Induction of apoptosis/necrosis in various human cell lineages by Haemophilus ducreyi cytolethal distending toxin.
We investigated the impact of highly purified Haemophilus ducreyi cytolethal distending toxin (HdCDT) on the apoptosis and necrosis of various human cells; including myeloid cells, epithelial cells, keratinocytes, and primary fibroblasts. The levels of apoptosis and necrosis induced in these cells were compared to those induced by HdCDT in human T cells and in the Jurkat T cell line. Levels of caspase-3 activity were measured, and membrane changes like phosphatidylserine (PS) translocation was evaluated after double-staining with the fluorescein isothiocyanate (FITC)-labeled annexin V and propidium iodide (PI) using flow cytometry. HdCDT induced various degrees of apoptosis and necrosis in dose- and time-dependent manners in cells of various lineages. Early and late apoptosis (annexin V-stained cells) were induced in more than 90% of T cells and monocytes after treatment with 100 ng/ml HdCDT for 24 and 48 h, respectively. The corresponding numbers for epithelial cells, keratinocytes, and fibroblasts were 26-32% after treatment with 100 ng/ml HdCDT for 48 h. HdCDT appears to eliminate effectively by inducing apoptosis those cells that are involved in immune responses. Epithelial cells, keratinocytes and fibroblasts, which are important for the healing of chancroid ulcers, are eliminated by apoptosis or necrosis after contact with HdCDT, albeit slower and to a lesser extent than T cells.
Wising
Catharina
C
Department of Medical Microbiology and Immunology, The Sahlgrenska Academy, Göteborg University, Box 435, 40530 Göteborg, Sweden.
Azem
Jozef
J
Zetterberg
Madeleine
M
Svensson
Liselott A
LA
Ahlman
Karin
K
Lagergård
Teresa
T
D017304:Annexin A5; D017209:Apoptosis / Q000187:drug effects*; D001427:Bacterial Toxins / Q000633:toxicity*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D005434:Flow Cytometry; D006191:Haemophilus ducreyi / Q000737:chemistry*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects; D022423:Myeloid Cells / Q000187:drug effects; D009336:Necrosis; D010718:Phosphatidylserines / Q000378:metabolism; D011419:Propidium; D021381:Protein Transport / Q000187:drug effects; D018709:Statistics, Nonparametric
D017304:Annexin A5; D001427:Bacterial Toxins; D010718:Phosphatidylserines; C067044:cytolethal distending toxin; D011419:Propidium; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases