Advertisement

Parasitology Research

, Volume 101, Issue 3, pp 547–555 | Cite as

Immunohistochemical study of the local immune response in lambs experimentally infected with Dicrocoelium dendriticum (Digenea)

  • M. Carmen Ferreras-EstradaEmail author
  • R. Campo
  • C. González-Lanza
  • V. Pérez
  • J. F. García-Marín
  • M. Y. Manga-González
Original Paper

Abstract

Phenotypic expression of inflammatory cells in liver and hepatic lymph nodes (HLN) has been examined in lambs experimentally infected with Dicrocoelium dendriticum using immunohistochemical techniques. Thirty-two lambs, 12 infected with 1,000 D. dendriticum metacercariae, 12 with 3,000, and 8 controls were used. Half the lambs in each group were slaughtered on days 60 and 180 post-infection (p.i.), respectively. Primary antibodies (Abs) against T cell epitopes (CD3+, CD4+, CD8+ and WC1+ γδ), B cell epitopes (CD79αcy+, CD45R+), immunoglobulin (IgG)-bearing plasma cells, macrophages (CD14+, VPM32+) and major histocompatibility complex (MHC) class IIβ antigen were used. T lymphocytes (CD3+, CD4+ and CD8+) and B lymphocytes (CD79αcy+ and CD45R+) with diffuse pattern or forming lymphoid aggregates and follicles surrounded the septal bile ducts (SBD) and inter-lobular bile ducts, whereas the WC1 γδ T cells were scattered. Numerous IgG+ plasma cells were observed around SBD. CD14 and VPM32+ macrophages intermingled with lymphocytes were immunostained by the anti-MHC class IIβ. This Ab also reacted with lymphoid cells. Likewise, increased positive immunostaining for all Abs used was observed in the HLN of infected lambs. There was no qualitative difference regarding the phenotype expression of inflammatory cells between the lambs infected with D. dendriticum. The humoral and cell-mediated local immune responses observed were similar in the two groups of lambs infected with different doses.

Keywords

Bile Duct Major Histocompatibility Complex Class Cell Epitope Lymphoid Follicle Small Round Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We would like to thank M. P. del Pozo, C. Espiniella and M. L. Carcedo of the Estación Agrícola Experimental (CSIC, León, Spain) and S. Liébana and J. Benavides of the Departamento de Sanidad Animal de la Universidad de León (Spain) for their technical assistance. We also wish to thank P. Díez, L. Pérez and H. Fidalgo for taking care of and handling the animals (CSIC, León). This study was supported by the Spanish CICYT (Projects No. AGF92-0588 and AGF96-0416) and by the “Junta de Castilla y León” (Project No. CSIC01A06). The experiments carried out with lambs in this study comply with current Spanish law on animal experimentation.

References

  1. Aould J (1994) Antigen unmasking in routinely processed paraffin sections by pressure cooking. United Kingdom National External Quality Assessment Schemes—ICC, pp 6–9Google Scholar
  2. Berthon P, Bernard F, Olivier M, Bernard S, Lantier F (1996) Immunohistochemical study on the reactivity of Workshop monoclonal antibodies with sheep lymph nodes. Vet Immunol Immunopathol 52:393–401PubMedCrossRefGoogle Scholar
  3. Campo R, Manga-González MY, González-Lanza C (2000) Relationship between egg output and parasitic burden in lambs experimentally infected with different doses of Dicrocoelium dendriticum (Digenea). Vet Parasitol 87:139–149PubMedCrossRefGoogle Scholar
  4. Chauvin A, Boulard C (1996) Local immune response to experimental Fasciola hepatica infection in sheep. Parasite 3:209–215PubMedGoogle Scholar
  5. Chauvin A, Moreau E, Boulard C (2001) Responses of Fasciola hepatica infected sheep to various infection levels. Vet Res 32:87–92PubMedCrossRefGoogle Scholar
  6. Corpa JM, Juste RA, García Marín JF, Reyes LE, González J, Pérez V (2001) Distribution of lymphocyte subsets in the small intestine lymphoid tissue of 1-month-old lambs. Anat Histol Embryol 30:121–127PubMedCrossRefGoogle Scholar
  7. Gershwin LJ, Krakowka S, Olsen RG (1995) Immunology and immunopathology of domestic animals. Mosby, St. Louis, MO, pp 62–67Google Scholar
  8. González L, Anderson I, Deane D, Summers C, Buxton D (2001) Detection of immune system cells in paraffin wax-embedded ovine tissues. J Comp Path 125:41–47PubMedCrossRefGoogle Scholar
  9. González-Lanza C, Manga-González MY, Campo R, Del-Pozo P, Sandoval H, Oleaga A, Ramajo V (2000) IgG antibody response to ES or somatic antigens of Dicrocoelium dendriticum (Trematoda) in experimentally infected sheep. Parasitol Res 86:472–479PubMedCrossRefGoogle Scholar
  10. González-Lanza C, Manga-González MY, Revilla-Nuín B (2006) Preliminary protective capacity study of a Dicrocoelium dendriticum antigenic protein in hamsters. Parasitol Res 99:749–752PubMedCrossRefGoogle Scholar
  11. Isse K, Harada K, Sato Y, Nakanuma Y (2006) Characterization of biliary intra-epithelial lymphocytes at different anatomical levels of intrahepatic bile ducts under normal and pathological conditions: Numbers of CD4+CD28-intra-epithelial lymphocytes are increased in primary biliary cirrhosis. Pathol Int 56:17–24PubMedCrossRefGoogle Scholar
  12. Jankovic D, Liu Z, Gause WC (2001) Th1-and Th2-cell commitment during infectious disease: asymmetry in divergent pathways. Trends Immunol 22(8):450–457PubMedCrossRefGoogle Scholar
  13. Manga-González MY, González-Lanza C, Cabanas E, Campo R (2001) Contributions to and review of dicrocoeliosis, with special reference to the intermediate hosts of Dicrocoelium dendriticum. Parasitology 123:S91–S114PubMedCrossRefGoogle Scholar
  14. Manga-González MY, Ferreras MC, Campo R, González-Lanza C, Pérez V, García Marín JF (2004) Hepatic marker enzymes, biochemical parameters and pathological effects in lambs experimentally infected with Dicrocoelium dendriticum. Parasitol Res 93:344–355PubMedCrossRefGoogle Scholar
  15. Martínez-Moreno A, Jiménez-Luque V, Moreno T, Redondo ES, de las Mulas JM, Pérez J (1999) Liver pathology and immune response in experimental Fasciola hepatica infections of goats. Vet Parasitol 82(1):19–33PubMedCrossRefGoogle Scholar
  16. Mateo A, Garrido JJ, Pérez de la Lastra J, Martín de las Mulas J, Moreno A, Pintado CO, Llanes (1999) A new epitope on sheep CD45R molecule detected by a monoclonal antibody. Comp Immunol Microbiol Infect Dis 22:125–136PubMedCrossRefGoogle Scholar
  17. Meeusen E, Lee CS, Rickard MD, Brandon MR (1995) Cellular responses during liver fluke infection in sheep and its evasion by the parasite. Parasite Immunol 17:37–45PubMedCrossRefGoogle Scholar
  18. Molina EC, Skerratt LF (2005) Cellular and humoral responses in liver of cattle and buffaloes infected with a single dose of Fasciola gigantica. Vet Parasitol 131:157–163PubMedCrossRefGoogle Scholar
  19. Mosmann TR, Coffman RL (1989) Th1 and Th2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol 7:145–173PubMedCrossRefGoogle Scholar
  20. Mulcahy G, Dalton JP (2001) Cathepsin L proteinases as vaccines against infection with Fasciola hepatica (liver fluke) in ruminants. Res Vet Sci 70(1):83–86PubMedCrossRefGoogle Scholar
  21. Mwangi DM, Hopkins J, Luckins AG (1990) Cellular phenotypes in Trypanosoma congolense infected sheep: the local skin reaction. Parasite Immunol 12:647–658PubMedCrossRefGoogle Scholar
  22. Otranto D, Traversa D (2003) Dicrocoeliosis of ruminants: a little known fluke disease. Trends Parasitol 19:2–15CrossRefGoogle Scholar
  23. Pérez J, Martín de las Mulas J, Chacón-M De Lara F, Gutiérrez-Palomino PN, Becerra-Martel C, Martínez-Moreno A (1998) Immunohistochemical study of the local immune response to Fasciola hepatica in primarily and secondarily infected goats. Vet Immunol Immunopathol 64:337–348PubMedCrossRefGoogle Scholar
  24. Pérez J, Martín de las Mulas J, Carrasco L, Gutiérrez PN, Martínez-Cruz MS, Martínez-Moreno A (1999) Pathological and immunohistochemical study of the liver and hepatic lymph nodes in goats infected with one or more doses of Fasciola hepatica. J Comp Path 120:199–210PubMedCrossRefGoogle Scholar
  25. Pérez J, Ortega J, Moreno T, Morrondo P, López-Sánchez C, Martínez-Moreno A (2002) Pathological and immunohistochemical study of the liver and hepatic lymph nodes of sheep chronically reinfected with Fasciola hepatica, with or without triclabendazole treatment. J Comp Path 127:30–36PubMedCrossRefGoogle Scholar
  26. Pérez J, Ortega J, Bravo A, Díez-Baños P, Morrondo P, Moreno T, Martínez-Moreno A (2005) Phenotype of hepatic infiltrates and hepatic lymph nodes of lambs primarily and challenge infected with Fasciola hepatica, with and without triclabendazole treatment. Vet Res 36:1–12PubMedCrossRefGoogle Scholar
  27. Rack J, Adusu E, Jelinek T (2004) Human infection with Dicrocoelium dendriticum. Dtsch Med Wochenschr 129:2538–2540PubMedCrossRefGoogle Scholar
  28. Rimmelzwann GF, Osterhaus ADME (1999) The immune response. In: Pastoret PP, Blancou J, Vannier P, Verschueren C (eds) Veterinary vaccinology. Elsevier, Amsterdam, pp 55–64Google Scholar
  29. Timoteo O, Maco V Jr, Maco V, Neyra V, Yi PJ, Leguía G, Espinoza JR (2005) Characterization of the humoral immune response in alpacas (Lama pacos) experimentally infected with Fasciola hepatica against cysteine proteinases Fas1 and Fas2 and histopathological findings. Vet Immunol Immunopathol 106:77–86PubMedGoogle Scholar
  30. Tliba O, Sibille P, Boulard C, Chauvin A (2000) Local hepatic immune response in rats during primary infection with Fasciola hepatica. Parasite 7(1):9–18PubMedGoogle Scholar
  31. Tliba O, Moire N, Le Vern Y, Boulard C, Chauvin A, Sibille P (2002) Early hepatic immune response in rats infected with Fasciola hepatica. Vet Res 33(3):261–270PubMedCrossRefGoogle Scholar
  32. Vray B (2002) Macrophages in parasitic infection. In: Burke B, Lewis CE (eds) The macrophage. Oxford Univ. Press, Oxford, UK pp 253–304Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • M. Carmen Ferreras-Estrada
    • 1
    Email author
  • R. Campo
    • 2
  • C. González-Lanza
    • 2
  • V. Pérez
    • 1
  • J. F. García-Marín
    • 1
  • M. Y. Manga-González
    • 2
  1. 1.Departamento de Sanidad Animal (Histología y Anatomía Patológica), Facultad de VeterinariaUniversidad de LeónLeonSpain
  2. 2.Consejo Superior de Investigaciones Científicas (CSIC)Estación Agrícola ExperimentalLeonSpain

Personalised recommendations