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Several carcinoembryonic antigens (CD66) serve as receptors for gonococcal opacity proteins.

Chen T, Grunert F, Medina-Marino A, Gotschlich EC - J. Exp. Med. (1997)

Bottom Line: This conclusion was based on the following observations.We also compared the adherence and invasion by Opa+ bacteria of CD66 transfected HeLa cells: HeLa-BGPa, HeLa-CGM6, HeLa-NCA, HeLa-CGM1a, HeLa-CEA, and HeLa-Neo serving as negative control.Among the Opa proteins tested, OpaC proved to be bifunctional, able to mediate adherence to both syndecan receptors and to CD66 antigens.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, NY 10021-6399, USA.

ABSTRACT
Neisseria gonorrhoeae (GC) is a human pathogen that adheres to and invades genital surfaces. Although pili are required for the initial adherence, the interaction of GC with epithelial cells is also promoted by a family of outer membrane proteins, the opacity (Opa) proteins such as OpaA protein from strain MS11. Studies have demonstrated that the interaction of the OpaA GC with epithelial cells involves binding to heparan sulfate attached to syndecan receptors. However, other Opa proteins interact with CEA gene family member 1 (CGM1) or biliary glycoprotein (BGP), members of the CD66 antigen family. In this study, we demonstrate that, in addition, the 180-kD carcinoembryonic antigen (CEA) is a receptor for Opa proteins. This conclusion was based on the following observations. First, transfected HeLa cells expressing CEA (HeLa-CEA) and the CEA-expressing colon cancer cell line (LS 174T) bound and subsequently engulfed the Opa+ bacteria. These interactions were inhibited by anti-CEA antibody, but could not be inhibited by addition of heparin. Furthermore, OpaI E. coli directly bound purified CEA. We also compared the adherence and invasion by Opa+ bacteria of CD66 transfected HeLa cells: HeLa-BGPa, HeLa-CGM6, HeLa-NCA, HeLa-CGM1a, HeLa-CEA, and HeLa-Neo serving as negative control. Using OpaI as the prototype, the relative ability of the transfected HeLa cell lines to support adherence was (CEA = BGPa >CGM1a >NCA >CGM6 = Neo). The ability to mediate invasion of the transfectant cells was (CGM1a >CEA >BGPa >NCA >CGM6 = Neo). Among the Opa proteins tested, OpaC proved to be bifunctional, able to mediate adherence to both syndecan receptors and to CD66 antigens.

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Adherence and internalization of pEXI by HeLa  cells transfected with various  CD66 antigens. All the cell lines  were grown to confluence in 24well culture plates containing  RPMI medium 1640 and were  incubated with pEXI for 4.5 h.  The adherent and intracellular E.  coli were distinguished by incubation with gentamicin. (A)  The ability of these cell lines to  bind pEXI: HeLa-CEA >HeLaBGP >HeLa-CGM1a >NCA  >>HeLa-CGM6 = HeLa-Neo  = 0. (B) The number of intracellular bacteria. Although HeLaCGM1a showed the highest  internalization of pEXI, HeLaCEA could also engulf pEXI. The  internalization of pEXI into HeLaBGP was negligible. Bars, SEM.
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Figure 1: Adherence and internalization of pEXI by HeLa cells transfected with various CD66 antigens. All the cell lines were grown to confluence in 24well culture plates containing RPMI medium 1640 and were incubated with pEXI for 4.5 h. The adherent and intracellular E. coli were distinguished by incubation with gentamicin. (A) The ability of these cell lines to bind pEXI: HeLa-CEA >HeLaBGP >HeLa-CGM1a >NCA >>HeLa-CGM6 = HeLa-Neo = 0. (B) The number of intracellular bacteria. Although HeLaCGM1a showed the highest internalization of pEXI, HeLaCEA could also engulf pEXI. The internalization of pEXI into HeLaBGP was negligible. Bars, SEM.

Mentions: The HeLa-Neo, HeLa-BGPa (CD66a), HeLaCGM6 (CD66b), HeLa-NCA (CD66c), HeLa-CGM1a (CD66d), and HeLa-CEA (CD66e) were examined for their ability to support adherence and invasion of OpaI bacteria. The internalization of Opa+ bacteria into HeLa-CD66 transfectants was measured by gentamicin killing assay and the results were confirmed by electron microscopy. pEXI adhered poorly to HeLa-Neo and CGM6, but attached very well to HeLa-CEA and HeLa-BGP (Fig. 1 A). The adherence of pEXI to HeLa-CGM1a and HeLa-NCA was intermediate.


Several carcinoembryonic antigens (CD66) serve as receptors for gonococcal opacity proteins.

Chen T, Grunert F, Medina-Marino A, Gotschlich EC - J. Exp. Med. (1997)

Adherence and internalization of pEXI by HeLa  cells transfected with various  CD66 antigens. All the cell lines  were grown to confluence in 24well culture plates containing  RPMI medium 1640 and were  incubated with pEXI for 4.5 h.  The adherent and intracellular E.  coli were distinguished by incubation with gentamicin. (A)  The ability of these cell lines to  bind pEXI: HeLa-CEA >HeLaBGP >HeLa-CGM1a >NCA  >>HeLa-CGM6 = HeLa-Neo  = 0. (B) The number of intracellular bacteria. Although HeLaCGM1a showed the highest  internalization of pEXI, HeLaCEA could also engulf pEXI. The  internalization of pEXI into HeLaBGP was negligible. Bars, SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196295&req=5

Figure 1: Adherence and internalization of pEXI by HeLa cells transfected with various CD66 antigens. All the cell lines were grown to confluence in 24well culture plates containing RPMI medium 1640 and were incubated with pEXI for 4.5 h. The adherent and intracellular E. coli were distinguished by incubation with gentamicin. (A) The ability of these cell lines to bind pEXI: HeLa-CEA >HeLaBGP >HeLa-CGM1a >NCA >>HeLa-CGM6 = HeLa-Neo = 0. (B) The number of intracellular bacteria. Although HeLaCGM1a showed the highest internalization of pEXI, HeLaCEA could also engulf pEXI. The internalization of pEXI into HeLaBGP was negligible. Bars, SEM.
Mentions: The HeLa-Neo, HeLa-BGPa (CD66a), HeLaCGM6 (CD66b), HeLa-NCA (CD66c), HeLa-CGM1a (CD66d), and HeLa-CEA (CD66e) were examined for their ability to support adherence and invasion of OpaI bacteria. The internalization of Opa+ bacteria into HeLa-CD66 transfectants was measured by gentamicin killing assay and the results were confirmed by electron microscopy. pEXI adhered poorly to HeLa-Neo and CGM6, but attached very well to HeLa-CEA and HeLa-BGP (Fig. 1 A). The adherence of pEXI to HeLa-CGM1a and HeLa-NCA was intermediate.

Bottom Line: This conclusion was based on the following observations.We also compared the adherence and invasion by Opa+ bacteria of CD66 transfected HeLa cells: HeLa-BGPa, HeLa-CGM6, HeLa-NCA, HeLa-CGM1a, HeLa-CEA, and HeLa-Neo serving as negative control.Among the Opa proteins tested, OpaC proved to be bifunctional, able to mediate adherence to both syndecan receptors and to CD66 antigens.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, NY 10021-6399, USA.

ABSTRACT
Neisseria gonorrhoeae (GC) is a human pathogen that adheres to and invades genital surfaces. Although pili are required for the initial adherence, the interaction of GC with epithelial cells is also promoted by a family of outer membrane proteins, the opacity (Opa) proteins such as OpaA protein from strain MS11. Studies have demonstrated that the interaction of the OpaA GC with epithelial cells involves binding to heparan sulfate attached to syndecan receptors. However, other Opa proteins interact with CEA gene family member 1 (CGM1) or biliary glycoprotein (BGP), members of the CD66 antigen family. In this study, we demonstrate that, in addition, the 180-kD carcinoembryonic antigen (CEA) is a receptor for Opa proteins. This conclusion was based on the following observations. First, transfected HeLa cells expressing CEA (HeLa-CEA) and the CEA-expressing colon cancer cell line (LS 174T) bound and subsequently engulfed the Opa+ bacteria. These interactions were inhibited by anti-CEA antibody, but could not be inhibited by addition of heparin. Furthermore, OpaI E. coli directly bound purified CEA. We also compared the adherence and invasion by Opa+ bacteria of CD66 transfected HeLa cells: HeLa-BGPa, HeLa-CGM6, HeLa-NCA, HeLa-CGM1a, HeLa-CEA, and HeLa-Neo serving as negative control. Using OpaI as the prototype, the relative ability of the transfected HeLa cell lines to support adherence was (CEA = BGPa >CGM1a >NCA >CGM6 = Neo). The ability to mediate invasion of the transfectant cells was (CGM1a >CEA >BGPa >NCA >CGM6 = Neo). Among the Opa proteins tested, OpaC proved to be bifunctional, able to mediate adherence to both syndecan receptors and to CD66 antigens.

Show MeSH
Related in: MedlinePlus