Rabies virus glycoprotein. II. Biological and serological characterization

Abstract

Purified rabies virus glycoprotein (G) was shown by complement fixation and immunodiffusion tests to be a second distinct antigen of the virus. It it the only structural protein of the virus that induces the formation of virus-neutralizing antibodies and which confers immunity to animals. When the G protein is taken as antigen, the complement fixation test can be used for the assay of virus-neutralizing antibodies. The total protective activity of the virus was recovered in the purified G protein preparation. The protective activity of G protein increased with purification: 9 ng of G protein was required to protect 50% of the mice as compared to 1.63 micrograms of the virus. Selective immunofluorescent membrane staining and immunocytolysis of rabies virus-infected cells were shown to be G protein specific. Due to its purity and potency, the G protein preparation can be considered the ideal human antirabies vaccine.

Rabies DNA vaccine: No impact of MHC Class I and Class II targeting sequences on immune response and protection against lethal challenge

“…These facts indicate the need of more faithful candidate vaccines which must be capable of inducing strong immune response to protect from infection. More importantly, the candidate immunogen must be able to induce a strong Th2 immunity as it has been established that Th2; that is, the humoral immune response plays a predominant role in induction of protective immunity against rabies virus [32][33][34]. Rabies virus glycoprotein is the main antigen responsible for inducing the production of rabies virus-neutralizing antibodies and for conferring immunity against lethal rabies infection.…”

A rapid immunochromatographic test strip for detecting rabies virus antibody

“…It was found that rabies virus G protein could be isolated from Triton X-100-disrupted rabies virus, without losing its biological and serological properties (Atanasiu et al, 1974(Atanasiu et al, , 1976Cox et al, 1977;Smith et al, 2006). In this study, the inactivated rabies virus was purified by sucrose gradient centrifugation, then soluble glycoprotein was dissociated from the virus by incubation with 1% Triton X-100 and purified by sucrose centrifugation and Sephadex G-50 filtration.…”

“…Rabies virus glycoprotein is the only structural protein of the virus that induces the formation of virus-neutralizing antibodies, which confer immunity to animals. The protective activity was shown to be associated only with the G protein and not with the membrane associated protein M and P, although they contain residual traces of G protein (Cox et al, 1977). Competitive ELISA that utilizes a cell line stably expressing the rabies virus G protein as the coating antigen has higher specificity and sensitivity than that of ELISA based on whole rabies virus antigen (Bahloul et al, 2005;Servat and Cliquet, 2006).…”

“…In the study presented here, the virus was cultured at 32 • C for 4 d, and this was sufficient to obtain more G protein than virus grown at 37 • C. In order to reduce the false-positive results, sucrose gradient centrifugation was used to purify the virus. Subsequently the G protein was dissociated from the purified virus by incubation with Triton X-100, which could have a higher biological and protective activity than that solubilized by deoxycholate or by Nonidet P-40 (Cox et al, 1977). The sensitivity and specificity of ICTS in this study did not produce any difference between serum samples derived from dogs vaccinated with inactivated Pasteur virus (PV) strain, inactivated Evelyn-Rokitnicki-Abelseth (ERA) strain, Flury-LEP vaccines and commercial vaccines (Intervet Nobivac) (Table 1), and there was also an excellent correlation with the FAVN test.…”

“…The rabies virus glycoprotein (RABV G) was prepared by a modification of a technique which was described previously (Barton and Campbell, 1988;Cox et al, 1977;Smith et al, 2006). MNA cells were infected with CVS-11 at a multiplicity of infection (MOI) of 0.1, and incubated at 32 • C for 4 d. Culture fluid was collected, cells were removed by centrifugation (1500 × g, 20 min, 4 • C), and the virus was pelleted by ultracentrifugation at 45,000 × g for 2 h at 4 • C. Pellets were resuspended in PBS and left overnight at 4 • C. Betapropiolactone (BPL, Sigma, Inc.) was added to the virus solution to a final BPL dilution of 1:3000.…”