Author:
- INTRODUCTION
- MATERIALS AND METHODS
- Animals.
- Parasites and parasite antigens.
- Clinical assessment.
- Parasitologic evaluation and infectivity for vectors.
- Parasite-specific antibody determinations.
- Culture of PBMCs for lymphoproliferation and cytokine analysis.
- RNA isolation.
- Determination of cytokine expression by RT-real time-PCR.
- Pathologic evaluation.
- Statistical analyses.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Desjeux P, 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27 :305–318.
-
2
Valladares JE, Riera C, Gonzalez-Ensenyat P, Diez-Cascon A, Ramos G, Solano-Gallego L, Gallego M, Portus M, Arboix M, Alberola J, 2001. Long term improvement in the treatment of canine leishmaniosis using an antimony liposomal formulation. Vet Parasitol 97 :15–21.
-
3
Baneth G, Shaw SE, 2002. Chemotherapy of canine leishmaniosis. Vet Parasitol 106 :315–324.
-
4
Koutinas AF, Saridomichelakis MN, Mylonakis ME, Leontides L, Polizopoulou Z, Billinis C, Argyriadis D, Diakou N, Papadopoulos O, 2001. A randomised, blinded, placebo-controlled clinical trial with allopurinol in canine leishmaniosis. Vet Parasitol 98 :247–261.
-
5
Vexenat JA, Olliaro PL, Fonseca de Castro JA, Cavalcante R, Furtado Campos JH, Tavares JP, Miles MA, 1998. Clinical recovery and limited cure in canine visceral leishmaniasis treated with aminosidine (paromomycin). Am J Trop Med Hyg 58 :448–453.
-
6
Campino L, Santos-Gomes G, Rica Capela MJ, Cortes S, Abranches P, 2000. Infectivity of promastigotes and amastigotes of Leishmania infantum in a canine model for leishmaniosis. Vet Parasitol 92 :269–275.
-
7
Leandro C, Santos-Gomes GM, Campino L, Romao P, Cortes S, Rolao N, Gomes-Pereira S, Rica Capela MJ, Abranches P, 2001. Cell mediated immunity and specific IgG1 and IgG2 antibody response in natural and experimental canine leishmaniosis. Vet Immunol Immunopathol 79 :273–284.
-
8
Martinez-Moreno A, Moreno T, Martinez-Moreno FJ, Acosta I, Hernandez S, 1995. Humoral and cell-mediated immunity in natural and experimental canine leishmaniasis. Vet Immunol Immunopathol 48 :209–220.
-
9
Santos-Gomes GM, Rosa R, Leandro C, Cortes S, Romao P, Silveira H, 2002. Cytokine expression during the outcome of canine experimental infection by Leishmania infantum. Vet Immunol Immunopathol 88 :21–30.
-
10
Killick-Kendrick R, Killick-Kendrick M, Pinelli E, Del Real G, Molina R, Vitutia MM, Canavate MC, 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.
-
11
Paranhos-Silva M, Oliveira GG, Reis EA, de Menezes RM, Fernandes O, Sherlock I, Gomes RB, Pontes-de-Carvalho LC, dos-Santos WL, 2003. A follow-up of Beagle dogs intradermally infected with Leishmania chagasi in the presence or absence of sand fly saliva. Vet Parasitol 114 :97–111.
-
12
Santos-Gomes GM, Campino L, Abranches P, 2000. Canine experimental infection: intradermal inoculation of Leishmania infantum promastigotes. Mem Inst Oswaldo Cruz 95 :193–198.
-
13
Pinelli E, Killick-Kendrick R, Wagenaar J, Bernadina W, del Real G, Ruitenberg J, 1994. Cellular and humoral immune responses in dogs experimentally and naturally infected with Leishmania infantum. Infect Immun 62 :229–235.
-
14
Lemesre JL, Holzmuller P, Cavaleyra M, Goncalves RB, Hottin G, Papierok G, 2005. Protection against experimental visceral leishmaniasis infection in dogs immunized with purified excreted secreted antigens of Leishmania infantum promastigotes. Vaccine 23 :2825–2840.
-
15
Poot J, Spreeuwenberg K, Sanderson SJ, Schijns VE, Mottram JC, Coombs GH, Vermeulen AN, 2006. Vaccination with a preparation based on recombinant cysteine peptidases and canine IL-12 does not protect dogs from infection with Leishmania infantum. Vaccine 24 :2460–2468.
-
16
Ramiro MJ, Zarate JJ, Hanke T, Rodriguez D, Rodriguez JR, Esteban M, Lucientes J, Castillo JA, Larraga V, 2003. Protection in dogs against visceral leishmaniasis caused by Leishmania infantum is achieved by immunization with a heterologous prime-boost regime using DNA and vaccinia recombinant vectors expressing LACK. Vaccine 21 :2474–2484.
-
17
Riera C, Valladares JE, Gallego M, Aisa MJ, Castillejo S, Fisa R, Ribas N, Carrio J, Alberola J, Arboix M, 1999. Serological and parasitological follow-up in dogs experimentally infected with Leishmania infantum and treated with meglumine antimoniate. Vet Parasitol 84 :33–47.
-
18
Travi BL, Tabares CJ, Cadena H, Ferro C, Osorio Y, 2001. Canine visceral leishmaniasis in Colombia: relationship between clinical and parasitologic status and infectivity for sand flies. Am J Trop Med Hyg 64 :119–124.
-
19
Saldarriaga OA, Travi BL, Melby PC, 2006. Quantification of canine cytokines using real time reverse transcriptase polymerase chain reaction. Biomedica (Bogota) 26 (Suppl 1): 254–263.
-
20
Saldarriaga OA, Travi BL, Park W, Perez LE, Melby PC, 2006. Immunogenicity of a multicomponent DNA vaccine against visceral leishmaniasis in dogs. Vaccine 24 :1928–1940.
-
21
Leutenegger CM, Mislin CN, Sigrist B, Ehrengruber MU, Hofmann-Lehmann R, Lutz H, 1999. Quantitative real-time PCR for the measurement of feline cytokine mRNA. Vet Immunol Immunopathol 71 :291–305.
-
22
Gradoni L, 2001. An update on antileishmanial vaccine candidates and prospects for a canine Leishmania vaccine. Vet Parasitol 100 :87–103.
-
23
Martinez-Moreno A, Martinez-Cruz MS, Blanco A, Hernandez-Rodriguez S, 1993. Immunological and histological study of T- and B-lymphocyte activity in canine visceral leishmaniosis. Vet Parasitol 51 :49–59.
-
24
Solano-Gallego L, Llull J, Ramos G, Riera C, Arboix M, Alberola J, Ferrer L, 2000. The Ibizian hound presents a predominantly cellular immune response against natural Leishmania infection. Vet Parasitol 90 :37–45.
-
25
Solano-Gallego L, Morell P, Arboix M, Alberola J, Ferrer L, 2001. Prevalence of Leishmania infantum infection in dogs living in an area of canine leishmaniasis endemicity using PCR on several tissues and serology. J Clin Microbiol 39 :560–563.
-
26
Cabral M, O’Grady J, Alexander J, 1992. Demonstration of Leishmania specific cell mediated and humoral immunity in asymptomatic dogs. Parasite Immunol 14 :531–539.
-
27
Dye C, Killick-Kendrick R, Vitutia MM, Walton R, Killick-Kendrick M, Harith AE, Guy MW, Canavate MC, Hasibeder G, 1992. Epidemiology of canine leishmaniasis: prevalence, incidence and basic reproduction number calculated from a cross-sectional serological survey on the island of Gozo, Malta. Parasitology 105 :35–41.
-
28
Paranhos M, dos Santos WC, Sherlock I, Oliveira GG, de Carvalho LC, 1993. Development of eosinophilia in dogs intradermically inoculated with sand fly saliva and Leishmania (Leishmania) chagasi stationary-phase promastigotes. Mem Inst Oswaldo Cruz 88 :249–251.
-
29
Oliveira GG, Santoro F, Sadigursky M, 1993. The subclinical form of experimental visceral leishmaniasis in dogs. Mem Inst Oswaldo Cruz 88 :243–248.
-
30
Sanchez MA, Diaz NL, Zerpa O, Negron E, Convit J, Tapia FJ, 2004. Organ-specific immunity in canine visceral leishmaniasis: analysis of symptomatic and asymptomatic dogs naturally infected with Leishmania chagasi. Am J Trop Med Hyg 70 :618–624.
-
31
Tafuri WL, Barbosa AJ, Michalick MS, Genaro O, Franca-Silva JC, Mayrink W, Nascimento E, 1996. 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. Rev Inst Med Trop Sao Paulo 38 :81–89.
-
32
Ferreira JH, Gentil LG, Dias SS, Fedeli CE, Katz S, Barbieri CL, 2008. Immunization with the cysteine proteinase Ldccys1 gene from Leishmania (Leishmania) chagasi and the recombinant Ldccys1 protein elicits protective immune responses in a murine model of visceral leishmaniasis. Vaccine 26 :677–685.
-
33
Silvestre R, Cordeiro-Da-Silva A, Santarem N, Vergnes B, Sereno D, Ouaissi A, 2007. SIR2-deficient Leishmania infantum induces a defined IFN-gamma/IL-10 pattern that correlates with protection. J Immunol 179 :3161–3170.
-
34
Belkaid Y, Mendez S, Lira R, Kadambi N, Milon G, Sacks D, 2000. A natural model of Leishmania major infection reveals a prolonged “silent” phase of parasite amplification in the skin before the onset of lesion formation and immunity. J Immunol 165 :969–977.
-
35
Alves CF, de Amorim IF, Moura EP, Ribeiro RR, Alves CF, Michalick MS, Kalapothakis E, Bruna-Romero O, Tafuri WL, Teixeira MM, Melo MN, 2009. Expression of IFN-gamma, TNF-alpha, IL-10 and TGF-beta in lymph nodes associates with parasite load and clinical form of disease in dogs naturally infected with Leishmania (Leishmania) chagasi. Vet Immunol Immunopathol 128 :349–358.
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36
Kenney RT, Sacks DL, Gam AA, Murray HW, Sundar S, 1998. Splenic cytokine responses in Indian kala-azar before and after treatment. J Infect Dis 177 :815–818.
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37
Karp CL, el-Safi SH, Wynn TA, Satti MM, Kordofani AM, Hashim FA, Hag-Ali M, Neva FA, Nutman TB, Sacks DL, 1993. In vivo cytokine profiles in patients with kala-azar. Marked elevation of both interleukin-10 and interferon-gamma. J Clin Invest 91 :1644–1648.
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38
Rhalem A, Sahibi H, Guessous-Idrissi N, Lasri S, Natami A, Riyad M, Berrag B, 1999. Immune response against Leishmania antigens in dogs naturally and experimentally infected with Leishmania infantum. Vet Parasitol 81 :173–184.
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39
Lachaud L, Chabbert E, Dubessay P, Dereure J, Lamothe J, Dedet JP, Bastien P, 2002. Value of two PCR methods for the diagnosis of canine visceral leishmaniasis and the detection of asymptomatic carriers. Parasitology 125 :197–207.
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40
Leontides LS, Saridomichelakis MN, Billinis C, Kontos V, Koutinas AF, Galatos AD, Mylonakis ME, 2002. A cross-sectional study of Leishmania spp. infection in clinically healthy dogs with polymerase chain reaction and serology in Greece. Vet Parasitol 109 :19–27.
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41
Quinnell RJ, Courtenay O, Davidson S, Garcez L, Lambson B, Ramos P, Shaw JJ, Shaw MA, Dye C, 2001. Detection of Leishmania infantum by PCR, serology and cellular immune response in a cohort study of Brazilian dogs. Parasitology 122 :253–261.
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42
Solano-Gallego L, Riera C, Roura X, Iniesta L, Gallego M, Valladares JE, Fisa R, Castillejo S, Alberola J, Ferrer L, Arboix M, Portus M, 2001. Leishmania infantum-specific IgG, IgG1 and IgG2 antibody responses in healthy and ill dogs from endemic areas. Evolution in the course of infection and after treatment. Vet Parasitol 96 :265–276.
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43
Bourdoiseau G, Bonnefont C, Hoareau E, Boehringer C, Stolle T, Chabanne L, 1997. Specific IgG1 and IgG2 antibody and lymphocyte subset levels in naturally Leishmania infantum-infected treated and untreated dogs. Vet Immunol Immunopathol 59 :21–30.
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44
Deplazes P, Smith NC, Arnold P, Lutz H, Eckert J, 1995. Specific IgG1 and IgG2 antibody responses of dogs to Leishmania infantum and other parasites. Parasite Immunol 17 :451–458.
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45
Cordeiro-da-Silva A, Cardoso L, Araujo N, Castro H, Tomas A, Rodrigues M, Cabral M, Vergnes B, Sereno D, Ouaissi A, 2003. Identification of antibodies to Leishmania silent information regulatory 2 (SIR2) protein homologue during canine natural infections: pathological implications. Immunol Lett 86 :155–162.
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46
Iniesta L, Gallego M, Portus M, 2005. Immunoglobulin G and E responses in various stages of canine leishmaniosis. Vet Immunol Immunopathol 103 :77–81.
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47
Courtenay O, Quinnell RJ, Garcez LM, Shaw JJ, Dye C, 2002. Infectiousness in a cohort of Brazilian dogs: why culling fails to control visceral leishmaniasis in areas of high transmission. J Infect Dis 186 :1314–1320.
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48
Sherlock IA, 1996. Ecological interactions of visceral leishmaniasis in the state of Bahia, Brazil. Mem Inst Oswaldo Cruz 91 :671–683.
-
49
Michalsky EM, Rocha MF, da Rocha Lima AC, Franca-Silva JC, Pires MQ, Oliveira FS, Pacheco RS, dos Santos SL, Barata RA, Romanha AJ, Fortes-Dias CL, Dias ES, 2007. Infectivity of seropositive dogs, showing different clinical forms of leishmaniasis, to Lutzomyia longipalpis phlebotomine sand flies. Vet Parasitol 147 :67–76.
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50
Guarga JL, Moreno J, Lucientes J, Gracia MJ, Peribanez MA, Alvar J, Castillo JA, 2000. Canine leishmaniasis transmission: higher infectivity amongst naturally infected dogs to sand flies is associated with lower proportions of T helper cells. Res Vet Sci 69 :249–253.
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Clinical, Parasitologic, and Immunologic Evolution in Dogs Experimentally Infected with Sand Fly-Derived Leishmania chagasi Promastigotes
Bruno L. Travi
Bruno L. Travi
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Elvia Yaneth Osorio
Elvia Yaneth Osorio
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Omar A. Saldarriaga
Omar A. Saldarriaga
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Horacio Cadena
Horacio Cadena
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Carlos Javier Tabares
Carlos Javier Tabares
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Alex Peniche
Alex Peniche
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Shuko Lee
Shuko Lee
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Peter C. Melby
Peter C. Melby
Research Service, Department of Veterans Affairs Medical Center, South Texas Veterans Health Care System, San Antonio, Texas; Department of Medicine, The University of Texas Health Science Center, San Antonio, Texas; Centro Internacional de Entrenamiento e Investigaciones Medicas-CIDEIM, AA5390, Cali, Colombia
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Experimental infection of dogs with Leishmania infantum has yielded heterogeneous clinical, parasitologic, and immunologic results. We studied dogs infected with 105 or 104 sand fly-derived promastigotes delivered by the intradermal (ID) or intravenous (IV) routes. Total mortality over 1 year post-infection reached 23.8%. The mortality and proportion of sustained polysymptomatic dogs was highest in the IV-105 group. The early appearance of polysymptoms was associated with an increased risk of progression to death. Dissemination of the parasite to lymph nodes was faster, and the subsequent infectivity to sand flies higher, in the IV compared with ID-infected dogs. Parasite-specific IgG1 or IgG2 production was similar among the groups, but higher interferon-γ (IFN-γ) and interleukin-10 (IL-10) expression was associated with polysymptomatic dogs. On the basis of the data obtained from this study, a sample size analysis using different endpoints for future vaccine trials is described.
- INTRODUCTION
- MATERIALS AND METHODS
- Animals.
- Parasites and parasite antigens.
- Clinical assessment.
- Parasitologic evaluation and infectivity for vectors.
- Parasite-specific antibody determinations.
- Culture of PBMCs for lymphoproliferation and cytokine analysis.
- RNA isolation.
- Determination of cytokine expression by RT-real time-PCR.
- Pathologic evaluation.
- Statistical analyses.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Desjeux P, 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27 :305–318.
-
2
Valladares JE, Riera C, Gonzalez-Ensenyat P, Diez-Cascon A, Ramos G, Solano-Gallego L, Gallego M, Portus M, Arboix M, Alberola J, 2001. Long term improvement in the treatment of canine leishmaniosis using an antimony liposomal formulation. Vet Parasitol 97 :15–21.
-
3
Baneth G, Shaw SE, 2002. Chemotherapy of canine leishmaniosis. Vet Parasitol 106 :315–324.
-
4
Koutinas AF, Saridomichelakis MN, Mylonakis ME, Leontides L, Polizopoulou Z, Billinis C, Argyriadis D, Diakou N, Papadopoulos O, 2001. A randomised, blinded, placebo-controlled clinical trial with allopurinol in canine leishmaniosis. Vet Parasitol 98 :247–261.
-
5
Vexenat JA, Olliaro PL, Fonseca de Castro JA, Cavalcante R, Furtado Campos JH, Tavares JP, Miles MA, 1998. Clinical recovery and limited cure in canine visceral leishmaniasis treated with aminosidine (paromomycin). Am J Trop Med Hyg 58 :448–453.
-
6
Campino L, Santos-Gomes G, Rica Capela MJ, Cortes S, Abranches P, 2000. Infectivity of promastigotes and amastigotes of Leishmania infantum in a canine model for leishmaniosis. Vet Parasitol 92 :269–275.
-
7
Leandro C, Santos-Gomes GM, Campino L, Romao P, Cortes S, Rolao N, Gomes-Pereira S, Rica Capela MJ, Abranches P, 2001. Cell mediated immunity and specific IgG1 and IgG2 antibody response in natural and experimental canine leishmaniosis. Vet Immunol Immunopathol 79 :273–284.
-
8
Martinez-Moreno A, Moreno T, Martinez-Moreno FJ, Acosta I, Hernandez S, 1995. Humoral and cell-mediated immunity in natural and experimental canine leishmaniasis. Vet Immunol Immunopathol 48 :209–220.
-
9
Santos-Gomes GM, Rosa R, Leandro C, Cortes S, Romao P, Silveira H, 2002. Cytokine expression during the outcome of canine experimental infection by Leishmania infantum. Vet Immunol Immunopathol 88 :21–30.
-
10
Killick-Kendrick R, Killick-Kendrick M, Pinelli E, Del Real G, Molina R, Vitutia MM, Canavate MC, 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.
-
11
Paranhos-Silva M, Oliveira GG, Reis EA, de Menezes RM, Fernandes O, Sherlock I, Gomes RB, Pontes-de-Carvalho LC, dos-Santos WL, 2003. A follow-up of Beagle dogs intradermally infected with Leishmania chagasi in the presence or absence of sand fly saliva. Vet Parasitol 114 :97–111.
-
12
Santos-Gomes GM, Campino L, Abranches P, 2000. Canine experimental infection: intradermal inoculation of Leishmania infantum promastigotes. Mem Inst Oswaldo Cruz 95 :193–198.
-
13
Pinelli E, Killick-Kendrick R, Wagenaar J, Bernadina W, del Real G, Ruitenberg J, 1994. Cellular and humoral immune responses in dogs experimentally and naturally infected with Leishmania infantum. Infect Immun 62 :229–235.
-
14
Lemesre JL, Holzmuller P, Cavaleyra M, Goncalves RB, Hottin G, Papierok G, 2005. Protection against experimental visceral leishmaniasis infection in dogs immunized with purified excreted secreted antigens of Leishmania infantum promastigotes. Vaccine 23 :2825–2840.
-
15
Poot J, Spreeuwenberg K, Sanderson SJ, Schijns VE, Mottram JC, Coombs GH, Vermeulen AN, 2006. Vaccination with a preparation based on recombinant cysteine peptidases and canine IL-12 does not protect dogs from infection with Leishmania infantum. Vaccine 24 :2460–2468.
-
16
Ramiro MJ, Zarate JJ, Hanke T, Rodriguez D, Rodriguez JR, Esteban M, Lucientes J, Castillo JA, Larraga V, 2003. Protection in dogs against visceral leishmaniasis caused by Leishmania infantum is achieved by immunization with a heterologous prime-boost regime using DNA and vaccinia recombinant vectors expressing LACK. Vaccine 21 :2474–2484.
-
17
Riera C, Valladares JE, Gallego M, Aisa MJ, Castillejo S, Fisa R, Ribas N, Carrio J, Alberola J, Arboix M, 1999. Serological and parasitological follow-up in dogs experimentally infected with Leishmania infantum and treated with meglumine antimoniate. Vet Parasitol 84 :33–47.
-
18
Travi BL, Tabares CJ, Cadena H, Ferro C, Osorio Y, 2001. Canine visceral leishmaniasis in Colombia: relationship between clinical and parasitologic status and infectivity for sand flies. Am J Trop Med Hyg 64 :119–124.
-
19
Saldarriaga OA, Travi BL, Melby PC, 2006. Quantification of canine cytokines using real time reverse transcriptase polymerase chain reaction. Biomedica (Bogota) 26 (Suppl 1): 254–263.
-
20
Saldarriaga OA, Travi BL, Park W, Perez LE, Melby PC, 2006. Immunogenicity of a multicomponent DNA vaccine against visceral leishmaniasis in dogs. Vaccine 24 :1928–1940.
-
21
Leutenegger CM, Mislin CN, Sigrist B, Ehrengruber MU, Hofmann-Lehmann R, Lutz H, 1999. Quantitative real-time PCR for the measurement of feline cytokine mRNA. Vet Immunol Immunopathol 71 :291–305.
-
22
Gradoni L, 2001. An update on antileishmanial vaccine candidates and prospects for a canine Leishmania vaccine. Vet Parasitol 100 :87–103.
-
23
Martinez-Moreno A, Martinez-Cruz MS, Blanco A, Hernandez-Rodriguez S, 1993. Immunological and histological study of T- and B-lymphocyte activity in canine visceral leishmaniosis. Vet Parasitol 51 :49–59.
-
24
Solano-Gallego L, Llull J, Ramos G, Riera C, Arboix M, Alberola J, Ferrer L, 2000. The Ibizian hound presents a predominantly cellular immune response against natural Leishmania infection. Vet Parasitol 90 :37–45.
-
25
Solano-Gallego L, Morell P, Arboix M, Alberola J, Ferrer L, 2001. Prevalence of Leishmania infantum infection in dogs living in an area of canine leishmaniasis endemicity using PCR on several tissues and serology. J Clin Microbiol 39 :560–563.
-
26
Cabral M, O’Grady J, Alexander J, 1992. Demonstration of Leishmania specific cell mediated and humoral immunity in asymptomatic dogs. Parasite Immunol 14 :531–539.
-
27
Dye C, Killick-Kendrick R, Vitutia MM, Walton R, Killick-Kendrick M, Harith AE, Guy MW, Canavate MC, Hasibeder G, 1992. Epidemiology of canine leishmaniasis: prevalence, incidence and basic reproduction number calculated from a cross-sectional serological survey on the island of Gozo, Malta. Parasitology 105 :35–41.
-
28
Paranhos M, dos Santos WC, Sherlock I, Oliveira GG, de Carvalho LC, 1993. Development of eosinophilia in dogs intradermically inoculated with sand fly saliva and Leishmania (Leishmania) chagasi stationary-phase promastigotes. Mem Inst Oswaldo Cruz 88 :249–251.
-
29
Oliveira GG, Santoro F, Sadigursky M, 1993. The subclinical form of experimental visceral leishmaniasis in dogs. Mem Inst Oswaldo Cruz 88 :243–248.
-
30
Sanchez MA, Diaz NL, Zerpa O, Negron E, Convit J, Tapia FJ, 2004. Organ-specific immunity in canine visceral leishmaniasis: analysis of symptomatic and asymptomatic dogs naturally infected with Leishmania chagasi. Am J Trop Med Hyg 70 :618–624.
-
31
Tafuri WL, Barbosa AJ, Michalick MS, Genaro O, Franca-Silva JC, Mayrink W, Nascimento E, 1996. 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. Rev Inst Med Trop Sao Paulo 38 :81–89.
-
32
Ferreira JH, Gentil LG, Dias SS, Fedeli CE, Katz S, Barbieri CL, 2008. Immunization with the cysteine proteinase Ldccys1 gene from Leishmania (Leishmania) chagasi and the recombinant Ldccys1 protein elicits protective immune responses in a murine model of visceral leishmaniasis. Vaccine 26 :677–685.
-
33
Silvestre R, Cordeiro-Da-Silva A, Santarem N, Vergnes B, Sereno D, Ouaissi A, 2007. SIR2-deficient Leishmania infantum induces a defined IFN-gamma/IL-10 pattern that correlates with protection. J Immunol 179 :3161–3170.
-
34
Belkaid Y, Mendez S, Lira R, Kadambi N, Milon G, Sacks D, 2000. A natural model of Leishmania major infection reveals a prolonged “silent” phase of parasite amplification in the skin before the onset of lesion formation and immunity. J Immunol 165 :969–977.
-
35
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