Production and Characterization of Monoclonal Antibodies Against the Lethal Factor Component of Bacillus Anthracis Lethal Toxin

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 1990 Jun 1

PMID 2111283

Citations 42

Authors

S F Little

S H Leppla

A M Friedlander

Affiliations

Soon will be listed here.

Abstract

The lethal toxin of Bacillus anthracis consists of two components, protective antigen and lethal factor. Protective antigen is cleaved after binding to cell receptors, yielding a receptor-bound fragment that binds lethal factor. Sixty-one monoclonal antibodies to the lethal factor protein have been characterized for specificity, antibody subtype, and ability to neutralize lethal toxin. Three monoclonal antibodies (10G3, 2E7, and 3F6) neutralized lethal toxin in Fisher 344 rats. However, in a macrophage cytolysis assay, monoclonal antibodies 10G3, 2E7, 10G4, 10D4, 13D10, and 1D8, but not 3F6, were found to neutralize lethal toxin. Binding studies showed that five of the monoclonal antibodies that neutralized lethal toxin in the macrophage assay (10G3, 2E7, 10G4, 10D4, and 13D10) did so by inhibiting the binding of lethal factor to the protective antigen fragment bound to cells. Monoclonal antibody 1D8, which was also able to neutralize lethal toxin activity after lethal factor was prebound to cell-bound protective antigen, only partially inhibited binding of lethal factor to protective antigen. Monoclonal antibody 3F6 did not inhibit the binding of lethal factor to protective antigen. A competitive-binding enzyme-linked immunosorbent assay showed that at least four different antigenic regions on lethal factor were recognized by these seven neutralizing hybridomas. The anomalous behavior of 3F6 suggests that it may induce a conformational change in lethal factor. Differences in neutralizing activity of monoclonal antibodies were related to their relative affinity and epitope specificity and the type of assay.

Citing Articles

Aerosolized Intratracheal Inoculation of Recombinant Protective Antigen (rPA) Vaccine Provides Protection Against Inhalational Anthrax in B10.D2-Hc Mice.

Song X, Zhang W, Zhai L, Guo J, Zhao Y, Zhang L Front Immunol. 2022; 13:819089.

PMID: 35154137 PMC: 8826967. DOI: 10.3389/fimmu.2022.819089.

Evaluation of immune response to recombinant LFD1-PA4 chimeric protein.

Mirhaj H, Honari H, Zamani E Iran J Vet Res. 2019; 20(2):112-119.

PMID: 31531033 PMC: 6716276.

Biochip for the Detection of Bacillus anthracis Lethal Factor and Therapeutic Agents against Anthrax Toxins.

Silin V, Kasianowicz J, Michelman-Ribeiro A, Panchal R, Bavari S, Robertson J Membranes (Basel). 2016; 6(3).

PMID: 27348008 PMC: 5041027. DOI: 10.3390/membranes6030036.

A Novel Chimeric Anti-PA Neutralizing Antibody for Postexposure Prophylaxis and Treatment of Anthrax.

Xiong S, Tang Q, Liang X, Zhou T, Yang J, Liu P Sci Rep. 2015; 5:11776.

PMID: 26134518 PMC: 4488766. DOI: 10.1038/srep11776.

Immunization of Mice with Anthrax Protective Antigen Limits Cardiotoxicity but Not Hepatotoxicity Following Lethal Toxin Challenge.

Devera T, Prusator D, Joshi S, Ballard J, Lang M Toxins (Basel). 2015; 7(7):2371-84.

PMID: 26120785 PMC: 4516918. DOI: 10.3390/toxins7072371.

References

PAPPENHEIMER Jr A, Uchida T, Harper A . An immunological study of the diphtheria toxin molecule. Immunochemistry. 1972; 9(9):891-906. DOI: 10.1016/0019-2791(72)90163-2. View

Stanley J, Smith H . Purification of factor I and recognition of a third factor of the anthrax toxin. J Gen Microbiol. 1961; 26:49-63. DOI: 10.1099/00221287-26-1-49. View

Leppla S . Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci U S A. 1982; 79(10):3162-6. PMC: 346374. DOI: 10.1073/pnas.79.10.3162. View

Luthy P, Braun D . Specificities of monoclonal antibodies against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis. Infect Immun. 1983; 40(2):608-12. PMC: 264897. DOI: 10.1128/iai.40.2.608-612.1983. View

Van Heyningen V, Brock D, van Heyningen S . A simple method for ranking the affinities of monoclonal antibodies. J Immunol Methods. 1983; 62(2):147-53. DOI: 10.1016/0022-1759(83)90000-5. View

Mosmann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2):55-63. DOI: 10.1016/0022-1759(83)90303-4. View

Green L, Reade J, Ware C . Rapid colorimetric assay for cell viability: application to the quantitation of cytotoxic and growth inhibitory lymphokines. J Immunol Methods. 1984; 70(2):257-68. DOI: 10.1016/0022-1759(84)90190-x. View

Ray K, Vockley J, Harris H . Epitopes of human intestinal alkaline phosphatases, defined by monoclonal antibodies. FEBS Lett. 1984; 174(2):294-9. DOI: 10.1016/0014-5793(84)81176-x. View

Ezzell J, Ivins B, Leppla S . Immunoelectrophoretic analysis, toxicity, and kinetics of in vitro production of the protective antigen and lethal factor components of Bacillus anthracis toxin. Infect Immun. 1984; 45(3):761-7. PMC: 263363. DOI: 10.1128/iai.45.3.761-767.1984. View

10.

BUSSARD A . How pure are monoclonal antibodies?. Dev Biol Stand. 1984; 57:13-5. View

11.

Redinbaugh M, Campbell W . Adaptation of the dye-binding protein assay to microtiter plates. Anal Biochem. 1985; 147(1):144-7. DOI: 10.1016/0003-2697(85)90020-x. View

12.

Carbone F, Paterson Y . Monoclonal antibodies to horse cytochrome c expressing four distinct idiotypes distribute among two sites on the native protein. J Immunol. 1985; 135(4):2609-16. View

13.

Vaidya H, Dietzler D, Ladenson J . Inadequacy of traditional ELISA for screening hybridoma supernatants for murine monoclonal antibodies. Hybridoma. 1985; 4(3):271-6. DOI: 10.1089/hyb.1985.4.271. View

14.

Wnek A, Strouse R, McClane B . Production and characterization of monoclonal antibodies against Clostridium perfringens type A enterotoxin. Infect Immun. 1985; 50(2):442-8. PMC: 261972. DOI: 10.1128/iai.50.2.442-448.1985. View

15.

Machuga E, Calton G, Burnett J . Purification of Bacillus anthracis lethal factor by immunosorbent chromatography. Toxicon. 1986; 24(2):187-95. DOI: 10.1016/0041-0101(86)90121-2. View

16.

Chia J, Pollack M, Avigan D, Steinbach S . Functionally distinct monoclonal antibodies reactive with enzymatically active and binding domains of Pseudomonas aeruginosa toxin A. Infect Immun. 1986; 52(3):756-62. PMC: 260923. DOI: 10.1128/iai.52.3.756-762.1986. View

17.

Friedlander A . Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem. 1986; 261(16):7123-6. View

18.

Van Heyningen V . A simple method for ranking the affinities of monoclonal antibodies. Methods Enzymol. 1986; 121:472-81. DOI: 10.1016/0076-6879(86)21046-0. View

19.

Adam E, Nasz I, Lengyel A, Erdei J, FACHET J . Determination of different antigenic sites on the adenovirus hexon using monoclonal antibodies. Arch Virol. 1987; 93(3-4):261-71. PMC: 7086970. DOI: 10.1007/BF01310979. View

20.

Little S, Leppla S, Cora E . Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin. Infect Immun. 1988; 56(7):1807-13. PMC: 259481. DOI: 10.1128/iai.56.7.1807-1813.1988. View