A novel cell adhesive protein engineered by insertion of the Arg-Gly-Asp-Ser tetrapeptide

This research was originally published in the Journal of Biological Chemistry. T Maeda, R Oyama, K Ichihara-Tanaka, F Kimizuka, I Kato, K Titani and K Sekiguchi. A novel cell adhesive protein engineered by insertion of the Arg-Gly-Asp-Ser tetrapeptide. J. Biol. Chem. 1989; 264: 15165-15168 © the American Society for Biochemistry and Molecular Biolog

In Vivo Safety and Persistence of Endoribonuclease Gene-Transduced CD4+ T Cells in Cynomolgus Macaques for HIV-1 Gene Therapy Model

BACKGROUND: MazF is an endoribonuclease encoded by Escherichia coli that specifically cleaves the ACA sequence of mRNA. In our previous report, conditional expression of MazF in the HIV-1 LTR rendered CD4+ T lymphocytes resistant to HIV-1 replication. In this study, we examined the in vivo safety and persistence of MazF-transduced cynomolgus macaque CD4+ T cells infused into autologous monkeys. METHODOLOGY/PRINCIPAL FINDINGS: The in vivo persistence of the gene-modified CD4+ T cells in the peripheral blood was monitored for more than half a year using quantitative real-time PCR and flow cytometry, followed by experimental autopsy in order to examine the safety and distribution pattern of the infused cells in several organs. Although the levels of the MazF-transduced CD4+ T cells gradually decreased in the peripheral blood, they were clearly detected throughout the experimental period. Moreover, the infused cells were detected in the distal lymphoid tissues, such as several lymph nodes and the spleen. Histopathological analyses of tissues revealed that there were no lesions related to the infused gene modified cells. Antibodies against MazF were not detected. These data suggest the safety and the low immunogenicity of MazF-transduced CD4+ T cells. Finally, gene modified cells harvested from the monkey more than half a year post-infusion suppressed the replication of SHIV 89.6P. CONCLUSIONS/SIGNIFICANCE: The long-term persistence, safety and continuous HIV replication resistance of the mazF gene-modified CD4+ T cells in the non-human primate model suggests that autologous transplantation of mazF gene-modified cells is an attractive strategy for HIV gene therapy

Role of type III homology repeats in cell adhesive function within the cell-binding domain of fibronectin

This research was originally published in the Journal of Biological Chemistry. F Kimizuka, Y Ohdate, Y Kawase, T Shimojo, Y Taguchi, K Hashino, S Goto, H Hashi, I Kato, K Sekiguchi and K Titani. Role of type III homology repeats in cell adhesive function within the cell-binding domain of fibronectin. J. Biol. Chem. 1991; 266: 3045-3051 © the American Society for Biochemistry and Molecular Biolog

The Double Polymerase Chain Reaction with Consensus Primers Permits Rapid and Sensitive Detection of Genital Human Papillomavirus Oncogenes

We have developed a sensitive procedure for the detection of relatively low copy numbers of multiple genital human papillomaviruses (HPVs) using the polymerase chain reaction (PCR). HPV DNAs were detected by agarose gel electrophoresis and ethidium bromide staining after 2 rounds of PCR amplification (double PCR) with outer and inner consensus primer pairs for HPV-6, 11, 16, 18, 31, 33, 52, and 58. The detection limit of this method (i. e., 10?? copy of HPV DNA per cell in 1 μg cell DNA) was sufficient for analysis of cervical intraepithelial neoplasia (CIN) specimens. Overall prevalence rate of HPV was 100% in 20 cases of CIN specimens. HPV typing by restriction enzyme analysis revealed that HPV-16 sequence was present in 11 cases, HPV-18 in 1 case, HPV-31 in 4 cases, HPV-33 in 1 case, HPV-52 in 2 cases, HPV-58 in 3 cases, and an unidentified type(s) in 3 cases. There were 4 cases of mixed infections. This procedure obviates the use of hybridization- based for-mat for identification of at least 8 types of HPV sequences present in a small fraction of cells within a heterogeneous population

DNA Microarray Analysis of the Expression Profile of Escherichia coli in Response to Treatment with 4,5-Dihydroxy-2-Cyclopenten-1-One

We carried out DNA microarray-based global transcript profiling of Escherichia coli in response to 4,5-dihydroxy-2-cyclopenten-1-one to explore the manifestation of its antibacterial activity. We show that it has widespread effects in E. coli affecting genes encoding proteins involved in cell metabolism and membrane synthesis and functions. Genes belonging to the regulon involved in synthesis of Cys are upregulated. In addition, rpoS and RpoS-regulated genes responding to various stresses and a number of genes responding to oxidative stress are upregulated