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PubMed2005

Clinical and epidemiologic characteristics of viral infections in a neonatal intensive care unit during a 12-year period.

BACKGROUND: The incidence of viral infections in patients treated in the neonatal intensive care unit (NICU) is not well-known. We summarized the data of all patients with laboratory-confirmed viral infections admitted at the NICU of our hospital during the period of 1992-2003. OBJECTIVES: To determine the incidence of viral infections among infants hospitalized in a NICU, the associated clinical manifestations and their outcome. METHODS: Retrospective analysis of epidemiologic, virologic and clinical data from infants with proven viral infection. The diagnosis viral infection was confirmed by positive viral culture and/or polymerase chain reaction from clinical samples. RESULTS: Viral infection was confirmed in 51 of 5396 infants (1%) admitted to the NICU; 20 (39%) had an enterovirus and parechovirus (EV/PEV) infection, 15 (29%) a respiratory syncytial virus (RSV) infection, 5 (10%) a rotavirus infection, 3 (6%) a cytomegalovirus (CMV) infection, 2 (4%) an adenovirus infection, 2 (4%) a parainfluenza virus infection, 2 (4%) a herpes simplex virus infection, 1 (2%) a rhinovirus infection and 1 (2%) a rubella virus infection. Three of the infants presented at birth with symptomatic rubella virus, CMV or herpes simplex virus infection. RSV infection developed mostly in hospitalized infants (60%), and 93% of infections occurred during the winter (November-March). The clinical presentations of EV/PEV disease were sepsis-like illness, prolonged seizures in term infants and gastrointestinal disease in preterm infants. RSV, parainfluenza virus, rhinovirus and CMV caused respiratory disease, predominantly in preterm infants. Gastrointestinal disease was seen only in preterm infants with adenovirus, rotavirus or EV/PEV infection. Mortality and serious sequelae were high in patients infected with EV/PEV (10 and 15%, respectively). CONCLUSIONS: The incidence of viral infection in the NICU was 1%. Enteroviral infections were the most frequently diagnosed infections, occurred often in term infants born at home and presented with sepsis-like illness or seizures. Preterm infants hospitalized from birth mainly developed gastrointestinal disease caused by rotavirus and adenovirus infection or respiratory disease caused by RSV, parainfluenza and CMV infection. Enteroviruses were responsible for the highest mortality and development of serious sequelae.

Enterovirus↗
PubMed1991

The 10,400- and 14,500-dalton proteins encoded by region E3 of adenovirus form a complex and function together to down-regulate the epidermal growth factor receptor.

In adenovirus-infected cells, the epidermal growth factor receptor (EGF-R) is internalized from the cell surface via endosomes and is degraded, and the E3 10,400-dalton protein (10.4K protein) is required for this effect (C. R. Carlin, A. E. Tollefson, H. A. Brady, B. L. Hoffman, and W. S. M. Wold, Cell 57:135-144, 1989). We now report that both the E3 10.4K and E3 14.5K proteins are required for this down-regulation of EGF-R in adenovirus-infected cells. Down-regulation of cell surface EGF-R was demonstrated by results from several methods, namely the absence of EGF-R autophosphorylation in an immune complex kinase assay, the inability to iodinate EGF-R on the cell surface, the formation of endosomes containing EGF-R as detected by immunofluorescence, and the degradation of the metabolically [35S]Met-labeled fully processed 170K species of EGF-R. No effect on the initial synthesis of EGF-R was observed. This down-regulation was ascribed to the 10.4K and 14.5K proteins through the analysis of cells infected with rec700 (wild-type), dl748 (10.4K-, 14.5K+), or dl764 (10.4K+, 14.5K-) or coinfected with dl748 plus dl764. Further evidence that the 10.4K and 14.5K proteins function in concert was obtained by demonstrating that the 10.4K protein was coimmunoprecipitated with the 14.5K protein by using three different antisera to the 14.5K protein, strongly implying that the 10.4K and 14.5K proteins exist as a complex. Together, these results indicate that the 10.4K and 14.5K proteins function as a complex to stimulate endosome-mediated internalization and degradation of EGF-R in adenovirus-infected cells.

Adenoviridae↗
PubMed2000

Fatal pneumonia in an AIDS patient coinfected with adenovirus and Pneumocystis carinii.

BACKGROUND: Adenovirus infections are common in immunocompromised hosts. However, pulmonary adenovirus infections rarely cause significant morbidity in HIV-infected patients. PATIENT: Here we describe a 27-year-old man with AIDS who presented with tachypnea, hypoxemia and an infiltrate in the upper left lobe on chest X-ray. Bronchoscopy was performed and Pneumocystis carinii was detected in bronchoalveolar lavage (BAL) fluid. Treatment with cotrimoxazole and prednisone initially resulted in improvement, but after 10 days the patient's clinical condition deteriorated rapidly and he died after 23 days due to respiratory failure. RESULTS: On autopsy histopathologic examination showed abundant "smudge cells," suggestive of adenoviral infection. Electron microscopy revealed adenovirus-like particles arrayed in a paracrystalline manner. Subsequent immunohistochemistry confirmed the extensive presence of adenovirus in addition to P. carinii. CONCLUSION: This case demonstrates a pathogenetic role for adenovirus coinfection in P. carinii pneumonia (PCP). Earlier diagnosis, e.g. by PCR analysis of the BAL fluid or transbronchial biopsy, might have led to the consideration of ribavirin treatment.

AIDS-Related Opportunistic Infections↗
PubMed1975

Cytoplasmic inclusions in canine cells infected with infectious canine laryngotracheitis (ICL) adenovirus.

Spherical dark inclusions were observed in both the cytoplasm and nuclei of infectious canine laryngotracheitis (ICL) adenovirus infected MDCK (Madin-Darby Canine Kidney) cells. The distribution of these inclusions in the cells appeared to indicate that they were formed in both the cytoplasm and in the nucleus at about the same time and there did not appear to be movement of these inclusions between the cytoplasm and the nucleus during the early stages of infection. Morphological appearance, 3H-leucine autoradiography, and immunoferritin labelling showed that the cytoplasmic inclusions were similar in nature to the dark nuclear inclusions, and contained the adenovirus hexon antigen, but not the penton base and fiber antigens.

Adenoviruses, Canine↗
PubMed2000

[Bacterial infection in the conjunctiva of patients with adenoviral conjunctivitis].

PURPOSE: We evaluate the microbiological features of mixed infection in adenovirus-infected conjunctiva. SUBJECTS: Isolation of bacteria was performed in 82 samples of adenoviral conjunctivitis at six eye clinics in Japan. METHODS: For microbiological diagnosis, we performed immunochromatography (IC) and polymerase chain reaction--restriction fragment length polymorphism (PCR-RFLP) analysis for detection and serotyping of adenovirus, and PCR for detection of herpes simplex virus (HSV) and Chlamydia trachomatis antigens out of 82 samples with adenoviral conjunctivitis. RESULTS: Pathogenic bacteria were isolated in 6 samples out of 82. Out of these 6 cases, 5 samples were gram-negative rods and most of them were Flavobacterium meningosepticum (4 samples). Adenovirus type 8 was isolated from all these mixed infection cases. However, HSV-1 and Chlamydia trachomatis were not found. CONCLUSIONS: From these results, it is suggested that gram-negative rods, especially F. meningosepticum, are the most common bacteria causing mixed bacterial infection in adenoviral conjunctivitis.

Adenovirus Infections, Human↗
PubMed1998

Differential transmission of adenovirus in a twin pregnancy.

BACKGROUND: Intrauterine adenovirus infection has been associated with nonimmune hydrops. We describe a twin pregnancy in which adenovirus was demonstrated by polymerase chain reaction in the amniotic fluid of one twin afflicted by hydrops. Adenovirus was absent in the amniotic fluid of the other twin. CASE: A 25-year-old, gravida 3, para 1, with twin gestation at 20 weeks' gestation was noted on ultrasound anomaly screen to have hydrops associated with twin B. Polymerase chain reaction analysis of amniotic fluid revealed adenovirus in twin B with none noted in twin A. At 26 weeks' gestation both infants died. CONCLUSION: This case demonstrates differential transmission of adenovirus in a twin pregnancy. One twin developed hydrops fetalis possibly secondary to adenovirus infection.

Adenovirus Infections, Human↗
PubMed2000

The growth inhibitory effect of p21 adenovirus on human bladder cancer cells.

PURPOSE: To evaluate whether p21 (WAF-1/CIP1) should be considered a potential candidate for human bladder cancer gene therapy, we determined: (1) the basal level of p21 expression in bladder cancer cell lines, (2) the response of bladder cancer cells to increased p21 expression following p21 adenovirus infection, and (3) the mechanism of growth inhibition produced by p21 overexpression. MATERIALS AND METHODS: Five established human bladder cancer cell lines and one primary culture derived from an invasive transitional cell carcinoma were used in this study. To examine the effect of p21 protein on the growth of human bladder cancer cells, a recombinant adenovirus vector system containing p21 cDNA, under the control of cytomegalovirus promoter, was constructed. A control virus containing p21 in an antisense orientation was used to eliminate potential artifacts caused by viral toxicity. RESULTS: Human bladder cancer cell lines exhibit variable endogenous p21 levels which correlate with the in vitro growth status. Significant, but highly variable increases in the steady-state level of p21 were detected in p21 adenovirus infected cells. Human bladder cancer cell lines responded heterogeneously to p21 adenovirus infection. Growth of the WH cell line was substantially inhibited in a dose and time-course dependent fashion. The mechanism of p21 growth inhibition was found to be due to G0/G1 arrest and not the induction of apoptosis. In contrast, p21 adenovirus failed to inhibit the growth of T24 bladder cancer cells because T24 cells were resistant to viral infection. The 253J bladder cancer cells exhibited marked sensitivity to adenovirus; substantial growth inhibition was seen with both sense and antisense p21 very early in the time course of infection. CONCLUSIONS: We found significant variation in the basal level of p21 protein expression in several human bladder cancer cell lines. Increased p21 expression as a result of adenoviral infection may be a potent growth suppressor in some human bladder cancer because it elicits cell cycle arrest in G0/G1 stage, but not the induction of apoptosis. Bladder cancer cells exhibit a wide spectrum of sensitivity to adenoviral infection that may be caused by the presence of viral receptor heterogeneity. This wide spectrum of sensitivity has significant basic scientific and clinical implications and warrants further study.

Adenoviridae↗
PubMed2003

Lipoproteins produced by ApoE-/- astrocytes infected with adenovirus expressing human ApoE.

We have developed an astrocyte cell culture system that is attractive for the study of apoE structure and its impact on astrocyte lipoproteins and neuronal function. Primary astrocytes from apoE-/- mice were infected with adenovirus expressing apoE3 or apoE4 and the nascent lipoproteins secreted were characterized. The nascent apoE-containing astrocyte particles were predominantly the size of plasma high density lipoprotein (HDL). ApoE4, in contrast to apoE3, appeared to be distributed in two distinct lipoprotein peaks and the apoE4-containing lipoproteins contained significantly more radiolabeled triglyceride. On electron micrographs the astrocyte particles were both discoidal and spherical in shape with a prevalence of stacked discs in apoE3 particles, but single discs and larger spheres in apoE4 particles. The apoE4 discs were significantly wider than apoE3 discs. These properties of the astrocyte lipoproteins are similar to those obtained from apoE isoform transgenic mice. Astrocyte lipoproteins containing apoE3, but not apoE4, stimulated neurite outgrowth in Neuro-2a cells. These studies suggest that the isoform-specific effects of apoE lipoproteins may involve differences in particle size and composition. Finally we demonstrate the usefulness of this system by expressing a truncated apoE3 (delta202-299) mutant and show preliminary data indicating that a liver X receptor agonist promotes HDL output by the astrocytes without an increase in apoE in the media. This cell culture system is more flexible and allows for more rapid expression of apoE mutants.

Adenoviridae↗
PubMed1998

Enhancement of adenovirus-mediated gene transfer to human bone marrow cells.

Adenovirus infection of CD34+ hematopoietic stem/progenitor cells is dependent on the multiplicity of infection (MOI), time of incubation, the volume in which the co-incubation occurs and the presence or absence of growth factors. Studies revealed that a brief co-incubation (1-8 hours), resulted in low levels of transgene expression, suggesting that adenovirus infection of CD34+ cells occurs slowly, and optimal transduction requires a 24 hour exposure to adenovirus. Infection by Ad/beta-gal or Ad/p53 at a MOI of 500:1 provided a high transduction efficiency but inhibited hematopoietic function. However, treatment at a MOI of 50-100 resulted in efficient transduction (10.7-15.7% positive) without detectable toxicity. Secondary proof of adenovirus transgene expression was demonstrated by detection of mRNA for p53 in Ad/p53 infected stem cells. We conclude that a 24 hour exposure to recombinant adenovirus encoding p53 or beta-gal, at a MOI of 50-100 is optimal for in vitro gene transfer to BM cells and has no significant effect on hematopoietic function. Adenovirus-mediated transduction of BM cells can also be modulated by growth factors (IL-3, GM-CSF and G-CSF) with improved gene delivery and maintenance of hematopoietic function. In summary, adenovirus vectors can be used to transiently transduce stem cells, and conditions have been defined to maximize expression and limit inhibitory effects on CD34+ cells. These data support continued investigation of this vector for local cytokine delivery and purging of stem cell products.

Adenoviruses, Human↗
PubMedDate not supplied

A fatal case of viral pneumonia in a child infected with adenovirus type 3.

This report describes the clinical, roentgenologic, pathologic, and virologic findings in a 2 years and ten months old girl who died from a severe pneumonia. Initially, the patient presented with fever and cough for 2 days. Physically, the patient appeared lethargic, and breathing sounds revealed diffuse rales and wheezing. Hemogram showed mild leukocytosis and lymphocyte predominant. Chest X ray revealed diffuse interstitial infiltration of the right upper lung, left upper and left lower lung field. Bacteria infection was first impressed. Although treated with several antibiotics (ampicillin, cefuroxime, amikacin, penicillin, cephazolin, imipenem and vancomycin) in three different hospitals, the patient's condition went downhill and the patient died 2 weeks later. Finally, adenoviurs type 3 was isolated from sputum specimen taken before death and necropsy lung tissue. The lung pathology showed diffuse necrotizing inflammation with fibrinopurulent exudate, and eosinophilic intranuclear inclusion bodies were also noted in the alveolar cells. These data proved the diagnosis of adenovirus pneumonia. This case adds to the evidence that adenovirus type 3 infection during childhood may cause fatal disease.

Adenovirus Infections, Human↗
PubMed2005

Quantitation of adenovirus genome during acute infection in normal children.

BACKGROUND: Adenovirus infection causes a wide range of clinical illness in normal children. New molecular techniques allow quantitation of viral genome to study the natural history of adenovirus infection and viral load in normal children. METHODS: Clinical samples were collected from 38 previously healthy, febrile children, and viral cultures were performed. Quantitative polymerase chain reaction (PCR) was used to detect adenovirus genome and to determine viral load. Adenovirus isolates were genotyped with a PCR-based assay. RESULTS: Adenovirus culture was positive in 6 children who were diagnosed with acute adenovirus infection. Throat swabs contained high copy numbers of adenovirus genome (1.6 x 10(6)-6 x 10(7) copies/swab) from 4 of 4 adenovirus culture-positive children. Only 2 of 32 adenovirus culture-negative children had detectable adenovirus genome from throat swabs, but with a lower copy number (8 x 10(2) copies/swab). Adenovirus genome was not detected in blood samples from 5 of 6 adenovirus culture-positive children with uncomplicated upper respiratory tract infection and from all adenovirus culture-negative children. High level viremia (1.8 x 10(8)/ml) was detected in an adenovirus culture-positive 6-month-old infant with fever, pneumonia, conjunctivitis and hepatitis. Subsequent reduction in viral load paralleled her clinical recovery. Adenovirus viruria (1 x 10(9) copies/ml) with normal urinanalysis was detected in another adenovirus culture-positive child. All 6 adenovirus isolates were genotyped as adenovirus type 7h. CONCLUSION: Viral load assessment in clinical samples determined by quantitative PCR can be useful in the diagnosis of adenovirus infection in immunocompetent, febrile children.

Acute Disease↗
PubMed1990

E4F and ATF, two transcription factors that recognize the same site, can be distinguished both physically and functionally: a role for E4F in E1A trans activation.

Previous experiments have identified an element in the adenovirus E4 promoter that is critical for E1A-dependent trans activation and that can confer inducibility to a heterologous promoter. This DNA element is a recognition site for multiple nuclear factors, including ATF, which is likely a family of DNA-binding factors with similar DNA recognition properties. However, ATF activity was found not to be altered in any demonstrable way as a result of adenovirus infection. In contrast, another factor that recognizes this element, termed E4F, was found at only very low levels in uninfected cells but was increased markedly upon adenovirus infection, as measured in DNA-binding assays. Although both the ATF activity and the E4F activity recognized and bound to the same two sites in the E4 promoter, they differed in their sequence recognition of these sites. Furthermore, E4F bound only to a small subset of the ATF recognition sites; for instance, E4F did not recognize the ATF sites in the E2 or E3 promoters. Various E4F and ATF binding sites were inserted into an expression vector and tested by cotransfection assays for responsiveness to E1A. We found that a sequence capable of binding E4F could confer E1A inducibility. In contrast, a sequence that could bind ATF but not E4F did not confer E1A inducibility. We also found that E4F formed a stable complex with the E4 promoter, whereas the ATF DNA complex was unstable and rapidly dissociated. We conclude that the DNA-binding specificity of E4F as well as the alterations in DNA-binding activity of E4F closely correlates with E1A stimulation of the E4 promoter.

Activating Transcription Factors↗
PubMed1978

Stimulation of antibodies to Epstein-Barr virus (EBV) in acute viral infections.

Acute and convalescent sera from 77 patients with serologically confirmed influenza, measles and adenovirus infections and from 36 healthy controls were tested for the level of antibodies to Epstein-Barr (EB) virus. In the three groups of patients significantly higher titers of antibodies to EB viral capsid antigen (VCA) were found as compared to the controls. In 19 patients twofold or higher rise in antibody titers between the first and second blood sample was demonstrated. It is suggested that in patients with influenza, measles or adenovirus infections, involvement of lymphocytes leads to reactivation of EB virus and antibody formation is stimulated.

Acute Disease↗
PubMed1979

Cell surface location of simian virus 40-specific proteins on HeLa cells infected with adenovirus type 2-simian virus 40 hybrid viruses Ad2+ND1 and Ad2+ND2.

HeLa cells infected with the nondefective adenovirus type 2-simian virus 40 hybrid viruses Ad2+ND1 or Ad2+ND2 were analyzed for cell surface location of the SV40-specific hybrid virus proteins by indirect immunofluorescence microscopy. Two different batches of sera from SV40 tumor-bearing hamsters, serum from SV40 tumor-bearing mice, or two different antisera prepared against purified sodium dodecyl sulfate-denatured SV40 T-antigen, respectively, were used. All sera were shown to exhibit comparable T- and U-antibody titers and to specifically immunoprecipitate the SV40-specific proteins from cell extracts of Ad2+ND2-infected cells. Whereas analysis of living, hybrid virus-infected HeLa cells did not yield conclusive results, analysis of Formalin-fixed cells resulted in positive cell surface fluorescence with both Ad2+ND1- and Ad2+ND2-infected HeLa cells when antisera prepared against sodium dodecyl sulfate-denatured SV40 T-antigen were used as first antibody. In contrast, sera from SV40 tumor-bearing animals were not or only very weakly able to stain the surfaces of these cells. The fact that the tumor sera had comparable or even higher T- and U-antibody titers than the antisera against sodium dodecyl sulfate-denatured T-antigen but were not able to recognize SV40-specific proteins on the cell surface suggests that SV40 tumor-specific transplantation antigen may be an antigenic entity different from T- or U-antigen.

Adenoviruses, Human↗
PubMed1984

Disseminated adenovirus serotype 31 infection in an immunocompromised host.

Adenoviruses are ubiquitous organisms but only infrequently have been implicated as a cause of hepatitis and rarely as a cause of fatal disease. A fatal case of disseminated adenovirus serotype 31 infection is described. It occurred in a patient with severe combined immunodeficiency who received a fetal liver transplant and subsequently developed massive hepatic necrosis. To the best knowledge of the authors, this is the first reported case of disseminated disease due to serotype 31.

Adenoviridae Infections↗