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J C Hierholzer

Publications and source records attributed to J C Hierholzer.

121 records · Page 7Linked to original sources

Production and evaluation of a purified adenovirus group-specific (hexon) antigen for use in the diagnostic complement fixation test.

A simple procedure for the production of large volumes of purified adenovirus group-specific complement-fixing (CF) (hexon) antigen by selective adsorption to and elution from CaHPO(4) is described. Results of immunodiffusion tests, electrophoresis, electron microscopy, and tests for hemagglutination and infectivity indicate that the purified antigen consisted of a single virus component (hexon). The purified product contained little host materials. Unlike the crude virus harvest usually employed for serodiagnostic CF tests, the purified antigen demonstrated no anticomplementary activity and did not develop such activity during storage. The purified antigen was equal to or slightly more sensitive than crude virus harvests for serodiagnosis of adenovirus infections.

Adenoviridae↗

Nonspecific precipitation of serum proteins by sodium lauryl sulfate in agar diffusion and immunoelectrophoresis.

The anionic detergent sodium lauryl sulfate (SLS), in a final concentration of 0.1% and greater, reacted with whole serum in agar diffusion and immunoelectrophoresis to form artifactual precipitin lines. These lines occurred when either Ionagar or agarose was used as the supporting gel and were not affected by the presence of urea and 2-mercaptoethanol. Analytic chemical tests confirmed that the precipitating agent is SLS, and staining techniques showed that the detergent precipitates both protein and lipoprotein components of whole serum. Multiple artifactual precipitin lines occurred with a wide variety of animal sera, and a single line formed with human 7S immunoglobulin. Hence, in agar diffusion studies in which SLS is present in the test system, these artifactual lines may be easily misinterpreted as true antigen-antibody precipitin reactions.

Agar↗

Immunological basis of the adenovirus 8-9 cross-reaction.

The dedecon and hexon components of adenovirus types 8 and 9 have been extensively purified for use in establishing the basis of the cross-reaction between these types. Dodecons, the complete hemagglutinins, were purified 304- to 362-fold by fluorocarbon extraction, calcium phosphate batch chromatography, and ion-exchange column chromatography. Hexons, the group complement-fixation (CF) antigens, were purified 230- to 240-fold by erythrocyte adsorption, ion-exchange chromatography, and exclusion chromatography. Component antisera prepared in rabbits were tested in reciprocal fashion with crude virus and dodecon and hexon components. By hemagglutination-inhibition (HI), the dodecons of types 8 and 9 demonstrated the same predominantly one-sided relationship characteristic of the crude antigens. Some neutralizing activity was associated with both dodecons and hexons of each type. However, combining anti-dodecon and anti-hexon sera or producing antisera against the combined dodecon-hexon components resulted in neutralizing titers which were identical to titers obtained with antisera against the crude virus harvests. Dedecons of each type appear to share at least one antigenic determinant with hexons of the same type, and this determinant may reside on the vertex capsomere. Hexons possess group- and type-specific determinants, as shown by CF, neutralization, and immunodiffusion tests, and may exhibit some minor relationship between types 8 and 9. The results with the purified components are consistent with the predominantly one-sided antigenic relationship between types 8 and 9 in the conventional HI tests and the largely type-specific relationship by neutralization tests.

Adenoviridae↗

Standardized viral hemagglutination and hemagglutination-inhibition tests. I. Standardization of erythrocyte suspensions.

A spectrophotometric procedure for standardizing red blood cell suspensions for use in viral hemagglutination tests is described. The procedure is based on highly reproducible cyanmethemoglobin absorbance readings at 540 nm on any suitable spectrophotometer. Target values for milligrams of cyanmethemoglobin per 100 ml are given for the red cell suspensions of all mammalian and avian species employed in viral hemagglutination tests. By use of these values and a cyanmethemoglobin standard curve for a particular photometer, approximate 4% erythrocyte suspensions can be diluted to any lesser concentration. Coefficients of variation for the various diluted suspensions range from 4 to 7%.

Animals↗

Standardized viral hemagglutination and hemagglutination-inhibition tests. II. Description and statistical evaluation.

Standardized hemagglutination and hemagglutination-inhibition procedures are described and statistically evaluated for all animal viruses where applicable, except for rubella and the arbovirus group. The standardized tests employ a constant phosphate-buffered saline diluent and constant volumes of serum, antigen, and standardized erythrocyte suspension. The standardized hemagglutination test has a reproducibility of 84 to 96% with adenoviruses, rubeola, and the myxoviruses, and 78 to 93% with reoviruses; the standardized hemagglutination-inhibition test has a reproducibility of 95 to 100% with all viruses tested.

Adenoviridae↗