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

Huan, Jianya; Meza-Romero, Roberto; Mooney, James; Chou, Yuan K.; Edwards, David M.; Rich, Cathleen; Link, Jason M.; Vandenbark, Arthur A.; Bourdette, Dennis; Bächinger, Hans Peter; Burrows, Gregory G.

doi:10.1002/jctb.1086

Cited by 13 publications

(18 citation statements)

References 51 publications

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“…These single chain rTCR ligands (RTLs) (18 -20) refold in a manner that allows binding of allele-specific epitopes (21). In this study, we present data characterizing HLA-DP-derived RTLs, including direct measurement of beryllium binding to rHLA-DP2.…”

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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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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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“…In our previous studies, we demonstrated that both monomeric DR2-derived and I-A s -derived RTLs could protect and treat the clinical and histological signs of EAE in DR2 Tg mice (39,41) and SJL mice (40), respectively. In the current study, we used a similar approach, constructing monomeric I-A q -derived RTLs covalently linked with bCII257-270 peptide (RTL2001) (US patent no.…”

Section: Discussionmentioning

confidence: 99%

“…General methods for the design, cloning and expression of RTLs have been described previously (37,39,40). In brief, mRNA was isolated from the splenocytes of DBA1/LacJ mice using an Oligotex Direct mRNA mini kit (Qiagen).…”

Section: Rtl Construction Modification and Productionmentioning

confidence: 99%

MHC Class II Derived Recombinant T Cell Receptor Ligands Protect DBA/1LacJ Mice from Collagen-Induced Arthritis

Huan¹,

Kaler²,

Mooney³

et al. 2008

The Journal of Immunology

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We previously demonstrated the therapeutic effects of MHC class II derived recombinant T cell receptor ligands (RTL), single-chain two domain complexes of the α1 and β1 domains of MHC class II molecules genetically linked with an immunodominant peptide, in experimental autoimmune encephalomyelitis. In the current study, we produced a monomeric murine I-Aq-derived RTL construct covalently linked with bovine collagen type II peptide (bCII257–270) suitable for use in DBA/1LacJ mice that develop collagen-induced arthritis (CIA), an animal model of human rheumatoid arthritis, after immunization with bCII protein in CFA. In this study, we demonstrate that the I-Aq-derived RTLs reduced the incidence of the disease, suppressed the clinical and histological signs of CIA and induced long-term modulation of T cells specific for arthritogenic Ags. Our results showed that the I-Aq/bCII257–270 molecule could systemically reduce proinflammatory IL-17 and IFN-γ production and significantly increase anti-inflammatory IL-10, IL-13, and FoxP3 gene expression in splenocytes. Moreover, I-Aq/bCII257–270 molecule could also selectively inhibit IL-1β, IL-6, and IL-23 expression in local joint tissue. This is the first report demonstrating effective prevention of joint inflammation and clinical signs of CIA with an I-Aq-derived RTL, thus supporting the possible clinical use of this approach for treating rheumatoid arthritis in humans.

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“…Work in my laboratory has taken a reductionist approach, focused on engineering the smallest possible molecule that retains the peptide binding/TCR recognition features of the class II molecule. It appears we have achieved that goal in the form of single chain class II-derived Recombinant TCR Ligands (RTLs) consisting of the α1 and β1 domains of MHC class II molecules genetically linked into a single polypeptide chain [68][69][70][71]. The design of the RTLs was based on extensive molecular modeling studies of MHC class II proteins including crystal structures of human DR [72][73][74][75][76] and murine I-A k with covalently bound single peptides [77], and modeling studies predicted that the antigen binding domain would remain stable in the absence of the α2 and β2 Ig-fold domains [69,71,78].…”

Section: Structure Of the Mhc Class II Molecule And Design Of Rtlsmentioning

confidence: 99%

“…Along with "empty" RTLs that can be loaded exogenously with synthetic peptides, we have produced a variety of genetically-encoded variants with antigenic peptide attached by a linker to the N-terminus of the beta-chain (Table I) [64,69,79,80]. RTL molecules have been used for studying binding specificity in vitro [71,81,82], for exploring primary TCR signaling events independent of co-stimulatory input associated with the MHC II α2 and β2 domains or with other molecules expressed by antigen presenting cells [83], and for treating CD4 + T cell-mediated autoimmune disease in an MHC II/epitope-specific manner [68,70,80,83,84].…”

Section: Structure Of the Mhc Class II Molecule And Design Of Rtlsmentioning

confidence: 99%

Systemic Immunomodulation of Autoimmune Disease Using MHC-Derived Recombinant TCR Ligands

Burrows¹

2005

CDTIA

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Human autoimmune disease involves local activation of antigen-specific CD4 + T cells that produce inflammatory Th1 cytokines leading to the further recruitment and activation of lymphocytes and monocytes, resulting ultimately in the destruction of target tissue. Antigen presenting cells (APCs) initiate activation of CD4 + T cells in a multistep process that minimally involves co-ligation of the TCR and CD4 by the MHC class II/peptide complex and costimulation through additional T cell surface molecules such as CD28. Disruption of this highly orchestrated series of events can result in the direct modulation of CD4 + T cell behavior. The interaction between MHC and TCR holds unique promise as a focal point for therapeutic intervention in the pathology of CD4 + T cell-mediated diseases, and MHC class II-derived Recombinant TCR Ligands ("RTLs") have emerged as a new class of therapeutics with potent clinical efficacy in a diverse set of animal models for multiple sclerosis. Here I review the systemic effect that RTL therapy has on the intact immune system and present an overview of a molecular mechanism by which RTL therapy could induce these systemic changes.

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

Cited by 13 publications

References 51 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

MHC Class II Derived Recombinant T Cell Receptor Ligands Protect DBA/1LacJ Mice from Collagen-Induced Arthritis

Systemic Immunomodulation of Autoimmune Disease Using MHC-Derived Recombinant TCR Ligands

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