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Dissimilar Properties of Two Recombinant Lactobacillus acidophilus Strains Displaying Salmonella FliC with Different Anchoring Motifs

Kajikawa, Akinobu, Nordone, Shila K., Zhang, Lin, Stoeker, Laura L., LaVoy, Alora S., Klaenhammer, Todd R., Dean, Gregg A.
Applied and environmental microbiology 2011 v.77 no.18 pp. 6587-6596
Lactobacillus acidophilus, Salmonella, bicarbonates, binding proteins, buffers, cell walls, dendritic cells, digestive juices, enzymes, flagellin, humans, immunization, juices, mucus, recombinant vaccines, soybeans, surface antigens, trypsin inhibitors, viability
Display of heterologous antigens on the cell surface is considered a useful technique for vaccine delivery by recombinant lactobacilli. In this study, two recombinant Lactobacillus acidophilus derivatives displaying Salmonella flagellin (FliC) were constructed using different anchor motifs. In one instance, the FliC protein was fused to the C-terminal region of a cell envelope proteinase (PrtP) and was bound to the cell wall by electrostatic bonds. In the other case, the same antigen was conjugated to the anchor region of mucus binding protein (Mub) and was covalently associated with the cell wall by an LPXTG motif. These two recombinant L. acidophilus cell surface displays resulted in dissimilar maturation and cytokine production by human myeloid dendritic cells. The surface-associated antigen was highly sensitive to simulated gastric and small intestinal juices. By supplementation with bicarbonate buffer and soybean trypsin inhibitor, the cell surface antigen was protected from proteolytic enzymes during gastric challenge in vitro. The protective reagents also increased the viability of the L. acidophilus cells upon challenge with simulated digestive juices. These results demonstrate the importance of protecting cells and their surface-associated antigens during oral immunization.