Soluble, High-Affinity Dimers of T-Cell Receptors and Class II Major Histocompatibility Complexes: Biochemical Probes for Analysis and Modulation of Immune Responses

Lebowitz, Michael; O'Herrin, Sean M.; Hamad, Abdel Rahim A.; Fahmy, Tarek M.; Marguet, Didier; Barnes, Nicholas C.; Pardoll, Drew M.; Bieler, Joan Glick; Schneck, Jonathan P.

doi:10.1006/cimm.1999.1441

Cited by 45 publications

(33 citation statements)

References 42 publications

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“…Acidic and basic leucine zippers are attached to the extracellular domain of the b-and a-chains of the class II molecules; a linker region is attached, which includes a thrombin cleavage site, and is inserted between the class II chain and leucine zipper and a biotinylation sequence attached to the C-termini of the acidic zipper. This and the use of chimeric IgG Fc domains promote assembly and stability of the heterodimer [25,27,28].…”

Section: Mhc Class II Tetramersmentioning

confidence: 99%

MHC–peptide tetramers for the analysis of antigen-specific T cells

Sims

Willberg

Klenerman

2010

Expert Review of Vaccines

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Section: Mhc Class II Tetramersmentioning

confidence: 99%

MHC–peptide tetramers for the analysis of antigen-specific T cells

Sims

Willberg

Klenerman

2010

Expert Review of Vaccines

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“…To isolate peptide-reactive T cells, divalent MHC class II (I-E d ) molecules covalently bound to human IgG1-Fc (generous gift of K. Karjalainen, Institute for Research in Biomedicine, Bellinzona, Switzerland) were loaded at a molar ratio of 1:10 with peptide at pH 4.8 (6) before incubation for 30 min with purified splenocytes. The multimeric nature of the MHC class II complex significantly increases the avidity of this chimeric molecule for cognate ligands, thus overcoming the low-affinity constraint of the I-E d /TCR interaction and enabling direct isolation of Agreactive T cells (7,8). After wash and mouse Fc␥R block, FITC-conjugated goat anti-human IgG1-Fc and fluorochrome-labeled Ab to relevant surface markers or control isotype-matched fluorochrome-labeled Ab were added for 20 -30 min at 4°C in PBS/2% FCS.…”

Section: Treatment Of Micementioning

confidence: 99%

Ig-Reactive CD4+CD25+ T Cells from Tolerized (New Zealand Black × New Zealand White)F1 Mice Suppress In Vitro Production of Antibodies to DNA

Cava

Ebling

Hahn

2004

The Journal of Immunology

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We have recently shown that tolerogenic administration of an artificial peptide (pConsensus) that is based on sequences within the VH regions of several murine anti-dsDNA Ig delays appearance of autoantibodies in female (New Zealand Black (NZB) × New Zealand White (NZW))F1 (NZB/W F1) mice and significantly prolongs their survival. The aim of this study was to characterize the T cell population(s) involved in pConsensus-induced down-regulation of autoimmune responses in tolerized NZB/W F1 mice. Using MHC class II dimers loaded with tolerogenic peptide, we found that pCons favored expansion of peptide-reactive CD4+CD25+ regulatory T cells (TR) that inhibited in vitro production of anti-dsDNA Ab-forming cells. Suppression by TR was abrogated by the presence in culture of Ab to glucocorticoid-induced TNFR family member 18 or to TGFβ latency-associated protein. These findings suggest possible relevance of Ag specificity in the mechanism of TR-mediated immune tolerance to Ig-derived peptides in NZB/W F1 mice.

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“…Two potent classes of reagents have been developed to bind to such complexes. Specific TCR, engineered from cloned T cells of known pMHC specificity, have been used to visualize cell surface pMHC complexes (4,5), and MHC-restricted, peptide-specific mAb, identified by various immunization and screening schemes, have been isolated and used similarly (5)(6)(7)(8)(9)(10)(11)(12). However, it has recently been suggested that in vitro selection of phage libraries displaying Ab fragments could be one of the most efficient ways to select specific pMHC binders (3).…”

mentioning

confidence: 99%

TCR-Like Human Antibodies Expressed on Human CTLs Mediate Antibody Affinity-Dependent Cytolytic Activity

Chames

Willemsen

Rojas

et al. 2002

The Journal of Immunology

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The permanent genetic programming via gene transfer of autologous T cells with cell surface receptors directed toward tumor-related Ags holds great promise for the development of more-specific tumor therapies. In this study we have explored the use of Abs directed to MHC-peptide complexes (or TCR-like Abs) to engraft CTLs with exquisite specificity for cancer cells. First, we affinity matured in vitro a previously selected TCR-like Ab, Fab-G8, which is highly specific for the peptide melanoma-associated Ag-A1 presented by the HLA-A1 molecule. A combination of L chain shuffling, H chain-targeted mutagenesis, and in vitro selection of phage display libraries yielded a Fab-G8 Ab derivative, Fab-Hyb3, with an 18-fold improved affinity yet identical peptide fine specificity. Fab-G8 and Fab-Hyb3 were expressed on primary human T lymphocytes as cell surface-anchored Fab, demonstrating that T cells expressing the high-affinity Fab-Hyb3 molecule eradicate tumor cells much more effectively. Furthermore, the gain in ligand-binding affinity resulted in a 2-log improvement in the detection of peptide/MHC complexes on melanoma-associated Ag-A1 peptide-loaded cells. In summary, an affinity-matured Ab specifically recognizing a cancer-related peptide/MHC complex was generated and used to improve the tumor cell killing capacity of human T cells. This strategy, based on engraftment of T cells with in vitro engineered Abs, is an attractive alternative to the laborious, and in many cases unsuccessful, generation of highly potent tumor-specific T lymphocytes.

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Soluble, High-Affinity Dimers of T-Cell Receptors and Class II Major Histocompatibility Complexes: Biochemical Probes for Analysis and Modulation of Immune Responses

Cited by 45 publications

References 42 publications

MHC–peptide tetramers for the analysis of antigen-specific T cells

MHC–peptide tetramers for the analysis of antigen-specific T cells

Ig-Reactive CD4+CD25+ T Cells from Tolerized (New Zealand Black × New Zealand White)F1 Mice Suppress In Vitro Production of Antibodies to DNA

TCR-Like Human Antibodies Expressed on Human CTLs Mediate Antibody Affinity-Dependent Cytolytic Activity

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