Leishmania (Viannia) braziliensis: Immunoblotting analysis for the detection of IgG subclasses in the diagnosis of symptomatic and asymptomatic dogs

Zanini, Marcos Santos; Viana, Kelvinson Fernandes; Reis, Alexandre Barbosa; Campos, Diefrey Ribeiro; Mussi, Jamili Maria Suhet; Zanini, Surama Freitas; Lemos, Elenice Moreira

doi:10.1016/j.vetpar.2010.06.002

Cited by 3 publications

(8 citation statements)

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“…For immunoblotting, a peroxidase-conjugated secondary anti-total IgG was used to differentiate the antibody panel of vaccinated and positive control dogs in the presence of the antigens of L. (V.) braziliensis. Immunoblotting analysis was performed according to the method described by Zanini et al (2010) with minor modifications. Briefly, 10 µg of the soluble antigens of L. braziliensis (one in each lane) and SDS-PAGE Molecular Weight Standards (Biorad Laboratories;…”

Section: Immunoblotting Analysismentioning

confidence: 99%

“…Previous findings indicate that indirect ELISA and immunoblotting have high sensitivity for detecting L. (V.) braziliensis infections and are considered a valuable tool for screening and diagnosing this disease, given that the association of two or more indirect methods helps increase the sensitivity of diagnosis (RIBEIRO et al, 2007;SZARGIKI et al, 2009;ZANINI et al, 2010). Conventionally, diagnostic serological tests such as enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA) have a sensitivity of 91.8% and 90.8% and a specificity of 83.4% and 53.4%, respectively (ARRUDA et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen

Pinheiro

Giori

Almeida

et al. 2018

SCA

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American cutaneous leishmaniasis (ACL) is a zoonosis caused by Leishmania, a protozoan. Common antigens occur in the strains found in America, which allow antigenic cross-reactivity. Therefore, multivalent vaccines can be used for this pathogen. In this study, we investigated the efficacy of two different commercial vaccines for visceral leishmaniasis to induce an immune response to the soluble L. (Viannia) braziliensis antigens. In 2014, 70 seronegative dogs from the municipality of Iúna (Espírito Santo State, Brazil) were vaccinated and serologically evaluated by ELISA and immunoblotting by using the soluble antigen of L. braziliensis. Of the 121 dogs initially selected, only 70 received vaccination because 51 dogs tested positive by ELISA, yielding a positive frequency of 42.14% in the asymptomatic group. These 70 dogs were divided into two equal groups and administered three doses of each vaccine, according to the manufacturers’ instructions. We found that the sera of dogs immunized with three doses of both vaccines A and B had antibodies against the soluble antigens of L. (V.) braziliensis, as determined by ELISA and immunoblotting 120 days post vaccination. Antibodies produced in response to vaccines A and B were found in 22/35 and 18/35 serum samples, respectively, at T1 (120 days), while 7/35 and 4/35 serum samples tested positive at T2 (240 days). Furthermore, immunoblotting allowed us to differentiate between vaccinated and asymptomatic dogs.

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Section: Immunoblotting Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen

Pinheiro

Giori

Almeida

et al. 2018

SCA

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“…Apesar da importância do efeito protetor da resposta imune mediada por Th1 ainda não ser bem esclarecido, sabe-se que o aparecimento dos sinais clínicos característicos das formas tegumentar e visceral da LC está associado com o desenvolvimento da imunidade humoral mediada por anticorpos (Reis et al, 2006;Zanini et al, 2010 (Freitas et al, 2012b). Esse achado difere do observado por Reis et al (2006).…”

Section: Estudo Das Imunoglobulinas Nas Formas Tegumentar E Visceral unclassified

“…Esse achado difere do observado por Reis et al (2006). Portanto, fica demonstrada a necessidade de estudos mais aprofundados, com atenção toda especial para os animais assintomáticos, para que se chegue a um consenso da participação dessas classes de imunoglobulinas (Zanini et al, 2010).…”

Section: Estudo Das Imunoglobulinas Nas Formas Tegumentar E Visceral unclassified

“…Trabalhos realizados por diversos pesquisadores mostram que a presença de anticorpos IgG1 e IgG2 está relacionada com a progressão da doença (Solano-Gallego et al, 2001;Quinnell et al, 2003a;Reis et al, 2006;Teixeira-Neto et al, 2010;Zanini et al, 2010). Entretanto, a participação diferenciada de cada imunoglobulina, em relação ao curso clínico, durante o processo infeccioso, ainda não está claramente elucidada.…”

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Leishmaniosis: an approach about the immunoglobulins and cytokines involved in infection and vaccination

Carneiro

Nunes-Pinheiro

2013

AVB

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RESUMO:Objetivou-se revisar os principais trabalhos científicos, discutindo a participação das imunoglobulinas e as citocinas detectadas nas leishmanioses caninas (LCs) e na vacinação de cães sadios. Nas leishmanioses a resposta imune protetora é regulada por células Th1 e Th17, sendo INF-γ, IL-2, TNF-α, IL-17, IL-21, IL-22, IL-6 e TGF-β, as citocinas características, com consequente aumento na produção de IgG2. Por outro lado, a resposta imune mediada por Th2 e Treg, apresenta padrão de citocinas associadas a IL-4, IL-5, IL-13 e IL-10, as quais inibem a resposta de Th1, aumentando a produção de IgG1, IgA e IgE e o desenvolvimento sintomático de uma severa doença. O perfil de anticorpos associados à produção de IgG1 e IgG2 ainda é conflitante entre os pesquisadores. Sendo assim, a interpretação das imunoglobulinas não poderá funcionar como marcador, quer seja no diagnóstico quer seja no tratamento de animais doentes. A vacinação surgiu como nova estratégia de controle das LCs. As formulações vacinais de primeira e terceira gerações são capazes de induzir uma resposta mediada tanto por INF-γ quanto IL-4, enquanto as de segunda geração são caracterizadas pelo aumento da produção de INF-γ e NO. Nestas situações, há aumento na capacidade de resposta imune celular e na produção de anticorpos. Ressalta-se que antígenos vacinais disponíveis não são capazes de desencadear uma resposta satisfatória mediada por linfócitos T CD4 + de memória. Pode-se concluir que as imunoglobulinas e as citocinas podem ser utilizadas como marcadores das LCs e do efeito vacinal, contudo seu uso na clínica deve ser resguardado.Palavras-chave: leishmanioses canina; biomarcadores; vacinas. ABSTRACT:It was aimed to review the main scientific work, discussing the participation of immunoglobulins and cytokines detected in canine leishmaniosis (CLs) and in the vaccination of healthy dogs. In the leishmaniosis the protective immune response is regulated by Th1 and Th17 cells, being INF-γ, IL-2, TNF-α, IL-17, IL-21, IL-22, IL-6 and TGF-β, the cytokines features, with a consequent increase in the production of IgG2. Moreover, the immune response mediated by Th2 and Treg presents pattern associated of cytokines IL-4, IL-5, IL-13 and IL-10, which inhibit Th1 response, increasing the production of IgG1, IgA and IgE and development of severe symptomatic disease. The antibody profile associated with the production of IgG1 and IgG2 is still conflict among researchers. As such, the interpretation of immunoglobulins can not function as marker, either in the diagnosis whether the treatment of sick animals. Vaccination has emerged as a new control strategy of CLs. The vaccine formulations of the first and third generations are capable of inducing a response mediated both for IFN-γ as IL-4, while the second generation are characterized by the increased production of IFN-γ and NO. In these situations, there is an increase in the capacity of cellular immune response and the production of antibodies. It is highlighted that available vaccine antigens are not able t...

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The spiny rat Proechimys guyannensis (Rodentia: Echimydae) fails to respond to intradermal inoculation with Leishmania (Viannia) braziliensis

et al. 2015

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Leishmaniasis a disease of worldwide occurrence is caused by protozoa of the Leishmania genus. In Brazil, Leishmania (Viannia) braziliensis is the main parasite responsible for the American cutaneous leishmaniasis. Main hosts of this protozoa are small wild mammals particularly marsupials and rodents. The aim of this study was to evaluate if spiny rat Proechimys guyannensis (Rodentia: Echimydae) has role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis. Thus, promastigotes (the flagellate stage) of Leishmania (Viannia) braziliensis were used to inoculate seven spiny rats (Proechimys guyannensis). After inoculated intradermal at the ear pinna, nose and plantar pad, the rats were monitored for 180 days. Tissue samples collected at 90 and 180 days from the rats proved to be negative for the presence of genetic material from the parasite. After euthanasia, the protozoa also failed to growth in culture medium containing tissue samples collected from the rats showing that there was no infection. These results fail to prove that spiny rat has a role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis. KEYWORDS: Leishmaniasis, American cutaneous leishmaniasis, rodents, reservoirs O rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) falhou para inoculação intradérmica com Leishmania (Viannia) braziliensis. RESUMOA leishmaniose é uma doença de ocorrência mundial causada por protozoários do gênero Leishmania. No Brasil, a Leishmania (Viannia) braziliensis é o principal parasita responsável pela leishmaniose tegumentar americana. Os principais hospedeiros deste protozoário são pequenos mamíferos selvagens em particular marsupiais e roedores. O objetivo deste estudo foi avaliar o papel do rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) no ciclo da leishmaniose tegumentar americana. Para isto, formas promastigotas (estágio flagelado) de Leishmania (Viannia) braziliensis foram inoculadas em sete ratos espinhosos (Proechimys guyannensis). Após a inoculação intradérmica no pavilhão auricular, focinho e área plantar, os ratos foram monitorizados durante 180 dias. Amostras de tecido colhidas aos 90 e 180 dias dos ratos revelaram-se negativas para a presença de material genético do parasita. Após eutanásia, tecidos coletados dos ratos também falharam para crescimento em meio de cultura demonstrando que não houve infecção. Estes resultados demonstram que o rato espinhoso não tem papel no ciclo da leishmaniose tegumentar americana causada por L. (V.) braziliensis. PALAVRAS-CHAVE: Leishmaniose, leishmaniose tegumentar americana, roedores, reservatórios. ) in association. A fragment of 1 cm 2 was collected from the lesion edge. The fragment was divided into two parts -one for performing the PCR, and the other for culture in NNN-LIT (Novy MacNeal Nicolle -Liver Infusion Tryptose) medium, increased 10% Fetal Bovine Serum, which served as a source of promastigotes and culture for the parasite. In their stationary growth phase the two strains...

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Leishmania (Viannia) braziliensis: Immunoblotting analysis for the detection of IgG subclasses in the diagnosis of symptomatic and asymptomatic dogs

Cited by 3 publications

References 16 publications

Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen

Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen

Leishmaniosis: an approach about the immunoglobulins and cytokines involved in infection and vaccination

The spiny rat Proechimys guyannensis (Rodentia: Echimydae) fails to respond to intradermal inoculation with Leishmania (Viannia) braziliensis

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