MHC class II tetramers identify peptide-specific human CD4+ T cells proliferating in response to influenza A antigen

Novak, Erik J.; Liu, Andrew W.; Nepom, Gerald T.; Kwok, William W.

doi:10.1172/jci8476

Cited by 354 publications

(358 citation statements)

References 23 publications

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“…For functional studies, NKT cells were purified from PBMCs or organs by flow cytometry cell sorting or the MACS bead system based on staining with the -GalCer-loaded CD1d tetramer (17,18). The -GalCerloaded CD1d tetramers were synthesised and labelled as previously described (19,20). All mAbs were purchased from R&D Systems Europe Ltd. (Abingdon, UK), except that the anti-V11 mAb and anti-V24 mAb were from CoulterImmunoTech (Marseille, France) and the anti-CD161 pAb was from Santa Cruz Biotechnology (Santa Cruz, CA).…”

Section: Patients Cells and Tetramermentioning

confidence: 99%

EBV-Induced Human CD8+ NKT Cells Synergise CD4+ NKT Cells Suppressing EBV-Associated Tumours Upon Induction of Th1-Bias

Xiao

Zhou

et al. 2009

Cell Mol Immunol

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Section: Patients Cells and Tetramermentioning

confidence: 99%

EBV-Induced Human CD8+ NKT Cells Synergise CD4+ NKT Cells Suppressing EBV-Associated Tumours Upon Induction of Th1-Bias

Xiao

Zhou

et al. 2009

Cell Mol Immunol

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“…The production of MHC class II tetramers is described elsewhere [1]. Briefly, DRB4*0101-or DRB1*0405-containing HLA class II molecules were produced in a Cu-inducible Drosophila expression vector, enzymatically biotinylated, and loaded with specific peptide using a detergent-facilitated exchange reaction for 48-72 hours at 37° C. These peptide-loaded monomers were assembled into tetramers by addition of phycoerythrin (PE)-labeled streptavidin.…”

Section: Tetramer Analysismentioning

confidence: 99%

Recurrence of autoreactive antigen-specific CD4+ T cells in autoimmune diabetes after pancreas transplantation

Laughlin

Burke

Pugliese

et al. 2008

Clinical Immunology

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CD4+ T cells specific for the diabetes-associated autoantigen GAD65 were analyzed using peripheral blood samples after pancreas transplantation in subjects with T1D with clinical evidence of recurrent autoimmune diabetes. MHC class II tetramers facilitated the identification and cloning of antigenspecific autoreactive cells, which were found at several time points over a multiyear span, in spite of chronic immunosuppression of the subjects. Comparisons of TCR clonotypes by cDNA sequencing revealed that identical T cells were present in the circulation, separated by long time intervals, consistent with a persistent memory response associated with recurrent autoimmunity.

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“…Biotinylated soluble HLA class II monomers were purified as described [22]. The resulting biotinylated heterodimers were loaded with TT 947-966 or HA 307-319 peptide for 3 days at 37 C to generate DR1101/TT 947-966 and DR0401/HA 307-319 monomers.…”

Section: Hla Class II Tetramersmentioning

confidence: 99%

“…The resulting biotinylated heterodimers were loaded with TT 947-966 or HA 307-319 peptide for 3 days at 37 C to generate DR1101/TT 947-966 and DR0401/HA 307-319 monomers. The procedures for peptide loading were identical to those described earlier [22]. Fluorescent peptide-loaded tetramers were produced by cross-linking the biotinylated monomers with streptavidin-allophycocyanin (BD Biosciences, San Jose, CA) overnight at room temperature.…”

Section: Hla Class II Tetramersmentioning

confidence: 99%

“…Peptides representing MHC class II-restricted epitopes from tetanus toxoid (TT) and influenza hemagglutinin (HA) were used to stimulate human CD4 + T cells from pre-immunized donors in vitro; antigen specificity of proliferating cells was confirmed by fluorescent MHC class II tetramers [19][20][21][22]. Proliferation of antigen-specific CD4 + T cells was found to be enhanced by Hsp70:peptide complexes as compared to peptide alone.…”

Section: Introductionmentioning

confidence: 99%

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The heat shock protein Hsp70 enhances antigen-specific proliferation of human CD4+ memory T cells

et al. 2005

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Heat shock proteins (HSP) can interact with a wide variety of peptides and the resulting HSP:peptide complexes are known to be highly immunogenic. The ability of HSP:peptide complexes to elicit CD8 + T cell responses by cross-presentation of exogenous antigen via MHC class I is well known. In contrast, their role in the activation of CD4 + T cells is less clearly defined, although several recent studies in mice and T cell lines suggest an involvement of HSP in the presentation of antigenic peptides via MHC class II. In this study we have investigated the potential of antigenic peptides from tetanus toxin and influenza hemagglutinin complexed to the human stress-inducible Hsp70 to enhance activation and proliferation of human memory CD4 + T cells. Hsp70:peptide complexes were found to amplify the proliferation of antigen-specific CD4 + T cells as confirmed by HLA-DR tetramer staining. Complex formation of the antigenic peptide with Hsp70 was absolutely required to elicit an antigen-specific amplification. This effect was most pronounced at low doses of antigen and decreasing APC/CD4 + T cell ratios. Taken together, we show the potential of Hsp70 to enhance antigen-specific CD4 + T cell proliferation and to increase the immunogenicity of presented peptides in human CD4 + T cells.

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MHC class II tetramers identify peptide-specific human CD4+ T cells proliferating in response to influenza A antigen

Cited by 354 publications

References 23 publications

EBV-Induced Human CD8+ NKT Cells Synergise CD4+ NKT Cells Suppressing EBV-Associated Tumours Upon Induction of Th1-Bias

EBV-Induced Human CD8+ NKT Cells Synergise CD4+ NKT Cells Suppressing EBV-Associated Tumours Upon Induction of Th1-Bias

Recurrence of autoreactive antigen-specific CD4+ T cells in autoimmune diabetes after pancreas transplantation

The heat shock protein Hsp70 enhances antigen-specific proliferation of human CD4+ memory T cells

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