Interestingly, our results show an efficient transfer of cellular material from influenza-treated B cells (flu-B cells) to PDC, in an actin and Ca2+ dependent manner.
This increase was not due to direct T cell activation by flu-B cells, as flu-B cells alone did not induce any IFNγ increase.
We evaluate the biological power of the model in two yeast systems (gene perturbations and cell cycle), and a human dataset (response of epithelial cells to flu infection).
Cell-based flu vaccines offer a number of advantages over the traditional method: (a) cell lines are fully characterized and in compliance with regulatory guidelines [2]; (b) the raw materials for production are defined and can be easily produced in a short period [2], [9].
But Novartis is building a cell culture flu vaccine factory in Holly Springs, N.C., which might be ready for use in 2010 or 2011.
S anofi-Aventis and GlaxoSmithKline both have cell-based flu vaccines in early stages of clinical development.
DLD1 cells, DLD1/Sv-Flu previously cultured and DLD1/SV40-rLuc transfected with Renilla luciferase (rLuc) GL4.82 reporter gene under the control of Simian Virus 40 GL4.18 promoter, cells were maintained in complete RPMI 1640 medium and incubated in humidified incubator at 37°C and 5% CO2 level.
Cross-presentation of viral antigens by PDC was then examined by incubating GEN3 or primary PDC (both HLA-A2pos) with B cells or flu-B cells (HLA-A2neg), prior to addition of T cells.
In summary, the current study has demonstrated that: (i) SF medium (Plus MDCK) could support both the cell-growth in the solid microcarriers and H5N1 virus-replication in the well controlled and scalable bioreactors; (ii) the cell-based flu vaccines could be manufactured in a safe and user-friendly process with fewer contaminants and consistently high yield.
Gating was established for G1, G2/M, labeled undivided (flu) cells, and labeled divided (fld) cells.
