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Infection and Immunity, September 2001, p. 5671-5678, Vol. 69, No. 9
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.9.5671-5678.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Expression of a Mycobacterium tuberculosis Arabinomannan Antigen In Vitro and In Vivo

J. Reid Schwebach,1 Arturo Casadevall,1,2 Rachel Schneerson,3 Zhongdong Dai,3 Xiaojuan Wang,4 John B. Robbins,3 and Aharona Glatman-Freedman5,*

Department of Microbiology and Immunology,1 Department of Medicine,2 Department of Pediatrics-Children's Hospital at Montefiore,5 and Howard Hughes Medical Institute,4 Albert Einstein College of Medicine, Bronx, New York 10461, and National Institute of Child Health and Human Development and Biotechnology Unit, National Institutes of Health, Bethesda, Maryland 208923

Received 3 January 2001/Returned for modification 26 March 2001/Accepted 7 June 2001

The outermost layer of Mycobacterium tuberculosis contains two major polysaccharides, arabinomannan (AM) and glucan (GC). We studied the in vitro and in vivo expression of an M. tuberculosis AM antigen using monoclonal antibody (MAb) 9d8 (2a), an isotype-switched variant of the immunoglobulin G3 (IgG3) MAb 9d8. MAb 9d8 had been previously shown to bind M. tuberculosis AM and the M. tuberculosis surface. Our in vitro experiments showed that MAb 9d8(2a) bound strongly to whole-cell M. tuberculosis Erdman but not to the CDC 1551 strain grown in medium for an extended period. However, AM antigen was detected in the culture supernatant of both strains, and its concentration increased in a time-dependent manner. The detection of AM antigen from both strains was decreased in the presence of Tween 80. In mice infected with M. tuberculosis Erdman, AM antigen accumulated in organ homogenates concomitant to an increase in bacterial organ burden and an increase in IgG and IgM titer to AM. These results (i) indicate that the surface expression of AM during in vitro growth changes with culture age, is strain dependent, and is affected by the presence of Tween 80 in the culture media; (ii) show that AM is produced by bacteria growth in vivo; and (iii) demonstrate that the amount of in vivo-detected AM can be dependent on the number of bacteria in the infected organ.

* Corresponding author. Mailing address: 702 Golding Building, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-3768. Fax: (718) 430-8701. E-mail: afreedma{at}aecom.yu.edu.

Infection and Immunity, September 2001, p. 5671-5678, Vol. 69, No. 9
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.9.5671-5678.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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