Table 1.
Percentage of Subjects Developing Serum Hemagglutination-inhibition Antibody Titers ≥1∶40 (or 1∶32) by Vaccine HA Dosage after Vaccinations with Monovalent Inactivated Influenza A Virus Vaccines.1
Figure 1.
Evaluation sequence of antigen uptake, processing and presentation of influenza A vaccine and HA antigens to human T cells.
Figure 2.
Percent of human dendritic cells maturing after influenza A antigen loading.
Data are before and after dialysis with 10 and 20% of the initial vaccine or HA amount used so as to reduce any inhibitory effect. The vaccine concentration should not affect maturation; maturation in the presence of vaccine should be similar to the PBS control. The PBS control is also before and after dialysis. Available vaccine was insufficient for testing all four variables for the HA of Hong Kong/G1/99 (H9N2).
Table 2.
Dendritic Cell Maturation and Cytokine Secretion by T Cells Stimulated in vitro with Influenza A Virus Vaccines and Hemagglutinin Proteins.1
Figure 3.
Native and unfoldon ELISA assays for viruses and vaccines using polyclonal antisera and monoclonal antibodies.
The viruses and vaccines were diluted to equivalent hemagglutination units (HAU) although not comparable for detergent-released HA in vaccines versus virus particles and the H7 vaccine had no HAU. Analysis of the binding curves gives total binding sites (Bmax) (Figure 3A) and overall avidity (Kd) (Figure 3B).
Figure 4.
Polyacrylamide gels run under reducing and nonreducing conditions for trimeric HA and HA1 and HA2 subunits bound and separate.
Coomassie blue stain was used for protein and western blots with polyclonal antisera for protein identity. Gels are for vaccines (Figure 4A) and viruses (Figure 4B). Baculovirus expressed H3 HA was used as control (not shown).
Table 3.
Hemagglutination Titers of Monovalent Inactivated Influenza A Avian and Pandemic 2009 Influenza A Vaccines.
Figure 5.
Selected electron micrographs of vaccines illustrating the morphologic structures described in Table 4.
Figure 5A shows intact and split virus particles (asterisks) in the influenza A/Cal/04 (H1N1) subunit vaccine (CSL) of Table 1 along with structures of indistinct morphology (arrows). Also shown in Figure 5A is an EM of the A/Vietnam/04 (H5N1) subunit vaccine (SP) of references 5, 6, 10 in Table 1 that was selected to show a large number of stellate structures (arrows point to examples) although indistinct structures similar to those in the H1N1 vaccine were dominant in the H5N1 vaccine (not shown). Figure 5B is an EM of the influenza A/CK/G9 (H9N2) vaccine (Novartis) of Table 1 that shows the predominant varying size particles of membrane with external projections as well as a number of stellates, one apparently within an empty particle (arrow). Also in Figure 5B is an EM of the influenza A/Mallard (H7N7) vaccine (SP, ref. 12 in Table 1) that primarily showed small (5 to 20 nm) round and elongated structures (arrows point to examples). Some stellates and a rare intact particle were also seen.
Table 4.
Relative Proportions of Different Morphologic Units Seen in Electron Micrographs of Monovalent Influenza Vaccines.