Skip to main content

Advertisement

Log in

Setting New Immunobiological Parameters in the Hamster Model of Visceral Leishmaniasis for In Vivo Testing of Antileishmanial Compounds

  • Published:
Veterinary Research Communications Aims and scope Submit manuscript

Abstract

To establish suitable immunobiological parameters for in vivo testing of new antileishmanial compounds in the golden hamster model of visceral leishmaniasis, two groups of 8 animals were infected each with 105 or 107 stationary promastigotes by the intracardiac route and the clinical and immunoparasitological features were monitored up to day 155 after infection. All animals became infected at both doses, although significant differences were observed between parasite burdens in liver and spleen. The mean number of parasites in animals infected with 107 promastigotes increased by 9.5 times in liver and by 43.1 times in spleen compared with those infected with 105 promastigotes. In animals given the higher dose, the outcome of the disease occurred between days 75 and 90 after infection, whereas no signs of disease were apparent in those given the lower infecting dose. Positive antibody (IgG) responses were detected earlier (week 5–7 after infection) in animals infected with the higher dose than in those infected with the lower dose (week 8–10 after infection), but these responses did not correlate with individual parasitological loads in liver and spleen. An inverse correlation was observed between infecting doses and in vitro spleen lymphocyte proliferation against mitogens (ConA). The proportion of CD4+ and CD19+ spleen cell increased in animals given the higher infection, whereas it decreased in those given the lower infection compared to naive controls.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

CSE:

crude saline extract

Con A:

concanavalin A

EDTA:

ethylenediaminetetraacetic acid

ELISA:

enzyme-linked immunosorbent assay

FACS:

fluorescence-activated cell sorter

FCS:

fetal calf serum

FELASA:

Federation of European Laboratory Animal Associations

FITC:

fluorescein isothiocyanate

IC:

intracardiac

NNN:

Novy, Nicolle and McNeal

OD:

optical density

OPD:

orthophenylenediamine

PBS:

phosphate-buffered saline

Phyt:

phytohaemagglutinin

R-PE:

(R)-phycoerythrin

SI:

stimulation index

References

  • Alvar, J., Cañavate, C., Gutierrez-Solar, B, Jimenez, M., Laguna, F., Lopez-Velez, R., Molina, R. and Moreno, J., 1997. Leishmania and human immunodeficiency virus coinfection: the first 10 years. Clinical Microbiology Reviews, 10, 298–319

    PubMed  CAS  Google Scholar 

  • Alvar, J., Cañavate, C., Molina, R., Moreno, J. and Nieto, J., 2004. Canine leishmaniasis. Advances in Parasitology, 57, 1–88

    Article  PubMed  Google Scholar 

  • Belkaid,Y., Kamhawi. S., Modi, G., Valenzuela, J., Noben-Trauth, N., Rowto, E., Ribeiro, J. and Sacks, D.L., 1998. Development of a natural model of cutaneous leishmaniasis: powerful effects of vector saliva and saliva pre-exposure on the long-term outcome of Leishmania major infection in the mouse ear dermis. Journal of Experimental Medicine, 188, 1941–1953

    Article  PubMed  CAS  Google Scholar 

  • Binzahim, A.A., Chapman, W.L. Jr., Shin, S.S. and Hanson, W.L., 1993. Determination of virulence and pathogenesis of a canine strain of Leishmania infantum in hamsters and dogs. American Journal of Veterinary Research, 54, 113–121

    Google Scholar 

  • Bourdoiseau, G., Bonnefont, C., Hoareau, E., Boehringer, C., Stolle, T. and Chabanne, L., 1997. Specific IgG1 and IgG2 antibody and lymphocyte subsets levels in naturally Leishmania infantum-infected treated and untreated dogs. Veterinary Immunology. Immunopathology, 59, 21–30

    Article  CAS  Google Scholar 

  • Bradford, M., 1976. A rapid and sensitive method for quantification of microgramquantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–252

    Article  PubMed  CAS  Google Scholar 

  • Bryceson, A., 2001. Current issues in the treatment of visceral leishmaniosis. Medical Microbiology Immunology, 190, 81–84

    CAS  Google Scholar 

  • Cabral, M., McNerney, R., Gomes, S., O'Grady, J., Frame, I., Sousa, J.C., Miles, M.A. and Alexander, J., 1993. Demonstration of natural Leishmania infection in asymptomatic dogs in the absence of specific humoral immunity. Archives del Institut Pasteur de Tunis, 70, 473–479

    CAS  Google Scholar 

  • Carrera, L., Balaña-Fouce, R. and Alunda, J.M., 1994. Polyamine content and drug sensitivities of different clonal lines of Leishmania infantum promastigotes. Parasitology Research, 80, 203–207

    Article  PubMed  CAS  Google Scholar 

  • Carrio, J., de Colmenares, M., Riera, C., Gállego, M., Arboix, M. and Portús, M., 2000. Leishmania infantum: Stage-specific activity of pentavalent antimony related with the assay conditions. Experimental Parasitology, 95, 209–214

    Article  PubMed  CAS  Google Scholar 

  • Croft, S., 2001. Monitoring drug resistance in leishmaniosis. Tropical Medicine and International Health, 6, 899–905

    Article  PubMed  CAS  Google Scholar 

  • De Luna, R., Vuotto, M.L., Lelpo, M.T.L., Ambrosio, L., Piantedosi, D., Moscatiello, V., Ciaramella, P., Scalone, A., Gradoni, L. and Mancino, D., 1999. Early suppression of lymphoproliferative responses in dogs with natural infection by Leishmania infantum. Veterinary Immunology Immunopathology, 70, 95–1103

    Article  Google Scholar 

  • Dedet, J.P. and Pratlong, F., 2000. Leishmania, Trypanosoma and monoxenous trypanosomatids as emerging opportunistic agents. Journal of Eukaryotic Microbiology, 47, 37–39

    Article  PubMed  CAS  Google Scholar 

  • Desjeux, P. 1995. Leishmania/HIV co-infections. African Health, 18, 20–22

    CAS  Google Scholar 

  • Fernández-Pérez, F.J., Gomez-Muñoz, M.T., Méndez, S. and Alunda, J.M., 2003. Leishmania-specific lymphoproliferative responses and IgG1/IgG2 immunodetection patterns by Western blot in asymptomatic, symptomatic and treated dogs. Acta Tropica, 86, 83–91

    Article  PubMed  CAS  Google Scholar 

  • Gifawesen, C. and Farrell, J.P., 1989. Comparison of T-cell responses in self-limiting versus progressive visceral Leishmania donovani infections in golden hamsters. Infection and Immunity, 57, 3091–3096

    PubMed  CAS  Google Scholar 

  • Gonzalez, J.L., Rollan, E., Novoa, C. and Castaño, M., 1988. Structural and ultrastructural hepatic changes in experimental canine leishmaniasis. Histology and Histopathology, 3, 323–329

    PubMed  CAS  Google Scholar 

  • Gradoni, L. 1999. Epizootiology of canine leishmaniasis in southern Europe. In: R. Killick-Kendrick (ed.), Canine Leishmaniasis: An update, (Hoechst Roussel Vet, Wiesbaden), 32–39

    Google Scholar 

  • Guarga, J.L., Moreno, J., Lucientes, J., Gracia, M.J., Peribañez, M.A., Alvar, J. and Castillo, J.A., 2000. Canine leishmaniasis transmission: higher infectivity amongst naturally infected dogs to sand flies is associated with lower proportions of T helper cells. Research in Veterinary Science, 69, 249–253

    Article  PubMed  CAS  Google Scholar 

  • Hill, J.O., North, R.J. and Collins, F.M., 1983. Advantages of measuring changes in the number of viable parasites in murine models of experimental cutaneous leishmaniasis. Infection and Immunity, 39, 1087–1094

    PubMed  CAS  Google Scholar 

  • Keithly, J.S., 1976. Infectivity of Leishmania donovani amastigotes and promastigotes for golden hamster. Journal of Protozoology, 23, 244–245

    PubMed  CAS  Google Scholar 

  • Leclercq, V., Lebastard, M., Belkaid, Y., Lousi, J. and Milon, G.,1996. The outcome of the parasite process initiated by Leishmania infantum in laboratory mice. A tissue-dependent pattern controlled by the Lsh and MHC loci. Journal of Immunology, 157, 4537–4545

    CAS  Google Scholar 

  • Killick-Kendrick, R., Killick-Kendrick, M., Pinelli, E., Del Real, G., Molina, R., Vitutia, M., Cañavate, M.C. and Nieto. J., 1994. A laboratory model of canine leishmaniasis: the inoculation of dogs with Leishmania infantum promastigotes from midguts of experimentally infected phlebotomine sandflies. Parasite, 1, 311–318

    PubMed  CAS  Google Scholar 

  • Martínez-Moreno, A., Moreno, T., Martínez-Moreno, F.J., Acosta, I. and Hernández, S., 1995. Humoral and cell-mediated immunity in natural and experimental leishmaniasis. Veterinary Immunology Immunopathology, 48, 209–220

    Article  Google Scholar 

  • Mauricio. I.L., Stothard, J.R. and Miles, M.A., 2000. The strange case of Leishmania chagasi. Parasitology Today, 16, 188–189

    Article  PubMed  CAS  Google Scholar 

  • Melby, P.C., Tryon, V.V., Chandraseka, B. and Freeman, G.L., 1998. Cloning of Syrian hamster (Mesocricetus auratus) cytokine cDNAs and analysis of cytokine mRNA expression in experimental visceral leishmaniasis. Infection and Immunity, 66, 2135–2142

    PubMed  CAS  Google Scholar 

  • Melby, P.C., Chandrasekar, B., Zhao, W. and Coe, J.E., 2001. The hamster as a model of human visceral leishmaniasis: progressive disease and impaired generation of nitric oxide in the face of a prominent Th1-like cytokine response. Journal of Immunology, 166, 1912–1920

    CAS  Google Scholar 

  • Mendez, S., Nell, M. and Alunda, J.M., 1996. Leishmania infantum: infection of macrophages in vitro with promastigotes. International Journal of Parasitology, 26, 619–622

    Article  PubMed  CAS  Google Scholar 

  • Mendez, S., Gurunathan, S., Kamhawi, S., Belkaid, Y., Moga, M.A., Skeiky, Y.A., Campos-Neto. A., Reed, S., Seder, R.A. and Sacks, D., 2001. The potency and durability of DNA- and protein-based vaccines against Leishmania major evaluated using low-dose, intradermal challenge. Journal of Immunology, 166, 5122–5128

    CAS  Google Scholar 

  • Moreno, J., Nieto, J., Chamizo, C., Gonzalez, F., Blanco, F., Barrer, D.C. and Alvar, J., 1999. The immune response and PBMC subsets in canine visceral leishmaniasis before, and after chemotherapy. Veterinary Immunology Immunopathology, 71, 181–195

    Article  CAS  Google Scholar 

  • Mukherjee, P., Ghosh, A.K. and Ghose, A.C. 2003. Infection pattern and immune response in the spleen and liver of BALB/c mice intracardially infected with Leishmania donovani amastigotes. Immunological Letters, 86, 131–138

    Article  CAS  Google Scholar 

  • Pearson, R.D. and de Queiroz-Sousa, A., 1996. Clinical spectrum of leishmaniasis. Clinical Infection Diseases, 22, 1–13

    CAS  Google Scholar 

  • Nickol, A.D. and Bonventre, P.F., 1985. Visceral leishmaniasis in congenic mice of susceptible and resistant phenotypes: immunosuppression by adherent spleen cells. Infection and Immunity, 50, 160–168

    PubMed  CAS  Google Scholar 

  • Noli, C. and Auxilia, S.T., 2005. Treatment of canine Old World visceral leishmaniasis: a systematic review. Veterinary Dermatology, 16, 213–232

    Article  PubMed  Google Scholar 

  • Requena, J.M., Soto, M., Doria, M.D. and Alonso, C., 2000. Immune and clinical parameters associated with Leishmania infantum infection in the golden hamster model. Veterinary Immunology Immunopathology, 76, 269–281

    Article  CAS  Google Scholar 

  • Rica-Capela, M.J., Cortes, S., Leandro, C., Peleteiro, M.C., Santos-Gomes, G. and Campino, L., 2003. Immunological and histopathological studies in a rodent model infected with Leishmania infantum promastigotes or amastigotes. Parasitology Research, 89,163–169

    PubMed  CAS  Google Scholar 

  • Rosypal, A.C., Zajac, A.M., Troy, G.C. and Lindsay, D.S., 2005. Infection in immunocompetent and immune-deficient mice with promastigotes of a North-American isolate of Leishmania infantum. Veterinary Paarsitology, 130, 19–27

    Article  Google Scholar 

  • Rousseau, D., Demartino, S., Anjuere, F., Ferrua, B., Fragak, K., Le Fichoux, Y. and Kubar, J., 2001. Sustained parasite burden in the spleen of Leishmania infantum-infected BALB/c mice is accompanied by expression of MCP-1 transcripts and lack of protection against challenge. European Cytokine Network, 12, 340–347

    PubMed  CAS  Google Scholar 

  • Schnurr, L., Zuckerman, A. and Montilio, B., 1973. Dissemination of leishmanias to the organs of Syrian hamsters following intrasplenic inoculation of promastigotes. Experimental Parasitology, 34, 432–437

    Article  Google Scholar 

  • Stauber, L.A., 1966. The origin and significance of the distribution of parasites in visceral leishmaniasis. Transactions of the New York Academy of Sciences, 28, 635–643.

    PubMed  CAS  Google Scholar 

  • Titus, R.G., Marchand, M., Boon, T. and Louis, J.A., 1985. A limiting dilution assay for quantifying Leishmania major in tissues of infected mice. Parasite Immunology, 7, 545–555

    PubMed  CAS  Google Scholar 

  • Vercammen, F., Fernandez-Perez, F.J., del Amo, C. and Alunda J.M., 2002. Follow-up of Leishmania infantum naturally infected dogs treated with allopurinol: immunofluorescence antibody test, ELISA and Western blot. Acta Tropica, 84, 175–181

    Article  PubMed  CAS  Google Scholar 

  • Wyllie, S. and Fairlamb, A.H., 2006. Refinement of techniques for the propagation of Leishmania donovani in hamsters. Acta Tropica, 97, 364–369

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to F. Bolás-Fernandez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dea-Ayuela, M.A., Rama-Íñiguez, S., Alunda, J.M. et al. Setting New Immunobiological Parameters in the Hamster Model of Visceral Leishmaniasis for In Vivo Testing of Antileishmanial Compounds. Vet Res Commun 31, 703–717 (2007). https://doi.org/10.1007/s11259-007-0040-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11259-007-0040-5

Keywords

Navigation