Selection of phage-displayed human antibody fragments specific for CD1b presenting the Mycobacterium tuberculosis glycolipid Ac2SGL

Int J Mycobacteriol. 2016 Jun;5(2):120-7. doi: 10.1016/j.ijmyco.2015.12.002. Epub 2016 Feb 17.

Abstract

Objective/background: The development of new tools capable of targeting Mycobacterium tuberculosis (Mtb)-infected cells have potential applications in diagnosis, treatment, and prevention of tuberculosis. In Mtb-infected cells, CD1b molecules present Mtb lipids to the immune system (Mtb lipid-CD1b complexes). Because of the lack of CD1b polymorphism, specific Mtb lipid-CD1b complexes could be considered as universal Mtb infection markers. 2-Stearoyl-3-hydroxyphthioceranoyl-2'-sulfate-α-α'-d-trehalose (Ac2SGL) is specific for Mtb, and is not present in other mycobacterial species. The CD1b-Ac2SGL complexes are expressed on the surface of human cells infected with Mtb. The aim of this study was to generate ligands capable of binding these CD1b-Ac2SGL complexes.

Methods: A synthetic human scFv phage antibody library was used to select phage-displayed antibody fragments that recognized CD1b-Ac2SGL using CD1b-transfected THP-1 cells loaded with Ac2SGL.

Results: One clone, D11-a single, light-variable domain (kappa) antibody (dAbκ11)-showed high relative binding to the Ac2SGL-CD1b complex.

Conclusion: A ligand recognizing the Ac2SGL-CD1b complex was obtained, which is a potential candidate to be further tested for diagnostic and therapeutic applications.

Keywords: Antibodies; CD1b; Lipids; Tuberculosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Bacterial / genetics
  • Antibodies, Bacterial / immunology*
  • Antigens, CD1 / genetics
  • Antigens, CD1 / immunology*
  • Bacteriophages / genetics
  • Bacteriophages / metabolism
  • Gene Expression
  • Glycolipids / immunology*
  • Humans
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / immunology*
  • Single-Chain Antibodies / genetics*
  • Single-Chain Antibodies / immunology
  • Tuberculosis / immunology*
  • Tuberculosis / microbiology

Substances

  • Antibodies, Bacterial
  • Antigens, CD1
  • CD1b antigen
  • Glycolipids
  • Single-Chain Antibodies