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Expression of adenovirus type 2 late genes in HeLa cells infected with adenovirus type 2 or adenovirus type 2-simian virus 40 hybrid viruses.

The relative levels of expression of the five 3'-coterminal mRNA families encode in the 16.4 to 99 map unit transcription unit of adenovirus type 2 (Ad2) are determined in part by the frequency of utilization of five possible polyadenylation sites. The possibility that polyadenylation frequencies at these sites may be regulated was tested by examining the expression of the fiber (polypeptide IV) gene in cells infected with nondefective (ND) Ad2-simian virus 40 hybrid viruses which express a sixth late mRNA family. Both the relative rate of fiber protein synthesis and the relative abundance of fiber mRNA were normal in cells infected with the hybrid viruses Ad2+ND1, Ad2+ND2, or Ad2+ND4.

Adenoviruses, Human↗

Persistent infection with adenovirus types 5 and 6 in lymphoid cells from humans and woolly monkeys.

Chronic infection with adenovirus types 5 and 6 was established in primary mononuclear leukocytes from human umbilical cord blood and in Epstein-Barr virus (EBV)-transformed B lymphocytes from human umbilical cord blood and from woolly monkey blood. Adenovirus could be recovered from cultures of primary leukocytes and of EBV-transformed lymphocytes for two and three months, respectively, without visible alteration of cell growth. Infection in cultures of EBV-transformed lymphocytes from woolly monkey blood was obliterated by exposure to antibody, but EBV-transformed lymphocytes from human umbilical cord blood contained small amounts of virus for prolonged periods that restored infection in the culture when antibody was removed. Thus, chronic infection of lymphoid cells by some adenoviruses is maintained by at least two mechanisms: cell-to-cell spread of virus in the absence of antibody and intracellular persistence of infectious virus in the presence of antibody.

Adenoviridae Infections↗

The induction of cellular DNA synthesis in G1-arrested BKH21 cell infected with adenovirus type 2.

G1-arrested BHK21 cells infected with adenovirus type 2 were studied to determine the effect of infection on host DNA synthesis in a productive cycle of infection. At various intervals following infection, analysis of the intracellular DNA was carried out by zonal sedimentation centrifugation through alkaline sucrose gradients. In addition to material which sedimented to the position of the virus markers (34S), a class of high mol. wt. DNA (40 to 100S) was found to be synthesized beginning approx. 13 h p.i. and continuing up to about 35 h p.i. DNA--DNA hybridization studies on this newly synthesized DNA showed it to be cellular DNA. When this material was density labelled with 5-bromodeoxyuridine and centrifuged to equilibrium through alkaline CsCl gradients, it was found to be the product of semi-conservation replication and not of repair synthesis.

Adenoviruses, Human↗

An epidemic of adenovirus 7a infection in a neonatal nursery: course, morbidity, and management.

An epidemic of adenovirus 7a in our neonatal intensive care nursery and intermediate care nursery in July and August 1987 caused the death of two patients. Significant symptomatic infection possibly due to the virus occurred in nine patients, ten staff, and three parents, of whom three patients, three staff, and one parent were positive by culture. As a direct consequence of the outbreak, 58 staff days of work were lost; the intensive care nursery had to be closed to admissions for 19 days and the intermediate care nursery for 14 days. Seventeen newborns were transferred to other hospitals and four mothers were sent elsewhere for delivery. Control measures, which included cohorting of patients, use of gloves, gowns and goggles, and exclusion of symptomatic staff from the unit, appeared effective. Rapid immunofluorescence testing of virological specimens was of little use in monitoring the outbreak, largely because of poor specimen quality. This outbreak further underlines the ease of transmission and high morbidity of neonatal adenovirus infection.

Adenoviridae Infections↗

[Research of modulation of CD95-mediated apoptosis in lymphoblastic MP-1 and BJAB cells infected by adenovirus and Epstein-Barr virus].

Model systems of infecting limphoblastic MP-1 and BJAB cells by Epstein-Barr virus, 5 serotype adenovirus and double infection are developed. A rather high level of accumulation of DNA of these viruses in the cells in dynamics at monoinfection and inhibition interference at multi-infection was shown by PCR method. The influence of virus infection on proliferative activity was studied. The stimulation of cells growth in the system BJAB + EBV was detected, and double infecting inhibited the process by 50%. The 25% difference in development of apoptosis process between cells infected by adenovirus and EBV was established when defining CD95-mediated apoptosis in infected MP-1 cells. The infecting of BJAB cells by viruses had a scarce effect on the processes of spontaneous apoptosis, but the data on CD95-mediated apoptosis at EBV infection testify to inhibition of this process both at a monoinfection, and at a double infection. The work was performed in the framework of the fundamental agreement of Ministry of Education and Science of Ukraine F7/366-2001, and grant INTAS N011-2382.

Adenoviridae↗

Reciprocal regulation of plasmacytoid dendritic cells and monocytes during viral infection.

Reciprocal regulation of opposing functions characterizes biological systems. We now show that adenovirus-infected plasmacytoid dendritic cells (PDC) inhibit monocyte to myeloid dendritic cell (MDC) differentiation and function, and that adenovirus-infected monocytes inhibit PDC type I interferon secretion. Adenovirus-infected PDC secreted IFN-alpha, beta and omega in an 86:2:1 ratio. PDC type I interferons inhibited MDC differentiation and function (reduced IL-12 secretion, IFN-gamma induction, MLR and CD40 expression, and increased CD1a(+)CD14(+) cells). Type I interferon receptor blocking antibody reversed all PDC effects, and recombinant IFN-alpha, beta or omega replicated all effects, except reduced CD40. Adenovirus-infected monocytes suppressed PDC type I interferon secretion, which was reversed with anti-IL-10 neutralizing antibodies. Exogenous IL-10 suppressed PDC type I interferon secretion without reducing PDC viability. Therefore, monocyte IL-10 regulates PDC type I interferon secretion, and PDC type I interferons inhibit MDC differentiation and function. Such reciprocal regulation of potentially opposing influences may help modulate anti-pathogen immunity.

Adenovirus Infections, Human↗

Serological detection of adenoviruses in non-human primates maintained in a colony in Kenya.

BACKGROUND: Adenoviruses are known to cause several human diseases including acute febrile respiratory syndromes, epidemic conjunctivitis and gastroenteritis. These diseases associated with adenovirus infection affect adults and are usually more severe in infants and children. Forty-seven human adenoviruses serotypes have so far been identified adenovirus. The diversity of these viruses has delayed progress on vaccine development due to difficulties in identifying appropriate vaccine targets. To date, limited studies have been done to determine the prevalence of adenovirus infection in non-human primates with the goal of developing a non-human primate model that can be used to study the mechanisms of infection. OBJECTIVE: To determine the prevalence of enteric adenovirus infection in Kenyan non-human primates. DESIGN: A prospective study to investigate the prevalence of enteric andenovirus infection in captive non-human primates maintained in a colony. SETTING: Faecal samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. SUBJECTS: Ninety four faecal samples were collected from three species of non-human primates consisting of various ages and sex. Samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. All the faecal samples were screened for presence of adenoviruses using a commercial antigen-capture enzyme immunoassay (EIA) kit, this is an enzyme-linked immunosorbent assay (ELISA) kit designed for diagnosis of human enteric adenoviruses in stool samples. RESULTS: The highest prevalence of adenoviruses, detected by EIA kit, was in olive baboons (Papio anubis, 52.9%), followed by vervet monkeys (Cercopithecus aethiops, 48.9%) and the yellow baboons (Papio cynocephalus, 18.8%). Sub-grouping within each species (based on age and sex) indicated no significant differences (p > 0.05) in adenovirus infection signifying equal susceptibility. The prevalence of adenoviruses in vervet monkeys that were also Simian Immunodeficiency virus (SIV) seropositive was determined and shown to be 63.2%. CONCLUSION: The results of this study indicate that adenovirus infection is prevalent among non-human primates in Kenya. These findings suggest that cross species transmission in Kenyan non-human primates may be a common occurrence and there is a possibility of zoonotic transmission of adenoviruses. Furthermore, our results highlight the potential of using these non-human primates as models for testing safety and efficacy of candidate adenovirus vaccines prior to clinical trials in humans.

Adenoviridae↗

A model of latent adenovirus 5 infection in the guinea pig (Cavia porcellus).

A model of adenovirus 5 (Ad5) infection was developed in guinea pigs to begin to study its role in the pathogenesis of peripheral lung inflammation. Forty animals were inoculated intranasally with 10(7.0) pfu of Ad5/animal, and 15 animals inoculated with sterile culture media served as controls. Viral titres were 10(4.4), 10(6.1), 10(5.2), and 10(2.9) pfu/animal, on days 1, 3, 4, and 7 after infection, respectively. In situ hybridization to viral DNA and immunocytochemistry for Ad5 E1A protein localized the virus to airway and alveolar epithelial cells. Histologic examination showed an extensive inflammatory cell infiltration around the airways, with epithelial necrosis and an alveolar exudate that caused localized alveolar collapse in the infected areas. Immunocytochemistry identified the cells in the infiltrate as cytotoxic T cells. Although all animals 20 and 47 days after infection had seroconverted to Ad5, virus was not detected in these groups either by viral plaque assay or in situ hybridization. Ad5 E1A DNA was detected by polymerase chain reaction in five of six animals 20 days after infection and in five of five animals 47 days after infection. In these same animals, E1A protein was detected 20 days after infection in two and 47 days after infection in one while persistent bronchiolitis was observed in four and three animals 20 and 47 days after infection, respectively. These results demonstrate that the guinea pig provides a useful model to study the role of Ad5 infection in chronic airway inflammation.

Adenovirus E1A Proteins↗

Detection of virus-specific DNA and RNA base-sequences in individual cells transformed or infected by adenovirus type 2.

By means of 3H thymidine-labelled adenovirus 2 DNA, adenovirus-specific DNA sequences have been localized in individual nuclei of adenovirus 2- or 12-infected cells, and adenovirus-specific RNA sequences have been detected in the permissive cells as well as in cells transformed by adenovirus type 2. The ability to detect such specific virus RNA base-sequences in particular, by in situ hybridization, should be useful in studying the transcriptional specificities of tumours and neoplasms.

Adenoviridae↗

An interaction between penton base and alpha v integrins plays a minimal role in adenovirus-mediated gene transfer to hepatocytes in vitro and in vivo.

Studies in cultured cell lines have shown that adenovirus infection involves binding of adenovirus fiber to its cell surface receptor and binding of penton base to alpha v integrins. However, much less is known about the role of these interactions in cells that are targets for adenovirus-mediated gene transfer. Earlier work showed that hepatocytes are readily infected by adenovirus, making them an attractive target for gene therapy in several diseases. We found that addition of fiber protein blocked adenovirus infection of primary cultures of hepatocytes. This suggests an important role for fiber and its receptor. However, mutation of the integrin-binding motif in penton base did not inhibit infection of hepatocytes, even though the mutation impaired infection of HeLa cells. Hepatocytes had undetectable amounts of alpha v integrins on their cell surface and showed no specific adherence to vitronectin, the natural substrate of alpha v integrins. Adenovirus with an intact penton base enhanced infection of liver following intravenous injection, but only by three-fold as compared with virus in which the integrin-binding motif was disrupted. These studies suggest that interactions between cell surface integrins and penton base are not required for adenovirus infection of hepatocytes in vitro, but the interaction enhances infection to a small degree in vivo.

Adenoviridae↗

Immunofluorescence of green monkey kidney cells infected with adenovirus 12 and with adenovirus 12 plus simian virus 40.

Malmgren, Richard A. (National Cancer Institute; Bethesda, Md.), Alan S. Rabson, Paula G. Carney, and Frances J. Paul. Immunofluorescence of green monkey kidney cells infected with adenovirus 12 and with adenovirus 12 plus simian virus 40. J. Bacteriol. 91:262-265. 1966.-Immunofluorescence studies of the viral antigens and tumor (T) antigens of adenovirus 12 and simian virus 40 (SV40) in green monkey kidney (GMK) cells infected with adenovirus 12 alone or in combination with the SV40 virus showed that the adenovirus 12 viral antigen was produced in detectable amounts only in the cells infected with both viruses. The adenovirus 12 T antigen, on the other hand, was formed in the GMK cells infected with the adenovirus 12 only. This antigen was formed as early as 18 hr after viral infection, and persisted for at least 48 hr after virus infection. There was a correlation between the appearance of the immunofluorescent T antigen in the nucleus and the electron microscope appearance of "nuclear stippling," which developed in the nuclei of GMK cells after infection with adenovirus 12 only, as well as after infection with both viruses.

Journal Article↗

Enhanced infectivity of adenovirus type 2 DNA and a DNA-protein complex.

The infectivity of adenovirus type 2 DNA and a DNA-protein complex was studied in 293 cells, a human embryonic kidney cell line transformed by sheared adenovirus type 5 DNA, and in human KB cells. Adenovirus type 2 DNA was more infectious (up to about 40-fold) in 293 cells than in KB cells, whereas a DNA-protein complex (prepared by a rapid procedure) had about the same infectivity in both cell lines. These data may mean that a factor present in 293 cells (perhaps a viral-coded protein) enhances the infectivity of free viral DNA. The infectivity of DNA and the DNA-protein complex was increased up to fivefold by brief treatment of cell monolayers with 25% dimethyl sulfoxide after transfection. Under these conditions, (i) the infectivity of native adenovirus type 2 DNA ranged from 400 to 1,300 PFU/microgram of DNA in 293 cells and from about 9 to 14 PFU/microgram of DNA in KB cells, and (ii) the infectivity of the DNA-protein complex was 6 X 10(3)to 2 X 10(4) PFU/microgram in 293 cells and 1.4 X 10(4) to 1.6 X 10(4) PFU/microgram in KB cells.

Adenoviruses, Human↗

[Characteristics of several indices of the mitotic regime of clone strains of HeLa cells infected with adenovirus type 7].

The effect of adenovirus type 7 on the mitotic activity, the level and qualitative characteristics of pathological mitoses was studied in mycoplasma-free and latently mycoplasma-infected cells of clone HeLa cell lines with different sensitivity to this virus. The level and the extent of changes of the mitotic regimen in adenovirus-infected cells depend upon their susceptibility to the virus and initial characteristics of the mitotic regimen of the culture which is somehow related to the presence or absence of mycoplasmal contaminants in the culture. Successful analysis of the virus effect on the mitotic regimen of one or another cell system requires the use of a culture with minimal pathology of the mitotic apparatus and careful tests for the lack of mycoplasmal contaminants.

Adenoviridae↗

[Laboratory procedures in adenoviruses. VIII. Immunoglobulin class-specific ELISA for the demonstration of antibodies against adenovirus hexon].

Serum pairs from 114 patients with adenovirus infections (mostly ocular disease in adults) were investigated by an ELISA, with purified adenovirus 5 hexon coated to the solid phase, diluted sera, and peroxydase-labeled antihuman IgG, IgA, and IgM antibodies. Serum dilutions of 1:3200, 1:200, and 1:100 respectively proved to be optimal to demonstrate antibody rises. Sera from infants served as negative controls. Antibody rises were found in the IgG and/or IgA test 85% of the patients, but only in 56% by complement fixation. Antibody rises in all three ELISA tests occurred mostly around the 10th day after onset of disease. Fewer antibody rises and weak reactions were found in IgM test. Moreover, IgM antibodies were present also in persons without fresh adenovirus infection. Hence, this test is not suitable for an early diagnosis. This observation is probably due to the frequent stimulation of the immune system by the genusspecific determinant alpha of the hexon by repeated adenovirus infections during life.

Adenoviridae Infections↗

p53 status does not determine outcome of E1B 55-kilodalton mutant adenovirus lytic infection.

The ability of the adenovirus type 5 E1B 55-kDa mutants dl1520 and dl338 to replicate efficiently and independently of the cell cycle, to synthesis viral DNA, and to lyse infected cells did not correlate with the status of p53 in seven cell lines examined. Rather, cell cycle-independent replication and virus-induced cell killing correlated with permissivity to viral replication. This correlation extended to S-phase HeLa cells, which were more susceptible to virus-induced cell killing by the E1B 55-kDa mutant virus than HeLa cells infected during G1. Wild-type p53 had only a modest effect on E1B mutant virus yields in H1299 cells expressing a temperature-sensitive p53 allele. The defect in E1B 55-kDa mutant virus replication resulting from reduced temperature was as much as 10-fold greater than the defect due to p53 function. At 39 degreesC, the E1B 55-kDa mutant viruses produced wild-type yields of virus and replicated independently of the cell cycle. In addition, the E1B 55-kDa mutant viruses directed the synthesis of late viral proteins to levels equivalent to the wild-type virus level at 39 degreesC. We have previously shown that the defect in mutant virus replication can also be overcome by infecting HeLa cells during S phase. Taken together, these results indicate that the capacity of the E1B 55-kDa mutant virus to replicate independently of the cell cycle does not correlate with the status of p53 but is determined by yet unidentified mechanisms. The cold-sensitive nature of the defect of the E1B 55-kDa mutant virus in both late gene expression and cell cycle-independent replication leads us to speculate that these functions of the E1B 55-kDa protein may be linked.

Adenovirus E1B Proteins↗

Persistent enteral infections with adenovirus types 1 and 2 in infants: no evidence of reinfection.

Isolates of adenovirus types 1 and 2, obtained from 11 infants with prolonged faecal excretion (up to 515 days), were compared by DNA restriction analysis with seven standard endonucleases which recognize hexanucleotides and two additional endonucleases which recognize tetranucleotides. In all instances identical genome types were identified in isolates obtained early and late after infection. Our interpretation of these data is that a chronic persistent infection occurred in these children, and not a reinfection with the same serotype.

Adenoviridae Infections↗

Flow cytophotometric and time-lapse cinematographic study of human cells infected by adenovirus type 2 wild-type and two DNA-negative temperature-sensitive mutants.

Flow cytophotometry and time-lapse cinematography were used to study viral DNA synthesis and alteration of the cellular DNA content of human cells infected by adenovirus type 2 wild-type and two DNA-negative temperature-sensitive mutants. Cell populations with DNA contents greater than 4n (where n represents the normal haploid DNA content of cells) were found after infection and their origin is discussed with regard to cell cycle changes, viral DNA replication and cell fusion or endomitosis.

Adenoviridae Infections↗