Incubation of intact Ty21a, serovar Typhimurium () aroA or DH5α with membrane vesicles (MVs) derived from either M90T or dsp89 resulted in a significant incorporation of vesicle antigens into the outer membrane of the bacteria; each recipient strain possessed a surface mosaic of new and antigens intermixed with the native antigens of the strains. Electron microscopy of preparations during the integration of vesicle antigens revealed that the MVs rapidly fused with the outer membrane of the host strains. Western blot analysis of host bacteria confirmed the integration of foreign antigens. Quantitative analysis for binding and fusion of antigens using an ELISA showed that approximately 78·7 ± 12·8 ng of the and 67·5 ± 13·8 ng of the LPSs (μg host protein) were integrated into the strain. Similar integrations of the or vesicles were found with the or strains. There was no loss of viability in the recipient bacteria after incorporation of the MV's, although vesicle antigens became diluted during continued growth as daughter cells shared the vesicle antigens. The new antigens were highly stable after being incorporated into recipient strains, being able to withstand storage of several months at 4°C as well as several cycles of freezing and thawing. Since the recipient bacteria are common vaccine strains, the procedure described here offers a simple efficient means of introducing exogenous surface antigens, in their native form, into the outer membranes of Gram-negative bacteria for possible vaccine use.


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