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HIGH RATES OF APOPTOSIS IN HUMAN MYCOBACTERIUM ULCERANS CULTURE-POSITIVE BURULI ULCER SKIN LESIONS

Buruli ulcer, a disease caused by Mycobacterium ulcerans, causes ulcerative skin disease likely generated by a toxin that mediates apoptosis. We analyzed paraffin-embedded sections of surgically excised Buruli ulcer lesions (two ulcers and one edematous plaque) and adjacent non-lesional skin samples (n = 9) for apoptosis by an indirect immunofluorescent terminal deoxynucleotide transferase–mediated dUTP-digoxigenin nick end labeling (TUNEL) assay. All samples were stained for acid-fast bacilli (AFB) and cultured for mycobacteria, and most were analyzed with an M. ulcerans-specific diagnostic polymerase chain reaction (PCR). TUNEL (+) bodies were numerous in both ulcers and the plaque, and sparse or absent in adjacent non-lesional skin. The AFB tissue stains and cultures for M. ulcerans were positive only in the three lesions. The result of the PCR for M. ulcerans was positive in all three lesions and in four of six non-lesional tissue samples; three contained sparse TUNEL (+) bodies. An abundance of TUNEL (+) bodies in the three AFB stain (+), culture (+), and PCR (+) Buruli ulcer lesional samples, but not in nearby AFB stain (–), culture (–), and PCR (+) non-lesional skin samples, strengthen the evidence that apoptosis is an important tissue destruction mechanism in human lesions closely associated with viable M. ulcerans.

Received February 10, 2005. Accepted for publication March 31, 2005.

Acknowledgments: We thank Sister Julia Aguiar and Dr. Lucie Avoaka-Cisse for providing the tissue samples, and Dr. Moo Hwang for photomicrography. This work was presented in part at the 53rd Annual Meeting of the American Society of Tropical Medicine and Hygiene, November 7–11, 2004, Miami, Florida (Abstract # 597). The American Committee on Clinical Tropical Medicine and Travelers’ Health (ACCTMTH) assisted with publication expenses.

Financial support: This work was supported in part by the Damien Foundation (Brussels) and the American Registry of Pathology (Washington, DC).

Disclaimer: The opinions or assertions presented herein are the private views of the authors and are not to be construed as official or as reflecting the views of the U.S. Departments of the Army or Defense.

Disclosure: There are no known commercial interests or other associations that might pose a conflict of interest.

^* Address correspondence to Dr. Douglas S. Walsh, Dermatology Service, Eisenhower Army Medical Center, Fort Gordon, GA, 30905. E-mail: douglas.walsh{at}se.amedd.army.mil

Authors’ addresses: Douglas S. Walsh, Dermatology Service, Eisenhower Army Medical Center, Fort Gordon, GA 30905, Telephone: 706-787-1472, Fax: 706-787-1354, E-mail: douglas.walsh{at}se.amedd.army.mil. Wayne M. Meyers, Department of Microbiology, Armed Forces Institute of Pathology, Washington, DC 20306-6000, E-mail: wmekmeyers{at}erols.com. Francoise Portaels, Mycobacteriology Unit, Institute of Tropical Medicine, Antwerp, Belgium, E-mail: portaels{at}itg.be. Joshua E. Lane, Department of Medicine, Section of Dermatology, The Medical College of Georgia, Augusta, GA 30912, E-mail: joshua.lane{at}lycos.com. Duangrat Mongkolsirichaikul, Kittinun Hussem, and Khin Saw Aye Myint, Department of Virology, United States Army Medical Component, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, E-mail: MyintK{at}afrims.org. Panita Gosi, Department of Immunology and Medicine, United States Army Medical Component, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, E-mail: GosiP{at}afrims.org.

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