Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/15394
Title: Application of rapid in vitro co-culture system of macrophages and T-cell subsets to assess the immunogenicity of dogs vaccinated with live attenuated Leishmania donovani centrin deleted parasites (LdCen−/−).
Authors: Viana, Kelvinson Fernandes
Fiuza, Jacqueline Araújo
Gannavaram, Sreenivas
Dey, Ranadhir
Selvapandiyan, Angamuthu
Bartholomeu, Daniella Castanheira
Lemos, Denise da Silveira
Bueno, Lilian Lacerda
Dutra, Walderez Ornelas
Fujiwara, Ricardo Toshio
Nakhasi, Hira L.
Giunchetti, Rodolfo Cordeiro
Affilliation: Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Morfologia. Laboratório de Interações de células. Belo Horizonte, MG, Brasil/Universidade Federal de Tocantins. Departamento de Ciencias Agrarias e Tecnologia. Laboratorio de Biomoleculas e Vacinas. Gurupi, TO, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, MG, Brasil.
US Food and Drug Administration. Center for Biologics Evaluation and Research. Office of Blood Research and Review. Division of Emerging and Transfusion Transmitted Diseases. Silver Spring, MD, USA.
US Food and Drug Administration. Center for Biologics Evaluation and Research. Office of Blood Research and Review. Division of Emerging and Transfusion Transmitted Diseases. Silver Spring, MD, USA.
Institute of Molecular Medicine. New Delhi, India.
Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitas. Belo Horizonte, MG, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Laboratorio de Diagnostico de Monitoramento e Biomarcadores. Belo Horizonte, MG, Brasil.
Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitas. Belo Horizonte, MG, Brasil.
Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Morfologia. Laboratório de Interações de células. Belo Horizonte, MG, Brasil.
Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitas. Belo Horizonte, MG, Brasil.
US Food and Drug Administration. Center for Biologics Evaluation and Research. Office of Blood Research and Review. Division of Emerging and Transfusion Transmitted Diseases. Silver Spring, MD, USA.
Universidade Federal de Minas Gerais. Instutito de Ciencias Biologicas. Departamento de Morfologia. Laboratório de Interações de células. Belo Horizonte, MG, Brasil.
Abstract: Background: Live attenuated Leishmania donovani parasites as LdCen−/− were shown to confer protective immunity against Leishmania infection in mice, hamsters, and dogs. Strong immunogenicity in dogs vaccinated with LdCen−/− has been previously reported, including increased antibody response favoring Th1 response lymphoproliferative responses, CD4+ and CD8+ T-cells activation, increased levels of Th1 and reduction of Th2 cytokines, in addition to a significant reduction in parasite burden after 18 and 24 months post virulent parasite challenge. Methods: Aimed at validating a new method using in vitro co-culture systems with macrophages and purified CD4+or CD8+ or CD4+:CD8+ T-cells of immunized dogs with both LdCen−/− and Leishmune® to assess microbicide capacity of macrophages and the immune response profile as the production of IFN-γ, TNF-α, IL-12, IL-4 and IL-10 cytokines. Results and discussion: Our data showed co-cultures of macrophages and purified T-cells from dogs immunized with LdCen−/− and challenged with L. infantum were able to identify high microbicidal activity, especially in the co-culture using CD4+ T-cells, as compared to the Leishmune® group. Similarly, co-cultures with CD8+ T-cells or CD4+:CD8+ T-cells in both experimental groups were able to detect a reduction in the parasite burden in L. infantum infected macrophages. Moreover, co-cultures using CD4+ or CD8+ or CD4+:CD8+ T-cells from immunized dogs with both LdCen−/− and Leishmune® were able to identify higher levels of IFN-γ and IL-12 cytokines, reduced levels of IL-4 and IL-10, and a higher IFN-γ/IL-10 ratio. While the highest IFN-γ levels and IFN-γ/IL-10 ratio were the hallmarks of LdCen−/− group in the coculture using CD4+ T-cells, resulting in strong reduction of parasitism, the Leishmune® immunization presented a differential production of TNF-α in the co-culture using CD4+:CD8+ T-cells. Conclusion: The distinct conditions of co-culture systems were validated and able to detect the induction of immune protection. The method described in this study applied a new, more accurate approach and was able to yield laboratory parameters useful to test and monitor the immunogenicity and efficacy of Leishmania vaccines in dogs.
Keywords: Researc Vaccine
In vitro co-culture
CD4+ and CD8+ T-cells
Canine visceral leishmaniasis LdCen−/−
Leishmune®
Issue Date: 2016
Publisher: BioMed Central
Citation: VIANA , Kelvinson Fernandes et al. Application of rapid in vitro co-culture system of macrophages and T-cell subsets to assess the immunogenicity of dogs vaccinated with live attenuated Leishmania donovani centrin deleted parasites (LdCen−/−). Parasites & Vectors, v. 9, p.250, 2016
DOI: 10.1186/s13071-016-1528-z
ISSN: 1756-3305
Copyright: open access
Appears in Collections:MG - IRR - Artigos de Periódicos

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