BD DimerX Antigen Presentation Reagents

For direct detection of antigen specific T cells and epitope mapping

BD™ DimerX reagents are MHC-immunoglobulin fusion proteins developed to detect antigen-specific T-cells. Three extracellular domains of MHC class I molecules are fused to the N terminal of VH region of the mouse IgG1 through recombinant DNA technology. The expression vector containing the fusion protein is then co-transfected with genes containing human β2-microglobulin (β2m) into a myeloma cell line deficient in immunoglobulin heavy chain but retains the expression of immunoglobulin light chain (lambda). The secreted molecule is a three-chain complex molecule consisting of a recombinant heavy chain of MHC-Ig fusion chain, an immunoglobulin light chain disulphide bonded to the heavy chain, and a non-covalently associated human β2m molecule.

The bivalent nature of peptide-binding sites of the DimerX molecules increases the avidity and results in stable binding to antigen-specific T-cells. Furthermore, the hinge region in the immunoglobulin scaffold of DimerX provides a more flexible access for T-cell binding.

References

1. Wang Y.D. and Chen W.F. Detecting specific cytotoxic T lymphocytes against SARS-coronavirus with DimerX HLA-A2:Ig fusion protein. Clin Immunol, 2004; 113(2): 151
2. Furuichi Y. et al. Depletion of CD25+CD4+T cells (Tregs) enhances the HBV-specific CD8+ T cell response primed by DNA immunization. World J Gastroenterol, 2005; 11(24): 3772
3. Oki S. et al. The clinical implication and molecular mechanism of preferential IL-4 production by modified glycolipid-stimulated NKT cells. J. Clin. Invest., Jun 2004; 113: 1631
4. Chen Y.T. and Kung J.T.CD1d-Independent Developmental Acquisition of Prompt IL-4 Gene Inducibility in Thymus CD161(NK1)–CD44^lowCD4⁺CD8^– T Cells Is Associated with 5. Complementarity Determining Region 3-Diverse and Biased V2/V7/V8/V3.2 T Cell Receptor Usage. J. Immunol., 2005; 175: 6537
5. Mayr C. et al. Fibromodulin as a novel tumor-associated antigen (TAA) in chronic lymphocytic leukemia (CLL), which allows expansion of specific CD8⁺ autologous T lymphocytes. Blood, Feb 2005; 105: 1566
6. Curtsinger J.M. et al. Signal 3 Tolerant CD8 T Cells Degranulate in Response to Antigen but Lack Granzyme B to Mediate Cytolysis.  J. Immunol., Oct 2005; 175: 4392
7. Curtsinger J.M. et al. Signal 3 Determines Tolerance versus Full Activation of Naive CD8 T Cells: Dissociating Proliferation and Development of Effector Function. J. Exp. Med., May 2003; 197: 1141.
8. Oki S. et al. Preferential Th2 polarization by OCH is supported by incompetent NKT cell induction of CD40L and following production of inflammatory cytokines by bystander cells in vivo. Int. Immunol., Dec 2005; 17: 1619
9. Chang D.H. et al. Sustained expansion of NKT cells and antigen-specific T cells after injection of -galactosyl-ceramide loaded mature dendritic cells in cancer patients. J. Exp. Med., May 2005; 201: 1503
10. Valenzuela J.O. et al. Cutting Edge: Bcl-3 Up-Regulation by Signal 3 Cytokine (IL-12) Prolongs Survival of Antigen-Activated CD8 T Cells. J. Immunol., Jan 2005; 174: 600
11. Crough T. et al. Granulocyte Colony-Stimulating Factor Modulates -Galactosylceramide-Responsive Human V24+V11+ NKT Cells. J. Immunol., Oct 2004; 173: 4960
12. Maeda M. et al. CD1d-Independent NKT Cells in 2-Microglobulin-Deficient Mice Have Hybrid Phenotype and Function of NK and T Cells. J. Immunol., May 2004; 172: 6115
13. Chen H. et al. Response of Memory CD8⁺ T Cells to Severe Acute Respiratory Syndrome (SARS) Coronavirus in Recovered SARS Patients and Healthy Individuals. J. Immunol., Jul 2005; 175: 591
14. Rechtsteiner G. et al.Cutting Edge: Priming of CTL by Transcutaneous Peptide Immunization with Imiquimod. J. Immunol., Mar 2005; 174: 2476
15. Dal Porto, J. D., T. E. Johansen, B. Catipovic, D. J. Parfiit, D. Tuveson, U. Gether, S. Kozlowski, D. T. Fearon, and J. P. Schneck. (1993). A soluble divalent class I major histocompatibility complex molecule inhibits alloreactive T cells at nanomolar concentrations. Proc. Natl. Acad. Sci. USA. 90:6671-6675.
    
16. Khilko, S.N., Jelonek, M.t., Corr, M., Boyd, L.F., Bothwell, L.M., and Margulies, D.H. (1995). Measuring interactions of MHC class I molecules using surface plasmon resonance. J. of Immunol. Methods. 183: 77-94
17. Parker, K.c., Carreno, B.M., Sestak, L., Utz, U., Biddison, W.E., and Coligan, J.E. (1992). J. Biol. Chem. 267(8): 5451-5459.Parker, K.C., Bednarek, M.A., Hull, L.K., Utz, U., Cunningham, B., Zweerink, H.J., Biddison, W.E., and Coligan, J.e. 1992. J of Immunol. 149: 2580- 3587.
18. Pharmingen unpublished results.
19. Schneck, J. P., J. E. Slansky, S. M., O‘Herrin, and T. F. Greten. Monitoring antigen-specific T cells using MHC-Ig dimers. Cur. Protocols Immunol. In Press.
20. Sun, D. et al. CD8⁺ Encephalitogenic T cells in press
21. Welsch, R. (2001). Assessing CD8 T Cell Number and Dysfunction in the Presence of Antigen. J. Exp. Med. 193:5:F19-F22