Abstract
The development of molecules specific for M. tuberculosis-infected cells has important implications, as these tools may facilitate understanding of the mechanisms regulating host pathogen interactions in vivo. In addition, development of new tools capable to targeting M. tuberculosis-infected cells may have potential applications to diagnosis, treatment, and prevention of tuberculosis (TB). Due to the lack of CD1b polymorphism, M. tuberculosis lipid-CD1b complexes could be considered as universal tuberculosis infection markers. The aim of the present study was to display on the PIII surface protein of m13 phage, a human αβ single-chain T-cell receptor molecule specific for CD1b:2-stearoyl-3-hydroxyphthioceranoyl-2´-sulfate-α-α´-D-trehalose (Ac2SGL) which is a complex presented by human cells infected with M. tuberculosis. The results showed the pIII fusion particle was successfully displayed on the phage surface. The study of the recognition of the recombinant phage in ELISA and immunohistochemistry showed the recognition of CD1b:Ac2SGL complexes and cells in human lung tissue from a tuberculosis patient respectively, suggesting the specific recognition of the lipid-CD1b complex.
Highlights
TB, caused by M. tuberculosis (Mtb), remains a leading cause of morbidity and mortality, being responsible for more than 2 million deaths every year [1]
To design a ab scTCR the variable genes were linked by a flexible hydrophilic peptide consisting in glycine and serine (Gly4Ser)3, which make it flexible and resistant to proteases [6]
The phagemid pHEN1 was selected to display the scTCR linked to PIII protein of m13 phage
Summary
TB, caused by M. tuberculosis (Mtb), remains a leading cause of morbidity and mortality, being responsible for more than 2 million deaths every year [1]. Prevention of transmission remains one of the most effective strategies and new tools useful for early and rapid diagnosis are urgently needed. Assays currently used to detect specific T cell responses have shown several problems including lack of response to used antigens, variations due to concomitant infections and, last but not least, lack of evidence of current presence of the infectious agent [2]. Glycolipids and lipopeptides derived from Mtb are presented to T cells by non-polymorphic CD1 cell-surface molecules [3,4], expanding the possible targets present study was, to display a functional ab scTCR recognizing the CD1b:Ac2SGL complex from M. tuberculosis, on the PIII protein of m13 phage
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