title: A monoclonal antibody against the extracellular domain of mouse and human epithelial V-like antigen 1 reveals a restricted expression pattern among CD4- CD8- thymocytes.
abstract: Expression of transcripts for the homotypic adhesion protein epithelial V-like antigen 1 (EVA1), also known as myelin protein zero like-2 (Mpzl2), is known to be present in thymic stromal cells. However, protein expression within different thymic subsets, stromal and/or lymphoid, has not been characterized due a lack of specific reagents. To address this, we generated a hybridoma (G9P3-1) secreting a monoclonal antibody (G9P3-1Mab), reactive against both human and mouse EVA1. The G9P3-1Mab was generated by immunizing Mpzl2-deficient gene-targeted mice with the extracellular domain of EVA1, followed by a conventional hybridoma fusion protocol, illustrating the feasibility of using gene-targeted mice to generate monoclonal antibodies with multiple species cross-reactivity. We confirmed expression of EVA1 on cortical and medullary epithelial cell subsets and revealed a restricted pattern of expression on CD4- CD8- double negative (DN) cell subsets, with the highest level of expression on DN3 (CD44(low)CD25(+)) thymocytes. G9P3-1MAb is a valuable reagent to study thymic T cell development and is likely useful for the analysis of pathological conditions affecting thymopoiesis, such as thymic involution caused by stress or aging.
authors.affiliation: 1 Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) , Barcelona, Spain .
authors.forename: Nahir
authors.initials: N
authors.lastname: Garabatos
authors.forename: Jesus
authors.initials: J
authors.lastname: Blanco
authors.forename: Cesar
authors.initials: C
authors.lastname: Fandos
authors.forename: Elena
authors.initials: E
authors.lastname: Lopez
authors.forename: Pere
authors.initials: P
authors.lastname: Santamaria
authors.forename: Andrea
authors.initials: A
authors.lastname: Ruiz
authors.forename: Maria Laura
authors.initials: ML
authors.lastname: Perez-Vidakovics
authors.forename: Patricia
authors.initials: P
authors.lastname: Benveniste
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Galkin
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Zuñiga-Pflucker
authors.forename: Pau
authors.initials: P
authors.lastname: Serra
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D015815:Cell Adhesion Molecules / Q000276:immunology*; D003429:Cross Reactions / Q000276:immunology; D004847:Epithelial Cells / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL
chemical_list: D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D008565:Membrane Proteins