Recombinant measles AIK-C strain expressing current wild-type hemagglutinin protein.
We constructed a recombinant measles virus cDNA, pIC-MVAIK-H/87-K, in which the hemagglutinin (H) gene of the AIK-C vaccine strain was replaced by the wild-type (MVi/Tokyo.JPN/87-K: genotype D3) H gene and the remaining genes were the same as the AIK-C vaccine strain. To investigate the feasibility of the recombinant vaccine strain expressing wild-type H protein instead of the AIK-C H protein, we constructed two recombinant measles cDNA, having Leu (small plaque-type) and Phe (large plaque-type) at position 278 of the F protein. Infectious chimeric virus strains, MVAIK-H/87-K/S (small plaque-type) and MVAIK-H/87-K/L (large plaque-type), were recovered, which were designed to induce small (S) and large (L) plaques in Vero cells. The MVAIK-H/87-K/S and MVAIK-H/87-K/L did not grow at 39-40 degrees C, similar to the original AIK-C strain, and retained the temperature sensitivity (ts) characteristics. They did not induce cytopathic effect (CPE) in Vero cells but produced CPE in B95a cells, similar to the current wild-type measles MVi/Tokyo.JPN/87-K. From the results of Western blotting, the mobility of the H protein of MVAIK-H/87-K/S and MVAIK-H/87-K/L was similar to that of MVi/Tokyo.JPN/87-K. Hyper-immune sera raised by MVAIK-H/87-K/S neutralized all types of current wild strains. Thus, the chimeric measles virus expressing the current wild H protein demonstrated wild-type H properties with ts characteristics of the vaccine strain, indicating that the construction strategy of recombinant measles virus can cope with the hyper-mutated measles virus.
Kumada
Atsushi
A
Department of Pediatrics, Tokyo Medical College, 160-0023 Tokyo, Japan.
Komase
Katsuhiro
K
Nakayama
Tetsuo
T
D000818:Animals; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006388:Hemagglutinins / Q000096:biosynthesis*; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology; D006801:Humans; D008458:Measles Vaccine / Q000235:genetics* / Q000276:immunology*; D008459:Measles virus / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013696:Temperature; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic / Q000276:immunology; D014709:Vero Cells; D010948:Viral Plaque Assay
D018076:DNA, Complementary; D004279:DNA, Viral; D006388:Hemagglutinins; D006389:Hemagglutinins, Viral; D008458:Measles Vaccine; D011993:Recombinant Fusion Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; C032397:hemagglutinin protein G, measles virus