Preparation of monoclonal antibodies against mannosylated lipoarabinomannan (ManLAM), a surface antigen of BCG vaccine produced in Iran

Authors

1 Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Clinical Biochemistry, School of Medicine, The University of Tarbiat Modares, Tehran, Iran

Abstract

Background: Bacille Calmette–Guerin (BCG) vaccine is the only vaccine that is used against Mycobacterium tuberculosis, but its efficacy is limited in mycobacterium-endemic regions. One of the major antigens present on the cell envelope of the vaccine that suppresses the immune system is mannosylated lipoarabinomannan (ManLAM).
Materials and Methods: In this study, we immunized 4-week-old mice with sonicated BCG vaccine injected intraperitoneally two times at an interval of 2 weeks and with ManLAM antigen injected intravenously and then extracted the spleen cells of the immunized mice. They were fused with SP2/0 myeloma cells.
Results: Five cell line clones producing antibody against ManLAM antigens were prepared and each clone was tested for immunoreactivity against sonicated BCG and purified ManLAM by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The clones designated H13F33E11 and H23D91G4 reacted strongly with ManLAM. Immunoblotting using monoclonal antibodies (MAbs) H13F33E11 and H23D91G4 showed that these MAbs bind to ManLAM with a molecular weight of 35 kDa.
Conclusions: In this study, we produced a monoclonal antibody of immunoglobulin G3 (IgG3) subclass. This MAb can be used for purification of ManLAM in culture media and detection of the antigen in patient's urine and for increasing the efficacy of BCG vaccine.

Keywords

1.
Dye C, Scheele S, Dolin P, Pathania V, Raviglione MC. Consensus statement. Global burden of tuberculosis: Estimated incidence, prevalence, and mortality by country. WHO Global Surveillance and Monitoring Project. JAMA 1999;282:677-86.  Back to cited text no. 1
    
2.
Colditz GA, Brewer TF, Berkey CS, Wilson ME, Burdick E, Fineberg HV, et al. Efficacy of BCG vaccine in the prevention of tuberculosis. Meta-analysis of the published literature. JAMA 1994;271:698-702.  Back to cited text no. 2
    
3.
Fard RM, Moslemy M, Golshahi H. The history of modern biotechnology in Iran: A medical review. J Biotechnol Biomater 2013;3:1-5.  Back to cited text no. 3
    
4.
Glatman-Freedman A, Casadevall A. Serum therapy for tuberculosis revisited: Reappraisal of the role of antibody-mediated immunity against Mycobacterium tuberculosis. Clin Microbiol Rev 1998;11:514-32.  Back to cited text no. 4
    
5.
Guirado E, Amat I, Gil O, Díaz J, Arcos V, Caceres N, et al. Passive serum therapy with polyclonal antibodies against Mycobacterium tuberculosis protects against post-chemotherapy relapse of tuberculosis infection in SCID mice. Microbes Infect 2006;8:1252-9.  Back to cited text no. 5
    
6.
de Vallière S, Abate G, Blazevic A, Heuertz RM, Hoft DF. Enhancement of innate and cell-mediated immunity by antimycobacterial antibodies. Infect Immun 2005;73:6711-20.  Back to cited text no. 6
    
7.
Prinzis S, Chatterjee D, Brennan PJ. Structure and antigenicity of lipoarabinomannan from Mycobacterium bovis BCG. J Gen Microbiol 1993;139:2649-58.  Back to cited text no. 7
    
8.
Venisse A, Berjeaud JM, Chaurand P, Gilleron M, Puzo G. Structural features of lipoarabinomannan from Mycobacterium bovis BCG. Determination of molecular mass by laser desorption mass spectrometry. J Biol Chem 1993;268:12401-11.  Back to cited text no. 8
    
9.
Pethe K, Alonso S, Biet F, Delogu G, Brennan MJ, Locht C, et al. The heparin-binding hemagglutinin of M. uberculosis is required for extrapulmonary dissemination. Nature 2001;412:190-4.  Back to cited text no. 9
    
10.
Torrelles JB, Sieling PA, Zhang N, Keen MA, McNeil MR, Belisle JT, et al. Isolation of a distinct Mycobacterium tuberculosis mannose-capped lipoarabinomannan isoform responsible for recognition by CD1b-restricted T cells. Glycobiology 2012;22:1118-27.  Back to cited text no. 10
    
11.
Hamasur B, Haile M, Pawlowski A, Schroder U, Kallenius G, Svenson SB. A mycobacterial lipoarabinomannan specific monoclonal antibody and its F(ab') fragment prolong survival of mice infected with Mycobacterium tuberculosis. Clin Exp Immunol 2004;138:30-8.  Back to cited text no. 11
    
12.
Köhler G, Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol 1976;6:511-9.  Back to cited text no. 12
    
13.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5.  Back to cited text no. 13
[PUBMED]    
14.
Nigou J, Zelle-Rieser C, Gilleron M, Thurnher M, Puzo G. Mannosylated lipoarabinomannans inhibit IL-12 production by human dendritic cells: Evidence for a negative signal delivered through the mannose receptor. J Immunol 2001;166:7477-85.  Back to cited text no. 14
    
15.
Lawn SD, Frimpong EH, Nyarko E. Evalutoin of a commercial immunodiagnostic kit incorporating lipoarabinomannan in the serodiagnosis of pulmonary tuberculosis in Ghana. Trop Med Int Health 1997;2:978-81.  Back to cited text no. 15
    
16.
Schlesinger LS. Macrophage phagocytosis of virulent but not attenuated strains of Mycobacterium tuberculosis is mediated by mannose receptors in addition to complement receptors. J Immunol 1993;150:2920-30.  Back to cited text no. 16
    
17.
Hamasur B, Källenius G, Svenson SB. A new rapid and simple method for large-scale purification of mycobacterial lipoarabinomannan. FEMS Immunol Med Microbiol 1999;24:11-7.  Back to cited text no. 17
    
18.
Venisse A, Rivière M, Vercauteren J, Puzo G. Structural analysis of the mannan region of lipoarabinomannan from Mycobacterium bovis BCG. Heterogeneity in phosphorylation state. J Biol Chem 1995;270:15012-21.  Back to cited text no. 18
    
19.
Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry 1978;15:429-36.  Back to cited text no. 19
[PUBMED]    
20.
Briken V, Porcelli SA, Besra GS, Kremer L. Mycobacterial lipoarabinomannan and related lipoglycans: From biogenesis to modulation of the immune response. Mol Microbiol 2004;53:391-403.  Back to cited text no. 20
    
21.
Miller RA, Buchanan TM. Production and characterization of a murine monoclonal antibody recognizating a shared mycobacterial polysaccharide. Int J Lepr Other Mycobact Dis 1984;52:461-7.  Back to cited text no. 21
[PUBMED]