Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen
Por:
Ortega, Paola A., Silva-Miranda, Mayra, Torres-Larios, Alfredo, Campos-Chavez, Eduardo, Franken, Kees C. L. C. M., Ottenhoff, Tom H. M., Ivanyi, Juraj, Espitia, Clara
Publicada:
2 oct 2020
Resumen:
T cells recognizing epitopes on the surface of mycobacteria-infected
macrophages can impart protection, but with associated risk for
reactivation to lung pathology. We aimed to identify antibodies specific
to such epitopes, which carry potentials for development toward novel
therapeutic constructs. Since epitopes presented in the context of major
histocompatibility complex alleles are rarely recognized by naturally
produced antibodies, we used a phage display library for the
identification of monoclonal human single domain antibody producing
clones. The selected 2C clone displayed T cell receptor-like recognition
of an HLA-A*0201 bound(199)KLVANNTRL(207)peptide from the Ag85B
antigen, which is known to be an immunodominant epitope for human T
cells. The specificity of the selected domain antibody was demonstrated
by solid phase immunoassay and by immunofluorescent surface staining of
peptide loaded cells of the T2 cell line. The antibody affinity binding
was determined by biolayer interferometry. Our results validated the
used technologies as suitable for the generation of antibodies against
epitopes on the surface ofMycobacterium tuberculosisinfected cells. The
potential approaches forward the development of antibody in
immunotherapy of tuberculosis have been outlined in the discussion.
Filiaciones:
Ortega, Paola A.:
Univ Nacl Autonoma Mexico, Inst Invest Biomd, Dept Inmunol, Mexico City, DF, Mexico
Silva-Miranda, Mayra:
Univ Nacl Autonoma Mexico, Inst Invest Biomd, Dept Inmunol, Mexico City, DF, Mexico
Univ Nacl Autonoma Mexico, CONACyT Inst Invest Biomed, Mexico City, DF, Mexico
Torres-Larios, Alfredo:
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Dept Biochem & Struct Biol, Mexico City, DF, Mexico
Campos-Chavez, Eduardo:
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Dept Biochem & Struct Biol, Mexico City, DF, Mexico
Franken, Kees C. L. C. M.:
Univ Med Ctr Leiden, Dept Infect Dis, Leiden, Netherlands
Ottenhoff, Tom H. M.:
Univ Med Ctr Leiden, Dept Infect Dis, Leiden, Netherlands
Ivanyi, Juraj:
Kings Coll London, Ctr Host Microbiome Interact, London, England
Espitia, Clara:
Univ Nacl Autonoma Mexico, Inst Invest Biomd, Dept Inmunol, Mexico City, DF, Mexico
Kings Coll London, Ctr Host Microbiome Interact, London, England
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