Cell wall-associated protein A as a tool for immunolocalization of Staphylococcus aureus in infected human airway epithelium.
Staphylococcus aureus is a common human pathogen involved in non-bronchial diseases and in genetic and acquired bronchial diseases. In this study, we applied an immunolabeling approach for in vivo and in vitro detection of S. aureus, based on the affinity of staphylococcal protein A (SpA) for the Fc region of immunoglobulins, especially IgG. Most strains of S. aureus, including clinical strains, can be detected with this labeling technique. The approach can be used for detection and localization with transmission electron microscopy or light-fluorescence microscopy of S. aureus in infected tissues such as human bronchial tissue from cystic fibrosis (CF) patients. The methodology can also be applied to cell culture models with the aim of characterizing bacterial adherence to epithelial cells in backscattered electron imaging with scanning electron microscopy. Application to the study of S. aureus adherence to airway epithelium showed that the bacteria did not adhere in vivo to intact airway epithelium. In contrast, bacteria adhered to the basolateral plasma membrane of columnar cells, to basal cells, to the basement membrane and were identified beneath the lamina propria when the epithelium was injured and remodeled, or in vitro when the epithelial cells were dedifferentiated.
Mongodin
E
E
INSERM U514, IFR53, CHU Maison-Blanche, Reims, France.
Bajolet
O
O
Hinnrasky
J
J
Puchelle
E
E
de Bentzmann
S
S
D000328:Adult; D002473:Cell Wall; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology; D005260:Female; D006046:Gold; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000378:metabolism* / Q000382:microbiology; D008297:Male; D008854:Microscopy, Electron / Q000379:methods; D008856:Microscopy, Fluorescence; D013205:Staphylococcal Protein A / Q000378:metabolism*; D013211:Staphylococcus aureus
D013205:Staphylococcal Protein A; D006046:Gold