Exploring the Association of Surface Plasmon Resonance with Recombinant MHC:Ig Hybrid Protein as a Tool for Detecting T Lymphocytes in Mice Infected with Leishmania (Leishmania) amazonensis.
Lenilton Silva da Silveira-JúniorFranklin Souza-SilvaBernardo Acácio Santini PereiraLéa Cysne-FinkelsteinGeraldo Barroso Cavalcanti JúniorCarlos Roberto AlvesPublished in: BioMed research international (2017)
A surface plasmon resonance- (SPR-) based recognition method applying H-2 Ld:Ig/peptides complexes for ex vivo monitoring cellular immune responses during murine infection with Leishmania (Leishmania) amazonensis is described. Lymphocytes from lesion-draining popliteal lymph nodes were captured on a carboxylated sensor chip surface previously functionalized with H-2 Ld:Ig (DimerX) protein bound to synthetic peptides derived from the COOH-terminal region of cysteine proteinase B of L. (L.) amazonensis. In computational analysis, these peptides presented values of kinetic constants favorable to form complexes with H-2 Ld at neutral pH, with a Gibbs free energy ΔG° < 0. The assayed DimerX:peptide complexes presented the property of attaching to distinct T lymphocytes subsets, obtained from experimentally infected BALB/c mice, in each week of infection, thus indicating a temporal variation in specific T lymphocytes populations, each directed to a different COOH-terminal region-derived peptide. The experimental design proposed herein is an innovative approach for cellular immunology studies of a neglected disease, providing a useful tool for the analysis of specific T lymphocytes subsets.
Keyphrases
- amino acid
- lymph node
- peripheral blood
- immune response
- high fat diet induced
- protein protein
- high throughput
- binding protein
- type diabetes
- dendritic cells
- clinical trial
- randomized controlled trial
- metabolic syndrome
- early stage
- skeletal muscle
- mass spectrometry
- inflammatory response
- case control
- single cell
- rectal cancer
- study protocol
- endovascular treatment