Göran Wadell

Classification of human adenoviruses by SDS-polyacrylamide gel electrophoresis of structural polypeptides.

The virion polypeptide compositions of 15 human adenovirus types, representing all members of subgroups B and C and selected members of subgroups A and D, have been analyzed by SDS-PAGE. The capsid polypeptides II, III, IIIa and IV displayed a pattern of apparent molecular weights which was characteristic for each adenovirus serotype analyzed. Adenovirus serotypes belonging to the same subgroup were also demonstrated to share an apparent molecular weight pattern of the internal structural polypeptides in the following manner: Subgroup A:V-resolved into two bands of 46--48.5 K and 51--51.5 K; VI-25.5 K; and VII-18 K. Subgroup B:V-53.5--54.5 K; VI-24 K; and VII-18 K. Subgroup C:V-48.5 K; VI-24 K; and VII-18.5 K. Subgroup D:V-50.5 K; VI-23.5 K; and VII-18 K. Adenovirus type 4: V-48 K; VI-24.5 K; and VII-18 K. The structural polypeptides represent a major portion of the adenovirus gene products. It is therefore proposed that the apparent molecular weight differences of viral polypeptides be used (i) to confirm the identification of an adenovirus type, and (ii) for subgroup classification of adenoviruses and, consequently, that adenovirus type 4 should be classified as a member of a separate subgroup designated E.

Hemagglutination with Adenovirus Serotypes Belonging to Rosen's Subgroups II and III

Summary The hemagglutination by adenovirus types 1, 2, 4, 5, 6 (subgroup III), and 9, 15 (subgroup II) was studied. Serotype 9 displayed, as should be expected, a complete agglutination of human red cells. In addition these cells were agglutinated partially by all the remaining serotypes, although a considerable variation in agglutinability of human erythrocytes obtained from different donors was demonstrated. The hemagglutination of these red cells by serotype 9 was RDE sensitive, whereas the agglutination of human cells by the remaining serotypes was resistant to this treatment. The partial agglutination of both human O and rat red cells peculiar to unfractionated preparations of members of subgroup III was suggested to be caused by the presence of a relative excess of incomplete HAs (i.e., fibers and pentons), in unfractionated preparations of these types, which competitively interfered with the complete hemagglutinin. The following observations supported this explanation (a) isolated complete HAs, such as virions, displayed complete agglutination with both kinds of red cells; (b) adsorption experiments demonstrated a larger amount of virus specific receptors on rat red cells reacting with members of subgroup II than with members of subgroup III; and (c) in a model system the presence of fibers belonging to both subgroup II and III competitively inhibited the virion-mediated agglutination of both rat and human cells. The author would like to express his gratitude to Professor Sven Gard and Dr. Erling Norrby for valuable discussions and helpful criticism. The excellent performance of biological tests by Miss Marianne Peterson and Miss Ragnvi Svensson is also gratefully acknowledged.

Soluble components of adenovirus type 4.

Abstract Four different soluble components were identified by zonal centrifugation of adenovirus type 4 material. Listed in order of decreasing sedimentation rates they were: complete hemagglutinin (HA), group-specific complement-fixation (CF) antigen, rapidly and slowly sedimenting incomplete HA. The rapidly sedimenting incomplete HA was demonstrable in the presence of antisera against members of both Rosens subgroup I (types 3 and 11) and subgroup III (types 2 and 5). In contrast the slowly sedimenting incomplete HA was detectable only in the presence of antibodies against members of Rosens subgroup III. Incubation of complete HA at 56° led to a degradation into incomplete HAs of both kinds. Trypsin treatment did not affect complete HA and slowly sedimenting incomplete HA, but converted rapidly sedimenting incomplete HA into the slowly sedimenting form. The nature of the different components are discussed in the light of previously presented data on sedimentation characteristics of soluble components of adenovirus type 3 and (as a reference in the present study) adenovirus type 5. It is suggested that the complete HA might represent a symmetrical aggregate of 12 penton antigens, the group-specific CF component hexon antigen, the rapidly sedimenting incomplete HA-isolated penton antigen, and the slowly sedimenting incomplete HA fiber antigen.

Incomplete Virus Particles of Adenovirus Type 16

Summary Haemagglutinating populations of virus particles of adenovirus type 16 (I–VIII) were separated by isopycnic centrifuging in CsCl. One of these populations (V) contained complete virus particles. Fresh preparations of virus particles from all populations studied had a morphology indistinguishable from that of complete virus particles. The five lightest populations were shown to contain [3H]-thymidine and to be infective; dose-response relationships indicated the occurrence of a multiplicity-dependent infectivity of the incomplete virus particles. SDS-polyacrylamide gel electrophoresis of incomplete virus particles of the lightest population demonstrated that they were deficient in three polypeptides, but contained three other polypeptides which were not detectable in complete particles. One characteristic polypeptide of incomplete particles and another characteristic polypeptide of complete particles displayed high ratios of arginine over threonine.

A population-based study of human papillomavirus deoxyribonucleic acid testing for predicting cervical intraepithelial neoplasia

OBJECTIVE Our purpose was to determine the predictive values of primary or secondary screening for cervical human papillomavirus infection for cytologic detection of cervical intraepithelial neoplasia. STUDY DESIGN Most of the 254 women referred for colposcopy in Västerbotten County in Sweden during October 1993 through December 1995 and 320 age-matched women from the general population were screened for human papillomavirus deoxyribonucleic acid by nested general-primer polymerase chain reaction. RESULTS Ninety-six percent of women with high-grade cervical intraepithelial neoplasia had human papillomavirus, compared with 4% of women with normal findings (odds ratio 606; 95% confidence interval 137 to 5607). Thirty-seven percent of referred women and 48% of referred women >39 years old had mostly minor cytologic abnormalities with no human papillomavirus. The human papillomavirus-associated positive predictive value for cervical intraepithelial neoplasia was 76% in the colposcopy group and 11% in the general population, whereas the negative predictive value was >97% in both populations. CONCLUSION Testing for human papillomavirus deoxyribonucleic acid seems diagnostically useful among women referred for colposcopy.

DNA restriction site mapping of adenovirus type 16 with BamI, EcoRI, HpaI AND SalI

Human adenoviruses are divided into four subgroups. DNA restriction (RE) site mapping of Ad 2 (member of subgroup III) has been performed with &m, BarnI, EcoRI, Hi&III, H’aI and SaZI [l] . The availability of these maps has enabled localization of ts-mutations [2], determination of the fraction of integrated viral genome in transformed cells [3] and mapping of early and late Ad 2 transcripts [4]. The moderately oncogenic adenovirus types belonging to subgroup I differ in several biological aspects from members of other subgroups [S-7]. Furthermore, members of subgroup I, represented by Ad 16, produce more incomplete virus-particles (i.e., virusparticles with DNA of less than unit size) than members of other subgroups [8]. Access to DNA restriction site maps is a prerequisite for analysis of the structure of these DNA molecules. The aim of this communication is to present the RE site maps of Ad 16 with EcoRI, BamI, HpaI and Sal1 since no RE site map of Ad-types belonging to subgroup I has been reported.

Preparation of sera for typing of adenovirus infections by immunofluorescence

The conditions for preparation of adenovirus antisera with group, subgroup and type-specific reactivity to be used in the immunofluorescence technique have been evaluated. Group-specific antibodies were prepared by passing an antiserum against virions or hexons of one serotype through an immunosorbent column containing soluble viral components of an adenovirus type belonging to a different subgroup. The group-specific antibodies were recovered from the solid phase by elution at pH 2.8. Reagents specific for subgroup I and II were obtained by passing anti-dodecon sera through an immunosorbent column containing soluble viral components from heterologous subgroups. In this case antibodies not attaching to the absorbent were recovered. Sera against fibers of subgroup III members could be used as subgroupspecific sera without absorption. Type-specific antibodies were prepared by removal of antibodies of other specificities by passing anti-virion or anti-hexon sera through immunosorbent columns containing soluble viral components of an adenovirus type belonging to the same subgroup. Reagents specific for seven adenovirus types representing Rosens three subgroups were prepared according to the outlined procedures.

Isolation of incomplete adenovirus 16 particles containing viral and host cell DNA

Abstract Four populations of adenovirus type 16 incomplete particles with different buoyant densities in CsCl have been identified. They contain less DNA than complete particles. The standard viral genome has a molecular weight of 23 × 10 6 . The molecular weights of the incomplete DNA molecules range from 1 × 10 6 to 16 × 10 6 , increasing with increasing density of the particles in CsCl. Characterization of isolated DNA by equilibrium density centrifugation in CsCl, DNA-DNA hybridization, and analysis of DNA labeled with [ 3 H]thymidine before infection shows that both viral and host cell-specific DNAS are incorporated into incomplete virus particles.

The relationship between the soluble antigens and the virion of adenovirus type 3: VI. Further characterization of antigenic sites available at the surface of virions

Abstract Absorption experiments demonstrated the presence of antigenic sites at the surface of virions of adenovirus type 3 which were not carried by spontaneously occurring soluble components. These unique sites were identified by use of virion antigen in neutralization and hemagglutination-inhibition (HI) tests with rabbit hyperimmune sera in the presence of sheep anti-rabbit serum and in complement-fixation tests. Antibodies against vertex capsomers (represented by an antipenton serum absorbed with purified fibers) or against fibers carried negligible neutralizing activity. However, efficient neutralization occurred when both kinds of antibodies were present simultaneously, i.e., as in antipenton and antidodecon sera, or when anti-rabbit serum was added. The possible occurrence of cross-reacting antigenic sites at the surface of virions was examined in HI tests with purified virions and soluble hemagglutinin of eight different human adenovirus serotypes, including at least two types for each one of Rosens subgroups. Rabbit hyperimmune sera against virions and purified soluble components were employed, and a sheep anti-rabbit serum was added to the HI tests in order to increase their sensitivity. Low-degree cross-reactions of subgroup-specific nature were identified. They were demonstrated to be due to the occurrence of vertex capsomer-associated subgroup-specific antigen(s). An antiserum against type 11 dodecons was found to display some capacity to neutralize virions of type 3 in the presence of anti-rabbit serum. This heterotypic neutralization is proposed to be caused by the subgroup-specific antibodies against vertex capsomers demonstrated by HI tests.