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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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The binding of purified CEA to pEXI. Equal numbers of pGEM (Opa−) and pEXI  (OpaI) were suspended in RPMI  containing equal amounts of  CEA. After incubation, the bacteria were recovered by centrifugation, washed, and lysed, and  the lysates were subjected to  SDS-PAGE and transfered to  Immobilon-P membrane. The  CEA bound on the bacteria was  detected by anti-CD66 mAb  (COL-1). Much more CEA was  bound by pEXI than by pGEM.
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Figure 3: The binding of purified CEA to pEXI. Equal numbers of pGEM (Opa−) and pEXI (OpaI) were suspended in RPMI containing equal amounts of CEA. After incubation, the bacteria were recovered by centrifugation, washed, and lysed, and the lysates were subjected to SDS-PAGE and transfered to Immobilon-P membrane. The CEA bound on the bacteria was detected by anti-CD66 mAb (COL-1). Much more CEA was bound by pEXI than by pGEM.

Mentions: The HeLa-CEA cells supported the highest level of adherence with pEXI (Fig. 1 A), indicating that the CEA protein is a receptor for Opa protein. To demonstrate a direct binding of Opa+ bacteria with CEA, purified CEA was mixed with equal amounts of Opa negative pGEM or positive pEXI bacteria. The bacteria–CEA mixture was incubated for 1 h, pelleted, and solubilized. The amount of CEA antigen bound to the bacteria was detected with COL-1 antibody by Western blotting. As seen in Fig. 3, pEXI bound much more CEA than pGEM.


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

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

The binding of purified CEA to pEXI. Equal numbers of pGEM (Opa−) and pEXI  (OpaI) were suspended in RPMI  containing equal amounts of  CEA. After incubation, the bacteria were recovered by centrifugation, washed, and lysed, and  the lysates were subjected to  SDS-PAGE and transfered to  Immobilon-P membrane. The  CEA bound on the bacteria was  detected by anti-CD66 mAb  (COL-1). Much more CEA was  bound by pEXI than by pGEM.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 3: The binding of purified CEA to pEXI. Equal numbers of pGEM (Opa−) and pEXI (OpaI) were suspended in RPMI containing equal amounts of CEA. After incubation, the bacteria were recovered by centrifugation, washed, and lysed, and the lysates were subjected to SDS-PAGE and transfered to Immobilon-P membrane. The CEA bound on the bacteria was detected by anti-CD66 mAb (COL-1). Much more CEA was bound by pEXI than by pGEM.
Mentions: The HeLa-CEA cells supported the highest level of adherence with pEXI (Fig. 1 A), indicating that the CEA protein is a receptor for Opa protein. To demonstrate a direct binding of Opa+ bacteria with CEA, purified CEA was mixed with equal amounts of Opa negative pGEM or positive pEXI bacteria. The bacteria–CEA mixture was incubated for 1 h, pelleted, and solubilized. The amount of CEA antigen bound to the bacteria was detected with COL-1 antibody by Western blotting. As seen in Fig. 3, pEXI bound much more CEA than pGEM.

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