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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Elimination of erroneous results related to bovine mononuclear cell immunophenotyping by antibodies binding to Fc receptors

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Author(s):
Okino, Cintia Hiromi [1] ; Giglioti, Rodrigo [2] ; Bassetto, Cesar Cristiano [3] ; Silva, Pamella Cristini [1] ; de Oliveira, Henrique Nunes [3] ; de Sena Oliveira, Marcia Cristina [1]
Total Authors: 6
Affiliation:
[1] Embrapa Pecuaria Sudeste, Lab Anim Hlth, Rodovia Washington Luiz, Km 234, BR-13560970 Sao Carlos, SP - Brazil
[2] IZ, Ctr Pesquisa Genet & Reprod Anim, Rua Heitor Penteado 56, BR-13380011 Nova Odessa, SP - Brazil
[3] Univ Estadual Paulista, Dept Zootecnia, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY; v. 213, JUL 2019.
Web of Science Citations: 0
Abstract

Blocking immunoglobulin G (IgG) binding receptors on leukocytes is an established and highly recommended preventive procedure for immunological assays. Failing to prevent such nonspecific binding can lead to erroneous results. Several studies testing different blocking reagents have been performed in murine or human cells, however, there are no specific studies on bovine cells. Our study aimed to investigate the efficiency of blocking reagents to inhibit the nonspecific binding of mouse monoclonal antibodies (mAbs) to bovine peripheral blood cells. We observed nonspecific interactions of IgG2a and IgG2b negative isotypes with bovine leukocytes, but not IgG1. We found that these nonspecific bindings could be eliminated by blocking with purified mouse IgG, whereas little or no blocking effect was observed when bovine serum or Mouse Seroblock FcR were applied. Moreover, in the absence of an efficient blocking reagent, the percentage of CD335 positive cells was significantly higher than in the group previously blocked with mouse IgG. Based on these results, and due to the lack of specific commercial blocking reagents for bovine cells, our recommendation is to use purified mouse IgG as a blocking reagent for immune assays targeting bovine leukocytes in order to enhance the accuracy of the results. (AU)

FAPESP's process: 16/07216-7 - Genomic study and immunological characterization of cattle resistant to bovine babesiosis and analysis of the genetic diversity of Babesia bovis and Babesia bigemina
Grantee:Henrique Nunes de Oliveira
Support Opportunities: Research Projects - Thematic Grants