERGER, L.A.; HARDY, P.H.; CUCULITIS, J.J. & MEYER, H.G. The unlabelled antibody-enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (Horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J. Histochem. Cytochem., 18: 315-333, 1970.
16. THAMAVIT, W.; KONGKANUNTIN, R.; TIMAWECH, D. & MOORE, M.A. Level of Opisthorchis infestation and carcinogen dose-dependence of cholangiocarcinoma induction in Syrian golden hamsters. Virchows Arch. B, 54: 52-58, 1987.
17. VIANNA, M.R.; GAYOTTO, L.C.C.; SANTOS, R.T.M.; ALVES, V.A.F. & DE BRITO, T. Intrahepatic bile duct changes in human hepatosplenic schistosomiasis mansoni. Liver, 9: 100-109, 1989.
18. WARREN, K.S. The influence of treatment on the development and course of murine hepato-splenic schistosomiasis mansoni. Trans. roy. Soc. trop. Med. Hyg., 56: 510-519, 1962.
19. WARREN, K.S. The pathogenesis of "clay-pipe stem cirrhosis" in mice with chronic schistosomiasis mansoni, with a note on the longevity of the schistosomes. Amer. J. Path., 49: 477-485, 1966.

Address for correspondence:

Thales F. de Brito, M. D.

Faculdade de Medicina da Universidade de São Paulo, Departamento de Patologia

Av. Dr. Arnaldo, 455

CEP 01246 São Paulo, SP, Brasil

Publication Dates

Publication in this collection
12 Sept 2006
Date of issue
Oct 1990

History

Received
10 Aug 1990

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

[1] 1. ABDUL-ALL, G. & ATTALLAH, A.M. Immunopathology of experimental Schistosoma mansoni infection: immunohistochemical localization of parasite antigens in the host tissue. Int. Arch. Allergy, 82: 89-94, 1987.

[2] 2. ACOSTA-FERREIRA, W.; VERCELLI-RELTA, J. & FALCONI, L.M. Fasciola hepatica human infection. Histopathological study of sixteen cases. Virchows Arch. A Path. Anat. Histol., 383: 319-327, 1979.

[3] 3. ANDRADE, Z.A. & GRIMAUD, J.A. Evolution of the schistosomal hepatic lesions in mice after curative chemotherapy. Amer. J. Path., 124: 59-65, 1986.

[4] 4. ANDREWS, P.; DYCKA, J. & FRANK, G. Effects of praziquantel on clinical-chemical parameters in healthy and schistosome infected mice. Ann. trop. Med. Parasit., 74: 167-177, 1980.

[5] 5. CHOU, S.T. & GIBSON, J.B. The histochemistry of biliary mucins and the changes caused by infestation with Clonorchis sinensis. J. Path., 101: 185-197, 1970.

[6] 6. DA SILVA, L.C.; ALVES, V.A.F.; ABRANTES, C.P.; LIMA, D.M.; CHRISTO, C.H. & DE BRITO, T. Effects of chemotherapy on mice submitted to multiple Schistosoma mansoni infection. A controlled randomized prospective study. Tropenmed. Parasit., 36: 150-154, 1985.

[7] 7. GÖNNERT, R.V. Schistosomiasis studien IV Zur Pathologie der Schistosomiasis der Maus. Z. Tropenmed. Parasit., 6: 279-336, 1955.

[8] 8. HOU, P.C. The pathology of Clonorchis sinensis infestation of the liver. J. Path. Bact., 70: 53-64, 1955.

[9] 9. HOU, P.C. The relationship between primary carcinoma of the liver and infestation with Clonorchis sinensis. J. Path. Bact., 72: 239-246, 1956.

[10] 10. HOU, P.C. Primary carcinoma of bile duct of the liver of the cat (Felis catus) infested with Clonorchis sinensis. J. Path. Bact., 87: 239-244, 1964.

[11] 11. KAPLAN, E.L. & MEIER, P. Nonparametric estimation from incomplete observations. J. Amer. Statist. Ass., 53: 457-481, 1958.

[12] 12. KOZUKA, S.; KURASHINA, M.; TSUBONE, M.; HACHISUKA, K. & YASUI, A. Significance of intestinal metaplasia for the evolution of cancer in the biliary tract. Cancer, 54: 2277-2285, 1984.

[13] 13. KURASHINA, M.; KOZUKA, S.; NAKASIMA, N.; HIRABAYASI, N. & MASAFUMI, I. Relationship of intrahepatic bile duct hyperplasia to cholangiocellular carcinoma. Cancer, 61: 2469-2474, 1988.

[14] 14. PETO, R. & PETO, J. Asymptotically efficient rank invariant procedures. J. roy. Statist. Soc., Series A, 135: 185-207, 1972.

[15] 15. STERNBERGER, L.A.; HARDY, P.H.; CUCULITIS, J.J. & MEYER, H.G. The unlabelled antibody-enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (Horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J. Histochem. Cytochem., 18: 315-333, 1970.

[16] 16. THAMAVIT, W.; KONGKANUNTIN, R.; TIMAWECH, D. & MOORE, M.A. Level of Opisthorchis infestation and carcinogen dose-dependence of cholangiocarcinoma induction in Syrian golden hamsters. Virchows Arch. B, 54: 52-58, 1987.

[17] 17. VIANNA, M.R.; GAYOTTO, L.C.C.; SANTOS, R.T.M.; ALVES, V.A.F. & DE BRITO, T. Intrahepatic bile duct changes in human hepatosplenic schistosomiasis mansoni. Liver, 9: 100-109, 1989.

[18] 18. WARREN, K.S. The influence of treatment on the development and course of murine hepato-splenic schistosomiasis mansoni. Trans. roy. Soc. trop. Med. Hyg., 56: 510-519, 1962.

[19] 19. WARREN, K.S. The pathogenesis of "clay-pipe stem cirrhosis" in mice with chronic schistosomiasis mansoni, with a note on the longevity of the schistosomes. Amer. J. Path., 49: 477-485, 1966.

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