Growth and Attenuation of Rabies Virus in Cell Cultures of Reptilian Origin

1972 ◽  
Vol 139 (4) ◽  
pp. 1317-1325 ◽  
Author(s):  
H. F. Clark
Keyword(s):  
1972 ◽  
Vol 6 (6) ◽  
pp. 988-995 ◽  
Author(s):  
T. J. Wiktor ◽  
H F. Clark

Author(s):  
Chunfu Li ◽  
Yongzhi Wang ◽  
Huiting Liu ◽  
Xinghua Zhang ◽  
Dalai Baolige ◽  
...  

Lab-attenuated rabies virus (RABV) is a highly cellular adaptation and less pathogenic than wild-type RABV. However, the molecular mechanisms that regulate the cellular adaptation and pathogenicity remain obscure. In this work, we isolated a wild-type RABV (CNIM1701) from a rabid bovine in northern China. The original CNIM1701 was lethal in adult mice and restricted replication in cell cultures. After 20 serial passages in the brains of suckling mice, the virus was renamed CNIM1701-P20, which was safe in adult mice and replicated well in cell cultures. In addition, sequence comparison analysis of the original CNIM1701 and CNIM1701-P20 identified 2 amino acid substitutions on G protein (Lys83 → Arg83 and Pro367 → Ser 367) related to pathogenesis and cellular adaptation. Using site-directed mutagenesis to exchange Lys83 with Arg83 and Pro367 with Ser 367 in the G protein of the RABV SAD strain, the pathogenicity of rSAD-K83R was significantly decreased. Our data indicate that the decreased pathogenicity of rSAD-K83R is due to increasing the expression of RABV-G, which also induced a higher level of apoptosis in infected cells. Furthermore, the K83 mutation induced high expression of MMP-2 and MMP-9 on DCs and promoted blood–brain barrier (BBB) permeability. These results demonstrate that the pathogenesis of RABV is partially dependent on G expression and BBB permeability, which may help in the design and development of highly safe, live-RABV vaccines.


Intervirology ◽  
1978 ◽  
Vol 9 (6) ◽  
pp. 359-361 ◽  
Author(s):  
Abigail L. Smith ◽  
Gregory H. Tignor ◽  
Richard W. Emmons ◽  
James D. Woodie

1989 ◽  
Vol 35 (8) ◽  
pp. 811-813
Author(s):  
W. A. Webster ◽  
K. M. Charlton

Murine neuroblastoma (NA-C1300) and baby hamster kidney (BHK-21/C13) cell cultures were infected with the Canadian Arctic strain of rabies virus. Subcultures were passed following incubation for 3 to 4 days at 35 °C. The supernatant fluids from the BHK cultures demonstrated increasing infectivity in both NA and BHK cells concomitantly with an increase in the number of parent cells staining with an anti-glycoprotein stain. On the other hand, the supernatant fluids from the NA cultures initially showed higher infectivity in NA cells than in BHK cells. This feature was related to a low production of glycoprotein-staining cells in the parent NA cultures. The reduction of infectivity in NA cells of some NA supernatant fluids (and brain suspensions) by anti-nucleoprotein antibodies suggests that nucleocapsid material is, in some manner, capable of infecting NA cells. Infectivity of this virus strain in experimental mice is also related to the production of glycoprotein and may not be correlated with the degree of infection in NA cell cultures.Key words: rabies, nucleocapsid, infection, cells.


2021 ◽  
Vol 14 (4) ◽  
pp. 28-37
Author(s):  
Yu. Krasnopolsky ◽  

Rabies is a neurological disease of a viral nature, leading to death. Rabies virus is an RNA virus that invades the central nervous system, leading to neuronal dysfunction. Timely vaccination can prevent the diseases development. Aim. The article is devoted to immunobiotechnological research aimed at creating antirabic vaccines. Results. The history of the antirabic vaccines creation from the first inactivated vaccines obtained from nervous tissue to the cultivation of the virus on animal cell cultures is considered. The article presents commercially available anti-rabies vaccines: their composition, the used rabies virus strains, cell cultures, the methods of inactivation and purification. The technology of producing an anti-rabies vaccine based on a Pitman Moore virus strain and a chicken fibroblast cell culture is presented. The advantages of different vaccine types are considered: live attenuated, peptide, liposomal, RNA vaccines, vaccines based on viral vectors, transgenic plants and reverse genetics methods. Conclusions. The development of biotechnology, immunology and virology makes it possible to improve constantly vaccine preparations, including those against rabies, increasing their effectiveness and safety.


2014 ◽  
Vol 3 (4) ◽  
pp. 323
Author(s):  
E. P. Barkova ◽  
F. G. Nagieva ◽  
V. G. Nikulina ◽  
A. N. Lisakov

1991 ◽  
Vol 60 (2) ◽  
pp. 161-164
Author(s):  
V. Celer ◽  
G. Belay ◽  
V. Celer

Author(s):  
Sergey V. Generalov ◽  
Pavel S. Erokhin ◽  
Oleg S. Kuznetsov ◽  
Elena G. Abramova ◽  
Ivan M. Zhulidov ◽  
...  

Background: Mouse neutralization test is widely used to determine the level of anti-rabies antibodies, but it is labor-intensive and time consuming. Alternative methods for determining the neutralizing activity of anti-rabies sera and immunoglobulin in cell cultures are also known. Methods such as FAVN and RFFIT involve the use of fluorescent diagnostics. Determination of Cytopathic Effect (CPE) is often complicated due to features of rabies virus replication in cells. Atomic Force Microscopy (AFM) is able to detect the interaction of the virus with the cell at an early stage. Therefore, in this study, a method has been developed for determining the specific activity of anti-rabies sera and immunoglobulin using AFM of cell cultures.   Methods: The method is based on the preliminary interaction of rabies virus with samples of rabies sera or immunoglobulin drug, adding the specified reaction mixture to cell culture (Vero or BHK-21), and then measuring the surface roughness of the cells using AFM. AFM was carried out in the intermittent contact mode by the mismatch method in the semi-contact mode. The results were compared with the values obtained in the mouse neutralization test. The consistency of the results obtained by both methods was evaluated by Bland-Altman method.   Results: The increment in the surface roughness of the cells is a consequence of the damaging effect of the virus, which is weakened as a result of its neutralization by rabies antibodies. A dilution allowing 50% suppression of the increase in the surface roughness of cells was selected as the titer of rabies sera or immunoglobulin. In this case, the recommended range for determining the antibody titer is from 1:100 to 1:3000.   Conclusion: For the first time, a new methodological approach in virology and pharmaceutical research is presented in this study. The use of the proposed methodological technique will reduce the time from 21 to 2 days to obtain results in comparison with the mouse neutralization test; also, fewer laboratory animals are required in this approach which is in agreement with 3 R Principle.


2001 ◽  
Vol 92 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Neuza M Frazzati-Gallina ◽  
Rosana L Paoli ◽  
Regina M Mourão-Fuches ◽  
Soraia A.C Jorge ◽  
Carlos A Pereira

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