Current challenges in implementing cell-derived influenza vaccines: implications for production and regulation, July 2007, NIBSC, Potters Bar, UK.

A meeting was held at NIBSC, UK in July 2007 to discuss the implications of progress in the use of cell culture systems for the manufacture of vaccines against influenza. Issues discussed included the effect of using eggs and different cell types in strain selection, development of seed viruses to be used in production and the nature of the reagents to be used in determining vaccine potency. Future studies to progress the field were reviewed.

Timely production of A/Fujian-like influenza vaccine matching the 2003-2004 epidemic strain may have been possible using Madin-Darby canine kidney cells.

Timely production and antigenic match with those of the epidemic strains are required for influenza vaccines. A/Fujian/411/2002-like (H3N2) virus was the main epidemic influenza virus during the 2003/2004 season in the northern hemisphere. But A/Fujian-like reassortant viruses were not available until more than one year later. We evaluated the A/Kumamoto/102/2002 strain, an A/Fujian/411/2002-like strain isolated in 2002, as a potential vaccine. We compared A/Kumamoto/102/2002 viruses isolated from the same clinical sample in Madin-Darby canine kidney (MDCK) cells and eggs. Kumamoto/102/2002 isolated from eggs grew poorly and showed amino acid mutations of haemagglutinin. In contrast, A/Kumamoto/102/2002 isolated from MDCK cells grew well in MDCK suspension culture. The amino acid sequence of MDCK-derived A/Kumamoto virus was identical to that of A/Fujian/411/2002. These results suggest that culture in MDCK cells could have produced an influenza vaccine with a better antigenetic match to the predicted epidemic strain for the 2003/2004 season than the vaccine actually produced.

Sequential immunization with V3 peptides from primary human immunodeficiency virus type 1 produces cross-neutralizing antibodies against primary isolates with a matching narrow-neutralization sequence motif.

An antibody response capable of neutralizing not only homologous but also heterologous forms of the CXCR4-tropic human immunodeficiency virus type 1 (HIV-1) MNp and CCR5-tropic primary isolate HIV-1 JR-CSF was achieved through sequential immunization with a combination of synthetic peptides representing HIV-1 Env V3 sequences from field and laboratory HIV-1 clade B isolates. In contrast, repeated immunization with a single V3 peptide generated antibodies that neutralized only type-specific laboratory-adapted homologous viruses. To determine whether the cross-neutralization response could be attributed to a cross-reactive antibody in the immunized animals, we isolated a monoclonal antibody, C25, which neutralized the heterologous primary viruses of HIV-1 clade B. Furthermore, we generated a humanized monoclonal antibody, KD-247, by transferring the genes of the complementary determining region of C25 into genes of the human V region of the antibody. KD-247 bound with high affinity to the "PGR" motif within the HIV-1 Env V3 tip region, and, among the established reference antibodies, it most effectively neutralized primary HIV-1 field isolates possessing the matching neutralization sequence motif, suggesting its promise for clinical applications involving passive immunizations. These results demonstrate that sequential immunization with B-cell epitope peptides may contribute to a humoral immune-based HIV vaccine strategy. Indeed, they help lay the groundwork for the development of HIV-1 vaccine strategies that use sequential immunization with biologically relevant peptides to overcome difficulties associated with otherwise poorly immunogenic epitopes.