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Effects of the ligand sequence modifications on the retargeted transduction by the retroviral vector having a ligand-chimeric Env protein

Kei Miyakawa^1,

Masumi Katane^1,

¹ Discovery Research Institute, RIKEN, Wako, Saitama 351-0192, Japan
² Department of AIDS Research, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan

Correspondence
Hiroshi Amanuma
h-amanuma{at}nihs.go.jp

There have been various attempts to redirect the cell entry receptor tropism of the murine leukemia virus vectors. We have recently reported the successful retargeting of the ecotropic Moloney murine leukemia virus vector. This vector (S3-D84K) contains a viral envelope (Env) protein into which a full-length (68 aa) stromal cell-derived factor-1alpha (SDF-1) was inserted at Pro-79. The S3-D84K vector transduces a certain human cell line through the CXC chemokine receptor 4 (CXCR4) at a titre of about 10⁴ c.f.u. ml^–1. Here, the S3-D84K vector was found to transduce another human cell line through CXCR4 with a titre close to 10⁶ c.f.u. ml^–1. The SDF-1 ligand of the S3-D84K Env protein was modified in different ways. In one, C-terminal truncations (by 3–51 aa) with or without a Cys-to-Gly change were performed, and in the other, Cys-to-Ala changes of the disulfide-forming cysteines without truncation were made. Seven truncation and three alanine mutant chimeric Env proteins were examined for virion incorporation, and the retroviral vectors displaying the mutant protein were examined for CXCR4 binding and retargeted transduction. Two mutant vectors showed transduction through CXCR4 with titres not higher than those of the S3-D84K vector, while the other mutant vectors minimally transduced cells through CXCR4 either due to a defect in virion incorporation of the chimeric Env protein or an inability to bind to CXCR4. These results suggest that a full-length sequence that may fold into a distinct domain within the chimeric Env protein is preferable as a targeting ligand.

Present address: Department of Molecular Virology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan.

Present address: Laboratory of Biomolecular Science, School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.

Supplementary material is available with the online version of this paper.