Studies of the Preparation of Rubella Virus Vaccine and Rubella Hyperimmune gamma Globulin

摘要

With the intent to improve virus yields toward vaccine development, a number of factors were studied. Extraction of RK 13 cell-associated virus by Hallauer's pH 9.0 glycine buffer method proved to be less applicable to rubella than to entero or arboviruses. Up to 1.0 log more rubella virus accumulated in culture fluids than in extracts. Virus replication at 36C. was enhanced by daily harvests as compared with cumulative titers in the same medium; this relationship did not hold for 34C. On the other hand, cumulative virus yields were generally higher at 34C. than at 36C. both in the presence and absence of serum. Comparable titers obtained in media containing bovine fetal, normal or agamma rabbit sera or combinations of them. Virus passaged rapidly in RK 13 cells resulted in a 1.25 log increased titer. Considerable attention was given to neutralization tests in RK 13. Although operative for a year, the results have lately been erratic and the causes are not fully understood. Enhanced sensitivity of the assay by addition of guinea pig serum could not be corroborated and some lots were either inhibitory to virus or toxic to cells. Several lots of virus were prepared in primary AGMK cells in anticipation of UV and betaprone or formalin inactivation studies. One lot is now being processed. Earlier lots had to be discarded due to a rash of simian virus contaminants or loss of virus in serum-free media. Some improvement of yields was accomplished by rocking of the cultures. Further studies are underway such as alternating of serum and serum-free maintenance media, lower temperatures and other details in technic. The Carbowax method was found to permit quantitative concentration of virus. (Author)