Biomedical subjects
Publications and source records attributed to T Adrian.
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.
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.
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Familial medullary carcinoma of thyroid (MCT) can be diagnosed early by measuring stimulated plasma calcitonin following provocation with 50 ml of oral whisky, or 0.5 microgram kg-1 i.v. pentagastrin. The calcitonin release, however, can be variable following both stimuli. To determine whether a humoral mechanism might be involved in mediating this calcitonin release, nine gut peptides, histamine, alcohol, calcium, and calcitonin, were measured in the plasma of eight patients whose family was affected by multiple endocrine neoplasia Type II syndrome. Pentagastrin produced significant rises in plasma calcitonin, histamine and pancreatic polypeptide. Whisky produced a variable rise in plasma alcohol, but significant rises in plasma calcitonin, histamine and pancreatic polypeptide as well as insulin (P less than 0.05). This study suggests that histamine may be the mediator of calcitonin release following provocation by either pentagastrin or alcohol. Histamine infusion requires evaluation as a screening test for MCT in MEN II syndrome.
From the stool of a Dutch girl, a new virus (LJ) was isolated that by morphological, biological, and immunological criteria was a human adenovirus. Because of its hemagglutinating activity for rat and human red blood cells and its DNA restriction enzyme pattern, the virus was classified as a member of subgenus D. It was distinct in neutralization tests from all other human adenoviruses. The hemagglutinin was antigenically similar to that of adenoviruses 13 and 39, but the DNA differed from that of these two species. It is proposed to call virus LJ candidate Mastadenovirus h 38. Two other viruses belonging to the same new species were isolated in the USA.
The possibility of a rapid diagnosis of adenovirus conjunctivitis was studied by demonstrating genus-specific adenovirus hexon antigen or antihexon IgA antibodies in conjunctival secretions by an ELISA. Hexon was found in 44% of about 100 patients with adenovirus conjunctivitis, proven by virus isolation, antihexon in 53%, either one or both in 69% of the conjunctival swabs from the acute phase. Hexon was found in the acute phase more frequently, antihexon in the convalescent phase. In patients with conjunctivitis of different etiology no hexon was found, whereas antihexon IgA was found in a few cases.
Adenovirus 8 (Ad 8, strain 1127) has the general structural features of an adenovirus; it produces the genus-specific hexon antigen. The fiber length of 12 nm corresponds to that of other subgenus D adenoviruses. In hemagglutination-inhibition, Ad 8 is related to Ad9 and Ad 15/H9. The slow multiplication, compared to most other subgenus D viruses, was shown not to be caused by an extended growth cycle, but appears to be due to an inefficient virus release into the medium. After restriction analysis with the enzymes Sma I and Bgl II, the typical fragment patterns of species from subgenus D were obtained, while very different patterns emerged from the analysis with Hind III, Bam HI, and Bst E II endonucleases. Several strains of Ad 8 produced more penton-associated toxin than Ad 15/H9 used as reference virus from subgenus D, which could explain the "clumping CPE" caused by Ad 8. Ad 8 toxin was neutralized by antisera from many adenovirus species. The poor virus yield of Ad 8 was not improved by removing soluble viral or cellular substances; all other attempts to improve virus yield by varying the conditions of cell culture also failed.
Serum pairs from 114 patients with adenovirus infections (mostly ocular disease in adults) were investigated by an ELISA, with purified adenovirus 5 hexon coated to the solid phase, diluted sera, and peroxydase-labeled antihuman IgG, IgA, and IgM antibodies. Serum dilutions of 1:3200, 1:200, and 1:100 respectively proved to be optimal to demonstrate antibody rises. Sera from infants served as negative controls. Antibody rises were found in the IgG and/or IgA test 85% of the patients, but only in 56% by complement fixation. Antibody rises in all three ELISA tests occurred mostly around the 10th day after onset of disease. Fewer antibody rises and weak reactions were found in IgM test. Moreover, IgM antibodies were present also in persons without fresh adenovirus infection. Hence, this test is not suitable for an early diagnosis. This observation is probably due to the frequent stimulation of the immune system by the genusspecific determinant alpha of the hexon by repeated adenovirus infections during life.
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The DNA of 60 adenovirus type 8 (AV8) isolates (collected during the period 1961 to 1982, mostly in Western Germany) was analysed by 6 endonucleases and revealed 6 different genome types, thus implying that the variability of AV8 is relatively low. It was found that 45 isolates belonged to the genome type D1. Restriction site maps of a prototype D1 and of all deviating restriction variants were elaborated for enzymes BamHI, BglII, Hind-III and SalI.
We propose to classify "genome types" of human adenoviruses in the same way as the "genomic clusters" defined by Li and Wadell, i.e. groups of closely related DNA variants of a given serotype named by the use of indices, P (standing for prototype) a, b etc. Variants within the genome type are named by letter-number indices (p1, p2; a1, a2 etc.). The percentage of comigrating restriction fragments and common restriction sites is significantly higher in variants than between genome types. The new naming system is applicable to adenoviruses from 5 subgenera, but requires additional work for subgenus C strains. Renaming is required mainly for adenovirus 7.
The amino acid sequence of the adenovirus type 31 (subgenus A) fibre polypeptide was deduced from the nucleotide sequence of the fibre gene. The analysed peptide sequence showed an organization consistent with the structural domains described for other adenoviruses: an amino-terminal tail region, an intervening shaft region and a carboxy-terminal knob. The AV31 fibre shaft displayed 20 repeats of the 15-amino-acid segments in the shaft domain, which agreed with the reported length of the fibre. The predicted AV31 fibre polypeptide sequence was compared to fibre polypeptides of serotypes representing subgenera A to F. As expected, AV31 and AV12, both belonging to subgenus A, showed the highest overall homology (75.4%). When comparing the AV31 fibre to the fibre polypeptides of subgenera B to F, AV31 and AV41 (subgenus F) shared the highest overall homology (35.3%), followed by AV40 (34.8%). The lowest overall homology (20.3%) was found for the AV31 and Av3 fibres (subgenus B). From the data presented, it could be suggested that AV31 is more closely related to the enteric viruses of subgenus F than to the other adenoviruses analysed. Comparing the fibre polypeptides of 14 adenovirus serotypes, 10 conserved amino acid sequences were detected, 5 of which were in the knob region. Since the fibre knob interacts with the host cell during infection, these conserved amino acids might be important for virus attachment. The gastroenteritis-causing adenoviruses AV40 and AV41 shared 3 additional conserved amino acid residues with AV31 and AV12 in the knob region.
The hexon gene of human adenovirus (AV) type 7 (subgenus B) was sequenced. The determined nucleotide and the predicted amino acid sequences were compared to the corresponding sequences of AV3 and AV16. The hexons of AV7 and AV3 revealed an overall homology of 94.3% at the protein level, whereas the AV7 and AV16 hexons only showed an overall homology of 85.7%. Utilizing the three-dimensional model of the AV2 hexon, the structure of the AV7 hexon was predicted. The major differences between the three subgenus B hexon polypeptides were confined to the I1 and I2 surface loops. The AV7 I4 hexon loop was 100% identical to the other subgenus B I4 loops, but differed from the corresponding regions of other subgenera. This supports the idea that this loop carries a subgenus-specific determinant.