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Gene Synthesis> | Expression of HA Probes, Flow Cytometry, Cell Sorting, and Sequencing HA constructs consisting of the extracellular domain of HA modified to ablate sialic acid binding and C-terminally fused to (1) a T4-fibritin trimerization motif, (2) a biotinylatable AviTag sequence, and (3) a hexahistidine affinity tag were synthesized (Genscript), cloned into a CMV-expression plasmid, and ex- pressed as previously described (Whittle et al. | Get A Quote |
Antibodies capable of neutralizing divergent influenza A viruses could form the basis of a universal vaccine. Here, from subjects enrolled in an H5N1 DNA/MIV-prime-boost influenza vaccine trial, we sorted hemagglutinin cross-reactive memory B cells and identified three antibody classes, each capable of neutralizing diverse subtypes of group 1 and group 2 influenza A viruses. Co-crystal structures with hemagglutinin revealed that each class utilized characteristic germline genes and convergent sequence motifs to recognize overlapping epitopes in the hemagglutinin stem. All six analyzed subjects had sequences from at least one multidonor class, and-in half the subjects-multidonor-class sequences were recovered fr... More