Functionally Active Recombinant α and β Chain-Peptide Complexes of Human Major Histocompatibility Class II Molecules

Nag, Bishwajit; Arimilli, Subhashini; Mukku, Prabha V.; Astafieva, Irina

doi:10.1074/jbc.271.17.10413

Cited by 18 publications

(9 citation statements)

References 33 publications

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“…Soluble recombinant MHC-II molecules in various forms have been developed, from full-length detergent solubilized MHC-II heterodimers (51,52) to various forms of the extracellular a1a2/b1b2 heterodimeric domains (53)(54)(55), and recently the a1 and b1 domains genetically linked into a single polypeptide chain (18 -21, 56). The latter appears to be the smallest structure that retains the peptide binding/ TCR recognition features of MHC-II molecules (33).…”

Section: Discussionmentioning

confidence: 99%

Recombinant HLA-DP2 Binds Beryllium and Tolerizes Beryllium-Specific Pathogenic CD4+ T Cells

Fontenot

Keizer

McCleskey

et al. 2006

The Journal of Immunology

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Chronic beryllium disease is a lung disorder caused by beryllium exposure in the workplace and is characterized by granulomatous inflammation and the accumulation of beryllium-specific, HLA-DP2-restricted CD4+ T lymphocytes in the lung that proliferate and secrete Th1-type cytokines. To characterize the interaction among HLA-DP2, beryllium, and CD4+ T cells, we constructed rHLA-DP2 and rHLA-DP4 molecules consisting of the α-1 and β-1 domains of the HLA-DP molecules genetically linked into single polypeptide chains. Peptide binding to rHLA-DP2 and rHLA-DP4 was consistent with previously published peptide-binding motifs for these MHC class II molecules, with peptide binding dominated by aromatic residues in the P1 pocket. 9Be nuclear magnetic resonance spectroscopy showed that beryllium binds to the HLA-DP2-derived molecule, with no binding to the HLA-DP4 molecule that differs from DP2 by four amino acid residues. Using beryllium-specific CD4+ T cell lines derived from the lungs of chronic beryllium disease patients, beryllium presentation to those cells was independent of Ag processing because fixed APCs were capable of presenting BeSO4 and inducing T cell proliferation. Exposure of beryllium-specific CD4+ T cells to BeSO4-pulsed, plate-bound rHLA-DP2 molecules induced IFN-γ secretion. In addition, pretreatment of beryllium-specific CD4+ T cells with BeSO4-pulsed, plate-bound HLA-DP2 blocked proliferation and IL-2 secretion upon re-exposure to beryllium presented by APCs. Thus, the rHLA-DP2 molecules described herein provide a template for engineering variants that retain the ability to tolerize pathogenic CD4+ T cells, but do so in the absence of the beryllium Ag.

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Section: Discussionmentioning

confidence: 99%

Recombinant HLA-DP2 Binds Beryllium and Tolerizes Beryllium-Specific Pathogenic CD4+ T Cells

Fontenot

Keizer

McCleskey

et al. 2006

The Journal of Immunology

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“…Soluble recombinant MHC class II molecules in various forms have been developed, including full length detergent solubilized MHC class II heterodimers (26, 27), various forms of the extracellular α1α2 and β1β2 heterodimeric domains (28–31), and recently the β1 and α1 domains genetically linked into a single polypeptide chain (8–10, 17, 32). RTL appears to be the smallest structure that retains the peptide binding/TCR recognition features of MHC class II molecules (33).…”

Section: Discussionmentioning

confidence: 99%

Single-chain recombinant HLA-DQ2.5/peptide molecules block α2-gliadin-specific pathogenic CD4+ T-cell proliferation and attenuate production of inflammatory cytokines: a potential therapy for celiac disease

et al. 2011

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Celiac disease (CD) is a disorder of the small intestine caused by intolerance to wheat gluten and related proteins in barley and rye. CD4+ T cells play a central role in CD, recognizing and binding complexes of HLA-DQ2.5 bearing gluten peptides that have survived digestion and that are deamidated by tissue transglutaminase (TG2), propagating a cascade of inflammatory processes that damage and eventually destroy the villous tissue structures of the small intestine. Here we present data showing that recombinant DQ2.5-derived molecules bearing covalently tethered α2-gliadin-61-71 peptide have a remarkable ability to block antigen-specific T cell proliferation and inhibited pro-inflammatory cytokine secretion in human DQ2.5-restricted α2-gliadin specific T cell clones obtained from patients with celiac disease. The results from our in vitro studies suggest that HLA-DQ2.5 derived molecules could significantly inhibit and perhaps reverse the intestinal pathology caused by T cell mediated inflammation and the associated production of proinflammatory cytokines.

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“…Moreover, the β2 domain contains a CD4 binding region that co‐ligates CD4 when the α1 and β1 domains with associated antigenic peptide interact with the TCR αβ heterodimer, whereas the α2 domain appears to contribute to ordered oligomerization in T cell activation 15. Complexes of MHC/antigen have been purified as detergent extracts of lymphocyte membranes,16 and as associated recombinant proteins using baculovirus and bacterial expression systems 17–21. These two‐chain, four‐domain molecular complexes, after loading with selected peptide epitopes, have been demonstrated to interact with T cells in an antigen‐specific manner 13, 14, 19, 22–24.…”

Section: Introductionmentioning

confidence: 99%

“…These two‐chain, four‐domain molecular complexes, after loading with selected peptide epitopes, have been demonstrated to interact with T cells in an antigen‐specific manner 13, 14, 19, 22–24. In some cases, in the absence of co‐stimulation, these complexes induced antigen‐specific apoptosis rather than anergy 20. Desbarats et al showed that cross‐linking CD4 can enhance Fas (CD95) expression and activate apoptotic cell death 25.…”

Section: Introductionmentioning

confidence: 99%

Rationally designed mutations convert complexes of human recombinant T cell receptor ligands into monomers that retain biological activity

Huan

Meza-Romero

Mooney

et al. 2004

J of Chemical Tech & Biotech

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Single-chain human recombinant T cell receptor ligands derived from the peptide binding/TCR recognition domain of human HLA-DR2b (DRA*0101/DRB1*1501) produced in Escherichia coli with and without amino-terminal extensions containing antigenic peptides have been described previously. While molecules with the native sequence retained biological activity, they formed higher order aggregates in solution. In this study, we used site-directed mutagenesis to modify the β-sheet platform of the DR2-derived RTLs, obtaining two variants that were monomeric in solution by replacing hydrophobic residues with polar (serine) or charged (aspartic acid) residues. Size exclusion chromatography and dynamic light scattering demonstrated that the modified RTLs were monomeric in solution, and structural characterization using circular dichroism demonstrated the highly ordered secondary structure of the RTLs. Peptide binding to the `empty' RTLs was quantified using biotinylated peptides, and functional studies showed that the modified RTLs containing covalently tethered peptides were able to inhibit antigen-specific T cell proliferation in vitro, as well as suppress experimental autoimmune encephalomyelitis in vivo. These studies demonstrated that RTLs encoding the Ag-binding/TCR recognition domain of MHC class II molecules are innately very robust structures, capable of retaining potent biological activity separate from the Ig-fold domains of the progenitor class II structure, with prevention of aggregation accomplished by modification of an exposed surface that was buried in the progenitor structure.

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Functionally Active Recombinant α and β Chain-Peptide Complexes of Human Major Histocompatibility Class II Molecules

Cited by 18 publications

References 33 publications

Recombinant HLA-DP2 Binds Beryllium and Tolerizes Beryllium-Specific Pathogenic CD4+ T Cells

Recombinant HLA-DP2 Binds Beryllium and Tolerizes Beryllium-Specific Pathogenic CD4+ T Cells

Single-chain recombinant HLA-DQ2.5/peptide molecules block α2-gliadin-specific pathogenic CD4+ T-cell proliferation and attenuate production of inflammatory cytokines: a potential therapy for celiac disease

Rationally designed mutations convert complexes of human recombinant T cell receptor ligands into monomers that retain biological activity

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