Mutations responsible for attenuation in measles virus or human respiratory syncytial virus subgroup B
A technology of measles virus and syncytial virus, applied in the direction of viruses, viral peptides, antiviral agents, etc.
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Embodiment 1
[0183] measles virus
[0184] Moraten MV vaccine virus was grown for one passage before preparing RNA for sequence analysis, directly from Attenuvax TMVaccine vials (Lot#0716B), allowing Schwarz vaccine virus to grow one generation (Lot96G04 / M179 G41D), while Zagreb and Rubeovax TM Vaccine viruses were grown in Vero cells for two passages each. MV wild-type isolate Montefiore (57) was passaged in Vero cells for 5-7 passages, and then the RNA material was extracted. Similarly, MV wild-type isolates 1977, 1983 (14) were grown for 5-5 years before extracting material for analysis. 7 generations. From J. The Edmonston wild-type isolate from Dr. Beeler (CBER) (see Figure 1) was received before the original isolate had been passaged 7 times in human kidney cells, 3 times in Vero cells, and then in Vero before being used for sequence analysis. A virus that is passed once in a cell.
[0185] RNA was prepared by infecting Vero cells at a multiplicity of infect...
Embodiment 2
[0202] RSV subgroup B
[0203] The temperature-sensitive (ts) phenotype is closely related to in vivo attenuation; in addition, some non-temperature-sensitive mutations may also be attenuating. Identification of temperature-sensitive and non-temperature-sensitive attenuated mutants was achieved by sequence analysis and assessment of the temperature-sensitive, cold-adapted (ca), and in vivo growth phenotypes of RSV mutants and revertants.
[0204] The genomes of the following three RSV 2B strains have been completely sequenced: the 2B wild-type parent, its ts and ca derivatives, designated 2B33F, and a ts(+) revertant, designated 2B33F TS(+). The 2B33F strain is described in US Patent No. 08 / 059,444(88), incorporated herein by reference. After identifying the mutation region in 2B33F, it was passaged in vitro at 39°C, and after passage in African green monkeys or chimpanzees, another nine 2B33F "reverted strains" were obtained, and their previously identif...
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