Generation of rat monoclonal antibodies against murine LAIR-1.
The leukocyte-associated Ig-like receptor 1 (LAIR-1), an inhibitory receptor bearing two immunoreceptor tyrosine-based inhibitory motifs (ITIM), is expressed on the majority of peripheral leukocytes, including NK cells, T cells, B cells, monocytes, dendritic cells, granulocytes, and thymocytes and is involved in immunologic regulation and hematopoiesis. Murine LAIR-1 (mLAIR-1) is the homolog molecule of human LAIR-1. Using mLAIR-1-Fc as the immunogen and the technique of rat B lymphocyte hybridoma, we raised three hybridoma cell lines secreting monoclonal antibodies (MAb) to mLAIR-1, designated FMU-mLAIR-1.1, -1.2, and -1.3. Rat immunoglobulin class and subclass of the MAb FMU-mLAIR-1.1 approximately 3 were determined to be IgM, IgG1, and IgM, respectively. All these MAbs can bind the mLAIR-1 in immunocytochemistry and immunohistochemistry. FMU-m LAIR-1.2 worked well not only in Western blot assay but also in recognizing natural LAIR-1 molecules on the surface of P388D1, J774, and WEHI3 cells, and mLAIR-1 cDNA-transfected CHO cells detected by FCM. Thus, successful production of rat anti-murine LAIR-1 monoclonal antibodies provides a new powerful tool for investigation of murine LAIR-1 function in mouse model, both in vitro and in vivo.
Wang
Chun-Yan
CY
Department of Immunology, Fourth Military Medical University, Xi'an, China.
Zhang
Yuan
Y
Wang
Dong-Lin
DL
Song
Chao-Jun
CJ
Han
Wei-Ning
WN
Yang
Kun
K
Jin
Bo-Quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011817:Rabbits; D051381:Rats; D011971:Receptors, Immunologic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D011971:Receptors, Immunologic; C108073:leukocyte-associated immunoglobulin-like receptor 1