Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease

Olga V. Nikolskaia, Ana Paula C De A Lima, Yuri V. Kim, John D. Lonsdale-Eccles, Toshihide Fukuma, Julio Scharfstein, Dennis J. Grab

Research output: Contribution to journalArticle

Abstract

In this study we investigated why bloodstream forms of Trypanosoma brucei gambiense cross human brain microvascular endothelial cells (BMECs), a human blood-brain barrier (BBB) model system, at much greater efficiency than do T. b. brucei. After noting that T. b. gambiense displayed higher levels of cathepsin L-like cysteine proteases, we investigated whether these enzymes contribute to parasite crossing. First, we found that T. b. gambiense crossing of human BMECs was abrogated by N-methylpiperazine-urea-Phe-homopheylalanine-vinylsulfone- benzene (K11777), an irreversible inhibitor of cathepsin L-like cysteine proteases. Affinity labeling and immunochemical studies characterized brucipain as the K11777-sensitive cysteine protease expressed at higher levels by T. b. gambiense. K11777-treated T. b. gambiense failed to elicit calcium fluxes in BMECs, suggesting that generation of activation signals for the BBB is critically dependant on brucipain activity. Strikingly, crossing of T. b. brucei across the BBB was enhanced upon incubation with brucipain-rich supernatants derived from T. b. gambiense. The effects of the conditioned medium, which correlated with ability to evoke calcium fluxes, were canceled by K11777, but not by the cathepsin B inhibitor CA074. Collectively, these in vitro studies implicate brucipain as a critical driver of T. b. gambiense transendothelial migration of the human BBB.

Original languageEnglish (US)
Pages (from-to)2739-2747
Number of pages9
JournalJournal of Clinical Investigation
Volume116
Issue number10
DOIs
StatePublished - Oct 2 2006

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Calcium Signaling
Cysteine Proteases
Trypanosomiasis
Blood-Brain Barrier
Parasites
Cathepsin L
Endothelial Cells
Brain
Trypanosoma brucei gambiense
Calcium
Transendothelial and Transepithelial Migration
Cathepsin B
Conditioned Culture Medium
Benzene
Urea
N-pip-phenylalanine-homophenylalanine-vinyl sulfone phenyl
Enzymes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Nikolskaia, O. V., Lima, A. P. C. D. A., Kim, Y. V., Lonsdale-Eccles, J. D., Fukuma, T., Scharfstein, J., & Grab, D. J. (2006). Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease. Journal of Clinical Investigation, 116(10), 2739-2747. https://doi.org/10.1172/JCI27798

Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease. / Nikolskaia, Olga V.; Lima, Ana Paula C De A; Kim, Yuri V.; Lonsdale-Eccles, John D.; Fukuma, Toshihide; Scharfstein, Julio; Grab, Dennis J.

In: Journal of Clinical Investigation, Vol. 116, No. 10, 02.10.2006, p. 2739-2747.

Research output: Contribution to journalArticle

Nikolskaia, OV, Lima, APCDA, Kim, YV, Lonsdale-Eccles, JD, Fukuma, T, Scharfstein, J & Grab, DJ 2006, 'Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease', Journal of Clinical Investigation, vol. 116, no. 10, pp. 2739-2747. https://doi.org/10.1172/JCI27798
Nikolskaia, Olga V. ; Lima, Ana Paula C De A ; Kim, Yuri V. ; Lonsdale-Eccles, John D. ; Fukuma, Toshihide ; Scharfstein, Julio ; Grab, Dennis J. / Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease. In: Journal of Clinical Investigation. 2006 ; Vol. 116, No. 10. pp. 2739-2747.
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