CD4 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

CD4 (cluster of differentiation 4), also known as L3T4 or T4, is a 55 kDa single chain type I transmembrane glycoprotein belonging to the immunoglobin (Ig) superfamily. CD4 is predominantly expressed on most thymocytes, a subset of mature T lymphocytes, and weakly on monocytes, tissue macrophages, dendritic cells, and granulocytes. It is also expressed on neurons and glial cells in the brain (1). CD4 is expressed along with CD8 on double positive T cells during their development in the thymus. Either CD4 or CD8 expression is then lost giving rise to single positive (SP) CD4+ or CD8+ mature T cells. CD4+ SP cells (T helper cells) further differentiate into multiple subsets of CD4+ cells including Th1, Th2, Th17, Tfh, and Treg cells which regulate humoral and cellular immunity (2). The extracellular region of CD4 consists of 372 amino acids (aa) with four immunoglobin-like domains (D1-D4). The structures of D1 and D3 resemble variable (IgV) domains while D2 and D4 resemble constant (IgC) domains (3).

Given its critical role in T cell development, CD4 also has diverse immunology-related functions. CD4 acts as a coreceptor with the T-cell receptor (TCR) during T cell activation and thymic differentiation by binding directly to major histocompatibility complex (MHC) class II antigens and associating with the protein tyrosine kinase, Lck (4). This interaction contributes to the formation of the immunological synapse (5). Defects in antigen presentation cause dysfunction of CD4+ T cells and the almost complete loss of MHC II expression on B cells in peripheral blood, as observed in severe combined immunodeficiency (SCID) (6). CD4 also functions as a receptor for the human immunodeficiency virus (HIV) by binding to gp120, the envelope glycoprotein of HIV-1. It has been shown that the V-like domains are critical for binding to gp120 (7). In immune mediated and infectious diseases of the central nervous system, CD4 functions as an indirect mediator of neuronal damage (8).

References

1. Omri, B., Crisanti, P., Alliot, F., Marty, M., Rutin, J., Levallois, C., . . . Pessac, B. (1994). CD4 expression in neurons of the central nervous system. International Immunology, 6(3), 377-385. doi:10.1093/intimm/6.3.377

2. Wan, Y. Y., & Flavell, R. A. (2009). How diverse-CD4 effector T cells and their functions. Journal of Molecular Cell Biology, 1(1), 20-36. doi:10.1093/jmcb/mjp001

3. Wu, H., Myszka, D. G., Tendian, S. W., Brouillette, C. G., Sweet, R. W., Chaiken, I. M., & Hendrickson, W. A. (1996). Kinetic and structural analysis of mutant CD4 receptors that are defective in HIV gp120 binding. Proceedings of the National Academy of Sciences, 93(26), 15030-15035. doi:10.1073/pnas.93.26.15030

4. Doyle, C., & Strominger, J. L. (1987). Interaction between CD4 and class II MHC molecules mediates cell adhesion. Nature, 330, 256-259. doi:10.1038/330256a0

5. Vignali, D. A. (2010). CD4 on the road to coreceptor status. The Journal of Immunology, 184(11), 5933-5934. doi:10.4049/jimmunol.1090037

6. Tasher, D., & Dalal, I. (2012). The genetic basis of severe combined immunodeficiency and its variants. The Application of Clinical Genetics, 5, 67-80. doi:10.2147/tacg.s18693

7. Arthos, J., Deen, K. C., Chaikin, M. A., Fornwald, J. A., Sathe, G., Sattentau, Q. J., . . . Sweet, R. W. (1989). Identification of the residues in human CD4 critical for the binding of HIV. Cell, 57(3), 469-481. doi:10.1016/0092-8674(89)90922-7

8. Buttini, M., Westland, C. E., Masliah, E., Yafeh, A. M., Wyss-Coray, T., Mucke, L. (1998). Novel role of human cd4 molecule identified in neurodegeneration. Nature Medicine, 4(4), 441-446. doi:10.1038/nm0498-441

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-04168

• CD4 Antibodies
• CD4 Antibody Pair
• CD4 ELISA Kits
• CD4 Lysate
• CD4 Proteins
• CD4 Proteins and Enzymes