Biomedical subjects
Publications and source records attributed to R Wigand.
A new serotype, candidate adenovirus type 42, a member of subgenus D, was isolated from the feces of a healthy child. It shared its hemagglutinin with adenovirus type 15, but had no relationship in neutralization to other human adenoviruses. It also had a distinct DNA restriction pattern.
Ten serologically intermediate strains of subgenus D related to six different serotypes were characterized by neutralization and hemagglutination-inhibition and by DNA restriction analysis with the endonucleases BamHI, BglII, BstEII, HindIII, and SmaI. Each strain was a unique genome type. While the serologically identical strains showed only slight variations in their DNA, the comparison of the intermediate strains with the serologically related prototypes resulted in only moderate DNA relationship (53 to 73% comigrating fragments, with one exception). Hence the possibility that the intermediate strains have originated by recombination of two prototype-like strains is unlikely. One strain lost its cross-reactivity with AV19 in hemagglutination-inhibition after terminal dilution.
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Virological and serological data from 242 patients with conjunctivitis or keratoconjunctivitis, infected with adenovirus type 8, and of 109 patients infected with other adenovirus types were compared by means of an electronic data processing system. About 55% of the patients in both groups showed a diagnostic antibody rise in genus-specific (antihexon) complement-fixation. This rate is significantly lower than that obtained with genus-specific ELISA IgG or IgA, or with neutralization tests against the homologous virus, or with hemagglutination-inhibition in case of adenovirus 8 infection. All these tests showed titer rises in 72 to 85% of the patients; differences were not significant. Virus isolation from conjunctival swabs, taken up to 9 days after onset of disease, was successful in 80 to 90%, thereafter in 30%. The time-course of the various antibody titers showed that genus-specific (antihexon) antibodies mostly rise 9 to 12 days after onset, whereas type-specific neutralizing or hemagglutination-inhibiting antibodies rise 10 to 14 days after onset.
One hundred twenty strains of adenovirus types 1, 2, 5, and 6 (subgenus C), isolated at various times and places, were tested by neutralization and hemagglutination inhibition with two antisera of each type. Except for one type 5 strain they reacted largely type-specifically in both tests and neither qualitative nor quantitative differences were observed. Serologically intermediate strains were not found. The antigenic homogeneity is in contrast to the heterogeneity of the genomes of subgenus C adenoviruses.
DNA restriction patterns of all known human adenovirus prototypes (1 to 41), ordered according to subgenera A to F, are presented for restriction endonucleases BamHI, BglII, BstEII, HindIII, and SmaI. This catalogue is a prerequisite for typing of adenoviruses by DNA restriction analysis in diagnostic laboratories and for strain identification in reference laboratories. To determine the genetic relationship of human adenoviruses within a subgenus and between subgenera, a pairwise analysis of comigrating fragments was performed. Adenoviruses of subgenus C were closely related. Adenoviruses of subgenus B showed two related clusters of four types each, whereas the numerous serotypes of subgenus D did not show any corresponding clustering. Little comigration was observed between DNA restriction fragments from members of subgenus A or F respectively.
To demonstrate adenovirus hexon, four different arrangements of a four-layer ELISA, to be performed with commercially available reagents, were compared with a three-layer ELISA. After evaluation of specific and control reagents and their dilutions, the guinea pig/rabbit or the rabbit/chicken method were equally practicable and sensitive as the three-layer method with rabbit serum and superior to two other methods. By testing about 100 stool samples and 74 throat swabs containing adenoviruses, it turned out that the sensitivity (1 ng/ml hexon) appears satisfactory for stool specimens in adenovirus enteritis of infants, but is much less sensitive than isolation in cell culture for adenoviruses in throat swabs. As confirmatory test, specimen were incubated with an adenovirus rabbit antiserum or normal serum and then tested.
After a period of high prevalence in the early 1960s, adenovirus serotype 21 (Ad21) was identified in The Netherlands only very sporadically for more than 20 years. From December 1984 to July 1985, Ad21 was isolated relatively often from hospitalized children living in different parts of The Netherlands. The patients in question suffered from respiratory, gastrointestinal, meningeal, or ocular disorders. An increase in the incidence of Ad21 infections was also observed in the Federal Republic of Germany during this period. The DNAs of 93 isolates of Ad21 were subjected to restriction enzyme analysis with eight endonucleases. All 50 strains isolated in The Netherlands between 1960 and 1963 proved to be DNA variant Ad21/D2/20655/Netherlands/60. This variant has already been described (T. Adrian, R. Wigand, and J. C. Hierholzer, Arch. Virol. 84:79-89, 1985) as typical for the Ad21 strains circulating since 1960. Analysis of the DNAs of the 28 Ad21 strains isolated in The Netherlands or in the Federal Republic of Germany in 1984 and 1985 showed them to belong to two new, closely related DNA variants designated Ad21/D7/1857/Netherlands/84 and Ad21/D8/5398/Netherlands/85. The BglI and KpnI restriction profiles were characteristic for these recent DNA variants.
Adenovirus 3-7, strain Takeuchi was related to both AV3 and AV7 by neutralization, but only to AV3 by hemagglutination-inhibition. DNA restriction analysis with seven endonucleases showed about equal genetic distance from both AV3 and 7 prototypes.
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47 adenovirus strains related to Ad11, 14, 16, 21, 34, and 35 (including all subgenus B prototypes) were studied in neutralization and hemagglutination-inhibition with 40 antisera, prepared against 16 virus strains, to determine their antigenic relatedness. Wild strains of Ad11-14 (14), Ad14, and a group of 16 strains related to Ad21 proved to be identical within their respective groups, whereas strains related to Ad11, 34, and 35 showed serological differences. Prototype strains of Ad11, 21, 34, and 35 fit the species definition (22) if selected antisera of known specificity are used. All virus strains with Ad16 hemagglutinin showed a bilaterial cross-neutralization either with Ad4, Ad14, Ad3, or Ad21. Doubly neutralizable strains with one hemagglutinin were Ad11 + 35/H11, Ad3 + 16/H16, Ad21 + 16/H16 (H stands for 'hemagglutinin'). Preliminary identification of subgenus B viruses (others than Ad3 and 7) may be performed by HI.
47 adenovirus strains related to Ad11, 14, 16, 21, 34, and 35 (including all subgenus B prototypes) were analyzed by DNA restriction analysis with Bam HI, Bgl II, Hind III, and Sma I. Typical fragments occurring in most of the strains were observed only with Hind III. Besides Ad3 and 7, 20 different genome types were distinguished. A pairwise analysis of comigrating fragments was performed for all genome types and summarized for the four enzymes. A medium rate of comigrating fragments (50 to 60 per cent) was found between Ad11, 14, 34, and 35 strains; Ad16 and 21 had 35 to 50 percent comigrating fragments. Within each group of closely related strains, Ad16 strains, Ad21 strains, and Ad34 + 35 strains showed a high rate (greater than 70 per cent) of comigrating fragments. Ad3 and 7 showed more comigration with Ad16 and 21 than with the others. Sixteen strains related to Ad21 were identical in their DNA restriction pattern and by serology. The relatedness determined by fragment comigration and by serology was compared by a quantitative estimate. The DNA relationship between Ad11 + 14 and Ad34 + 35 was not corroborated by serology. Ad21 was serologically related to Ad34 and 35, but not to Ad16; the opposite was found for DNA. The intermediate Ad21-16 showed a unique DNA pattern with some relation to Ad16.
Five adenovirus 15/Hx and one Ad9/Hx strain with a common novel hemagglutinin were compared serologically and by DNA restriction analysis with six endonucleases with the related prototypes. The five Ad 15/Hx strains represented five different genome types.
PVP-iodine was found to be virucidal for adenoviruses and herpes simplex virus in a concentration of 0.5%, i.e., a 1:20 dilution of Betaisodona. Similar concentrations did not delay the healing of corneal epithelium of the rabbit after experimental lesions (Ullerich and Durchschlag, 1953), although the eyes were slightly irritated. However, a higher concentration (1.25%) caused considerable ocular irritation, and healing of the cornea was delayed or completely inhibited. Arginase, which inhibited the multiplication of adenoviruses and herpes simplex virus in cell culture, did not impair corneal healing in solutions of up to 5%.
A numerical code to denominate adenovirus (AV) genome types is proposed. Seven restriction endonuclease patterns are listed in alphabetical order (BamHI, BGlII, BstEII, EcoRI, HindIII, KpnI, SmaI); patterns deviating from those of the prototype (1) are named 2, 3 etc. depending on the chronological order of the respective isolates. Thus AV6/3:1231121 is a type 6 strain deviating from the prototype in three patterns, i.e. those of BglII, BstEII and KpnI. From 24 AV6 isolates, six were identical with the prototype, whereas the other strains represented 13 different genome types.
17 adenovirus strains from various parts of the United States and Europe were characterized by immunological and biochemical methods. They were antigenically related to adenovirus 15 (Ad15) in neutralization tests. In hemagglutination-inhibition tests, a relationship was seen to Ad9, but not to Ad15. These strains were named Ad15/H9 intermediate strains. The relatedness of prototypes Ad15 and Ad29 in neutralization tests and of Ad9 and Ad8 in hemagglutination-inhibition tests was also found in the strains studied. The DNAs of these strains were analyzed by use of restriction endonucleases. After cleavage with the enzymes BamHI, BglII, BstEII, HindIII, and SmaI, fragment patterns of Ad15/H9 strains were found to be different from the prototypes and were more closely related to Ad9 than to Ad15. Furthermore, the 17 strains could be classified into six different genome types, a-f.
To study the infection rate with adenoviruses (Ad) in infants, sera from 75 infants aged 8 to 26 months were tested for neutralizing antibodies against Ad 1 to 39. Antibodies against Ad 1, 2, 5, and 6 were frequently found, against Ad 7 more rarely, whereas one serum reacted with each Ad 3, 4, 18, 31, and 37 and none with all other types. A significant association of positive reactions against several or all of Ad 1, 2, 5, and 6 was observed in neutralization, but not in hemagglutination inhibition, probably due to a subgenus-specific antibody response. The titer of neutralizing antibodies in human gammaglobulin was determined as an estimate for the prevalence of Ad 1 to 39 in the general population.