Immunogenicity of Concentrated and Purified Rabies Vaccine of Tissue Culture Origin

Wiktor, T J; Sokol, F.; Kuwert, E.; Koprowski, Hilary

doi:10.3181/00379727-131-33981

Cited by 77 publications

(39 citation statements)

References 4 publications

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“…Concentrated, purified, (l-propiolactone-inactivated rabies vaccine was prepared from the ERA strain of virus according to reported techniques (8). The antigenicity of this vaccine preparation was 20 times the value of the standard WHO reference vaccine (9).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants.

Wiktor¹,

Koprowski²

1978

Proc. Natl. Acad. Sci. U.S.A.

201

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Somatic cell hybrids (hybridomas) between mouse myeloma cells and spleen cells derived from BALB/c mice immunized with inactivated rabies vaccine were found to produce antibodies to rabies virus. Monoclonal antibodies with different specificities were obtained either from the mass culture directly after fusion or from clones derived from a sinle-cell cloning procedure. Several strains of fixed or street raies virus were analyzed by virus neutralization procedures which demonstrated differences in their antigenic composition. Hybridoma antibodies were able to protect experimental animals from lethal rabies virus infection. Antibodies produced in animals immunized with the nucleocapsid fraction of rabies virus crossreact with nucleocapsids of all strains of rabies but not with coat proteins (1, 2). Antibodies produced by immunization of animals with whole virions or with the glycoprotein fraction can barely detect antigenic differences in coat proteins of different strains of rabies virus (2,3).Because monoclonal antibodies produced by somatic cell hybrids (hybridomas) against influenza A virus (4, 5) detect even minor antigenic differences among variants of the same strain of virus (5), it should be possible to produce hybridomas expressing different antibody specificities for various rabies strains.In the present study we have investigated the feasibility of production of antirabies antibodies by hybridomas formed by fusion of P3 X 63Ag8 myeloma (6) with splenocytes from rabies virus-immunized mice. Antirabies antibodies produced by hybridomas were analyzed for their specificities in various assays and used to protect mice infected with rabies virus. MATERIALS AND METHODSVirus Strains. Clone-purified ERA, PM, CVS, and Flury HEP strains of rabies virus were propagated in BHK-21 cell culture monolayers as described (7). Nine additional strains of rabies virus were obtained from different sources, as indicated in Table 3, and adapted by us to growth in BHK-21 cell cultures by described techniques (7).Vaccine. Concentrated, purified, (l-propiolactone-inactivated rabies vaccine was prepared from the ERA strain of virus according to reported techniques (8). The antigenicity of this vaccine preparation was 20 times the value of the standard WHO reference vaccine (9).Mice. Ten-to 12-week-old female BALB/c mice (Jackson Laboratories, Bar Harbor, ME) received a primary immunization of 0.1 ml of vaccine by intraperitoneal inoculation and a booster inoculation of the same vaccine diluted 1:5 3-4 months later by intravenous inoculation.Production of Hybrid Cells. Splenocytes from rabiesimmunized mice were fused with P3 X 63Ag8 mouse myeloma cells (6) as described (4, The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Methodsmentioning

confidence: 99%

Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants.

Wiktor¹,

Koprowski²

1978

Proc. Natl. Acad. Sci. U.S.A.

201

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“…In the sixties, human diploid fibroblast cells (WI-38 (Hayflick and Moorhead 1961), MRC-5 (Jacobs et al 1970)) were established and Wiktor et al (1964) could show that these cells could be used for the production of rabies virus for vaccine purposes. A larger production scale (use of 1 l bottles) was described and used for studying the immunogenicity of purified rabies vaccine in 1969 (Wiktor et al 1969) (This production method was later used for the industrial production of an inactivated rabies vaccine (Nicolas et al 1978)). In the same time (in the early sixties) BHK-21 (C13) cells were established and in 1964 the commercial production of inactivated FMD (Foot and Mouth Disease) vaccine was commenced by using a suspension process (based on the work by Capstick et al (1962)).…”

mentioning

confidence: 99%

Introduction to animal cell culture technology—past, present and future

Merten

2006

Cytotechnology

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Section: Discussionmentioning

confidence: 99%

“…This slowly sedimenting antigen was detectable even when treatment of the fractions with detergent was omitted, suggesting that it was associated with subviral particles. The presence of such 'soluble' antigen in tissue culture fluids from which rabies virus has been pelleted has been described previously (Wiktor et al, 1969), but its relevance to vaccine efficacy requires further study.Evidence that the glycoprotein antigen was quantitatively removed from rabies virus particles by treatment with Mulgofen (2 %) was obtained from experiments in which virus particle antigen was detergent-treated and sedimented on sucrose gradients containing 1% Mulgofen. In these studies, all antigen activity detected by SRD resided in fractions 22 to 27.…”

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confidence: 99%

A Single-Radial-Immunodiffusion Technique for the Assay of Rabies Glycoprotein Antigen: Application for Potency Tests of Vaccines Against Rabies

Ferguson

Schild

1982

Journal of General Virology

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SUMMARYAn assay for rabies glycoprotein antigen based on single-radial-immunodiffusion (SRD) is described. Rabies glycoprotein antigen at concentrations of 0.7/lg/ml or greater (approx. l international unit, IU) produced well-defined SRD reaction zones in immunoplates containing antibody to purified glycoprotein. Plots of zone area against relative antigen concentration were linear. The method was found to be of suitable sensitivity for in vitro potency assays of inactivated cell culture rabies vaccines. Qualitative differences were detected between rabies vaccines prepared by two different methods when these were analysed in sucrose gradients for glycoprotein antigen associated with intact virions or in 'soluble' form associated with subviral structures. In vaccines prepared by zonal ultracentrifugation the glycoprotein was totally associated with intact virus, whilst in those prepared by ultrafiltration comparable quantities of subviral antigen were also detected. The SRD test appears to have considerable potential for assays of the antigenic content of rabies vaccines and has the advantage of reducing reliance on conventional in vivo tests for immunogenicity which employ infectious virus.The glycoprotein is the only antigen of the rabies virus particle which is important in stimulating virus-neutralizing antibodies (Cox et al., 1977). Methods to quantify this protein are therefore important for the control and standardization of inactivated rabies vaccines for use in man and animals. Currently, in vivo assays are used routinely to standardize these vaccines. The tests most commonly used are the NIH test (Seligman, 1973) and the Habel test (Habel, 1973). The former involves the immunization of mice with test and reference antigens followed by intracerebral challenge with a standard strain (CVS) of rabies virus. The latter requires the titration of a virus preparation in mice immunized with vaccine and in control mice. These tests are time consuming and have the disadvantage of employing infectious virus, necessitating the use of special containment facilities in certain countries. In addition, potency estimates derived from them show considerable variability Barth & Jaeger, 1979). There is general agreement (Barth & Jaeger, 1979) that more satisfactory and rapid tests are required for in process control and standardization of rabies vaccines. Barth et al. (1981) have described an antibody-binding test for rabies antigen which gave more reproducible results than in vivo tests but still requires infectious virus.SRD tests have been used for the assay of other inactivated virus vaccines, including influenza (Schild et aL, 1975;Wood et al., 1977) and poliovirus (Schild et al., 1980). SRD tests have been adopted internationally for the assay of influenza vaccines enabling reproducible and accurate estimates (coefficient of variation, 5 %) of their haemagglutinin antigen content to be made (Wood et al., 1981). These tests thus appeared to be worthy of evaluation for rabies antigens.For SRD assays with rabies antige...

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Immunogenicity of Concentrated and Purified Rabies Vaccine of Tissue Culture Origin

Cited by 77 publications

References 4 publications

Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants.

Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants.

Introduction to animal cell culture technology—past, present and future

A Single-Radial-Immunodiffusion Technique for the Assay of Rabies Glycoprotein Antigen: Application for Potency Tests of Vaccines Against Rabies

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