• Title, Summary, Keyword: adenovirus

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Characteristics of Oncolytic Adenovirus Replication and Gene Expression in Hypoxic Condition

  • Kim, Hong-Sung
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.185-190
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    • 2011
  • Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

Helper-Independent Live Recombinant Adenovirus Vector Expressing the Hemagglutinin-Esterase Membrane Glycoprotein

  • YOO, DONGWAN;ICK-DONG YOO;YOUNG-HO YOON;FRANK L GRAHAM;LORNE A. BABIUK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.174-182
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    • 1992
  • The hemagglutinin-esterase glycoprotein (HE) gene of bovine coronavirus, coupled with a simian virus 40 early promoter and polyadenylation signal, was inserted into a human adenovirus transfer vector. The transfer vector was used to co-transfect 293 cells along with adenovirus genomic DNA. The hemagglutinin-esterase transcription unit was rescued into the adenovirus genome by homologous in vivo DNA recombination between the vector plasmid DNA and the adenovirus genomic DNA, and a recombinant adenovirus was isolated by several rounds of plaque assays. Thus the recombinant adenovirus carries the hemagglutinin-esterase gene in the early transcription region 3 (E3) of the adenovirus genome in the parallel orientation to the E3 transcription. The recombinant adenovirus synthesized the HE polypeptide in HeLa cells as demonstrated by immunoprecipitation with anti-coronavirus rabbit antisera. The recombinant HE polypeptide could be labelled by $[^3H]$glucosamine, demonstrating that the recombinant HE was glycosylated. Cells expressing the HE polypeptide exhibited hemadsorption activity when incubated with mouse erythrocytes. The HE was transported to the plasma membrane as shown by the cell surface immunofluorescence, indicating that the recombinant HE polypeptide retained its biological activities. Potential for the use of infectious recombinant adenovirus as a live virus-vectored vaccine candidate for bovine coronavirus disease is discussed.

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Effect of Caffeine on Transformation Induced by Adenovirus type 12 (Adenovirus type 12에 의해 유발된 Transformation에 미치는 Caffeine의 영향)

  • Choi, Sung-Bae
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.269-272
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    • 1996
  • Adenovirus group consists of over 100 related viruses which have been isolated from respiratory or gastro-intestinal tract of primate, cattle, dog and mice. Approximately 40 serologic types of adenovirus producing a variety of human respiratory and conjunctival infections were identified. Adenoviruses are icosahedral virions containing double-stranded linea DNA. They are 70nm to 90nm in diameter and each of capsid is composed of 252 capsomeres. Several numbers of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be on-cogenic in young rat. Previous report involving effect of Hormone on replication of adenovirus(9) has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the effect of caffeine on the transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigation of the effect of caffeine on the adenovirus type 12-induced transformation in L cell. Results were as follows; 1. Adenovirus type 12-induced transformation was inhibited in the presence of caffeine. 2. Yields of adeoovirus type 12 in L cell were slightly inhibited by treatment of caffeine.

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Detection of Adenovirus from Respiratory and Alimentary Tract in Pusan, 1999

  • Cho, Kyung-Soon;Kim, Young-Hee
    • Journal of Life Science
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    • v.10 no.2
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    • pp.17-20
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    • 2000
  • Adenovirus which is an important infectious viral agent in respiratory and alimentary tract was investigated in Pusan, 1999. Fifteen cases of adenovirus were detected from stools and throat swabs of suspected patients. Two cases of enteric adenovirus were detected from a 5 years old boy and a 6-month-old boy. Thirteen cases of respiratory adenoviruses were detected from children aged under 10 years old and one adult. From respiratory specimens, 1 case of adenovirus type 2, 1 case of type 5, and 11 cases of type 3 were found. Enterotype 41 was detected from fecal preparations. Adenoviruses appeared mostly during winter months, January, February and December. Adenovirus showed a slowly progressive cytopathic effect on HEp-2 cells, Vero cells and BGM cells at 37$^{\circ}C$, in a 5-7% $CO_{2}$ incubation. An electron microscopic observation exhibited non-enveloped icosahedron with a diameter of 70nm. No significant differences on cytopathic effect and morphological features have been found from specimens of either alimentary tract or respiratory secretions.

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Enhancement of Adenovirus Type 12 Transformation by N-Methyl-N'-Nitro-N-Nitrosoguanidine (N-Methyl-N'-Nitro-N-Nitrosoguanidine에 의한 Adenovirus Type 12 Transformation의 증진)

  • Choi, Sung-Bae
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.257-260
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    • 1997
  • Adenoviruses are icosahedral virions containing double-stranded linea DNA. They are 70 nm to 90 nm in diameter and capsid is composed of 252 capsomeres. Several members of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be oncogenic in young rat. Previous report involving effect of caffein on transformation induced by Adenovirus type 12 [9] has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the effect of MNNG (N-methyl-N'-nitro-N-nitroguanidine) on transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigate the effect of MNNG on the adenovirus type 12 induced transformation in L cell. Results were as following 1. Adeno virus type 12 induced transformation was enhanced in the presence of MNNG. 2. Yields of adeno type 12 virus in L cell were slightly inhibited by treatment of MNNG.

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Human Papillomavirus E6 Knockdown Restores Adenovirus Mediated-estrogen Response Element Linked p53 Gene Transfer in HeLa Cells

  • Kajitani, Koji;Ken-Ichi, Honda;Terada, Hiroyuki;Yasui, Tomoyo;Sumi, Toshiyuki;Koyama, Masayasu;Ishiko, Osamu
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8239-8245
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    • 2016
  • The p53 gene is inactivated by the human papillomavirus (HPV) E6 protein in the majority of cervical cancers. Treatment of HeLa S3 cells with siRNA for HPV E6 permitted adenovirus-mediated transduction of a p53 gene linked to an upstream estrogen response element (ERE). Our previous study in non-siRNA treated HHUA cells, which are derived from an endometrial cancer and express estrogen receptor ${\beta}$, showed enhancing effects of an upstream ERE on adenovirus-mediated p53 gene transduction. In HeLa S3 cells treated with siRNA for HPV E6, adenovirus-mediated transduction was enhanced by an upstream ERE linked to a p53 gene carrying a proline variant at codon 72, but not for a p53 gene with arginine variant at codon 72. Expression levels of p53 mRNA and Coxsackie/adenovirus receptor (CAR) mRNA after adenovirus-mediated transfer of an ERE-linked p53 gene (proline variant at codon 72) were higher compared with those after non-ERE-linked p53 gene transfer in siRNA-treated HeLa S3 cells. Western blot analysis showed lower ${\beta}$-tubulin levels and comparatively higher p53/${\beta}$-tubulin or CAR/${\beta}$-tubulin ratios in siRNA-treated HeLa S3 cells after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with those in non-siRNA-treated cells. Apoptosis, as measured by annexin V binding, was higher after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with that after non-ERE-linked p53 gene transfer in siRNA-treated cells.

Disseminated adenovirus infection in a 10-year-old renal allograft recipient

  • Lee, Bora;Park, Eujin;Ha, Jongwon;Ha, Il Soo;Cheong, Hae Il;Kang, Hee Gyung
    • Kidney Research and Clinical Practice
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    • v.37 no.4
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    • pp.414-417
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    • 2018
  • Disseminated adenovirus infection can result in high mortality and morbidity in immunocompromised patients. Here, we report the case of a 10-year-old renal allograft recipient who presented with hematuria and dysuria. Adenovirus was isolated from his urine. His urinary symptoms decreased after intravenous hydration and reduction of immunosuppressants. However, 2 weeks later he presented with general weakness and laboratory tests indicated renal failure necessitating emergency hemodialysis. Adenovirus was detected in his sputum; therefore, intravenous ganciclovir and immunoglobulin therapy were initiated. Renal biopsy revealed diffuse necrotizing granulomatous tubulointerstitial nephritis compatible with renal involvement of the viral infection. Adenovirus was detected in his serum. Despite cidofovir administration for 2 weeks, adenovirus was also detected in the cerebrospinal fluid, resulting in generalized tonic-clonic seizure. The patient died 7 weeks after the onset of urinary symptoms. Adenovirus should be considered in screening tests for post-renal transplantation patients who present with hemorrhagic cystitis.

Discrimination of Kawasaki disease with concomitant adenoviral detection differentiating from isolated adenoviral infection

  • Kim, Jong Han;Kang, Hye Ree;Kim, Su Yeong;Ban, Ji-Eun
    • Korean Journal of Pediatrics
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    • v.61 no.2
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    • pp.43-48
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    • 2018
  • Purpose: Human adenovirus infection mimics Kawasaki disease (KD) but can be detected in KD patients. The aim of this study was to determine the clinical differences between KD with adenovirus infection and only adenoviral infection and to identify biomarkers for prediction of adenovirus-positive KD from isolated adenoviral infection. Methods: A total of 147 patients with isolated adenovirus were identified by quantitative polymerase chain reaction. In addition, 11 patients having KD with adenovirus, who were treated with intravenous immunoglobulin therapy during the acute phase of KD were also evaluated. Results: Compared with the adenoviral infection group, the KD with adenovirus group was significantly associated with frequent lip and tongue changes, skin rash and changes in the extremities. In the laboratory parameters, higher C-reactive protein (CRP) level and presence of hypoalbuminemia and sterile pyuria were significantly associated with the KD group. In the multivariate analysis, lip and tongue changes (odds ratio [OR], 1.416; 95% confidence interval [CI], 1.151-1.741; P=0.001), high CRP level (OR, 1.039; 95% CI 1.743-1.454; P= 0.021) and sterile pyuria (OR 1.052; 95% CI 0.861-1.286; P=0.041) were the significant predictive factors of KD. In addition, the cutoff CRP level related to KD with adenoviral detection was 56 mg/L, with a sensitivity of 81.8% and a specificity of 75.9%. Conclusion: Lip and tongue changes, higher serum CRP level and sterile pyuria were significantly correlated with adenovirus-positive KD.

Improvement of the Inferior Epigastric Artery Flap Viability Using Adenovirus-mediated VEGF and COMP-angiopoietin-1 (아래쪽배벽동맥피판의 생존향상을 위한 VEGF와 COMP-angiopoietin-1 유전자 치료)

  • Yoo, Eun Kyung;Son, Daegu;Kim, Hyung Tae;Lee, In Kyu;Choi, Taehyun;Kim, Junhyung;Han, Kihwan
    • Archives of Plastic Surgery
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    • v.36 no.1
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    • pp.1-10
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    • 2009
  • Purpose: Partial necrosis of skin flaps remains a substantial problem in reconstructive surgery. We investigated the potential use of an adenovirus vector encoding the VEGF, COMP-angiopoietin-1 gene in an attempt to promote the viability of the inferior epigastric artery flap in a rat model. Methods: Three by six cm lower abdominal transverse skin flaps, supplied only by the left inferior epigastric artery, were designed. After skin flap elevation, the adenovirus VEGF and adenovirus COMP-angiopoietin-1 were injected into the distal portion of the flap, which has a high tendency of developing flap ischemia. Control animals were injected with the same volume of normal saline. On 3, 7 and 14 days after the flap elevation, the flap survival and vascularization were assessed using Visitrak digital$^{(R)}$, CD31 immunohistochemistry in addition to evaluating the general histological characteristics. Results: There was a significant increase in the mean percentage of flap viability by 89.8%, 91.1% and 94.8% in flaps transfected with adenovirus VEGF, COMP-angiopoietin-1, coadministraion of VEGF and COMP-angiopoietin-1 at seven days, and by 95.6%, 94.8% and 96.3% at 14 days. Histological assessment revealed that there were more blood vessels formed after adenovirus with VEGF, COMP-angiopoietin-1 or VEGF plus COMP-angiopoietin-1 than with adenovirus Lac Z. Conclusion: The results of this study suggest that adenovirus-mediated VEGF, COMP-angiopoietin-1 gene therapy, promote therapeutic angiogenesis in patients that undergo reconstructive procedures.

Effect of Adenovirus-p53 to Non-Small Cell Lung Cancer Cell Lines (Adenovirus-p53이 비소세포폐암세포 성장에 미치는 영향에 관한 연구)

  • 박종호;이춘택;김주현
    • The Korean Journal of Thoracic and Cardiovascular Surgery
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    • v.31 no.12
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    • pp.1134-1146
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    • 1998
  • Background: The tumor suppressor gene p53 is one of the most frequently altered genes in human tumors, including those of the lung. There is now a compelling evidence that wild-type p53 can negatively influence cell growth by causing G1 arrest or by inducing apoptosis. The possibilities of using p53 for gene therapy are also gathering much interest. Material and Method: Our approach towards understanding p53 function would be to study the biological consequences of overexpression of wild-type p53 in normal and tumor cells by using adenovirus vectors capable of giving high levels of the p53 gene product in cells. We have used this vector containing wild-type p53 to infect tumor cells with different p53 status (null, mutant, or wild-type) to confirm that expression of p53 in null or mutant cell lines becomes possible by Adenovirus-p53 transduction, to examine the effects of high levels of p53 expression on the growth properties of tumor cells, to evaluate the role of apoptosis in p53-mediated biological effects, and to examine the effect of Adenovirus-p53 on the tumorigenicities of the lung cancer cell lines in vitro. Result: The results of our study showed that cells expressing endogenous mutant p53 and those devoid of p53 expression altogether were significantly more sensitive to Adenovirus-p53-mediated cytotoxicity compared to tumor cells expressing endogenous wild-type p53 and that overexpression of wild-type p53 induced programmed cell death. Also we knew that Adenovirus-p53 significantly reduced tumor colony formation of human non-small cell lung cancer cell lines, and decreased the growth of pre-formed colonies in vitro. Conclusion: These results suggest that adenovirus is an efficient vector for mediating transfer and expression of tumor suppressor genes in human non-small cell lung cancer cells and that the tumor cells null for p53 or expressing mutant p53 readily undergo apoptosis by Adenovirus-p53.

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