title: Obtaining anti-type 1 melatonin receptor antibodies by immunization with melatonin receptor-expressing cells.
abstract: Antibodies (Abs) specific to cell-surface receptors are attractive tools for studying the physiological role of such receptors or for controlling their activity. We sought to obtain such antibodies against the type 1 receptor for melatonin (MT1). For this, we injected mice with CHO cells transfected with a plasmid encoding human MT1 (CHO-MT1-h), in the presence or absence of an adjuvant mixture containing Alum and CpG1018. As we previously observed that the immune response to a protein antigen is increased when it is coupled to a fusion protein, called ZZTat101, we also investigated if the association of ZZTat101 with CHO-MT1-h cells provides an immunogenic advantage. We measured similar levels of anti-CHO and anti-MT1-h Ab responses in animals injected with either CHO-MT1-h cells or ZZTat101/CHO-MT1-h cells, with or without adjuvant, indicating that neither the adjuvant mixture nor ZZTat101 increased the anti-cell immune response. Then, we investigated whether the antisera also recognized murine MT1 (MT1-m). Using cloned CHO cells transfected with a plasmid encoding MT1-m, we found that antisera raised against CHO-MT1-h cells also bound the mouse receptor. Altogether our studies indicate that immunizing approaches based on MT1-h-expressing CHO cells allow the production of polyclonal antibodies against MT1 receptors of different origins. This paves the way to preparation of MT1-specific monoclonal antibodies.
authors.affiliation: CEA, institut de Biologie et de Technologie de Saclay (iBiTec-S), Service de Pharmacologie et d'immunoanalyse (SPI), 91191 Gif sur Yvette, France.
authors.forename: Nelia
authors.initials: N
authors.lastname: Cordeiro
authors.affiliation: University of Paris Diderot, Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Wijkhuisen
authors.affiliation: CEA, institut de Biologie et de Technologie de Saclay (iBiTec-S), Service de Pharmacologie et d'immunoanalyse (SPI), 91191 Gif sur Yvette, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Savatier
authors.affiliation: Institut de Recherches Servier, Division Biotechnologie, Pharmacologie Moléculaire et Cellulaire, 125 Chemin de Ronde, 78290 Croissy-Sur-Seine, France.
authors.forename: Natacha
authors.initials: N
authors.lastname: Moulharat
authors.affiliation: Institut de Recherches Servier, Division Biotechnologie, Pharmacologie Moléculaire et Cellulaire, 125 Chemin de Ronde, 78290 Croissy-Sur-Seine, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Ferry
authors.affiliation: CEA, institut de Biologie et de Technologie de Saclay (iBiTec-S), Service de Pharmacologie et d'immunoanalyse (SPI), 91191 Gif sur Yvette, France. Electronic address: michel.leonetti@cea.fr.
authors.forename: Michel
authors.initials: M
authors.lastname: Léonetti
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D016466:CHO Cells; D003412:Cricetulus; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization; D051379:Mice; D008810:Mice, Inbred C57BL; D044122:Receptor, Melatonin, MT1 / Q000096:biosynthesis* / Q000276:immunology*
chemical_list: D000906:Antibodies; D044122:Receptor, Melatonin, MT1
keywords: Antibody; Antibody response; Cellular immunization; Melatonin receptor