Production of monoclonal antibodies against canine leukocytes.
A panel of anti-canine leukocyte monoclonal antibodies (MAbs) was produced by immunizing BALB/c mice with canine peripheral blood mononuclear cells (PBMC), either resting or stimulated with concanavalin A (ConA). Three out of 28 clones-IH1, AB6, and HG6-screened by ELISA and producing antibody with the highest specificity for canine cell immunostaining, were subjected to three subsequent subcloning steps by limiting dilution, and selected for further characterization. These MAbs belonged to IgG1 (HG6 and IH1) and IgG2a (AB6) isotypes. The distribution of cell populations expressing the antigen recognized by the antibodies was identified by indirect immunoflorescence on canine PBMC and on tissue sections of lymph node, spleen, liver and skin. The possible crossreactivity with human PBMC was also examined in immunocytochemistry. One of the antibodies specifically recognized macrophages. The MAbs presented here can be foreseen as possible valuable diagnostic and research tools to study immune functions in dogs.
Aguiar
Paulo Henrique Palis
PH
Laboratory of Molecular and Cellular Immunology, Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Bahia, Brazil. phpa@ufba.br
Borges dos Santos
Roberto Robson
RR
Lima
Carla Andrade
CA
Rios de Sousa Gomes
Hilton
H
Larangeira
Daniela Farias
DF
Santos
Patrícia Meira
PM
Barrouin-Melo
Stella Maria
SM
Conrado dos-Santos
Washington Luis
WL
Pontes-de-Carvalho
Lain
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003208:Concanavalin A / Q000276:immunology; D004285:Dogs; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007150:Immunohistochemistry; D007962:Leukocytes / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D003208:Concanavalin A