A Monoclonal Antibody Against the Extracellular Domain of Mouse and Human Epithelial V-like antigen 1 (EVA1) reveals a restricted expression pattern among CD4- CD8- thymocytes

GARABATOS, NAHIR ; BLANCO, JESUS ; FANDOS, CESAR; LOPEZ, ELENA; SANTAMARIA, PERE; RUIZ, ANDREA; PEREZ VIDAKOVICS, MARIA LAURA; BENVENISTE, PATRICIA ; GALKIN, OLEKSANDR ; ZUÑIGA-PFLUCKER, JUAN CARLOS ; SERRA, PAU

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

Liebertpub

Año: 2014 vol. 33 p. 305 - 311

2167-9436

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 cells subsets, and revealed a restricted pattern of expression on CD4- CD8- double negative (DN) cell subsets, with the highest level of expression on DN3 (CD44lowCD25+) thymocytes. The G9P3-1Mab is a valuable reagent to study thymic T-cell development, and likely useful for the analysis of pathological conditions affecting thymopoiesis, such as thymic involution caused by stress or aging.