Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/6775
Title: Establishment of a microplate assay for flow cytometric assessment and it is use for the evaluation of age-related phenotypic changes in canine whole blood leukocytes
Authors: Reis, Alexandre Barbosa
Carneiro, Cláudia Martins
Carvalho, Maria das Graças
Carvalho, Andréa Teixeira de
Giunchetti, Rodolfo Cordeiro
Mayrink, Wilson
Genaro, Odair
Oliveira, Rodrigo Correa de
Martins Filho, Olindo Assis
Affilliation: Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, Brasil / Universidade Federal de Ouro Preto. Escola de Farmácia. Departamento de Analises Clínicas. Ouro Preto, MG, Brasil / Universidade Federal de Ouro Preto. Laboratório de Parasitologia e Histopatologia. Ouro Preto, MG, Brasil.
Universidade Federal de Ouro Preto. Escola de Farmácia. Departamento de Analises Clínicas. Ouro Preto, MG, Brasil / Universidade Federal de Ouro Preto. Instituto de Ciências Exatas e Biológicas. Núcleo de Pesquisas em Ciências Biológicas. Laboratório de Parasitologia e Histopatologia. Ouro Preto, MG, Brasil.
Universidade Federal de Minas Gerais. Faculdade de Farmácia. Laboratório de Hematologia Clínica. Belo Horizonte, MG, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, Minas Gerais, Brasil / Universidade Federal de Ouro Preto. Escola de Farmácia. Departamento de Analises Clínicas. Ouro Preto, MG, Brasil.
Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil.
Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil.
Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, Brasil.
Fundaçao Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratório de Doença de Chagas. Belo Horizonte, MG, Brasil.
Abstract: The effectiveness of flow cytometric assays for canine use is still requiring standardization. Despite several studies using purified mononuclear cells, no methodology or reference ranges are available for immunophenotyping of whole blood leukocytes (WBL). Fresh and pre-fixed WBL were used to identify cell-subsets, (Thy-1(+)/CD5(+)/CD4(+)/CD8(+)/CD21(+) and CD14(+)) and measure MHC-II, CD45RA/CD45RB expression. We described here an efficient method for fast quantification of canine-WBL, using pre-fix in a microplate assay, which allows long-term sample storage prior to phenotyping. Decreased percentage of CD5(+)-T-cells within the lymphocyte-gate and increased percentage of CD21(+)-B-cells were observed in young animals, which led to higher T/B cell ratios in middle-aged dogs. Lower numerical counts of Thy-1(+), CD4(+), CD8(+) and CD21(+) lymphocyte were observed when compared to young animals. In addition, we identified an age-related decline of MHC-II/CD45RA expression by lymphocytes. We proposed an improved method for phenotyping of canine peripheral blood mononuclear cells (PBMC) that has significant use for researchers and veterinary clinicians. The hematological changes of senescence previously identified on PBMC could be adequately reproduced on features identified by whole blood. Furthermore, this study supplies normal range references as baseline standards for clinical purposes, besides specific immunological parameters to monitor canine aging process.
Keywords: Flow cytometry
Canine
Peripheral blood
Lymphocyte subsets
Aging and sex
Issue Date: 2005
Citation: REIS, Alexandre Barbosa et al. Establishment of a microplate assay for flow cytometric assessment and it is use for the evaluation of age-related phenotypic changes in canine whole blood leukocytes. Vet Immunol Immunopathol., v. 103, n. 3-4, p. 173-185, Feb 10, 2005.
Copyright: restricted access
Appears in Collections:MG - IRR - Artigos de Periódicos

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