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J Med Microbiol 56 (2007), 579-586; DOI: 10.1099/jmm.0.46920-0
© 2007 Society for General Microbiology
ISSN 1473-5644

A peptide mimetic of the mycobacterial mannosylated lipoarabinomannan: characterization and potential applications

Ayelet Barenholz1, Avi-Hai Hovav1,{dagger}, Yolanta Fishman1, Galia Rahav2, Jonathan M. Gershoni3 and Hervé Bercovier1

1 Department of Clinical Microbiology, The Faculty of Medicine of the Hebrew University of Jerusalem, Jerusalem, Israel

2 Unit of Infectious Diseases, Sheba Medical Center, Ramat Gan, Israel

3 Department of Cell Research and Immunology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel

Correspondence
Ayelet Barenholz
ayeletba{at}ekmd.huji.ac.il

Received 29 August 2006
Accepted 5 January 2007

Mannosylated lipoarabinomannan (ManLAM), a complex lipoglycan, is a major component of Mycobacterium tuberculosis, the agent of tuberculosis (TB), and is an antigen used for serological diagnosis of TB. Screening random phage-display peptide libraries with anti-ManLAM mAb CS40 for peptide epitope mimics (mimotopes) led to the isolation of a panel of peptides. One of these peptides (B11) was characterized as a ManLAM mimotope: it bound the anti-ManLAM CS40 mAb and competed with ManLAM for antibody binding. Mice immunized with keyhole limpet haemocyanin-conjugated B11 peptide in a proper adjuvant developed antibodies that recognized ManLAM. Competition experiments demonstrated that the B11 peptide inhibited binding of mAb CS40 to ManLAM in a concentration-dependent manner. The data indicated that the affinity of CS40 mAb to B11 (KD 1.33x10–8) is higher than its affinity to ManLAM (KD 3.00x10–7). The sera of TB patients, as well as the sera of mice experimentally infected with M. tuberculosis, contained significant levels of antibodies that recognized both the B11 peptide and ManLAM. The specificity and sensitivity of the ELISA B11-based test were similar to those of the ELISA ManLAM-based test, indicating that the B11 antigen could be a good substitute for ManLAM serology for the diagnosis of TB.

Abbreviations: i.p., intraperitoneally; KLH, keyhole limpet haemocyanin; ManLAM, mannosylated lipoarabinomannan; MPL-TDM, monophosphoryl lipid A+trehalose dimycolate; s.c., subcutaneously; SPR, surface plasmon resonance; TB, tuberculosis.

{dagger}Present address: Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.






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