Retrovirus-Mediated Gene Transfer of PMP22 in Schwann Cells: Studies on Cell Growth

Background: We aimed to reveal if low dose X-rays would induce harmful or beneficial effect or dual response on biological cells and whether there are conditions the radiation can enhance gene transfer efficiency and promote cell growth but without damage to the cells. Method: A systematic study was performed on the effects of Kilo-V and Mega-V X-rays on the cell morphology, viability, membrane permeability, DNA damage, and gene transfection of 293 T and CHO cells. Results: The Kilo-V X-rays of very low doses from 0.01 to 0.04 Gray in principle didn’t induce any significant change in cell morphology, growth, membrane permeability, and cause DNA damage. The Mega-V X-ray had a damage threshold between 1.0 and 1.5 Gray. The 0.25 Gray Mega-V-X-ray could promote cell growth and gene transfer, while the 1.5 Gray Mega-V X-ray damaged cells. Conclusion: The very low dose of KV X-rays is safe to cells, while the effects of Mega-V-X-rays are dose-dependent. Mega-V-X-rays with a dose higher than the damage threshold would be harmful, that between 1.0 -1.5 Gray can evoke dual effects, whereas 0.25 Gray MV X-ray is beneficial for both cell growth and gene transfer, thus would be suitable for radiation-enhanced gene transfection.

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Effective gene transfer oflacZ andP0 into Schwann cells of P0-deficient mice

Glia ◽

10.1002/(sici)1098-1136(19990115)25:2<165::aid-glia7>3.0.co;2-l ◽

1999 ◽

Vol 25 (2) ◽

pp. 165-178 ◽

Cited By ~ 23

Author(s):

V�ronique Gu�nard ◽

Beat Schweitzer ◽

Eckhard Flechsig ◽

Silvio Hemmi ◽

Rudolf Martini ◽

...

Keyword(s):

Gene Transfer ◽

Schwann Cells ◽

Deficient Mice

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dsAAV8-mediated gene transfer and β-cell expression of IL-4 and β-cell growth factors are capable of reversing early-onset diabetes in NOD mice

Gene Therapy ◽

10.1038/gt.2011.181 ◽

2011 ◽

Vol 19 (8) ◽

pp. 791-799 ◽

Cited By ~ 11

Author(s):

D F Gaddy ◽

M J Riedel ◽

S Bertera ◽

T J Kieffer ◽

P D Robbins

Keyword(s):

Growth Factors ◽

Cell Growth ◽

Gene Transfer ◽

Early Onset ◽

Nod Mice ◽

Β Cell ◽

Onset Diabetes ◽

Cell Expression

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Dynamics of bacterial cell proliferation inhibited by specific proteins for effective horizontal transmission of the integrative and conjugative element ICEclc

10.1101/512285 ◽

2019 ◽

Author(s):

Sotaro Takano ◽

Akiko Koto ◽

Ryo Miyazaki

Keyword(s):

Cell Growth ◽

Gene Transfer ◽

Horizontal Gene Transfer ◽

Cellular Proliferation ◽

Recipient Cell ◽

Horizontal Transmission ◽

Mobile Dna ◽

Host Chromosome ◽

Growth Impairment ◽

Integrative And Conjugative Element

AbstractThe integrative and conjugative element ICEclc, a prevalent mobile element in proteobacteria, is one of the experimental models for horizontal gene transfer. ICEclc is usually retained in a bacterial chromosome, but can be excised and transferred from the donor to other bacterial lineages. The horizontal transmission is accomplished by developing specialized transfer competent (tc) cells in the donor population. The tc cells entirely dedicate to the ICE transmission by sacrificing their proliferation, which results in an increase in the transfer frequency. The cell growth impairment is mediated by two specific genes, parA and shi, on ICEclc, but its mechanistic details and cellular dynamics are still unknown. We here developed fluorescence reporter strains to monitor intracellular behavior of ParA and Shi proteins as well as host cellular proliferation at the single-cell level. Superresolution imaging revealed that ParA colocalized with the host nucleoid while Shi diffused in cytoplasm during the growth impairment. Mutations in the Walker A motif of ParA diminished the inhibitory effect. Combining quantitative time-lapse microscopy and numerical simulations using mathematical models, we found that ParA and Shi initially blocked cell division and then, as time elapsed, inhibited cellular elongation. The parA-shi locus is highly conserved in other ICEs, and the ParA-Shi-mediated growth inhibition was still observed in different proteobacterial species, suggesting that the ICEs have evolved the system to efficiently distribute themselves in the niche. The results of our study provide mechanistic insight into the novel and unique system on ICEs and help to understand such epistatic interaction between ICE genes and host physiology that entails efficient horizontal gene transfer.Author summaryHorizontal gene transfer is a major diving force for bacterial evolution, which is frequently mediated by mobile DNA vectors, such as plasmids and bacteriophages. Integrative and conjugative elements (ICEs) are a relatively new class of mobile vectors, which normally integrate in a host chromosome but under certain conditions can be excised and transferred from the host to a new recipient cell via conjugation. Recent genomic studies estimated that ICEs are more abundant than plasmids among bacteria. Why so prevalent? One of the characteristics of ICEclc, an ICE model in proteobacteria, is that it develops specialized cells which entirely dedicate to the ICE horizontal transmission by repressing their proliferation. Here, we qualitatively and quantitatively describe two proteins, which are expressed from ICEclc when it transfers, and how they actually inhibit the host cell growth. Our results suggest that the system has evolved for the efficient horizontal transmission and is widely conserved in the ICE family.

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Gene Transfer of CD40-Ligand to Human Dendritic Cells Induces Growth Arrest and Apoptosis of Tumor Cells Mediated by Up-Regulated TRAIL and its Receptors.

Blood ◽

10.1182/blood.v104.11.3454.3454 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3454-3454

Author(s):

Kei Tomihara ◽

Kazunori Kato ◽

Yukari Masuta ◽

Makoto Noguchi ◽

Hiroyoshi Hiratsuka ◽

...

Keyword(s):

Dendritic Cells ◽

T Cell ◽

Cell Growth ◽

Natural Killer Cells ◽

Gene Transfer ◽

Cell Surface ◽

Tumor Cell ◽

Tumor Cells ◽

Growth Arrest ◽

Tumor Cell Growth

Abstract Interaction of CD40 and its ligand CD40L is essential for the activation of dendritic cells (DC) followed by the induction of antigen-specific T-cell responses. Recent studies have also indicated that ligation of CD40 on tumor cells directly modulate tumor cell growth. In this study we examined whether gene transfer of CD40L to DC could exhibit direct anti-tumor effect on CD40-expressing tumors. First, we constructed a modified adenoviral vector driving human CD40L gene with CA promoter/enhancer and incorporating the integrin-binding motif, RGD, in the H1 loop of the fiber knob (AxCAhCD40L-F/RGD). At the same multiplicity of infection, cell surface expression of human CD40L on DC after infection with AxCAhCD40L-F/RGD (CD40L-DC) was induced to express superior higher levels of T-cell costimulatory molecules (CD80, CD86, and CD54), DC maturation markers (CD25 and CD83), MHC molecules and the production of IL-12 than soluble CD40L or TNF-a treated DC. In addition to the phenotypic alternation of CD40L-DC, we found that cultivation of tumor cells (including lymphoma, leukemia, glioma, head and neck carcinoma) with CD40L-DC significantly inhibited the expression of phospho-AKT and tumor cell growth in vitro. Importantly, these tumor cells in transwell cocultured with CD40L-DC, but not with TNF-a-treated DC or immature DC, were significantly induced to express apoptosis-inducing death receptors such as TRAIL-R1, TRAIL-R2 and Fas. The ability of CD40L-DC to inhibit tumor-cell growth and TRAIL receptor induction could not be abrogated by neutralizing antibodies specific for CD40L, IFN-b, IFN-g, and IL-12, indicating that these factors were not involved. Additionally, natural killer cells cocultured with CD40L-DC displayed marked up-regulation of TRAIL on their cell surface. To confirm the biological function of TRAIL and its receptors, we assessed the cytotoxic activity of NK cells to various tumors that were stimulated with CD40L-DC. Tumor cells stimulated with CD40L-DC were susceptible to apoptosis by recombinant TRAIL protein and natural killer cells expressing TRAIL. Overall, these results reveal that CD40L-transfected DC could induce direct tumor cell growth arrest mediated by AKT pathway and susceptibility of apoptosis by TRAIL, indicating that CD40L-DC have potential therapeutic implications which we were currently exploring.

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