Passage in Vero cells alters the characteristics of measles AIK-C vaccine strain.

Objective of this study is to evaluate the feasibility of measles vaccine production in Vero cell culture. We constructed the full-length cDNA, pIC-MVAIK-F278Leu (small plaque-type) and pIC-MVAIK-F278Phe (large plaque-type) from the AIK-C measles vaccine strain attenuated from the Edmonston wild-type. MVAIK-S/B2 was rescued from pIC-MVAIK-F278Leu after two passages in B95a cells and MVAIK-SL/B2V1 was obtained through large plaque cloning in Vero cells. MVAIK-SL/B2V8 was obtained after eight passages in Vero cells. It produced large plaques in Vero cells, grew well at 39 degrees C, and thus the characteristics of the AIK-C vaccine strain were lost. Thirteen amino acid changes were observed; one in the N, two in the P, one in the C, three in the F, one in the H, and five in the L protein regions. Twelve of these changes excluding one in the L gene were back mutated to the Edmonston strain. Change from Leu to Phe at position 278 of the F protein was an early event during adaptation to Vero cells and the P gene was back-mutated to the Edmonston wild-type. As for the control, MVAIK-L/B9 strain was obtained after passages in B95a cells from pIC-MVAIK-F278Phe (large plaque-type). It maintained the same temperature sensitivity as the AIK-C vaccine strain and only four amino acid changes, one in the N and three in the L protein region, were observed without any mutations in the P, C, M, F, and H genes. The passage of the measles vaccine AIK-C strain in Vero cells lost the characteristics of small plaque inducibility and temperature sensitivity (ts) phenotype.