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Infection and Immunity, July 1999, p. 3469-3480, Vol. 67, No. 7
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Neisseria gonorrhoeae Coordinately Uses Pili and Opa To Activate HEC-1-B Cell Microvilli, Which Causes Engulfment of the Gonococcidagger

J. McLeod Griffiss,1,2,* Claudia J. Lammel,2 Jun Wang,1,2,Dagger Nusi P. Dekker,2 and G. F. Brooks2

Centre for Immunochemistry1 and Department of Laboratory Medicine,2 University of California, San Francisco, California 94143

Received 7 December 1998/Returned for modification 29 January 1999/Accepted 6 April 1999

This study was undertaken to examine concomitant roles of pili and colony opacity-associated proteins (Opa) in promoting Neisseria gonorrhoeae adherence to and invasion of human endometrial HEC-1-B cells. Adherence of N. gonorrhoeae to cultured HEC-1-B cells was saturable, even though organisms adhered to <50% of the cells. During 4 to 6 h of incubation, adherent mono- and diplococci formed microcolonies on the surfaces of the cells. Microvilli of the HEC-1-B cells adhered by their distal ends to individual cocci within the microcolonies. When the microcolonies grew from isogenic pilus-negative (P-) Opa-, P- Opa+, or P+ Opa- gonococci, microvilli did not elongate, and the colonies were not engulfed. In contrast, the microvilli markedly elongated during exposure to P+ Opa+ gonococci. The microvilli adhered to the organisms along their full lengths and appeared to actively participate in the engulfment of the microcolonies. Internalized microcolonies, with P+ Opa+ gonococci, contained dividing cocci and appeared to be surrounded by cell membrane but were not clearly within vacuoles. In contrast, degenerate individual organisms were within vacuoles. Low doses of chloramphenicol, which inhibits protein synthesis by both prokaryotes and eukaryotes, prevented the microvillar response to and internalization of the P+ Opa+ gonococci; higher doses caused internalization without microvillus activation. Cycloheximide and anisomycin, which inhibit only eukaryotic protein synthesis, caused dose-dependent enhancement of uptake. Cytochalasins reduced engulfment; colchicine had no effect. These results show that gonococci must express both pili and Opa to be engulfed efficiently by HEC-1-B cells.

* Corresponding author. Mailing address: Department of Veterans Affairs Medical Center, 4150 Clement Street (111W1), San Francisco, CA 94121. Phone: (415) 476-5371. Fax: (415) 221-7542. E-mail: crapaud{at}vacom.ucsf.edu.

dagger Report no. 79 from the Centre for Immunochemistry, University of California, San Francisco.

Dagger Present address: Max-Planck-Institut für InfektionsbiologieAbteilung Molekular Biologie, D-10117 Berlin, Germany.

Infection and Immunity, July 1999, p. 3469-3480, Vol. 67, No. 7
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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