Construction Of LIGHT Transfected Cells And Preparation Of Mouse Monoclonal Antibodies Against Human LIGHT

Human LIGHT (TNFSF14), a member of the tumor-necrosis-factor ( TNF ) superfamily, is a type II membrane protein encoding 240 aa. Human LIGHT locus links to the TNF superfamily members CD27 ligand and 4-1BB ligand, and the third complement protein (C3),which positions LIGHT within the MHC paralog on chromosome 19p13.3. At a function level, FasL and LIGHT exhibit significant amino acid sequence homology in their ectodomains (31%), second only to the similarity between LIGHT and LT β (34%). This homology is reflected in the fact that LIGHT shares receptor-binding properties for the LT β R with heterotrimers composed of LT α₁β₂, HVEM binding with the homomeric forms of LT a (LTα₃), and soluble DcR3 binding with FasL. LIGHT mainly expressed on activated T cells and immature dendritic cells(DCs) generated from human PBMCs, its mRNA is highly expressed in splenocytes, activated PBL, CD8⁺ tumor infiltrating lymphocytes, granulocytes, and monocytes but not in the thymus and the tumor cells. Several studies indicate that LIGHT can trigger apoptosis of some tumor cells as well as cell activation, depending in part on expression of its receptors on the target cells. LIGHT is a CD28-independent co-stimulatory molecule for T-cell growth and differentiation. Accumulating data support an essential contribution of LIGHT in initiation, development, maintenance, and termination of immune responses.1 Cloning of human LIGHT geneTo explore the bionfunctions of human LIGHT, the cDNA of human LIGHT was obtained by RT-PCR from activated T cells, and then cloned into the cloning vector pMD18-T. Then identified by restriction enzyme digestion ,PCR and sequence analysis.2 Construction of LIGHT transfected cells...