ABSTRACT
Immunoelectron microscopy enables the detection and localization of bacterial antigens during in vitro infection (J.F.L. Weel and J.P.M. van Putten, Microb. Pathog. 4:213-222, 1988). In this study, we have used this method to get information on the role of lipooligosaccharides (LOS) in the pathogenesis of neisserial infections at the mucosal level. Ultrathin cryosections of Chang conjunctive epithelial cells infected with Neisseria gonorrhoeae (3 to 18 h) were incubated with LOS-specific monoclonal antibodies and gold-labeled protein A and viewed in the electron microscope. Our results demonstrate that the probed LOS determinants are stably expressed during the adherence, internalization, and intracellular processing of the bacteria. There was no indication of an adaptation of the gonococcal LOS expression to the host cell environment or of a degradation of the probed epitopes. The gold particles, representing LOS molecules, were predominantly located at the bacterial membranes, but sometimes the host cell plasma membrane was labeled as well, suggesting that LOS or LOS-containing membrane fragments interacted with the eucaryotic cells. This was confirmed when purified LOS was added to the cells. Two hours after LOS exposure, gold particles were observed at the plasma membrane of a subpopulation of the cells. After 18 h of LOS exposure, gold particles were also found in large vacuoles inside the cells, suggesting that LOS molecules were internalized by the cells. The function of observed LOS binding and endocytosis in the pathogenesis of neisserial infections remains to be defined.
