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Journal of Biomedicine and Biotechnology
Volume 2011, Article ID 578128, 11 pages
http://dx.doi.org/10.1155/2011/578128
Research Article

The Construction of Chimeric T-Cell Receptor with Spacer Base of Modeling Study of VHH and MUC1 Interaction

1Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran
2Department of Biochemistry, Biological Science, Tarbiat Modares University, Tehran, Iran
3National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
4Department of Medical Biotechnology, Tarbiat Modares University, Tehran, Iran

Received 19 March 2011; Revised 24 May 2011; Accepted 2 June 2011

Academic Editor: Ayman El-Kadi

Copyright © 2011 Nazanin Pirooznia et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Adaptive cell immunotherapy with the use of chimeric receptors leads to the best and most specific response against tumors. Chimeric receptors consist of a signaling fragment, extracellular spacer, costimulating domain, and an antibody. Antibodies cause immunogenicity; therefore, VHH is a good replacement for ScFv in chimeric receptors. Since peptide sequences have an influence on chimeric receptors, the effect of peptide domains on each other's conformation were investigated. CD3Zeta, CD28, VHH and CD8α, and FcgIIα are used as signaling moieties, costimulating domain, antibody, and spacers, respectively. To investigate the influence of the ligation of spacers on the conformational structure of VHH, models of VHH were constructed. Molecular dynamics simulation was run to study the influence of the presence of spacers on the conformational changes in the binding sites of VHH. Root mean square deviation and root mean square fluctuation of critical segments in the binding site showed no noticeable differences with those in the native VHH. Results from molecular docking revealed that the presence of spacer FcgIIα causes an increasing effect on VHH with MUC1 interaction. Each of the constructs was transformed into the Jurkat E6.1. Expression analysis and evaluation of their functions were examined. The results showed good expression and function.