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Dev Comp Immunol ; 100: 103417, 2019 11.
Article in English | MEDLINE | ID: mdl-31233758

ABSTRACT

CD200R1 and CD200R1-like are paired receptors which modulate activation of immune cells. Here, we describe the characterisation of their porcine homologues. Analysis of database porcine sequences shows an exceptionally high homology between the extracellular Ig-like domains of these receptors, being the rest more dissimilar. We have obtained two mAbs, PCT1 and PCT3, against a CD200R1-Fc recombinant protein, that bind on CHO cells expressing GFP-tagged CD200R1. The specificity of these mAbs was analysed on CD200R1 L, and also on a CD200R1 splicing variant that lacks the V-type Ig domain. PCT1 bound to both CD200R1 and CD200R1L, but not to the splicing variant, what suggests that recognises an epitope in the V-type Ig domain. PCT3 reacted with both CD200R1 variants, but not CD200R1L, probably binding to an epitope in the N-terminal sequence of CD200R1. Analysis of porcine cells with these mAbs showed expression of CD200R1/CD200R1L on B cells, monocytes and alveolar macrophages.


Subject(s)
Antibodies, Monoclonal/metabolism , Immunoglobulin Fc Fragments/metabolism , Orexin Receptors/metabolism , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , CHO Cells , Cricetulus , Epitopes/genetics , Epitopes/immunology , Epitopes/metabolism , Immunoglobulin Fc Fragments/immunology , Macrophages, Alveolar/immunology , Macrophages, Alveolar/metabolism , Monocytes/immunology , Monocytes/metabolism , Orexin Receptors/genetics , Orexin Receptors/immunology , Protein Domains/genetics , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , Sus scrofa
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