Histopathology and immunocytochemical study of type 3 and type 4 complement receptors in the liver and spleen of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi

Tafuri, Wagner Luiz; Tafuri, Washington Luiz; Barbosa, Alfredo José Afonso; Michalick, Marilene Susan Marques; Genaro, Odair; França-Silva, João Carlos; Mayrink, Wilson; Nascimento, Evaldo

doi:10.1590/S0036-46651996000200001

Abstracts

The objective of this study was to compare the histopathological changes and expression of CR3 and CR4 in the liver and spleen of dogs naturally and experimentally infected with L. chagasi. The basic histopathological lesions observed mainly in naturally infected dogs were: epithelioid hepatic granulomas, hyperplasia and hypertrophy of Kupffer cells, Malpigui follicles and mononucleated cells of the red pulp of the spleen. Sections from the liver and spleen by immunocytochemistry technique showed the presence of CD11b,c\CD 18 antigens in the control and infected animals and no qualitative or quantitative differences in the liver. Nevertheless, CD18 was always increased in the spleen of naturally and experimentally infected dogs. These results indicate that there is a difference in the activaton of CD 18 in both experimental and natural cases of canine visceral leishmaniasis that should play an important role in the immunological response to Leishmania chagasi infection.

Dogs; Leishmania chagasi; Histopathology; Immunocytology; Complement receptors

Os objetivos deste trabalho visaram uma análise comparativa das alterações histopatológicas e da expressão de CR3 e CR4 no fígado e baço de cães natural e experimentalmente infectados com L. chagasi. As lesões histopatológicas fundamentais observadas principalmente nos cães naturalmente infectados foram: os granulomas epitelióides hepáticos, a hiperplasia e a hipertrofia das células de Kupffer, dos folículos de Malpighi e das células mononucleadas da polpa vermelha do baço. Os cortes de fígado e baço corados pela técnica de imunocitoquímica mostraram a presença dos antígenos CD11b,c\CD18 nos animais controles e infectados, sem diferenças qualitativas e quantitativas no fígado. Entretanto, no baço dos cães natural e experimentalmente infectados a expressão de CD 18 (subunidade beta2 da molécula comum aos leucócitos) foi sempre aumentada. Em leishmaniose a resposta imune celular é considerada a mais importante na resolução da doença. A expressão de CD11 b,c\CD 18 pelos macrófagos deve representar um papel central na resposta celular. Estes resultados indicam que tanto na leishmaniose visceral canina experimental ou natural existe uma diferença na ativação de CD 18, o qual deve exercer uma importante função na resposta imunológica na infecção por Leishmania chagasi.

PARASITOLOGY

Histopathology and immunocytochemical study of type 3 and type 4 complement receptors in the liver and spleen of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi

Histopatologia e estudo imunocitoquímico dos receptores do tipo 3 e 4 do complemento no fígado e baço de cães natural e experimentalmente infectados com Leishmaniose Visceral

Wagner Luiz Tafuri; Washington Luiz Tafuri; Alfredo José Afonso Barbosa; Marilene Susan Marques Michalick; Odair Genaro; João Carlos França-Silva; Wilson Mayrink; Evaldo Nascimento

Department of General and Anatomical Pathology, UFMG

^{Correspondence to} Correspondence to: Wagner L. Tafuri Department of General Pathology, ICB/UFMG, Campus da Pampulha Av. Antônio Carlos 6627 30270-901 Belo Horizonte, Minas Gerais, Brasil

SUMMARY

The objective of this study was to compare the histopathological changes and expression of CR3 and CR4 in the liver and spleen of dogs naturally and experimentally infected with L. chagasi. The basic histopathological lesions observed mainly in naturally infected dogs were: epithelioid hepatic granulomas, hyperplasia and hypertrophy of Kupffer cells, Malpigui follicles and mononucleated cells of the red pulp of the spleen. Sections from the liver and spleen by immunocytochemistry technique showed the presence of CD11b,c\CD 18 antigens in the control and infected animals and no qualitative or quantitative differences in the liver. Nevertheless, CD18 was always increased in the spleen of naturally and experimentally infected dogs. These results indicate that there is a difference in the activaton of CD 18 in both experimental and natural cases of canine visceral leishmaniasis that should play an important role in the immunological response to Leishmania chagasi infection.

Keywords: Dogs; Leishmania chagasi; Histopathology; Immunocytology; Complement receptors.

RESUMO

Os objetivos deste trabalho visaram uma análise comparativa das alterações histopatológicas e da expressão de CR3 e CR4 no fígado e baço de cães natural e experimentalmente infectados com L. chagasi. As lesões histopatológicas fundamentais observadas principalmente nos cães naturalmente infectados foram: os granulomas epitelióides hepáticos, a hiperplasia e a hipertrofia das células de Kupffer, dos folículos de Malpighi e das células mononucleadas da polpa vermelha do baço. Os cortes de fígado e baço corados pela técnica de imunocitoquímica mostraram a presença dos antígenos CD11b,c\CD18 nos animais controles e infectados, sem diferenças qualitativas e quantitativas no fígado. Entretanto, no baço dos cães natural e experimentalmente infectados a expressão de CD 18 (subunidade b2 da molécula comum aos leucócitos) foi sempre aumentada. Em leishmaniose a resposta imune celular é considerada a mais importante na resolução da doença. A expressão de CD11 b,c\CD 18 pelos macrófagos deve representar um papel central na resposta celular. Estes resultados indicam que tanto na leishmaniose visceral canina experimental ou natural existe uma diferença na ativação de CD 18, o qual deve exercer uma importante função na resposta imunológica na infecção por Leishmania chagasi.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGMENTS

The authors wish to thank Professors Ivan Barbosa Machado Sampaio, Department of Zootechny, Veterinary School, UFMG, for assistance with the statistical analyses, and Professor Peter F. Moore, School of Veterinary Medicine, University of California - Los Angeles and Davis, USA, for kindly making available canine monoclonal antibodies. Financial support: FINEP, CNPq, PRPq-UFMG, PADCT/ CNPq/Biotecnologia.

Recebido para publicação em 27/09/95

Aceito para publicação em 21/03/96

1. ALENCAR, J.E. - Calazar canino. Contribuiçăo para o estudo da epidemiología no Brasil. Fortaleza, Imprensa Oficial, 1959. (Tese).
2. ANDERSON, D.C. & SPRINGER, T.A. - Leucocyte adhesion deficiency: an inherited defect in the Mac-1, LFA-1, and p150,55 glycoproteins. Ann. Rev. Med., 38: 175-194, 1987.
3. ANDRADE, Z.A. & ANDRADE, S.G. - Alguns novos aspectos da patologia do calazar (estudo morfológico de 13 casos necropsiados). Rev. Inst. Med. trop. S. Paulo, 8: 259-266, 1966.
4. ANOSA, V.O. & IDOWU, A.L. - The clinico-haematological features and pathology of leishmaniasis in a dog in Nigeria. Zbl. Vet. Med. B., 30: 600-608, 1983.
5. BAMBIRRA, E.A.; TAFURI, W.L. & RASO, P. - Alteraçőes ultra-estruturais do fígado na leishmaniose visceral (Calazar). Estudo de tręs casos. Rev. goiana Med., 26: 47-54, 1980.
6. BOGLIOLO, L. - Nova contribuiçăo ao conhecimento da anatomia patológica da leishmaniose visceral. A propósito de um caso brasileiro e com especial referęncia a fibrose hepática leishmaniótica. Hospital (Rio de J.), 3: 101-164, 1956.
7. CARVALHO, E.M.; BADARÓ, R.; REED, S.G. et al. - Absence of Gamma Interferon and Interleukin 2 production during active visceral leishmaniasis. J. clin. Invest.,76: 2066-2069, 1985.
8. CARVALHO, E.M.; BARRAL, A.; PEDRAL-SAMPAIO, D. et al. - Immunologic markers of clinical evolution in children recently infected with Leishmania donovani chagasi. J infect. Dis., 165: 535-540, 1992.
9. CERVIA, J.S.; ROSEN, H. & MURRAY, H.W. - Effector role of blood monocytes in experimental visceral leishmaniasis. Infect. Immun., 61: 1330-1333, 1993.
10. COOPER, A.; ROSEN, H. & BLACKWELL, J.M. - Monoclonal antibodies that recognize distinct epitopes of the macrophages type three complement receptor differ in their ability to inhibit binding of Leishmania promastigotes harvested at different phases of their growth cycle. Immunology, 65: 511-514, 1988.
11. De La HERA, A.; ALVAREZ-MON, M.; SANCHEZ-MADRID, F.; MARINEZ, A.C. & DURANTEZ, A. - Co-expression of Mac-1 and p150, 95 on CD5+ B cells. Structural and function characterization in a human chronic lymphocytic leukaemia. Europ. J. Immunol., 18: 1131-1134, 1988.
12. DEANE, L.M & DEANE, M.P. - Visceral leishmaniasis in Brazil. Geographical distribution and transmission. Rev. Inst. Med. trop. S. Paulo, 4: 198-212, 1962.
13. GENARO, O. - Leishmaniose visceral canina experimental., Belo Horizonte, 1993. (Tese de Doutoramento - Instituto de Cięncias Biológicas da Universidade Federal de Minas Gerais).
14. GEORGE, J.W.; NIELSEN, S.W.; SHIVELY, J.N. et al. - Canine leishmaniasis with amyloidosis. Vet. Path., 13: 363-373, 1976.
15. GRIMALDI, G.; TESH, R.B. & Mc MATHON-PRATT, D.A. - A review of the geographic distribution and epidemiology of leishmaniasis in the New World. Amer. J. trop. Med. Hyg., 41: 687-725, 1989.
16. GUTIERREZ, Y; MAKSEM, J.A. & REINER, N.E. - Pathological changes in murine leishmaniasis (Leishmania donovani) with special reference to the dynamics of granuloma formation in the liver. Amer. J. Path., 114: 222-230, 1983.
17. HO, M.K. & SPRINGER, T.A. - Mac-1 antigen: quantitative expression in macrophage populations and tissue, and immuno-fluorescent localization in spleen. J. Immunol., 128: 2281-2286, 1982.
18. HYNES, R.O. - Integrins: a family of cell surface receptors. Cell, 48: 549-554, 1987.
19. HYNES, R.O & LANDER, A.D. - Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons. Cell, 66: 303-322, 1992.
20. ISSEKUTZ, T.B. - The contributions of integrins to leukocyte infiltration in inflamed tissues. Curr. Top. Microbiol. Immunol., 184: 78-185, 1993.
21. KEENAN, C.N.; HENDRICKS, L.D.; LIGHTNER, L. & JOHNSON, A.J. - Visceral leishmaniasis in the German Shepherd Dog. II. Pathology. Vet. Path., 21: 80-86, 1984.
22. KISHIMOTO, T.K.; RICHARD, S.L.; CORBI, A.L. et al. - The leucocyte integrins. Advanc. Immunol., 46: 149-182, 1989.
23. KROEMER, G.; CUENDE, E. & MARTINEZ, A.C. - Compartimentalization of the peripheral immune system. Advanc. Immunol., 53: 157-215, 1993.
24. MARTINEZ-MORENO, A.; MARTINEZ-CRUZ, M.S. & HERNÁNDEZ-RODRIGUEZ, S. - Immunological and histological study of T- and B- Lymphocyte activity in canine visceral leishmaniosis. Vet. Parasit., 51: 49-59, 1993.
25. McELRATH, M.J.; MURRAY, H.W. & COHN, Z. A. - The dynamics of granuloma formation in experimental visceral leishmaniasis. J. exp. Med., 167: 1927-1937, 1988.
26. MILLER, L.J.; SCHWARTING, R. & SPRINGER, T.A. - Regulated expression of the Mac-1, LFA-1, p150,95 glycoprotein family during leukocyte differentiation. J. Immunol., 137: 2891-2900, 1986.
27. MINTER, D.M. - The leishmaniases. WHO/VBC/89.967: 93-106, 1989.
28. MOLYNEUX, D.H. & KILLICK-KENDRICK, R. - Morphology, ultraestructure and life cycles. In: PETERS, W. & KILLICK-KENDRICK, R., ed. The leishmaniasis in biology and medicine. London, Academic Press, 1987. v.1_x, p. 121-176.
29. MOORE, P.F.; ROSSITO, P.V. & DANILENKO, D.M. - Canine leukocyte integrins: characterization of a CDI8 homologue. Tissue Antigens, 36: 211-220, 1990.
30. MOSSER, D.M. & EDELSON, P.J. - The mouse macrophage receptor for C3bi (CR3) is a major mechanism in the phagocytosis of Leishmania promastigotes. J. Immunol., 135: 2785-2789, 1985.
31. MOSSER, D.M. & ROSENTHAL, L.A. - Leishmania-macrophage interactions: multiple receptors, multiple ligants and diverse cellular responses. Semin. Cell Biol., 4: 315-322, 1993.
32. MOSSER, D.M.; SPRINGER, T.A. & DIAMOND, M.S. - Leishmania promastigotes require opsonic complement to bind to the human leucocyte integrin Mac-1 (CD11b/CD18). J. Cell Biol., 116: 511-520, 1992.
33. NATTAN-LARRIER, L. - Les cirrhoses hépatiques due au Kalazar. Bull. Acad. Med., 98: 402-408, 1918.
34. OLIVEIRA, G.G.S.; SANTORO, F. & SADIGURSKY, M. - The subclinical form of experimental visceral leishmaniasis in dogs. Mem. Inst. Oswaldo Cruz, 88: 243-248, 1993.
35. PELLEGRINO, J. & BRENER, Z. - Reaçăo de fixaçăo de complemento com sangue dessecado no diagnóstico do calazar canino. Rev. bras. Malar., 10: 39-44, 1958.
36. RASO, P. & SIQUEIRA, J.T. - Subsídio ao conhecimento da anatomia patológica da leishmaniose visceral, com especial referęncia ŕs lesőes pulmonares e cardíacas. Hospital (Rio de J.), 65: 1291-1309, 1964.
37. ROGERS, L. - A peculiar intralobular cirrhosis of the liver produced by the protozoal parasite of Kala-azar. Ann. trop. Med. Parasit., 2: 147-152, 1908.
38. RUSSELL, D.G. & WRIGHT, S.D. - Complement receptor type 3 (CR3) binds to an Arg-Gly-Asp containing region of the major surface glycoprotein, gp63, of Leishmania promastigotes. J. exp. Med., 168: 279-292, 1988.
39. STELL, R.G.D. & TORRIE, J.H. - Principles and procedures statistics. New York, McGraw Hill, 1960.
40. STERNBERGER, L.A. - Immunocytochemistry. 2 ed. New York, John Wiley & Sons, 1986. p.20-35.
41. TALAMAS-ROHANA, P.; WRIGHT, S.D.; LENNARTZ, M.R. et al. - Lipophosphoglycan from Leishmania mexicana promastigotes binds to members of the CR3, p150,95 and LFA-1 family of leucocyte integrins. J. Immunol., 144: 4817-4824, 1990.
42. VAN DER WATER; DESTREE, A.T. & HYNES, R.O. - Fibronectin binds to some bacteria but not promote their uptake by phagocytic cells. Science, 220: 201-205, 1983.
43. VERESS, B.O.; OMER, A.; SATIR, A.A. & EL HASSAN, A.M. - Morphology of the spleen and lymph nodes in fatal visceral leishmaniasis. Immunology, 33: 605-610, 1977.
44. VIEIRA-DIAS, D.; TOLEDO, V.P.C.P.; MICHALICK, M.S.M. et al. - Intraocular parasitism by Leishmania chagasi in dogs with visceral leishmaniasis. Mem. Inst. Oswaldo Cruz, 88 (supl.): 124, 1993.
45. WRIGHT, S.D.; CRAIGMYLE, L.S. & SILVERSTEIN, S.C. - Fibronectin and serum amyloid P component stimulate C3b and C3bi-mediated phagocytosis in culture human monocytes. J. exp. Med., 158: 1338-1343, 1983.

Correspondence to:

Wagner L. Tafuri

Department of General Pathology, ICB/UFMG, Campus da Pampulha

Av. Antônio Carlos 6627

30270-901 Belo Horizonte, Minas Gerais, Brasil

Publication Dates

Publication in this collection
22 Sept 2006
Date of issue
Apr 1996

History

Received
27 Sept 1995
Accepted
21 Mar 1996

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

[1] 1. ALENCAR, J.E. - Calazar canino. Contribuiçăo para o estudo da epidemiología no Brasil. Fortaleza, Imprensa Oficial, 1959. (Tese).

[2] 2. ANDERSON, D.C. & SPRINGER, T.A. - Leucocyte adhesion deficiency: an inherited defect in the Mac-1, LFA-1, and p150,55 glycoproteins. Ann. Rev. Med., 38: 175-194, 1987.

[3] 3. ANDRADE, Z.A. & ANDRADE, S.G. - Alguns novos aspectos da patologia do calazar (estudo morfológico de 13 casos necropsiados). Rev. Inst. Med. trop. S. Paulo, 8: 259-266, 1966.

[4] 4. ANOSA, V.O. & IDOWU, A.L. - The clinico-haematological features and pathology of leishmaniasis in a dog in Nigeria. Zbl. Vet. Med. B., 30: 600-608, 1983.

[5] 5. BAMBIRRA, E.A.; TAFURI, W.L. & RASO, P. - Alteraçőes ultra-estruturais do fígado na leishmaniose visceral (Calazar). Estudo de tręs casos. Rev. goiana Med., 26: 47-54, 1980.

[6] 6. BOGLIOLO, L. - Nova contribuiçăo ao conhecimento da anatomia patológica da leishmaniose visceral. A propósito de um caso brasileiro e com especial referęncia a fibrose hepática leishmaniótica. Hospital (Rio de J.), 3: 101-164, 1956.

[7] 7. CARVALHO, E.M.; BADARÓ, R.; REED, S.G. et al. - Absence of Gamma Interferon and Interleukin 2 production during active visceral leishmaniasis. J. clin. Invest.,76: 2066-2069, 1985.

[8] 8. CARVALHO, E.M.; BARRAL, A.; PEDRAL-SAMPAIO, D. et al. - Immunologic markers of clinical evolution in children recently infected with Leishmania donovani chagasi. J infect. Dis., 165: 535-540, 1992.

[9] 9. CERVIA, J.S.; ROSEN, H. & MURRAY, H.W. - Effector role of blood monocytes in experimental visceral leishmaniasis. Infect. Immun., 61: 1330-1333, 1993.

[10] 10. COOPER, A.; ROSEN, H. & BLACKWELL, J.M. - Monoclonal antibodies that recognize distinct epitopes of the macrophages type three complement receptor differ in their ability to inhibit binding of Leishmania promastigotes harvested at different phases of their growth cycle. Immunology, 65: 511-514, 1988.

[11] 11. De La HERA, A.; ALVAREZ-MON, M.; SANCHEZ-MADRID, F.; MARINEZ, A.C. & DURANTEZ, A. - Co-expression of Mac-1 and p150, 95 on CD5+ B cells. Structural and function characterization in a human chronic lymphocytic leukaemia. Europ. J. Immunol., 18: 1131-1134, 1988.

[12] 12. DEANE, L.M & DEANE, M.P. - Visceral leishmaniasis in Brazil. Geographical distribution and transmission. Rev. Inst. Med. trop. S. Paulo, 4: 198-212, 1962.

[13] 13. GENARO, O. - Leishmaniose visceral canina experimental., Belo Horizonte, 1993. (Tese de Doutoramento - Instituto de Cięncias Biológicas da Universidade Federal de Minas Gerais).

[14] 14. GEORGE, J.W.; NIELSEN, S.W.; SHIVELY, J.N. et al. - Canine leishmaniasis with amyloidosis. Vet. Path., 13: 363-373, 1976.

[15] 15. GRIMALDI, G.; TESH, R.B. & Mc MATHON-PRATT, D.A. - A review of the geographic distribution and epidemiology of leishmaniasis in the New World. Amer. J. trop. Med. Hyg., 41: 687-725, 1989.

[16] 16. GUTIERREZ, Y; MAKSEM, J.A. & REINER, N.E. - Pathological changes in murine leishmaniasis (Leishmania donovani) with special reference to the dynamics of granuloma formation in the liver. Amer. J. Path., 114: 222-230, 1983.

[17] 17. HO, M.K. & SPRINGER, T.A. - Mac-1 antigen: quantitative expression in macrophage populations and tissue, and immuno-fluorescent localization in spleen. J. Immunol., 128: 2281-2286, 1982.

[18] 18. HYNES, R.O. - Integrins: a family of cell surface receptors. Cell, 48: 549-554, 1987.

[19] 19. HYNES, R.O & LANDER, A.D. - Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons. Cell, 66: 303-322, 1992.

[20] 20. ISSEKUTZ, T.B. - The contributions of integrins to leukocyte infiltration in inflamed tissues. Curr. Top. Microbiol. Immunol., 184: 78-185, 1993.

[21] 21. KEENAN, C.N.; HENDRICKS, L.D.; LIGHTNER, L. & JOHNSON, A.J. - Visceral leishmaniasis in the German Shepherd Dog. II. Pathology. Vet. Path., 21: 80-86, 1984.

[22] 22. KISHIMOTO, T.K.; RICHARD, S.L.; CORBI, A.L. et al. - The leucocyte integrins. Advanc. Immunol., 46: 149-182, 1989.

[23] 23. KROEMER, G.; CUENDE, E. & MARTINEZ, A.C. - Compartimentalization of the peripheral immune system. Advanc. Immunol., 53: 157-215, 1993.

[24] 24. MARTINEZ-MORENO, A.; MARTINEZ-CRUZ, M.S. & HERNÁNDEZ-RODRIGUEZ, S. - Immunological and histological study of T- and B- Lymphocyte activity in canine visceral leishmaniosis. Vet. Parasit., 51: 49-59, 1993.

[25] 25. McELRATH, M.J.; MURRAY, H.W. & COHN, Z. A. - The dynamics of granuloma formation in experimental visceral leishmaniasis. J. exp. Med., 167: 1927-1937, 1988.

[26] 26. MILLER, L.J.; SCHWARTING, R. & SPRINGER, T.A. - Regulated expression of the Mac-1, LFA-1, p150,95 glycoprotein family during leukocyte differentiation. J. Immunol., 137: 2891-2900, 1986.

[27] 27. MINTER, D.M. - The leishmaniases. WHO/VBC/89.967: 93-106, 1989.

[28] 28. MOLYNEUX, D.H. & KILLICK-KENDRICK, R. - Morphology, ultraestructure and life cycles. In: PETERS, W. & KILLICK-KENDRICK, R., ed. The leishmaniasis in biology and medicine. London, Academic Press, 1987. v.1_x, p. 121-176.

[29] 29. MOORE, P.F.; ROSSITO, P.V. & DANILENKO, D.M. - Canine leukocyte integrins: characterization of a CDI8 homologue. Tissue Antigens, 36: 211-220, 1990.

[30] 30. MOSSER, D.M. & EDELSON, P.J. - The mouse macrophage receptor for C3bi (CR3) is a major mechanism in the phagocytosis of Leishmania promastigotes. J. Immunol., 135: 2785-2789, 1985.

[31] 31. MOSSER, D.M. & ROSENTHAL, L.A. - Leishmania-macrophage interactions: multiple receptors, multiple ligants and diverse cellular responses. Semin. Cell Biol., 4: 315-322, 1993.

[32] 32. MOSSER, D.M.; SPRINGER, T.A. & DIAMOND, M.S. - Leishmania promastigotes require opsonic complement to bind to the human leucocyte integrin Mac-1 (CD11b/CD18). J. Cell Biol., 116: 511-520, 1992.

[33] 33. NATTAN-LARRIER, L. - Les cirrhoses hépatiques due au Kalazar. Bull. Acad. Med., 98: 402-408, 1918.

[34] 34. OLIVEIRA, G.G.S.; SANTORO, F. & SADIGURSKY, M. - The subclinical form of experimental visceral leishmaniasis in dogs. Mem. Inst. Oswaldo Cruz, 88: 243-248, 1993.

[35] 35. PELLEGRINO, J. & BRENER, Z. - Reaçăo de fixaçăo de complemento com sangue dessecado no diagnóstico do calazar canino. Rev. bras. Malar., 10: 39-44, 1958.

[36] 36. RASO, P. & SIQUEIRA, J.T. - Subsídio ao conhecimento da anatomia patológica da leishmaniose visceral, com especial referęncia ŕs lesőes pulmonares e cardíacas. Hospital (Rio de J.), 65: 1291-1309, 1964.

[37] 37. ROGERS, L. - A peculiar intralobular cirrhosis of the liver produced by the protozoal parasite of Kala-azar. Ann. trop. Med. Parasit., 2: 147-152, 1908.

[38] 38. RUSSELL, D.G. & WRIGHT, S.D. - Complement receptor type 3 (CR3) binds to an Arg-Gly-Asp containing region of the major surface glycoprotein, gp63, of Leishmania promastigotes. J. exp. Med., 168: 279-292, 1988.

[39] 39. STELL, R.G.D. & TORRIE, J.H. - Principles and procedures statistics. New York, McGraw Hill, 1960.

[40] 40. STERNBERGER, L.A. - Immunocytochemistry. 2 ed. New York, John Wiley & Sons, 1986. p.20-35.

[41] 41. TALAMAS-ROHANA, P.; WRIGHT, S.D.; LENNARTZ, M.R. et al. - Lipophosphoglycan from Leishmania mexicana promastigotes binds to members of the CR3, p150,95 and LFA-1 family of leucocyte integrins. J. Immunol., 144: 4817-4824, 1990.

[42] 42. VAN DER WATER; DESTREE, A.T. & HYNES, R.O. - Fibronectin binds to some bacteria but not promote their uptake by phagocytic cells. Science, 220: 201-205, 1983.

[43] 43. VERESS, B.O.; OMER, A.; SATIR, A.A. & EL HASSAN, A.M. - Morphology of the spleen and lymph nodes in fatal visceral leishmaniasis. Immunology, 33: 605-610, 1977.

[44] 44. VIEIRA-DIAS, D.; TOLEDO, V.P.C.P.; MICHALICK, M.S.M. et al. - Intraocular parasitism by Leishmania chagasi in dogs with visceral leishmaniasis. Mem. Inst. Oswaldo Cruz, 88 (supl.): 124, 1993.

[45] 45. WRIGHT, S.D.; CRAIGMYLE, L.S. & SILVERSTEIN, S.C. - Fibronectin and serum amyloid P component stimulate C3b and C3bi-mediated phagocytosis in culture human monocytes. J. exp. Med., 158: 1338-1343, 1983.