Analysis of the expression of porcine CD200R1 and CD200R1L by using newly developed monoclonal antibodies

• DOI: 10.1016/j.dci.2019.103417
• Publication Date: 2019-06-22T05:05:59Z
• Authors: T. Poderoso; P. Martínez de la...; H. Uenishi; B. Alvarez; D. Toki; E. Nieto-Pelegrín; F. Alonso; J. Domínguez; A. Ezquerra; C. Revilla
• Keywords: B-Lymphocytes; 0303 health sciences; Sequence Homology, Amino Acid; Sus scrofa; Antibodies, Monoclonal; CHO Cells; Monocytes; Recombinant Proteins; Immunoglobulin Fc Fragments; 3. Good health; Epitopes; 03 medical and health sciences; Cricetulus; Protein Domains; Orexin Receptors; Macrophages, Alveolar; Animals; Amino Acid Sequence

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.