| Texto completo | |
| Autor(es): Mostrar menos - |
Batista, Camila F.
[1]
;
Souza, Fernando N.
[1]
;
Santos, Kamila R.
[1]
;
Ramos Sanchez, Eduardo M.
[2]
;
Reis, Luiza Campos
[2]
;
Bertagnon, Heloisa G.
[1]
;
Blagitz, Maiara G.
[1]
;
Gomes, Renata C.
[1]
;
Lage, Andrey P.
[3]
;
Heinemann, Marcos B.
[4]
;
Della Libera, Alice M. M. P.
[1]
Número total de Autores: 11
|
| Afiliação do(s) autor(es): | [1] Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Clin Med, Vet Clin Immunol Res Grp, Av Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP - Brazil
[2] Univ Sao Paulo, Lab Sorol & Imunobiol, Inst Med Trop Sao Paulo, Av Prof Dr Eneas de Carvalho Aguiar 470, BR-05403000 Sao Paulo, SP - Brazil
[3] Univ Fed Minas Gerais, Escola Vet, Dept Med Vet Prevent, Av Presidente Antonio Carlos 6627, BR-31270010 Belo Horizonte, MG - Brazil
[4] Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Med Vet Prevent & Saude Anim, Av Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP - Brazil
Número total de Afiliações: 4
|
| Tipo de documento: | Artigo Científico |
| Fonte: | VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY; v. 196, p. 53-59, FEB 2018. |
| Citações Web of Science: | 0 |
| Resumo | |
The present study aimed to validate the use of R-phycoerythrin (R-PE)-labeled Mannheimia haemolytica to simultaneously stimulate phagocytosis and intracellular production of reactive oxygen species (ROS) by blood phagocytes in bronchoalveolar lavage (BAL) fluid. Initially, R-PE-labeled M. haemolytica was inactivated using a water bath at 60 degrees C for 60 min. Afterwards, R-PE labelling of bacteria was confirmed by flow cytometry. The geometric mean fluorescence intensity of R-PE-labeled bacteria (FL2 detector, 585 +/- 42 nm) was analyzed by flow cytometry and was 41.5-fold higher than the respective unlabeled controls, confirming the success of bacterial conjugation to R-PE. Phagocytosis and intracellular production of ROS by blood neutrophils and monocytes, and by BAL CD14 macrophages, in 12 healthy 6-month-old male calves were then performed using R-PE-labeled bacteria and 2',7'-dichlorofluoresceindiacetate (DCFH-DA) as probes. Confocal microscopy was used to confirm phagocytosis of R-PE-labeled M. haemolytica by phagocytes and to exclude erroneous measurements of bacteria adhering to the leukocyte membrane. The present study showed that there is no difference in the ROS production without stimulus and in the presence of M. haemolytica by peripheral blood neutrophils and monocytes, in contrast to the increased ROS production by local alveolar macrophages upon stimulation by M. haemolytica. This emphasizes the importance of alveolar macrophages in the maintenance of homeostasis and health of the respiratory system, which can be supported during the inflammatory process by the rapid recruitment of neutrophils with high microbicidal and phagocytic capacity. The method described here provides an easy and feasible tool to measure phagocytosis and intracellular ROS production by phagocytes, especially when commonly used probes for intracellular ROS production were used, such as DCFH-DA and dihydrorhodamine 123. (AU) | |
| Processo FAPESP: | 14/23189-4 - Interação das células epiteliais mamárias e das células mononucleares do sangue periférico na regulação da resposta imune e na infecção intramamária por Staphylococcus aureus, em bovinos |
| Beneficiário: | Fernando Nogueira de Souza |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 11/23936-6 - Dinâmica de fagócitos sanguíneos e alveolares em bezerros com mannheimiose |
| Beneficiário: | Camila Freitas Batista |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 12/00749-9 - Dinâmica de fagócitos sanguíneos e alveolares em bezerros com mannhemiose |
| Beneficiário: | Alice Maria Melville Paiva Della Libera |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |