RESUMO
In South America, visceral leishmaniasis is frequently caused by Leishmania infantum and, at an unknown frequency, by Leishmania amazonensis. Therefore, mixed infections with these organisms are possible. Mixed infections might affect the clinical course, immune response, diagnosis, treatment and epidemiology of the disease. Here we describe the clinical course of mixed infections with L. amazonensis and L. infantum in a hamster model. We show that mixed infections are associated with more severe clinical disease than infection with L. amazonensis or L. infantum alone. In spleens with mixed infections, L. infantum outcompeted L. amazonensis in the tissue, but not in culture from tissue. We found increased levels of IgG in animals infected with L. infantum. Although more than 30 bands were revealed in a Western blot, the highest immunogenicity was observed with proteins having molecular masses of 95 and 90 kDa, whereas proteins with molecular masses of lower than 50 kDa were reactive frequently with serum from hamsters infected with L. amazonensis, and proteins with molecular masses of 80 and 70 kDa were reactive only with serum from hamsters infected with L. infantum. This finding has important implications regarding the biology of Leishmania and humoral immune responses to infections with these organisms.
Assuntos
Imunidade Humoral , Leishmania infantum/imunologia , Leishmania/imunologia , Leishmaniose Visceral/imunologia , Leishmaniose/imunologia , Animais , Western Blotting , Coinfecção , Cricetinae , Modelos Animais de Doenças , Leishmaniose/parasitologia , Mesocricetus , América do Sul , Baço/parasitologiaRESUMO
The objective of the present study was to investigate the histopathological alterations and detection of parasites that occur in the eyes of dogs naturally infected with Leishmania chagasi. The eyes of 25 dogs with a positive parasitological and serological exam for L. chagasi were submitted to routine ophthalmic examination. Fragments were obtained from the eyeball and were analyzed by immunohistochemistry and histopathology. Among the structures studied, the conjunctiva of the third eyelid was the tissue most frequently stained for L. chagasi. Histopathological analysis revealed a predominantly mononuclear inflammatory infiltrate whose intensity ranged from discrete to intense in the various ocular structures. Vascular congestion and perivasculitis were also observed. The histopathological alterations that occur in the eyeball of dogs with visceral leishmaniasis might be related to the presence of antibodies.
O objetivo deste trabalho foi estudar as alterações histopatológicas e a detecção do parasito nos olhos de cães naturalmente infectados por Leishmania chagasi. Utilizaram-se os olhos de 25 cães com exame parasitológico e sorológico positivo para Leishmania chagasi. Os animais foram submetidos à semiotécnica oftálmica rotineira. Fragmentos do globo ocular foram coletados e destinados à imunoistoquímica e histopatologia. Entre as estruturas avaliadas, a conjuntiva da terceira pálpebra foi a que mais demonstrou imunomarcação da Leishmania chagasi. Na avaliação histopatológica, foi observado predominantemente infiltrado inflamatório mononuclear, variando de discreto a intenso nas diversas estruturas oculares. Congestão vascular e perivasculite também foram observadas. As alterações histopatológicas do bulbo do olho em cães com leishmaniose visceral podem estar relacionadas à presença do anticorpo.
RESUMO
The objective of the present study was to investigate the histopathological alterations and detection of parasites that occur in the eyes of dogs naturally infected with Leishmania chagasi. The eyes of 25 dogs with a positive parasitological and serological exam for L. chagasi were submitted to routine ophthalmic examination. Fragments were obtained from the eyeball and were analyzed by immunohistochemistry and histopathology. Among the structures studied, the conjunctiva of the third eyelid was the tissue most frequently stained for L. chagasi. Histopathological analysis revealed a predominantly mononuclear inflammatory infiltrate whose intensity ranged from discrete to intense in the various ocular structures. Vascular congestion and perivasculitis were also observed. The histopathological alterations that occur in the eyeball of dogs with visceral leishmaniasis might be related to the presence of antibodies.
O objetivo deste trabalho foi estudar as alterações histopatológicas e a detecção do parasito nos olhos de cães naturalmente infectados por Leishmania chagasi. Utilizaram-se os olhos de 25 cães com exame parasitológico e sorológico positivo para Leishmania chagasi. Os animais foram submetidos à semiotécnica oftálmica rotineira. Fragmentos do globo ocular foram coletados e destinados à imunoistoquímica e histopatologia. Entre as estruturas avaliadas, a conjuntiva da terceira pálpebra foi a que mais demonstrou imunomarcação da Leishmania chagasi. Na avaliação histopatológica, foi observado predominantemente infiltrado inflamatório mononuclear, variando de discreto a intenso nas diversas estruturas oculares. Congestão vascular e perivasculite também foram observadas. As alterações histopatológicas do bulbo do olho em cães com leishmaniose visceral podem estar relacionadas à presença do anticorpo.
RESUMO
The objective of the present study was to investigate the histopathological alterations and detection of parasites that occur in the eyes of dogs naturally infected with Leishmania chagasi. The eyes of 25 dogs with a positive parasitological and serological exam for L. chagasi were submitted to routine ophthalmic examination. Fragments were obtained from the eyeball and were analyzed by immunohistochemistry and histopathology. Among the structures studied, the conjunctiva of the third eyelid was the tissue most frequently stained for L. chagasi. Histopathological analysis revealed a predominantly mononuclear inflammatory infiltrate whose intensity ranged from discrete to intense in the various ocular structures. Vascular congestion and perivasculitis were also observed. The histopathological alterations that occur in the eyeball of dogs with visceral leishmaniasis might be related to the presence of antibodies.
O objetivo deste trabalho foi estudar as alterações histopatológicas e a detecção do parasito nos olhos de cães naturalmente infectados por Leishmania chagasi. Utilizaram-se os olhos de 25 cães com exame parasitológico e sorológico positivo para Leishmania chagasi. Os animais foram submetidos à semiotécnica oftálmica rotineira. Fragmentos do globo ocular foram coletados e destinados à imunoistoquímica e histopatologia. Entre as estruturas avaliadas, a conjuntiva da terceira pálpebra foi a que mais demonstrou imunomarcação da Leishmania chagasi. Na avaliação histopatológica, foi observado predominantemente infiltrado inflamatório mononuclear, variando de discreto a intenso nas diversas estruturas oculares. Congestão vascular e perivasculite também foram observadas. As alterações histopatológicas do bulbo do olho em cães com leishmaniose visceral podem estar relacionadas à presença do anticorpo.
RESUMO
In this study, we investigated in dogs the immunogenicity and protective immunity against Leishmania (Leishmania) chagasi infection induced by vaccination with a formulation containing the recombinant A2 protein, an amastigote specific antigen, and saponin. Vaccinated animals produced significantly increased levels of total IgG and IgG2, but not IgG1 anti-A2 antibodies, and remained negative in conventional leishmaniasis serodiagnostic methods. Significantly increased IFN-gamma and low IL-10 levels were detected in vaccinated animals before and after challenge, as compared to control animals. Importantly, while the symptoms onset appeared as early as three months after infection in most control dogs, 14 months after challenge, 5 out of 7 vaccinated dogs remained asymptomatic. Therefore, immunization with rA2 antigen was immunogenic and induced partial protection in dogs, and allowed the serological differentiation between vaccinated and infected animals, an important requirement for a canine visceral leishmaniasis (CVL) vaccine.
Assuntos
Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Doenças do Cão/imunologia , Doenças do Cão/prevenção & controle , Leishmania infantum/imunologia , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/veterinária , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Vacinas Protozoárias/imunologia , Vacinas Protozoárias/uso terapêutico , Animais , Células da Medula Óssea/imunologia , Separação Celular , Citocinas/biossíntese , DNA de Protozoário/análise , DNA de Protozoário/isolamento & purificação , Cães , Ensaio de Imunoadsorção Enzimática , Imunização , Imunoglobulina G/análise , Imunoglobulina G/biossíntese , Interferon gama/análise , Interferon gama/biossíntese , Interleucina-10/análise , Interleucina-10/biossíntese , Leishmaniose Visceral/prevenção & controle , Contagem de Leucócitos , Monócitos/imunologia , Monócitos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/uso terapêuticoRESUMO
Canine visceral leishmaniasis (CVL) is a zoonosis and a chronic systemic disease of the dog caused by a protozoan of the genus Leishmania. In the New World, the disease is caused by the species Leishmania (Leishmania) chagasi. There are only a few studies on the histopathology of lymph nodes in canine leishmaniasis. In the present paper, we report a histopathological description of lymph nodes considering animals with a defined clinical status and the parasite burden of lymph node tissues. Forty-eight mongrel dogs naturally infected with L. chagasi, were obtained from two endemic areas of Brazil. Cervical, axillary and popliteal lymph nodes were analyzed. The parasite burden, expressed as "Leishman-Donovan units", was variable among the defined types of clinical condition. Asymptomatic dogs can show higher parasitism than oligosymptomatic or symptomatic animals. Grossly, a generalized lymphadenopathy was found, but it was mainly observed in cervical and popliteal nodes. Histologically, the increased number and size of lymphoid follicles, and the marked hypertrophy and hyperplasia of medullary macrophages (cords and sinus) explained the lymphadenopathy. In addition, the clinical status or the tissue parasitism load might not be directly related to the intensity of the lesions.