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PubMed1989

Enzyme-linked immunosorbent assay (ELISA) for detecting antibodies in sera of patients with adenovirus infection.

The 41 distinct antigenic types of adenoviruses (Ads) are responsible for a broad spectrum of diseases in humans. We have developed an enzyme-linked immunosorbent assay (ELISA) using adenovirus (Ad) infected MRC-5 cells for detecting IgG and IgM antibodies to Ads. Using the ELISA, we detected IgG antibodies in 100% (20/20) of sera from normal adults (geometric mean titer, GMT = 1840.8, range = 40-20,480) and IgM antibodies in 3 of 20 sera (15%) with a GMT of 25.1. Our indirect immunofluorescence (IF) technique also detected IgG antibodies in 100% of these sera (GMT = 248.3, range = 40-5,120) and IgM antibodies in the 3 samples reactive in ELISA (GMT = 20.0, range = less than 5-40). In contrast, the complement fixation (CF) test detected antibodies to Ads in only 65% (13/20) of these sera (GMT = 10.9, range = less than 4-32). Moreover, IgG and IgM responses could not be distinguished using CF. Thus the sensitivity of these three techniques is greatest for ELISA. Additionally, a study of sequential sera from 3 patients with acute Ad infection disclosed seroconversion using all three methods. Both the ELISA and IF techniques permit the detection of transition from IgM to IgG, whereas CF only detects conversion from seronegativity to seropositivity. Finally, preliminary data suggest that the IgM response as measured by ELISA is specific for subgroups or types of Ad. This newly devised ELISA may be useful for detecting Ad infections.

Adenoviridae Infections↗
PubMed1977

Nuclear matrix of HeLa S3 cells. Polypeptide composition during adenovirus infection and in phases of the cell cycle.

A subnuclear fraction has been isolated from HeLa S3 nuclei after treatment with high salt buffer, deoxyribonuclease, and dithiothreitol. This fraction retains the approximate size and shape of nuclei and resembles the nuclear matrix recently isolated from rat liver nuclei. Ultrastructural and biochemical analyses indicate that this structure consists of nonmembranous elements as well as some membranous elements. Its chemical composition is 87% protein, 12% phospholipid, 1% DNA, and 0.1% RNA by weight. The protein constituents are resolved in SDS-polyacrylamide slab gels into 30-35 distinguishable bands in the apparent molecular weight range of 14,000 - 200,000 with major peptides at 14,000 - 18,000 and 45,000 - 75,000. Analysis of newly synthesized polypeptides by cylindrical gel electrophoresis reveals another cluster in the 90,000-130,000 molecular weight range. Infection with adenovirus results in an altered polypeptide profile. Additional polypeptides with apparent molecular weights of 21,000, 23,000, and 92,000 become major components by 22 h after infection. Concomitantly, some peptides in the 45,000-75,000 mol wt range become less prominent. In synchronized cells the relative staining capacity of the six bands in the 45,000-75,000 mol wt range changes during the cell cycle. Synthesis of at least some matrix polypeptides occures in all phases of the cell cycle, although there is decreased synthesis in late S/G2. In the absence of protein synthesis after cell division, at least some polypeptides in the 45,000-75,000 mol wt range survive nuclear dispersal and subsequent reformation during mitosis. The possible significance of this subnuclear structure with regard to structure-function relationships within the nucleus during virus replication and during the life cycle of the cell is discussed.

Adenoviridae↗
PubMedDate not supplied

Disseminated adenovirus infection in two premature infants.

We present two premature infants with disseminated neonatal adenovirus infection, whose epidemiology, clinical course and outcome differ to a great extent. The first infant, born vaginally at 35 weeks gestational age after premature rupture of membranes and maternal illness, developed pneumonia, hepatitis and coagulopathy and died of circulatory failure at the age of 17 days. The other infant, delivered by cesarean section at 36 weeks gestational age, did - in contrast to all documented cases in the literature - not show any signs of pneumonia and survived meningitis without sequelae. The mode of transmission of the viral infection may have been via the maternal birth canal in the first infant and transplacental in the second one. Diagnosis was obtained by direct immunofluorescent test and serology in the first patient and by maternal serology and the detection of viral antigen in tracheal aspirates (ELISA) in the second patient. Disseminated neonatal adenovirus infection has a high mortality and should be considered in the differential diagnosis of neonatal sepsis, especially when pneumonia, hepatitis and neurologic symptoms develop together with thrombocytopenia or disseminated intravascular coagulopathy.

Adenovirus Infections, Human↗
PubMed1977

Adenovirus infection in lambs. II. Experimental infection of lambs.

Adenovirus strain GY/14 isolated during a natural outbreak was used in experimental infection. Three weeks old lambs responded with temperature rise, respiratory symptoms and diarrhoea to the infection. Infection spread to a contact animal, too. Reisolation of the virus was successful from the nasal discharge and feces from the 3rd to 10th, and the 3rd to 5th day following infection, respectively. In the killed experimental animals the pathological and histological changes observed were similar to those observed in natural cases. On comparing the natural outbreaks with the experimental infection the only difference appeared in the severity of the changes. Following the experimental infection characteristic nuclear inclusions appeared in the nasal and bronchiolar epithelium, in the alveolar septal cells and in the reticular cells of the lymph nodes. Epizootiologic observations and experimental results confirm the assumption that our adenovirus strains isolated from natural cases are pathogenic for lambs.

Adenoviridae Infections↗
PubMed1964

Activity of aspartate transcarbamylase in uninfected and type 5 adenovirus-infected HeLa cells.

Consigli, Richard A. (University of Pennsylvania, Philadelphia), and Harold S. Ginsberg. Activity of aspartate transcarbamylase in uninfected and type 5 adenovirus-infected HeLa cells. J. Bacteriol. 87:1034-1043. 1964.-A two- to three-fold increase in aspartate transcarbamylase (ATCase) activity was observed in type 5 adenovirus-infected HeLa cells 18 hr after infection. The enhanced enzyme activity was virus-specific and dependent on biosynthesis of deoxyribonucleic acid and protein. When various characteristics as well as the kinetics of the enzymes from uninfected and infected cells were compared, ATCase from adenovirus-infected cells was shown to have an altered pH optimum, greater heat stability, increased maximal velocity, and increased K(m) value for aspartate.

Adenoviridae↗
PubMed2003

Transforming growth factor beta1 receptor II is downregulated by E1A in adenovirus-infected cells.

Transforming growth factor beta1 (TGF-beta1) signaling is compromised in many tumors, thereby allowing the tumor to escape the growth-inhibitory and proapoptotic activities of the cytokine. Human adenoviruses interfere with a number of cellular pathways involved in cell cycle regulation and apoptosis, initially placing the cell in a "tumor-like" state by forcing quiescent cells into the cell cycle and also inhibiting apoptosis. We report that adenovirus-infected cells resemble tumor cells in that TGF-beta1 signaling is inhibited. The levels of TGF-beta1 receptor II (TbetaRII) in adenovirus-infected cells were decreased, and this decrease was mapped, by using virus mutants, to the E1A gene and to amino acids 2 to 36 and the C-terminal binding protein binding site in the E1A protein. The decrease in the TbetaRII protein was accompanied by a decrease in TbetaRII mRNA. The decrease in TbetaRII protein levels in adenovirus-infected cells was greater than the decrease in TbetaRII mRNA, suggesting that downregulation of the TbetaRII protein may occur through more than one mechanism. Surprisingly in this context, the half-lives of the TbetaRII protein in infected and uninfected cells were similar. TGF-beta1 signaling was compromised in cells infected with wild-type adenovirus, as measured with 3TP-lux, a TGF-beta-sensitive reporter plasmid expressing luciferase. Adenovirus mutants deficient in TbetaRII downregulation did not inhibit TGF-beta1 signaling. TGF-beta1 pretreatment reduced the relative abundance of adenovirus structural proteins in infected cells, an effect that was potentiated when cells were infected with mutants incapable of modulating the TGF-beta signaling pathway. These results raise the possibility that inhibition of TGF-beta signaling by E1A is a means by which adenovirus counters the antiviral defenses of the host.

Adenovirus E1A Proteins↗
PubMed1971

Mechanism of inhibition of vaccinia virus replication in adenovirus-infected HeLa cells.

The ability of vaccinia virus to replicate in HeLa cells which had been previously infected with adenovirus type 2 (Ad2) was studied in order to gain insight into the mechanism by which adenovirus inhibits the expression of host cell functions. Vaccinia virus was employed in these studies because it replicates in the cytoplasm, whereas Ad2 replicates in the nucleus of the cell. It was found that vaccinia deoxyribonucleic acid (DNA) synthesis is greatly inhibited in adeno-preinfected HeLa cells provided that vaccinia superinfection does not occur before 18 hr after adeno infection. The inhibition of vaccinia DNA synthesis can be traced to an inhibition of vaccinia protein synthesis and viral uncoating. Vaccinia ribonucleic acid (RNA) synthesis is not inhibited in adeno-preinfected cells, but the vaccinia RNA does not become associated with polysomes.

Adenoviridae↗
PubMed1978

[Morphology of the adrenal cortex in infants with generalized adenovirus infections].

Morphometric, histological, and immunofluorescent studies were carried out with adrenal glands of 14 infants dying with generalized adenovirus infection sometimes concurrent with other acute viral respiratory diseases. Nonspecific changes were found: edema of the capsule and stroma, discomplexation of the trabeculae, circulatory disorders, delipoidization, an increase in the content of DNA and RNA in cells of the definitive cortex, resorption of the elements of the fetal cortex. The pattern of changes of morphometric values attests to increased functional activity of the adrenal cortex. The intensity of the observed changes was directly related to the duration of the disease. The results of clinico-morphological analysis confirm the importance of adrenal insufficiency in tanatogenesis in generalized adenovirus infection. In 10 cases epithelial and endothelial cells of the definitive cortex were found to have specific lesions, the so-called adenovirus cells.

Adenoviridae Infections↗
PubMed1996

mRNA export correlates with activation of transcription in human subgroup C adenovirus-infected cells.

To investigate the mechanisms by which viral mRNA species are distinguished from their cellular counterparts for export to the cytoplasm during the late phase of subgroup C adenovirus infection, we have examined the metabolism of several cellular and viral mRNAs in human cells productively infected by adenovirus type 5 (Ad5). Several cellular mRNAs that were refractory to, or could escape from, adenovirus-induced inhibition of export of mRNA from the nucleus have been identified. This group includes Hsp70 mRNAs synthesized upon heat shock of Ad5-infected 293 or HeLa cells during the late phase of infection. However, successful export in Ad5-infected cells is not a specific response to heat shock, for beta-tubulin and interferon-inducible mRNAs were also refractory to virus-induced export inhibition. The export of these cellular mRNAs, like that of viral late mRNAs, required the E1B 55-kDa protein. Export to the cytoplasm during the late phase of Ad5 infection of several cellular mRNAs, including members of the Hsp70 family whose export was inhibited under some, but not other, conditions, indicates that viral mRNA species cannot be selectively exported by virtue of specific sequence or structural features. Cellular and viral late mRNAs that can be exported from the nucleus to the cytoplasm were expressed from genes whose transcription was induced or activated during the late phase of Ad5 infection. Consistent with the possibility that successful export is governed by transcriptional activation in the late phase of adenovirus infection, newly synthesized viral early E1A mRNA was subject to export inhibition during the late phase of infection.

Adenovirus E1B Proteins↗
PubMed1977

[Morphological changes in the kidneys of pigs with an adenovirus infection].

Dystrophic and necrobiotic changes developed in the epithelium of the kidney tubuli of sucking pigs spontaneously and experimentally infected with adenoviruses. The epithelial cells showed hyperchromatosis of the nuclei, breaking up of the chromatin, and formation of nuclear function bodies. The swelling of the dystrophic cells produced certain amount of pressure on the adjacent capillaries. The pale colour of the kidney itselt was due to the dystrophy of the epithelium and the compression anemia. After the desquamation of the degenerated epithelial cells and their carrying away with the urine the capillaries became strongly dilatated. Groups of adjacent capillaries became macroscopically visible and assumed the appearance of a petechia. Edema in lymphoidcell proliferation were established in the interstitial tissue.

Adenoviridae Infections↗
PubMed1984

Association of genital adenovirus infection with urethritis in men.

Adenoviruses were isolated from the urethral swabs of 129 male patients in an STD clinic. After exclusion of patients with Chlamydia trachomatis or Neisseria gonorrhoea infections, 85 of the remaining 120 patients had urethritis, compared with 28 men with urethritis detected in a control group which was closely matched for age, sex, and date of specimen collection. This statistically significant difference suggests that genital adenovirus infection may be a cause of urethritis in some male patients.

Adenoviridae Infections↗
PubMed1997

Association of adenovirus infection with human obesity.

We previously reported that chickens infected with the avian adenovirus SMAM-1 developed a unique syndrome characterized by excessive intra-abdominal fat deposition accompanied by paradoxically low serum cholesterol and triglyceride levels. There have been no previous reports of avian adenoviruses infecting humans. We screened the serum of 52 humans with obesity in Bombay, India, for antibodies against SMAM-1 virus using the agar gel precipitation test (AGPT) method. Bodyweights and serum cholesterol and triglyceride levels were compared in SMAM-1-positive (P-AGPT) and SMAM-1-negative (N-AGPT) groups. Ten subjects were positive for antibodies to SMAM-1, and 42 subjects did not have antibodies. The P-AGPT group had a significantly higher bodyweight (p < 0.02) and body mass index (p < 0.001) (95.1 +/- 2.1 kg and 35.3 +/- 1.5 kg/m2, respectively) compared with the N-AGPT group (80.1 +/- 0.6 kg and 30.7 +/- 0.6 kg/m2, respectively). Also, the P-AGPT group had significantly lower serum cholesterol (p < 0.02) and triglyceride (p < 0.001) values (4.65 mmol/L and 1.45 mmol/L, respectively) compared with the N-AGPT group (5.51 mmol/L and 2.44 mmol/L, respectively). Two subjects positive for SMAM-1 antibodies had antibodies against each others' serum, suggesting the presence of antigens in one or both. When these two serum samples were inoculated into chicken embryos, macroscopic lesions compatible with SMAM-1 infection developed. The inoculation of serum from N-AGPT subjects did not produce such lesions. The presence of increased obesity, antibodies to SMAM-1, reduced levels of blood lipids, and viremia that produces a typical infection in chicken embryos suggests that SMAM-1, or a serologically similar human virus, may be involved in the cause of obesity in some humans.

Adenoviridae↗
PubMedDate not supplied

[Express diagnosis of eye adenovirus infection by fluorescent antibody method].

Adenoviruses often cause epidemic keratoconjunctivitis. The infection is highly contagious, often leads to corneal opacities, and is therefore to be timely diagnosed and properly treated. The authors propose a method for rapid diagnosis of adenovirus infection based on examination of a scraping off the conjunctiva by indirect immunofluorescence with polyvalent fluorescent rabbit serum and offer recommendations on the use of this method. Using this technique, adenovirus antigen was detected in 14 (61%) out of 23 patients with acute follicular conjunctivitis and in 19 (95%) out of 20 patients with keratoconjunctivitis. Despite clinical cure after antiviral therapy, the antigen persisted for 1 month in 22-30% patients and in some patients for 3 months (as a rule, it was in patients treated too late or ineffectively or after corticosteroids). That is why maintenance antivirus therapy is recommended for 2-3 months after clinical cure.

Adenovirus Early Proteins↗