Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Jul 15;102(2):347–360. doi: 10.1172/JCI2406

Pneumococcal trafficking across the blood-brain barrier. Molecular analysis of a novel bidirectional pathway.

A Ring 1, J N Weiser 1, E I Tuomanen 1
PMCID: PMC508893  PMID: 9664076

Abstract

Although Streptococcus pneumoniae is a major cause of meningitis in humans, the mechanisms underlying its traversal from the circulation across the blood-brain barrier (BBB) into the subarachnoid space are poorly understood. One mechanism might involve transcytosis through microvascular endothelial cells. In this study we investigated the ability of pneumococci to invade and transmigrate through monolayers of rat and human brain microvascular endothelial cells (BMEC). Significant variability was found in the invasive capacity of clinical isolates. Phase variation to the transparent phenotype increased invasion as much as 6-fold and loss of capsule approximately 200-fold. Invasion of transparent pneumococci required choline in the pneumococcal cell wall, and invasion was partially inhibited by antagonists of the platelet-activating factor (PAF) receptor on the BMEC. Pneumococci that gained access to an intracellular vesicle from the apical side of the monolayer subsequently were subject to three fates. Most opaque variants were killed. In contrast, the transparent phase variants were able to transcytose to the basal surface of rat and human BMEC in a manner dependent on the PAF receptor and the presence of pneumococcal choline-binding protein A. The remaining transparent bacteria entering the cell underwent a previously unrecognized recycling to the apical surface. Transcytosis eventually becomes a dominating process accounting for up to 80% of intracellular bacteria. Our data suggest that interaction of pneumococci with the PAF receptor results in sorting so as to transcytose bacteria across the cell while non-PAF receptor entry shunts bacteria for exit and reentry on the apical surface in a novel recycling pathway.

Full Text

The Full Text of this article is available as a PDF (374.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguzzi A., Kleihues P., Heckl K., Wiestler O. D. Cell type-specific tumor induction in neural transplants by retrovirus-mediated oncogene transfer. Oncogene. 1991 Jan;6(1):113–118. [PubMed] [Google Scholar]
  2. Andersson B., Eriksson B., Falsen E., Fogh A., Hanson L. A., Nylén O., Peterson H., Svanborg Edén C. Adhesion of Streptococcus pneumoniae to human pharyngeal epithelial cells in vitro: differences in adhesive capacity among strains isolated from subjects with otitis media, septicemia, or meningitis or from healthy carriers. Infect Immun. 1981 Apr;32(1):311–317. doi: 10.1128/iai.32.1.311-317.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bielecki J., Youngman P., Connelly P., Portnoy D. A. Bacillus subtilis expressing a haemolysin gene from Listeria monocytogenes can grow in mammalian cells. Nature. 1990 May 10;345(6271):175–176. doi: 10.1038/345175a0. [DOI] [PubMed] [Google Scholar]
  4. Birkness K. A., Swisher B. L., White E. H., Long E. G., Ewing E. P., Jr, Quinn F. D. A tissue culture bilayer model to study the passage of Neisseria meningitidis. Infect Immun. 1995 Feb;63(2):402–409. doi: 10.1128/iai.63.2.402-409.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bito H., Honda Z., Nakamura M., Shimizu T. Cloning, expression and tissue distribution of rat platelet-activating-factor-receptor cDNA. Eur J Biochem. 1994 Apr 1;221(1):211–218. doi: 10.1111/j.1432-1033.1994.tb18731.x. [DOI] [PubMed] [Google Scholar]
  6. Bito H., Honda Z., Nakamura M., Shimizu T. Cloning, expression and tissue distribution of rat platelet-activating-factor-receptor cDNA. Eur J Biochem. 1994 Apr 1;221(1):211–218. doi: 10.1111/j.1432-1033.1994.tb18731.x. [DOI] [PubMed] [Google Scholar]
  7. Cabellos C., MacIntyre D. E., Forrest M., Burroughs M., Prasad S., Tuomanen E. Differing roles for platelet-activating factor during inflammation of the lung and subarachnoid space. The special case of Streptococcus pneumoniae. J Clin Invest. 1992 Aug;90(2):612–618. doi: 10.1172/JCI115900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cundell D. R., Gerard N. P., Gerard C., Idanpaan-Heikkila I., Tuomanen E. I. Streptococcus pneumoniae anchor to activated human cells by the receptor for platelet-activating factor. Nature. 1995 Oct 5;377(6548):435–438. doi: 10.1038/377435a0. [DOI] [PubMed] [Google Scholar]
  9. Cundell D. R., Weiser J. N., Shen J., Young A., Tuomanen E. I. Relationship between colonial morphology and adherence of Streptococcus pneumoniae. Infect Immun. 1995 Mar;63(3):757–761. doi: 10.1128/iai.63.3.757-761.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Geelen S., Bhattacharyya C., Tuomanen E. The cell wall mediates pneumococcal attachment to and cytopathology in human endothelial cells. Infect Immun. 1993 Apr;61(4):1538–1543. doi: 10.1128/iai.61.4.1538-1543.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerard N. P., Gerard C. Receptor-dependent internalization of platelet-activating factor. J Immunol. 1994 Jan 15;152(2):793–800. [PubMed] [Google Scholar]
  12. Gibson R. L., Lee M. K., Soderland C., Chi E. Y., Rubens C. E. Group B streptococci invade endothelial cells: type III capsular polysaccharide attenuates invasion. Infect Immun. 1993 Feb;61(2):478–485. doi: 10.1128/iai.61.2.478-485.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huang S. H., Wass C., Fu Q., Prasadarao N. V., Stins M., Kim K. S. Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo: molecular cloning and characterization of invasion gene ibe10. Infect Immun. 1995 Nov;63(11):4470–4475. doi: 10.1128/iai.63.11.4470-4475.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hulse M. L., Smith S., Chi E. Y., Pham A., Rubens C. E. Effect of type III group B streptococcal capsular polysaccharide on invasion of respiratory epithelial cells. Infect Immun. 1993 Nov;61(11):4835–4841. doi: 10.1128/iai.61.11.4835-4841.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Isberg R. R., Falkow S. A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature. 1985 Sep 19;317(6034):262–264. doi: 10.1038/317262a0. [DOI] [PubMed] [Google Scholar]
  16. Juillerat-Jeanneret L., Aguzzi A., Wiestler O. D., Darekar P., Janzer R. C. Dexamethasone selectively regulates the activity of enzymatic markers of cerebral endothelial cell lines. In Vitro Cell Dev Biol. 1992 Jul-Aug;28A(7-8):537–543. doi: 10.1007/BF02634138. [DOI] [PubMed] [Google Scholar]
  17. Kim J. O., Weiser J. N. Association of intrastrain phase variation in quantity of capsular polysaccharide and teichoic acid with the virulence of Streptococcus pneumoniae. J Infect Dis. 1998 Feb;177(2):368–377. doi: 10.1086/514205. [DOI] [PubMed] [Google Scholar]
  18. Korth R., Benveniste J. BN 52021 displaces [3H]paf-acether from, and inhibits its binding to intact human platelets. Eur J Pharmacol. 1987 Oct 27;142(3):331–341. doi: 10.1016/0014-2999(87)90071-9. [DOI] [PubMed] [Google Scholar]
  19. Krivan H. C., Roberts D. D., Ginsburg V. Many pulmonary pathogenic bacteria bind specifically to the carbohydrate sequence GalNAc beta 1-4Gal found in some glycolipids. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6157–6161. doi: 10.1073/pnas.85.16.6157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. LACKS S., HOTCHKISS R. D. A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta. 1960 Apr 22;39:508–518. doi: 10.1016/0006-3002(60)90205-5. [DOI] [PubMed] [Google Scholar]
  21. Lamont R. J., Chan A., Belton C. M., Izutsu K. T., Vasel D., Weinberg A. Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun. 1995 Oct;63(10):3878–3885. doi: 10.1128/iai.63.10.3878-3885.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morrison D. A., Trombe M. C., Hayden M. K., Waszak G. A., Chen J. D. Isolation of transformation-deficient Streptococcus pneumoniae mutants defective in control of competence, using insertion-duplication mutagenesis with the erythromycin resistance determinant of pAM beta 1. J Bacteriol. 1984 Sep;159(3):870–876. doi: 10.1128/jb.159.3.870-876.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nizet V., Kim K. S., Stins M., Jonas M., Chi E. Y., Nguyen D., Rubens C. E. Invasion of brain microvascular endothelial cells by group B streptococci. Infect Immun. 1997 Dec;65(12):5074–5081. doi: 10.1128/iai.65.12.5074-5081.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pagano R. E., Sleight R. G. Defining lipid transport pathways in animal cells. Science. 1985 Sep 13;229(4718):1051–1057. doi: 10.1126/science.4035344. [DOI] [PubMed] [Google Scholar]
  25. Pitrak D. L., Tsai H. C., Mullane K. M., Sutton S. H., Stevens P. Accelerated neutrophil apoptosis in the acquired immunodeficiency syndrome. J Clin Invest. 1996 Dec 15;98(12):2714–2719. doi: 10.1172/JCI119096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ponpipom M. M., Hwang S. B., Doebber T. W., Acton J. J., Alberts A. W., Biftu T., Brooker D. R., Bugianesi R. L., Chabala J. C., Gamble N. L. (+/-)-trans-2-(3-Methoxy-5-methylsulfonyl-4-propoxyphenyl)-5-(3,4,5- trimethoxyphenyl)tetrahydrofuran (L-659,989), a novel, potent PAF receptor antagonist. Biochem Biophys Res Commun. 1988 Feb 15;150(3):1213–1220. doi: 10.1016/0006-291x(88)90758-9. [DOI] [PubMed] [Google Scholar]
  27. Prasadarao N. V., Wass C. A., Weiser J. N., Stins M. F., Huang S. H., Kim K. S. Outer membrane protein A of Escherichia coli contributes to invasion of brain microvascular endothelial cells. Infect Immun. 1996 Jan;64(1):146–153. doi: 10.1128/iai.64.1.146-153.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pucciarelli M. G., Finlay B. B. Polarized epithelial monolayers: model systems to study bacterial interactions with host epithelial cells. Methods Enzymol. 1994;236:438–447. doi: 10.1016/0076-6879(94)36032-4. [DOI] [PubMed] [Google Scholar]
  29. Quagliarello V. J., Long W. J., Scheld W. M. Morphologic alterations of the blood-brain barrier with experimental meningitis in the rat. Temporal sequence and role of encapsulation. J Clin Invest. 1986 Apr;77(4):1084–1095. doi: 10.1172/JCI112407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rosenow C., Ryan P., Weiser J. N., Johnson S., Fontan P., Ortqvist A., Masure H. R. Contribution of novel choline-binding proteins to adherence, colonization and immunogenicity of Streptococcus pneumoniae. Mol Microbiol. 1997 Sep;25(5):819–829. doi: 10.1111/j.1365-2958.1997.mmi494.x. [DOI] [PubMed] [Google Scholar]
  31. Spellerberg B., Prasad S., Cabellos C., Burroughs M., Cahill P., Tuomanen E. Penetration of the blood-brain barrier: enhancement of drug delivery and imaging by bacterial glycopeptides. J Exp Med. 1995 Oct 1;182(4):1037–1043. doi: 10.1084/jem.182.4.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stins M. F., Prasadarao N. V., Ibric L., Wass C. A., Luckett P., Kim K. S. Binding characteristics of S fimbriated Escherichia coli to isolated brain microvascular endothelial cells. Am J Pathol. 1994 Nov;145(5):1228–1236. [PMC free article] [PubMed] [Google Scholar]
  33. Talbot U. M., Paton A. W., Paton J. C. Uptake of Streptococcus pneumoniae by respiratory epithelial cells. Infect Immun. 1996 Sep;64(9):3772–3777. doi: 10.1128/iai.64.9.3772-3777.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tuomanen E. I. The biology of pneumococcal infection. Pediatr Res. 1997 Sep;42(3):253–258. doi: 10.1203/00006450-199709000-00001. [DOI] [PubMed] [Google Scholar]
  35. Vann J. M., Proctor R. A. Cytotoxic effects of ingested Staphylococcus aureus on bovine endothelial cells: role of S. aureus alpha-hemolysin. Microb Pathog. 1988 Jun;4(6):443–453. doi: 10.1016/0882-4010(88)90029-0. [DOI] [PubMed] [Google Scholar]
  36. Weiser J. N., Austrian R., Sreenivasan P. K., Masure H. R. Phase variation in pneumococcal opacity: relationship between colonial morphology and nasopharyngeal colonization. Infect Immun. 1994 Jun;62(6):2582–2589. doi: 10.1128/iai.62.6.2582-2589.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiser J. N., Markiewicz Z., Tuomanen E. I., Wani J. H. Relationship between phase variation in colony morphology, intrastrain variation in cell wall physiology, and nasopharyngeal colonization by Streptococcus pneumoniae. Infect Immun. 1996 Jun;64(6):2240–2245. doi: 10.1128/iai.64.6.2240-2245.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES