The inhibiting action of superhigh-frequency millimeter waves on adenovirus (author's transl)]

Virological and biochemical analysis was employed for comparative studies of the content of protein in continuous cultures of human embryo kidney cells (RH line) infected with adenovirus type 1, intact and irradiated with electromagnetic waves of UHF in the range of 5 divided by 8 mm at the density of power current of the order of 4 mW. When the cells were inoculated with intact virus, a cytopatic effect, a decrease in the content of protein by 4 hours postinfection and further decrease up to 72 hrs were observed. After infection with irradiated adenovirus the development of the infectious process and associated changes in the protein content were observed only by 24 hours and then up to 72 hours there was a delay in the development of virus infection as compared with controls. The experimental data indicate different response of RH cells to infection with irradiated and intact adenovirus, namely, that UHF of the millimeter range (lambda=6.50 mm) exert a specific effect on the infectious activity of adenovirus as manifested by changes in cell metabolism after infection with irradiated virus.     

Pathogenicity and complete genome sequence of a fowl adenovirus serotype 8 isolate

In this study we determined and analyzed the complete nucleotide sequence of the genome of a fowl adenovirus serotype 8 (FAdV-8) isolate and examined its pathogenicity in chickens. The full genome of FAdV-8 was 44,055 nucleotides in length with a similar organization to that of FAdV-1 and FAdV-9 genomes. No regions homologous to early regions E1, E3 and E4 of mastadenoviruses were recognized. Along with FAdV-9, FAdV-8 has only one fiber gene and with regard to sequence composition and genome organization, FAdV-8 is closer to FAdV-9 than to FAdV-1. Moreover, our findings suggest that FAdV-1 of species Fowl adenovirus A as the current type species despite its historical priority is not representative of the genus Aviadenovirus, and that FAdV-8 or FAdV-9 in species Fowl adenovirus E and Fowl adenovirus D, respectively, would be more suitable for that designation. Additionally, pathogenicity of FAdV-8 was studied in specific pathogen free chickens following oral and intramuscular inoculations. Despite lack of clinical signs and pathological changes virus was found in tissues and cloacal swabs of all birds with the highest viral copy numbers present in the cecal tonsils. The highest virus titers in the feces for orally and intramuscularly inoculated chickens were recorded at days 10 and 3 post-infection, respectively.     

Replication of human adenoviruses in guinea pig embryo cells: an ultrastructural study

Summary Guinea pig embryo (GPE) cells showed different degrees of susceptibility to human adenovirus types as determined by virus infectivity assay and electron microscopic examination. Adenovirus 2 and 5 induced extensive cellular changes and produced high titers of infectious virus in GPE cells as in human cells. Mature progeny virus and protein crystals were observed in both cell types. Adenovirus 7 induced some cellular changes in GPE cells but only a small number of cells yielded progeny virus as determined by electron microscopy. Adenovirus 3, 8 and 31 induced some cellular changes but no progeny virus was found under electron microscopic examination. Characteristic fibers were observed in nuclei of adenovirus 31 infected cells. The ability of human adenovirus 2 and 5 to replicate in GPE cells is an example of an unusual cross-species biological property of certain adenovirus types. This property may be useful as a biological marker for these virus types.With 8 Figures     

重组人干扰素α-2b喷雾剂预防SARS等呼吸道病毒感染的人群试验研究

目的评估重组人干扰素α2b喷雾剂(远策素喷雾剂)预防SARS等常见呼吸道病毒感染的效果。方法研究对象共14391人,用药剂量为90万IU/次,每日2次,连用5d,未次用药后15d取血,或用药前和用药3周后采取双份血清。采用随机、对照方法检测血清抗SARSCoVIgG抗体;采用双盲、随机、安慰剂对照方法测定血清抗常见呼吸道病毒(B型流感病毒、副流感病毒1～3型,呼吸道合胞病毒及腺病毒3、7型)的血清IgM抗体。结果两次实验中,干扰素组血清SARS病毒IgG抗体阳性率均较试验组高,但差异无统计学意义(P>0.05)。但用药组应用干扰素后副流感病毒1～3型,B型流感病毒,腺病毒3、7型和呼吸道合胞病毒IgM抗体阳性率(依次为6.45%、4.52%、4.30%和17.20%)均低于对照组(依次为19.40%、13.60%、7.12%和25.62%)。其中副流感病毒、B型流感病毒、腺病毒3种病毒IgM抗体阳性率差异均有统计学意义(P<0.01)。结论应用远策素喷雾剂鼻和咽部喷雾能不同程度地降低用药人群常见呼吸道病毒的感染率。     

A new subgroup 2 bovine adenovirus proposed as the prototype strain 10

Summary A slowly growing subgroup 2 bovine adenovirus (BAV) strain designated Ruakura 78-5371 was isolated from a yearling heifer with systemic adenovirus infection. Cross neutralization tests and restriction endonuclease analysis of the viral DNA showed the virus to be distinct from the other 9 recognised types of BAV. It is proposed that this strain should be regarded as the prototype strain of the new type BAV-10.     

Adenovirus 37: Identification and characterization of a medically important new adenovirus type of subgroup D

A new human adenovirus has been isolated from 62 eyes with (kerato)conjunctivitis and from nine genitourinary sites. The virus is closely related in haemagglutination inhibition tests to adenovirus type 19 (Ad 19) and Ad 10. Antiserum adsorption experiments demonstrated the presence of three haemag-glutinin antigens in the virus: One unique, another common to Ad 19, and a third common to Ad 10 and 19. In neutralization tests, the virus is distantly related to Ad 13, 30, 19, and 10. Despite this relationship, it is proposed to call the virus adenovirus 37, in agreement with current species definitions. It belongs to subgroup D of human adenoviruses. Antisera to the new virus show virtually no neutralization of other human adenovirus types. Only by use of this antiserum it is in practice possible to avoid wrong or indefinite typing, which has often occurred in the past.     

Structure,hemagglutinins, and subgroup position of adenovirus 20, 25, and 28

Summary By ultracentrifugation and red cell adsorption experiments it was proved, that hemagglutinating activity of adenovirus types 20, 25, and 28 against monkey blood cells is associated with the virus particles; while no soluble hemagglutinins were found for type 20 and 25, the soluble hemagglutinating activity of type 28 against rat blood cells was located in a dodecon. The length of the fibres, as observed on the virus surface or in pentons, was 115 to 120 Å for type 20, 25, and 28, coinciding with a reference virus (type 9–15) of subgroup II.These data support the previous suggestion, that adenovirus 20, 25, and 28 belong to subgroup II.Aided by grants of the Deutsche Forschungsgemeinschaft.     

Terminal dilution purification of adenovirus prototypes,and the antigenic relationship between types 4, 16 and 14

Two terminal dilution purifications in HEK cells were performed with adenovirus prototypes of types 1 to 7, 12, 16, 19 and 9-15, and one terminal dilution with a freshly isolated type 8 strain. The antisera prepared against viruses after terminal dilution neutralized the original and the purified virus to the same extent, indicating the antigenic purity of the respective prototypes. Cross-reactions in neutralization with other types within the subgroup were also identical. Only the prototype of type 16 (Ch 79) showed a cross-neutralization with type 4, while four type 16 wild strains showed cross-neutralization with type 14, but none with type 4. The prototype (CH 79) is a "prime strain" in relation to the type 16 wild strains.     

Infection and transformation of mouse cells by human adenovirus type 2

The susceptibilities of C57Bl/6J mouse embryo fibroblasts (MEF) and baby mouse kidney (BMK) cells to infection or transformation by adenovirus type 2 have been compared to those of rat embryo fibroblasts (REF). Both MEF and BMK cells were some 10-fold less permissive to replication of the virus than were REF cells, even though similar fractions of all three cell types, a maximum of 50-60%, produced viral tumor and structural antigens. This observation suggests that a very late step in adenovirus production, such as assembly or maturation, occurs much less efficiently in mouse cells than it does in rat cells. No significant differences in the frequencies of transformation, as assayed by the appearance of foci of morphologically transformed cells, were observed following transfection of adenovirus type 2 or type 5 DNA into the three cell types. However, it proved extremely difficult to establish permanent lines of adenovirus-transformed mouse cells: only 2 of more than 100 attempts were successful, compared to a success rate of close to 100% with adenovirus type 2-transformed REF or SV40-transformed MEF or BMK cells. The two lines of type 2 adenovirus-transformed MEF that were established have been shown to retain and express viral genetic information.     

Effect of Human Adenoviruses on the Response of Chickens to Sheep Erythrocytes

Human adenovirus types 6, 8, and 12 were immunosuppressive in chickens. A single intravenous injection of adenoviruses markedly depressed the 19S hemolytic plaque-forming cell response in the spleen to the immunization with sheep red blood cells. Hemagglutinin production was also decreased in adenovirus type 6-infected chickens. Adenoviruses caused a transient immunosuppression in chickens which could be detected 2 to 3 days after the virus infection, and no depressive effect was found 16 to 20 days after virus injection. The possible mechanism of immunosuppression observed is discussed.     

Quantitative Real-Time PCR Assay Panel for Detection and Type-Specific Identification of Epidemic Respiratory Human Adenoviruses

Outbreaks of human adenovirus (HAdV) acute respiratory illness (ARI) have been well documented among civilians and unvaccinated military recruits. Among the 7 recognized HAdV species (A to G), species B (particularly serotypes 3, 7, 11, 14, and 21) and E (serotype 4) have more often been associated with epidemic ARI. Rapid detection and type-specific identification of these viruses would enhance outbreak response and help guide prevention and control measures. To this end, we developed type-specific real-time quantitative PCR (qPCR) assays for HAdV types 3, 4, 7, 11, 14, 16, and 21 targeting the HAdV hexon gene. All type-specific qPCR assays reproducibly detected as few as 10 copies/reaction of quantified hexon recombinant plasmids with a linear dynamic range of 8 log units (10¹ to 10⁸ copies); in contrast, a generic qPCR assay that detects all HAdV types run concurrently detected between 10 and 100 copies/reaction, depending on the virus type. No nonspecific amplifications were observed with concentrated nucleic acid from 51 HAdV prototype strains or other common respiratory pathogens. All members of a panel of 137 previously typed HAdV field isolates and positive clinical specimens were correctly characterized by the type-specific qPCR assays; two different HAdV types were detected in three of the clinical specimens and confirmed by amplicon sequencing. The qPCR assays permit sensitive, specific, and quantitative detection and identification of seven clinically important respiratory HAdVs and should provide a convenient adjunct to classical typing methods for a rapid response to HAdV outbreaks.     

Monitoring of adenovirus from conjunctival scrapings in Japan during 2005--2006

A total of 189 conjunctival scrapings were collected from patients in Tokyo, Japan by monitoring adenovirus infection in community-based clinics during 2005 and 2006. Of the 189 samples, 155 (82%) had adenoviruses detected by polymerase chain reaction (PCR). The serotypes were determined by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, using a combination of endonucleases, such as HhaI, AluI, and HaeIII, and neutralization tests (NTs). PCR-RFLP identified five serotypes: serotype 3: 16.8%, serotype 4: 9.7%, serotype 8: 34.8%, serotype 11: 23.2%, and serotype 37: 15.5%. Adenovirus 8 was the most common serotype identified. A subset consisting of 25 isolates identified as adenovirus 8 from this study plus 25 isolates from Kawasaki were analyzed using PCR sequencing of the hexon gene. Compared with prototype adenovirus serotype 8 and serotype 9 derived in Tokyo and Kawasaki, these isolates shared 61.7-62.8% and 80.5-82.7% amino acid homology, respectively, suggesting that a variant adenovirus serotype 8 was involved in this outbreak, and is different from the prototype adenovirus 8 virus. This variant had not been detected in Japan prior to 1996 and appears to be the most common adenovirus type 8 involved in cases of epidemic keratoconjunctivitis in Japan at present.     

Serological and genetic characterisation of a unique strain of adenovirus involved in an outbreak of epidemic keratoconjunctivitis

AIMS: To characterise a novel strain of adenovirus (Ad) type Ad8 (genome type Ad8I) involved in an epidemic keratoconjunctivitis (EKC) outbreak in Hiroshima city using serological testing and sequence analysis of the fibre and hexon gene. METHODS: A neutralisation test (NT) was performed in microtitre plates containing a confluent monolayer of A549 cells using 100 tissue culture infectious doses of virus and type specific antisera. The haemagglutination inhibition test was also carried out in microtitre plates with rat erythrocytes using four haemagglutination units of virus and twofold dilutions of serum. The fibre gene was sequenced by generating overlapping polymerase chain reaction products or by direct sequencing of genomic DNA. Primer selection was based on alignment of the fibre genes of human adenovirus serotypes Ad8, Ad19, Ad37, Ad9, and Ad15 available from Gene Bank. RESULTS: The virus strain was specifically neutralised by anti-Ad8 antibodies, although there was a major crossreaction with anti-Ad9 antibodies. Haemagglutination was equally inhibited by anti-Ad8 and anti-Ad9 antibodies. The predicted amino acid sequences of the hypervariable regions (HVRs) of the Ad8I hexon gene showed higher homology with Ad9 (83.3%) than with Ad8 (62.0%). However, the Ad8I fibre knob was more homologous to Ad8 (94.4%) than to Ad9 (91.6%). CONCLUSIONS: Ad8I is a unique strain of adenovirus because of its lower genomic homology with Ad8, major crossreactivity with Ad9 in NT, and mixed genetic organisation of HVRs of the hexon gene. These factors may have enabled the virus to circumvent acquired immunity, resulting in the outbreak.     

Type-specific induction of interleukin-8 by adenovirus.

Infection with adenovirus (Ad) causes acute pneumonia in a type-specific fashion because type 7 but not type 5 Ad has been isolated as a causative agent. We postulated that the type specificity of induction of pneumonia may be related to type-specific cytokine induction in lung cells. To test this hypothesis, we infected human fetal lung fibroblasts and the lung epithelial cell line A549 with live type 5 and type 7 Ad. Virus inactivated by irradiation was used as a control. Type 7 but not type 5 Ad induced interleukin (IL)-8 protein production in both cell types in a dose- and time-dependent manner. Inactivated virus had no effect on the production of IL-8 protein. Type 7 but not type 5 virus also stimulated IL-8-specific messenger RNA (mRNA) production in these cells. Because half-life of IL-8 mRNA was prolonged in both type 5- and type 7-infected A549 cells, induction likely involves enhancement of message stability as well as other effects. Virus early gene expression did not consistently correlate with IL-8 message induction and followed induction in fibroblasts. These results suggest that there is type-specific induction of IL-8 production during infection of lung cells with Ad. Induction involves message stabilization and may not require viral gene expression. Because IL-8 is one of the important mediators of lung inflammation, type-specific induction of this and other cytokines may account for the different consequences of lung infection with different types of Ad.     

Latex agglutination test for adenovirus diagnosis in diarrheal disease

A commercial latex agglutination test for diagnosis of adenovirus in diarrheal disease (Adenolex, Orion Diagnostica, Finland) was evaluated by comparison with the results obtained by ELISA, electron microscopy (EM), and virus isolation. Fifty specimens originated from the diagnostic routine, and 50 were selected from a previous epidemiological study on the etiology of diarrheal disease in children. Thirteen of the 100 specimens reacted with the latex control, impairing interpretation of the results. Although the ELISA detected adenovirus antigen in 10(2) higher dilutions than the latex agglutination test, a total agreement was obtained between results by the two tests for 87 specimens including 42 positives. The two additional positives found by EM and virus isolation could not be diagnosed by the latex agglutination test. Of 37 specimens containing enteric adenoviruses (types 40 and 41), the agglutination test diagnosed all but 4 specimens containing type 41 virus. These four specimens were negative also by ELISA and adenovirus had been detected by virus isolation on the 293 cell line. The latex agglutination test gave positive results with nine specimens containing adenovirus types other than the enteric types 40 and 41. The latex agglutination test was found to be a rapid and simple method for the detection of adenovirus in diarrheal disease. Compared to ELISA and EM, the sensitivity was 100% and 95% respectively, and the specificity 100%.     

Effect of antioxidants on the HeLa cell infection process in culture

A certain role in the outcome of virus infection of cells was assumed to belong to cell membrane lipids peroxidation. The influence of known inhibitors of lipid peroxidation, beta-naphthol and ionol, on HeLa cells infection with human adenovirus type 2 was studied. These antioxidants in certain concentrations were found to be capable of effectively inhibiting virus infection of HeLa cell cultures. At the same time, beyond these concentration levels these antioxidants did not inhibit but stimulated virus infection of HeLa cell cultures. It is concluded that inhibition of virus infection of cells by antioxidants in moderate concentrations is due to their blocking of membrane lipids peroxidation. The opposite effect of antioxidants in much higher or lower concentrations may be due to their membranotropic effect which enhances the development of virus infection. The results of the study attest to the possible use of antioxidants in appropriate concentrations for control of virus infection.     

Adenovirus infection in patients with Kawasaki disease

Two outbreaks of Kawasaki disease at different times and areas (Kyoto in 1982 and Sapporo in 1985) of Japan were studied retrospectively for the presence of antibodies to adenoviruses and herpesviruses. Only 2 of 12 (16.7%) consecutive acute phase sera of patients from the outbreak in 1982 and 1 of 10 (10.0%) sera from 1985 showed positive antibodies for the common adenovirus antigen by a complement fixation (CF) test, whereas 10 of 16 (62.5%) age- and sex-matched controls during the outbreak of Kawasaki disease in 1985 were seropositive by the CF test. In contrast, using a recently developed enzyme-linked immunosorbent assay (ELISA), 9 patients (75.0%) in 1982 and 9 patients (90.0%) in 1985 had antibodies to adenovirus type 2. In addition, 5 of 10 (50.0%) of the 1982 and 6 of 9 (66.7%) of the 1985 patients who were seronegative for CF antibodies were positive for IgM antibodies to adenovirus type 2. Fifteen (93.8%) controls were positive for antibodies to adenovirus type 2 by ELISA and only two sera showed negative CF antibodies with positive IgM antibodies to adenovirus type 2. Sequential sera from 4 patients in 1985 had either IgM or IgG antibodies by ELISA and eventually three became seropositive by the CF test in time. Additionally, no significant difference was noted with antibody status to herpes simplex virus type 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), and cytomegalovirus (CMV) between patients and controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of adenovirus hexons by their epitope composition

Summary For the investigation of epitope composition of different adenovirus hexon types sixty-one mouse ascitic fluids containing monoclonal antibodies (MAbs) developed in three different panels were used. The distinction and marking of the different epitopes recognized by the MAbs were carried out by the determination of the composite cross-reactivity pattern, the titer and the correlation coefficient of all the 61 MAbs with 21 different hexon types representing all the six human subgenera, as well as different bovine and simian adenoviruses. The distinct epitopes were marked by two numbers refering the homologous hexon type to which the MAbs were directed and the serial number of the epitope specified by the different members of the given panel of the MAbs. The three panels of MAbs recognized 22 epitopes on the 21 hexon types among them a genus and three type specific ones and 18 different bi- and multilateral intertype (IT) specific epitopes that grouped adenoviruses within the genus, independently from the subgenus they belong to. Considering that the type specific epitope could be present only on the homologous hexon type, the largest number of the different epitopes distinguishable by the MAbs used could be 20 on the homologous hexon and 19 on the heterologous ones. It was found that the total number of IT specific epitopes on the hexons varied between 2 and 18. The distribution of the distinct specific epitopes on the different hexon types was different, as expected. The antigenic structure of the individual hexon types were characterized by the determination of their IT specific epitope spectrum. By pairwise analysis ten human hexon types formed three epitope clusters (types 4 and 19; types 8, 9, 9/13 and 10; as well as all types of subgenus C) showing identical epitope spectra. No clustering was found with human type 7, 12, 13, 18, 26, 27, 35 and 41, as well as with bovine and simian adenovirus hexons studied. However, they displayed a closer or looser antigenic relationship among each other and to members of the epitope clusters. The degree of antigenic relationship could be expressed by the similarity/dissimilarity percentage calculated from the number of the identical and different epitopes present on any two given hexon types.     

Mechanisms of changes in weight of the central lymphoid organs during adenovirus carcinogenesis

Transplantation of spleen cells of CBA mice at the 25th day of the latent period of carcinogenesis induced by SA7 (C8) virus into newborn syngeneic animals evoked a graft versus host reaction in them. Splenomegaly and a progressive decrease in the weight of the thymus were observed in the recipients. Similar changes in weight of the lymphoid organs were found in animals infected neonatally with oncogenic adenovirus SA7 (C8). The results show that adenovirus carcinogenesis has some manifestations of autoimmune disease.Laboratory of Virology, P. A. Gertsen Moscow Oncologic Research Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 9, pp. 356–358, September, 1978.