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PMID: 9712803 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Actinobacillus actinomycetemcomitans leukotoxin induces apoptosis in HL-60 cells.

Infection and immunity ·Vol. 66 ·No. 9 ·1998-09-00 ·Pages 4474-83

Korostoff J, Wang JF, Kieba I, Miller M, Shenker BJ, Lally ET

Abstract

Actinobacillus actinomycetemcomitans leukotoxin (Ltx) is a member of the repeats-in-toxin (RTX) family of pore-forming toxins and kills human immune cells. Currently, it remains unclear whether toxin-mediated killing of target cells involves the induction of necrosis or apoptosis. Therefore, the goal of this investigation was to determine whether Ltx is capable of causing apoptotic cell death in toxin-sensitive promyelocytic HL-60 cells. Multiparameter flow cytometric analysis of toxin-treated cells stained with Hoechst 33258 (or 33342) and 7-aminoactinomycin D allowed us to identify four populations: viable cells, early apoptotic cells, late apoptotic and/or secondarily necrotic cells, and a final population that was composed of cellular debris. Compared with control cells, HL-60 cells treated with Ltx exhibited a gradual decrease in forward light scatter with a coincident increase in side light scatter, indicative of a decrease in cell size and organelle condensation, respectively. Additional experiments demonstrated that Ltx-treated cells showed evidence of internucleosomal DNA fragmentation and phosphatidylserine translocation. The results of our studies clearly demonstrate that Ltx can kill HL-60 cells by inducing apoptosis. We hypothesize that elimination of acute inflammatory cells via this mechanism plays a critical role in the pathogenesis of diseases caused by A. actinomycetemeomitans.

MeSH Terms
Aggregatibacter actinomycetemcomitans/physiology Apoptosis Bacterial Toxins/pharmacology Cytotoxins/pharmacology DNA Fragmentation Dose-Response Relationship, Drug Exotoxins/pharmacology Flow Cytometry HL-60 Cells Humans Phosphatidylserines/metabolism Recombinant Fusion Proteins/pharmacology
Chemicals
Bacterial Toxins Cytotoxins Exotoxins Phosphatidylserines Recombinant Fusion Proteins leukotoxin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Korostoff J
Leon Levy Research Center for Oral Biology, University of Pennsylvania, Philadelphia, Pennsylvania, 19104-6002, USA. [email protected]
Wang J F
Kieba I
Miller M
Shenker B J
Lally E T
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.66.9.4474-4483.1998
Email
[email protected]
Published
1998-09-00
Pages
4474-83
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC108541
Subset
IM
Grants
NIDCR NIH HHS · R01 DE009517 · United States
NIDCR NIH HHS · R01 DE012305 · United States
NIDCR NIH HHS · DE09517 · United States
NIDCR NIH HHS · DE12305 · United States
External Links
PubMed source DOI: 10.1128/IAI.66.9.4474-4483.1998 PMC Full Text View journal details
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