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J C Hierholzer

Publications and source records attributed to J C Hierholzer.

At least 37 records · Page 2Linked to original sources

Astrovirus and adenovirus associated with diarrhea in children in day care settings.

The relative importance of astrovirus and adenoviruses as etiologic agents of diarrhea among children in day care was examined. Stool specimens from this prospective study were screened for both astrovirus and adenovirus hexon with two new indirect double-antibody assays and for enteric adenoviruses with an EIA specific for serotypes 40 and 41. Astrovirus was detected in a significantly greater percentage of children with diarrhea (4%, 21/524) than of those without (less than 1%, 1/138) (P less than .05); however, no difference between such such children with adenovirus infections was found (8%, 43/565, and 8%, 10/129, respectively). Overall, 30% (13/43) of all adenovirus hexon-positive specimens were enteric serotypes, and by extrapolation, enteric adenoviruses were identified in an equal percentage of children (2%) with and without diarrhea. This study documents the presence of astrovirus and enteric adenoviruses among children in day care in the United States, associates astrovirus with diarrhea in this setting, and suggests that viral agents may be the most common enteric pathogens among children with diarrhea in day care.

Adenovirus Infections, Human↗

NCI-H292 as an alternative cell line for the isolation and propagation of the human paramyxoviruses.

Primary rhesus monkey kidney (MK) cells have long been the cells of choice for isolation and propagation of the human paramyxoviruses (parainfluenza 1, 2, 3, 4A, 4B, and mumps). However, problems with the supply and cost of MK cells and the presence of endogenous viruses, including herpes B virus and SV-5, necessitated a search for an alternative cell line. Continuous cell cultures of human origin (L132, A-549, HuT-292, HEK, G-293, G-401, A-498, A-704, CAKI-1, RD) and simian origin (LLC-MK2, BSC-1, MA-104, Vero) were evaluated for their capacity to support the growth of the human paramyxoviruses, as followed by cytopathic effect, hemadsorption, hemagglutination, and EIA. NCI-H292 (HuT-292) human lung mucoepidermoid carcinoma cells (ATCC # CRL-1848) proved to be the most sensitive line for cultivating all serotypes and strains of the paramyxoviruses. These cells were also shown to be a suitable substitute for MK in primary isolation of paramyxoviruses from clinical specimens. RPMI-1640 with 1.5 micrograms/ml trypsin was the preferred maintenance medium; alternatively, Eagle's MEM supplemented with 1.5 micrograms/ml trypsin and 0.1% ITS was satisfactory. NCI-H292 cells are a continuous line with excellent growth characteristics, although the genetic polyploidy of the cells may limit the number of passages of usable cells.

Animals↗

Molecular epidemiology and restriction site mapping of adenovirus 7 genome types.

From the United States, the Federal Republic of Germany, and other regions, 168 strains of AV7, isolated between 1961 and 1985, were analyzed by six restriction endonucleases and nine genome types were identified. The enzymes BamHI and HindIII were most discriminative. The genome type D5 (or 7b) predominated with 120 isolates since 1970 in both countries. Strains of D2 (7a) and D4 (7c) were isolated for a limited time, D3 for an extended time period. Several clusters of infections with the same genome type were found. Differences in pathogenicity could not be derived from our data. On the basis of restriction site mapping, most other genome types were similar to D5, one to D2 and one to the prototype (D1). The genomic relation between AV7 and AV3 is discussed and shown by a dendrogram.

Adenoviridae↗

Comparison of monoclonal antibody time-resolved fluoroimmunoassay with monoclonal antibody capture-biotinylated detector enzyme immunoassay for respiratory syncytial virus and parainfluenza virus antigen detection.

An all-monoclonal antibody, time-resolved fluoroimmunoassay was compared with several enzyme immunoassays for the detection of respiratory syncytial virus and parainfluenza virus type 1, 2, and 3 antigens in clinical specimens. The most sensitive enzyme immunoassay for parainfluenza virus type 1 was an all-monoclonal antibody assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate, but for respiratory syncytial virus and parainfluenza virus types 2 and 3 the most sensitive assay was a polyclonal antibody assay with horse capture antibodies and bovine or rabbit detector antibodies with anti-species peroxidase. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illnesses and with cell culture harvests of multiple strains of each virus isolated over many years. The time-resolved fluoroimmunoassay detected respiratory syncytial virus antigen in 92% of the specimens positive by culture, which was a decidedly higher sensitivity than either the monoclonal or polyclonal antibody enzyme immunoassay format (62 and 76%, respectively). For the parainfluenza viruses the time-resolved fluoroimmunoassay detected type-specific antigen in 94 to 100% of culture-positive specimens and again was more sensitive than the all-monoclonal antibody enzyme immunoassays (75 to 89%) or all-polyclonal antibody enzyme immunoassays (66 to 95%). Combined with results from a previously reported adenovirus time-resolved fluoroimmunoassay, these tests identified respiratory antigens in large numbers of clinical specimens.

Antibodies, Monoclonal↗

Molecular epidemiology and restriction site mapping of adenovirus type 3 genome types.

One hundred thirty-eight strains of adenovirus type 3 isolated from patients in the United States, West Germany, and other regions between 1961 and 1984 were analyzed with six restriction endonucleases; 18 genome types were found. BglII was the most discriminative enzyme. Mapping of altered restriction sites was also performed for all six enzymes. The genome types D1 (like the prototype) and D3 prevailed among 45 and 47 strains, respectively. All genome types could be divided into two groups related to D1 or D3. Several clusters of infections by strains with the same genome type were observed. Only hints of differences were found in the pathogenicities of individual genome types. D1 strains were present in the United States and in Europe; group D3 prevailed almost exclusively in the United States.

Adenoviridae Infections↗

Analysis of antigenically intermediate strains of subgenus B and D adenoviruses from AIDS patients.

We carried out detailed antigenic and restriction enzyme (RE) analyses on the subgenus B and D adenovirus isolates from 48 AIDS patients. These isolates were unusual both in the diversity of serotypes and in the number of intermediate strains identified. All unusual isolates were strain-purified and tested by serum neutralization (SN) and hemagglutination-inhibition (HI) tests with reference horse and rabbit antisera to all the prototype human adenoviruses; conversely, rabbit antisera were prepared to 16 selected strains from this study and tested by SN and HI with all prototype viruses. Among subgenus B strains, 6 DNA variants of Ad 11 isolates were distinguished by endonucleases BamHI, BglII, BstEII, HindIII, and SmaI. The D 2 variant of Ad 11 was prominent with 5 isolates, and 7 other isolates differed from D 2 in only 1 or 2 enzymes. HindIII was the most discriminative endonuclease for Ad 11 and related strains. Within subgenus D, there were 16 isolates of intermediate strains, including 4 intermediate types related to new Ad types 43-47. The RE patterns of subgenus D strains showed fragment distributions typical of subgenus D, with various unique patterns. Particular care was taken to analyze multiple strains from the same patient, recovered as much as 8 months apart, for evidence of genetic changes. The possible long-term infections with these viruses may provide the opportunity for mutations and recombination to occur, with the resultant generation of a variety of new strains of adenoviruses.

Acquired Immunodeficiency Syndrome↗

Adenoviruses from patients with AIDS: a plethora of serotypes and a description of five new serotypes of subgenus D (types 43-47).

We analyzed 67 adenovirus isolates from 48 patients with AIDS. The isolates included 5 of subgenus A (all Ad31), 23 of subgenus B, 4 of subgenus C (all Ad5), and 35 of subgenus D. The subgenus A and C strains were from liver, lung, blood, urine, and stool and were unremarkable in their antigenic characteristics. The subgenus B strains were mostly from urine and were serotyped by neutralization (SN) and hemagglutination-inhibition (HAI) tests as Ad11 (12 isolates), Ad16 (1), Ad35 (3), and Ad21/H21 + 35 (1) plus new intermediate strains 34/H11 (3), 35/H11 (1), and 11 + 35/H11 (2). The subgenus D serotypes from 24 patients were mostly from stool specimens and consisted of 8 isolates of known serotypes, 16 of antigenically intermediate strains, and 11 of five new adenovirus types with distinct antigenic and restriction enzyme characteristics. The new serotypes were described as candidate types 43-47 in accordance with currently accepted taxonomic criteria. Ad45, 46, and 47 showed a major relation by HAI with Ad29, 24, and 20, respectively. Four intermediate types were related to the new serotypes. The plethora of serotypes from patients with AIDS suggests that no epidemiological significance can be attached to any one serotype.

Acquired Immunodeficiency Syndrome↗

Neutralization of respiratory syncytial virus by individual and mixtures of F and G protein monoclonal antibodies.

We identified several types of neutralization effected by F and G protein monoclonal antibodies (MAbs) reacted individually or as mixtures against respiratory syncytial virus (RSV). Neutralizing activity was identified by a microneutralization test in which virus replication was determined by enzyme immunoassay. Complete neutralization was seen only with MAbs against the F protein. Strain-specific neutralization, complete neutralization against some strains of RSV, and no neutralization against other strains were seen with an additional MAb against the F protein. Partial neutralization, virus replication significantly reduced but still present, and no neutralization were seen with MAbs against both the F and G proteins. Enhanced neutralization, enhanced efficacy of neutralization, or increased neutralizing titer with a mixture of two MAbs over that for the individual MAbs was seen with all MAbs against the F protein and all but three MAbs against the G protein. Most (10 of 13) of the MAbs that exhibited neutralizing activity reacted with some but not all strains of RSV in an enzyme immunoassay. The epitopes corresponding to these 10 MAbs probably contribute to the strain-specific component of the neutralizing antibody response to RSV. Our results suggest that interpretation of RSV neutralization with MAbs is complex and that studies of such neutralization should include mixtures of MAbs and multiple RSV strains.

Antibodies, Monoclonal↗

Evaluation of human adenoviruses 38, 39, 40, and 41 as new serotypes.

Four new serotypes of human adenovirus (Ad38-41) have been described in recent literature. We conducted a thorough evaluation of the prototype strains of these types by preparing rabbit and horse antisera to them and performing reciprocal neutralization and hemagglutination-inhibition (HI) tests with all 41 prototype viruses and antisera. Types 38 and 39 are in subgenus D, share hemagglutinins with Ad13 and each other in three-way HI tests, and have been rarely encountered thus far; types 40 and 41 are in subgenus F, are related to each other by serum neutralization and HI tests, and are well-established agents of acute gastroenteritis in infants. All four prototype strains meet the accepted criteria for new serotypes, although special mention is made of the serologic relationship between Ad40 and 41. The horse antisera have stable titers and can serve as international reference reagents for these viruses.

Adenoviruses, Human↗

The effect of centrifugation on the rapid detection of adenovirus in shell vials.

Monoclonal antibody, specific for the adenovirus group-reactive hexon antigen, was used for the detection of this agent by immunofluorescence 24 and 48 hours after inoculation of HEp-2 cell monolayers in 1-dram shell vials after low-speed centrifugation (700 X g, 30 minutes). Of 31 adenovirus isolates, 16 (sensitivity, 52%) and 30 (sensitivity, 97%) were detected after incubation for 24 hours and 48 hours, respectively. All adenovirus strains were detected in conventional tube cell cultures but required an average of four days incubation. The shell vial assay is rapid, sensitive, and specific and can be easily implemented in laboratories with cell culture experience.

Adenoviruses, Human↗

Comparison of monoclonal time-resolved fluoroimmunoassay with monoclonal capture-biotinylated detector enzyme immunoassay for adenovirus antigen detection.

A sensitive time-resolved fluoroimmunoassay (TR-FIA), adapted from TR-FIA procedures already described, was developed with monoclonal antibodies and compared with several enzyme immunoassays (EIAs) for detecting adenovirus antigens in clinical specimens. The most sensitive EIA was an all-monoclonal assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illness, with tissue homogenates from patients with systemic infection, and with stool specimens from gastrointestinal illnesses. For respiratory and tissue specimens, the TR-FIA detected adenovirus in 85% of the specimens positive by culture, which was a sensitivity similar to those of the all-monoclonal biotin-avidin EIA (79%) and the polyclonal-capture biotin-avidin EIA (88%). For stool specimens, the TR-FIA detected adenovirus in 100% of the specimens positive by culture, which was a decidedly higher sensitivity than either EIA format (78 and 75%, respectively). The TR-FIA was shown to be an efficient, flexible, and specific test for large numbers of clinical specimens.

Adenoviridae Infections↗

Freeze-drying of respiratory syncytial viruses for transportation and storage.

The stabilities of eight strains of respiratory syncytial virus were compared after the strains were freeze-dried in the presence and absence of the stabilizer SPGA, which contains 218 mM sucrose, 7.1 mM dipotassium hydrogen phosphate, 3.76 mM potassium dihydrogen phosphate, 4.9 mM sodium glutamate, and 1% (wt/vol) bovine albumin. Strains freeze-dried in the presence of SPGA showed only small-to-negligible losses at 4 degrees C and losses of approximately 2.0 log10 infectious units at 25 degrees C when held for 45 weeks. Losses at 37 degrees C for one strain were approximately 10-fold greater when the strain was freeze-dried in the absence of SPGA. These results indicate that respiratory syncytial virus strains freeze-dried in the presence of a suitable stabilizer can be transported as unrefrigerated samples without undue losses in infectivity.

Excipients↗

DNA restriction analysis of adenovirus prototypes 1 to 41.

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.

Adenoviruses, Human↗

Three adenovirus type 8 genome types defined by restriction enzyme analysis: prototype stability in geographically separated populations.

Adenovirus type 8 strains were collected over a 19-year period from eye specimens from patients with keratoconjunctivitis. These strains were divided by restriction enzyme analysis with the endonucleases SalI, HindIII, SacI, KpnI, and SmaI into three genotypic subgroups. The prototype strain (Trim) was found throughout the United States from 1966 through 1985 and also in Taiwan and Greece in the early 1980s. Genotype 8C was identified in an arc from Maryland to Missouri to Alabama from 1971 through 1974. Genotype 8D was found only in an epidemic of eye disease among Vietnamese refugees being resettled in northwest Florida in 1975. Genotypes 8C and D were distinct from genotypes 8A and B described from Japan in 1975 through 1981. Hemagglutination tests with a battery of avian and mammalian erythrocytes did not distinguish between the genotypic subgroups. Similarly, the genotypes could not be differentiated by hemagglutination inhibition or serum neutralization tests with reference prototype antisera. The long-term prevalence of the Trim strain suggests that adenovirus type 8 has greater genetic stability than the other adenovirus types studied to date.

Adenoviridae Infections↗

Identification of epitopes on respiratory syncytial virus proteins by competitive binding immunoassay.

To characterize the interrelationship of monoclonal antibodies (MAbs) against respiratory syncytial virus (RSV) and their respective epitopes, we developed a competitive binding assay based on the biotin-avidin system and a tissue culture enzyme-linked immunosorbent assay. The competitive binding assay clearly distinguished between competing and noncompeting MAbs. Eight MAbs against the fusion protein (F protein) demonstrated two blocking patterns consistent with two antigenic sites. MAbs reacting at one site neutralized the virus, while those reacting at the other site did not. Eight MAbs against the large glycoprotein (G protein) demonstrated five blocking patterns consistent with three antigenic sites, one with three epitopes and the other two with one each. None of the MAbs against G protein neutralized the virus. The reaction pattern of the MAbs against three strains of RSV identified three additional epitopes on the F protein and no additional epitopes on the G protein. The epitopes on G protein showed the greatest antigenic diversity among the three strains. These results help us better understand the functional and antigenic structure of the two surface glycoproteins of RSV.

Animals↗

Immunological and biochemical characterization of human adenoviruses from subgenus B. I. Antigenic relationships.

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.

Adenoviruses, Human↗