Recovery of Hepatitis Agents in the Marmoset from Human Cases Occurring in Costa Rica

Mascoli, Carmine C.; Ittensohn, Oswald L.; Villarejos, Víctor M.; Arguedas, Adriano; Provost, Philip J.; Hilleman, Maurice R.

doi:10.3181/00379727-142-37005

Cited by 63 publications

(14 citation statements)

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“…The CR326F strain of HAV was adapted to cell culture from a fecal specimen obtained during epidemiological studies in Costa Rica [5]. It was isolated into the diploid fetal rhesus monkey kidney cell line, FRhK-6 [6±8], and was later adapted to replicate in MRC-5 for 18 passages at 35°C and for 10 passages at 32°C.…”

Section: Hepatitis a Virus Master And Stock Seedmentioning

confidence: 99%

Development, preparation, and testing of VAQTA®, a highly purified hepatitis A vaccine

Hagen

Auniņš

DePhillips

et al. 2000

Bioprocess Engineering

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Manufacture of VAQTA Ò , an inactivated hepatitis A vaccine, uses state-of-the-art technologies in cell culture and bioprocessing science, which have made it possible to routinely produce the vaccine at manufacturing scale. VAQTA Ò consists of an attenuated strain of hepatitis A virus that is highly puri®ed and formaldehyde-inactivated, then formulated with an aluminum hydroxide adjuvant. Process development and scale-up have resulted in a well-characterized vaccine manufacturing process with appropriate in-process controls to assure consistent performance, and a reproducible, well-de®ned product. Results are presented from a series of manufacturing demonstration lots to show consistency, as well as comparability to clinical lots prepared at an earlier stage in development.

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Section: Hepatitis a Virus Master And Stock Seedmentioning

confidence: 99%

Development, preparation, and testing of VAQTA®, a highly purified hepatitis A vaccine

Hagen

Auniņš

DePhillips

et al. 2000

Bioprocess Engineering

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“…Natural hepatitis A and B cases in Costa Rica. These were cases of hepatitis A or B that occurred among persons who resided in the province of Alajuela, Costa Rica and who were subjects in the large-scale epidemiologic studies that were conducted there (2,5,7,8). They bear eight digit identification (e.g., 206-033-02).…”

mentioning

confidence: 99%

“…Preparation of CF antigen. The hepatitis A CF antigen preparations were made from livers of S. mystax marmosets infected intravenously with CR326 virus (2,5). The livers were collected from the animals at a time when the serum enzyme values (SGOT and SICD) were ele-vated.…”

mentioning

confidence: 99%

A Specific Complement-Fixation Test for Human Hepatitis A Employing CR326 Virus Antigen. Diagnosis and Epidemiology

Provost

Ittensohn

Villarejos

et al. 1975

Experimental Biology and Medicine

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Studies of human hepatitis A (infectious hepatitis) have been hampered by the lack of a simple assay for antibody against hepatitis A virus. Such a test has been developed as the present report will show.Historically, Deinhardt et al.(1) first presented evidence for propagation of hepatitis A virus in Saguinus nigricollis and Saguinus oedipus marmosets. Provost et al.(2) in our laboratories developed an assay for neutralizing antibody against hepatitis A virus in S. mystax marmosets and provided definitive proof of etiologic relationship between human hepatitis A isolate CR326 and human hepatitis A. The serum neutralization findings were confirmed by Holmes et al. (3). Using another approach, Feinstone et al. (4) later demonstrated a 27 nm diameter particle in the stool filtrate of a case of hepatitis A and the development of an antibody against it that was detected by immune electron microscopy.The present report describes the development of a specific complement-fixation (CF) test for human hepatitis A antibody employing, as antigen, liver extract of marmosets infected with the CR326 strain (2, 5 ) of human hepatitis A virus. Data relating to the development of CF antibody against the virus in hepatitis A cases are presented and discussed.Materials and Methods. Virus. The CR326 strain (5) of human hepatitis A virus isolated in marmosets from a case of hepatitis A in Costa Rica was used to prepare CF antigen. Preparation of CF antigen. The hepatitis A CF antigen preparations were made from livers of S. mystax marmosets infected intravenously with CR326 virus (2, 5). The livers were collected from the animals at a time when the serum enzyme values (SGOT and SICD) were ele-vated. The control CF antigens were made from livers of uninfected marmosets. All livers were perfused thoroughly with phosphate buffered saline (PBS), pH 7.2, minced with a scissors, and ground in a mortar with sterile alundum to give a 10% suspension by weight in PBS. The antigens consisted of the supernatant fluids obtained after low speed clarification. Both viral and control antigens were heated at 56" for 2 hr prior to use in the CF tests. Hepatitis A complement-fixation (CF) tests. Grid titrations were performed using serial dilutions of human hepatitis A convalescent serum and serial dilutions of antigen. The CF antigen titers were low and it was usually necessary to employ the infected liver preparation at a 1 :2 dilution (2 units of antigen) in the tests for hepatitis A antibody. The normal control CF antigen was used at the same dilution. Two and one half full units of guinea pig complement were employed in the tests. Incubation with complement was overnight at 4". The hemolytic system consisted of a 1 % suspension of sensitized sheep red blood cells. Antibody titers were expressed as the highest initial dilution of serum giving 50% or greater fixation of complement. Hepatitis B CF tests. Tests for the surface antigen (HB,Ag, Australia antigen) were performed according to Purcell et al. (6).Patients' sera. Natural hepatitis A and B ca...

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“…This was mainly because of the lack of a reliable method for detecting virus and viral antigens specifically. Thereafter, this situation has been improved to some extent by animal models employing marmoset monkeys or chimpanzees as susceptible hosts for the virus [Deinhardt et al, 1967;Lorenz et al, 1970;Mascoli et al, 1973;Maynard et al, 1975;Provost et al, 1975a;Deinhardt, 19763 by virtue of which fairly sufficient amounts of material were eventually provided to develop various Accepted for publication April 24, 1981. credible assays for HAV and the antibodies Provost et al, 1975b;Miller et al, 1975;Hollinger et al, 1975;Duermeyer et al, 19781. In fact, with the aid of the newly developed assays, Provost and Hilleman [1979] first reported authentic results for in vitro propagation of HAV by employing cell cultures of the marmoset liver and rhesus fetal kidney tissues.…”

Section: Introductionmentioning

confidence: 99%

Propagation of human hepatitis A virus in conventional cell lines

Kojima¹,

Shibayama²,

Sato³

et al. 1981

Journal of Medical Virology

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Fecal extracts of hepatitis A (HA) patients were selected for the presence of hepatitis A virus (HAV) by radioimmunoassay (RIA) and immune electron microscopy (IEM). When FL and Vero cells were inoculated with fecal extracts containing HAV, development of hepatitis A antigen (HAAg) was evident in the cytoplasm of the two cell lines by the indirect immunofluorescence (IF) test. The antigen was detectable in the cells 12 hr postinoculation (pi), and reached a plateau within two days pi. FL cell cultures inoculated with a specimen containing HAV were harvested and passaged four times. During the passages, efficient production of HAAg was confirmed in the infected cultures by three different serological tests: The indirect IF test, RIA using fixed cells, and RIA by the sandwich method. At the second and fourth passages, HAV particles were recovered in abundance from infected FL cell cultures by IEM. Throughout these experiments, no cytopathic effect (CPE) was discernible in the cultures.

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Recovery of Hepatitis Agents in the Marmoset from Human Cases Occurring in Costa Rica

Cited by 63 publications

References 0 publications

Development, preparation, and testing of VAQTA®, a highly purified hepatitis A vaccine

Development, preparation, and testing of VAQTA®, a highly purified hepatitis A vaccine

A Specific Complement-Fixation Test for Human Hepatitis A Employing CR326 Virus Antigen. Diagnosis and Epidemiology

Propagation of human hepatitis A virus in conventional cell lines

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