title: The construction of chimeric T-Cell receptor with spacer base of modeling study of VHH and MUC1 interaction.
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.
authors.affiliation: Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran.
authors.forename: Nazanin
authors.initials: N
authors.lastname: Pirooznia
authors.forename: Sadegh
authors.initials: S
authors.lastname: Hasannia
authors.forename: Majid
authors.initials: M
authors.lastname: Taghdir
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Rahbarizadeh
authors.forename: Morteza
authors.initials: M
authors.lastname: Eskandani
mesh_terms: D000818:Animals; D001665:Binding Sites; D004587:Electrophoresis, Agar Gel; D006801:Humans; D019169:Jurkat Cells; D056004:Molecular Dynamics Simulation; D018396:Mucin-1 / Q000737:chemistry* / Q000378:metabolism*; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry* / Q000378:metabolism*; D017452:Receptors, IgG / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000378:metabolism*; D055672:Static Electricity; D013696:Temperature
chemical_list: C080568:Fc gamma receptor IIA; D018396:Mucin-1; D011948:Receptors, Antigen, T-Cell; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies