This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Scheffold, C. Articles by Negrin, R. S. Search for Related Content PubMed PubMed Citation Articles by Scheffold, C. Articles by Negrin, R. S.

Visualization of Effective Tumor Targeting by CD8+ Natural Killer T Cells Redirected with Bispecific Antibody F(ab')₂HER2xCD3¹

Division of Bone Marrow Transplantation [C. S., M. K., Y. C. S., R. S. N.] and Department of Pediatrics [C. H. C.], Stanford University School of Medicine, Stanford, California 94305

HER2 is an attractive immunotherapeutic target for neoplastic disease because this cell surface molecule is overexpressed on a large fraction of malignant tumor cells. To directly assess therapeutic responses to targeted therapy by noninvasive in vivo imaging in small animals, human HER2-expressing ovarian carcinoma cells were genetically modified with a firefly luciferase gene, and light emission was used for visualization of tumor growth and response to therapy. This imaging approach was able to demonstrate in real-time tumor regression in a HER2 xenograft mouse model by adoptive transfer of in vitro induced and expanded cytotoxic CD8+ natural killer T (NKT) cells retargeted with a humanized bispecific antibody F(ab')₂HER2xCD3. Immunotherapy with effector cells alone or a humanized monoclonal antibody anti-p185^HER2 (4D5-8) resulted in significant but slower reduction in tumor burden. Long-term survival of tumor xenografts correlated inversely with visible residual tumor burden. In vitro, F(ab')₂HER2xCD3 substantially augmented cytotoxic activity of CD8+ NKT cells. By flow-sorting, CD8+ NKT cells coexpressing CD56 were found to have the highest redirected killing ability. Treatment with concanamycin A or EGTA abrogated CD8+ NKT cytotoxicity indicating that perforin is a major pathway of tumor cell lysis. In contrast, when CD8+ NKT cell were cross-linked with F(ab')₂HER2xCD3 neither the immunosuppressants cyclosporine A and FK506, nor the increase of intracellular cyclic AMP by dibutyryl cyclic AMP were able to inhibit cytotoxicity demonstrating that signaling via the CD3 antigen changes the biological activity of non-MHC-restricted effector cells. These studies have demonstrated that CD8+ NKT cells can be successfully redirected to tumor cells using bispecific antibodies and offer a promising strategy for adoptive immunotherapy of neoplastic diseases.

This article has been cited by other articles:

				D. Sangiolo, E. Martinuzzi, M. Todorovic, K. Vitaggio, A. Vallario, N. Jordaney, F. Carnevale-Schianca, A. Capaldi, M. Geuna, L. Casorzo, et al. Alloreactivity and anti-tumor activity segregate within two distinct subsets of cytokine-induced killer (CIK) cells: implications for their infusion across major HLA barriers Int. Immunol., July 1, 2008; 20(7): 841 - 848. [Abstract] [Full Text] [PDF]

				C. M. Lappas, Y.-J. Day, M. A. Marshall, V. H. Engelhard, and J. Linden Adenosine A2A receptor activation reduces hepatic ischemia reperfusion injury by inhibiting CD1d-dependent NKT cell activation J. Exp. Med., November 27, 2006; 203(12): 2639 - 2648. [Abstract] [Full Text] [PDF]

				J. K. Chan, C. A. Hamilton, M. K. Cheung, M. Karimi, J. Baker, J. M. Gall, S. Schulz, S. H. Thorne, N. N. Teng, C. H. Contag, et al. Enhanced killing of primary ovarian cancer by retargeting autologous cytokine-induced killer cells with bispecific antibodies: a preclinical study. Clin. Cancer Res., March 15, 2006; 12(6): 1859 - 1867. [Abstract] [Full Text] [PDF]

				M. R. Verneris, A. Arshi, M. Edinger, M. Kornacker, Y. Natkunam, M. Karami, Y.-a. Cao, N. Marina, C. H. Contag, and R. S. Negrin Low Levels of Her2/neu Expressed by Ewing's Family Tumor Cell Lines Can Redirect Cytokine-Induced Killer Cells Clin. Cancer Res., June 15, 2005; 11(12): 4561 - 4570. [Abstract] [Full Text] [PDF]

				W. J. Grossman, J. W. Verbsky, B. L. Tollefsen, C. Kemper, J. P. Atkinson, and T. J. Ley Differential expression of granzymes A and B in human cytotoxic lymphocyte subsets and T regulatory cells Blood, November 1, 2004; 104(9): 2840 - 2848. [Abstract] [Full Text] [PDF]

				A. Moore, J. Grimm, B. Han, and P. Santamaria Tracking the Recruitment of Diabetogenic CD8+ T-Cells to the Pancreas in Real Time Diabetes, June 1, 2004; 53(6): 1459 - 1466. [Abstract] [Full Text] [PDF]

				M. R. Verneris, M. Karami, J. Baker, A. Jayaswal, and R. S. Negrin Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells Blood, April 15, 2004; 103(8): 3065 - 3072. [Abstract] [Full Text] [PDF]