Recombinant Mouse CTLA-4 Fc Chimera Protein Summary
Details of Functionality
Measured by its ability to inhibit IL-2 secretion by stimulated Jurkat human acute T cell leukemia cells. Linsley, P.S. et al. (1991) J. Exp. Med. 174:561. The ED50 for this effect is 0.1-0.4 µg/mL when stimulated with 1 µg/mL Recombinant Human B7-1 Fc Chimera (Catalog # 140-B1) in the presence of PHA.
Mouse myeloma cell line, NS0-derived mouse CTLA-4 protein
>97%, by SDS-PAGE under reducing conditions and visualized by silver stain
<0.1 EU per 1 μg of the protein by the LAL method.
41 kDa (monomer). Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4
CTLA-4 (cytotoxic T-lymphocyte-4, designated CD152), is a type I transmembrane T cell inhibitory molecule that is a member of the Ig superfamily (1, 2). Human or mouse CTLA-4 cDNA encodes 223 amino acids (aa) including a 35 aa signal sequence, a 126 aa extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane (TM) sequence, and a 41 aa cytoplasmic sequence. It is found as a covalent homodimer of 41 ‑ 43 kDa (2) Within the ECD, mouse CTLA-4 shares 94% and 68 ‑ 71% aa sequence identity with rat and human/porcine/bovine/rabbit/feline/canine CTLA-4, respectively. A 174 aa form that lacks TM and cytoplasmic sequences (sCTLA-4) is possibly secreted (3 ‑ 5). Isoforms of 56 ‑ 79 aa that mainly contain parts of the cytoplasmic domain are reported. In mouse, an isoform lacking the Ig-like domain has ligand-independent inhibitory activity and is termed liCTLA-4 (6). CD28, which is structurally related to CTLA-4, is constitutively expressed on naïve T cells and promotes T cell activation when engaged by B7-2 on antigen-presenting cells (APC) within the immunological synapse (IS) (1, 7, 8). In contrast, CTLA-4 is recruited from intracellular vesicles to the IS beginning 1-2 days after T cell activation (2, 7, 8). It forms a linear lattice with B7-1 on APC, inducing negative regulatory signals and ending T cell activation (9). Abatacept, a therapeutic human CTLA-4-Ig fusion protein (trade name Orencia), competes with CD28 for B7-1 and B7-2 binding and has been used to antagonize T cell activation in autoimmune conditions and to enhance transplant survival (10). Mice deleted for CTLA-4 show no abnormalities until after birth, but then develop lethal autoimmune reactions due to continued T cell activation and poor control by regulatory T cells, which constitutively express CTLA-4 in wild-type mice and humans (11 ‑ 13).
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