Focus formation by rubella virus in rabbit kidney (RK-13) cell cultures and its application to the virus titration

1979 ◽  
Vol 60 (1) ◽  
pp. 75-78 ◽  
Author(s):  
M. Nawa
1969 ◽  
Vol 15 (1) ◽  
pp. 67-71 ◽  
Author(s):  
John Furesz ◽  
Pierre Moreau ◽  
Walter Yarosh

A simple and reproducible micro tissue culture assay has been devised in RK13 and LLC-RK1 rabbit kidney cells for the titration and neutralization of rubella virus. In this "one-step" assay all virus and serum dilutions were prepared with spiral loops in disposable microplates and tissue cultures suspended in medium 199 and 3% horse or fetal calf serum were added to the microcups simultaneously. Micro tissue cultures were kept in a humidified incubator (36 °C) under a constant pressure of 5% CO2 for 8 days and were read microscopically for viral cytopathic changes on the seventh and eighth day. The microneutralization test performed in LLC-RK1 cell cultures was shown to be a reliable method for the detection of small amounts of rubella antibodies in human sera.The micro assay may be also applied to the virus titration of live, attenuated rubella vaccines.


PEDIATRICS ◽  
1969 ◽  
Vol 44 (1) ◽  
pp. 5-23
Author(s):  
Harry M. Meyer ◽  
Paul D. Parkman ◽  
Hope E. Hopps

An attenuated strain of rubella virus was established by passage of the virus in primary African green monkey kidney cell cultures (GMK). In the first rubella vaccine clinical trial with seventy-seventh passage GMK material, none of eight inoculated children evidenced rubella-like illness and all developed antibody. The vaccine-induced infections were not transmitted to intimate contacts. Trials with large numbers of children confirmed the early findings of safety and immunogenicity of the vaccine. The second successfully attenuated virus, the Benoit strain, was developed by Hilleman and co-workers by adapting low GMK passages of virus to duck embryo cell cultures (DE). The Benoit strain, tested at several levels of tissue culture passage, designated A, B, C, D, and E, served as a clear demonstration of how rubella virus is modified by passage in mammalian or avian cells. Vaccines prepared from A and B level materials were not suitable, since they were either communicable or produced rubella-like symptoms. The C and D level materials were attenuated, but the higher passaged E level vaccine was termed "over-attenuated" since it induced antibody in only 60% of the vaccinees. More recently, the Cendehill strain, attenuated in primary rabbit kidney cell cultures (RK) and the RA 27/3 strain, attenuated in WI-38 cells, have been tested clinically. Vaccines which are now being produced by biologics manufacturers and considered candidates for license in the United States are: the HPV-77 strain passed 5 times in DE(HPV-77, DE5); the HPV-77 strain passed 12 times in dog kidney (DK) cell cultures (HPV-77, DK12), and the Cendehill strain passed 4 times in GMK and 53 times in RK (Cendehill GMK 4, RK 53). With each of these vaccines, antibody was detected in 90 to 100% of recipients. Fully attenuated rubella virus vaccines have not produced significant clinical reactions in children. However, mild rubella-like symptoms have been observed in adult women receiving vaccine; transient rashes, lymphadenopathy, arthralgia, and arthritis have been observed. Since the risk of spread of attenuated virus to the fetus in vaccinated women cannot be readily determined, the use of the vaccine during pregnancy would be potentially hazardous. Intranasal challenge of HPV-77 vaccinees with natural rubella virus demonstrated that vaccine-induced antibodies were protective against the clinical and virologic manifestations of rubella. Observation of vaccinated children over a 3-year period has demonstrated little evidence of a decline in the antibodies.


1970 ◽  
Vol 68 (3) ◽  
pp. 505-510 ◽  
Author(s):  
L. Grant ◽  
E. A. Belle ◽  
G. Provan ◽  
S. D. King ◽  
M. M. Sigel

SUMMARYThis report summarizes closed, family, and open studies conducted sequentially over a 10 month period with the Cendehill rubella virus vaccine in more than 16,000 children and adolescents. This strain of rubella was attenuated by serial propagation on primary rabbit kidney cell cultures. Inoculation of the Cendehill vaccine produced seroconversion in 97% of the 3589 susceptible (seronegative) vaccinated persons. There was no spread of the virus to susceptible controls living in close contact with those vaccinated. The vaccine was well tolerated. No arthritis or arthralgia occurred in 860 female subjects 13–18 years of age who were included in the study. The Cendehill vaccine would appear to meet the requirements of an acceptable vaccine.


1975 ◽  
Vol 9 (1) ◽  
pp. 61-68 ◽  
Author(s):  
T. Waller

Growth patterns of Nosema cuniculi ( Encephalitozoon cuniculi) in cell cultures of bovine kidney, canine kidney, feline lung, and rabbit kidney were studied. All cell cultures used were easy to manage and the last 3 are commercially-available established cell lines. The dog kidney cells were the most suitable for large-scale production of Nosema. When grown in plastic flasks with a bottom area of 75 cm2, the weekly yield from Nosema-infected canine kidney cells during the 10th to 17th week after inoculation was between 4·1 x 107 and 9·9 x 107 spores per flask. An equilibrium was obtained between the Nosema infection and the kidney cells during this time. A simple method for estimating the numbel of harvested spores is also described.


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