Using immunocytochemical techniques at the light and electron microscope levels, Legionella pneumophila and one of its extracellular proteases were located in the lungs of guinea pigs with experimental Legionnaires' disease (LD). L. pneumophila was immunostained by several peroxidase- and gold-labelling methods for light and electron microscopy. The protease was immunolabelled in tissue fixed in Carnoy's fluid at the light microscopical level and on broth-grown organisms at the ultrastructural level. It was not labelled in either formalin- or glutaraldehyde-fixed tissue. Using double-labelling techniques, L. pneumophila and protease were located in the same section and were shown to be intimately associated with pulmonary lesions, providing strong evidence for the role of this protease in LD pneumonia.
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