HLA class II tetramers: Tools for direct analysis of antigen-specific CD4+ T cells

Nepom, Gerald T.; Buckner, Jane H.; Novak, Erik J.; Reichstetter, Sandra; Reijonen, Helena; Gebe, John A.; Wang, Rongfang; Swanson, Eric S.; Kwok, William W.

doi:10.1002/1529-0131(200201)46:1<5::aid-art10063>3.0.co;2-s

Cited by 67 publications

(42 citation statements)

References 36 publications

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“…MHC tetramers have been used to detect autoreactive antigen-specific CD4+ T cells in human subjects associated with T1D, MS, pemphigus vulgaris, and celiac disease [2,4,6,11,[16][17][18][19]. Because construction of appropriate MHC tetramers requires knowledge both of the HLA restriction element and of a suitable peptide epitope, diseases that have dominant HLA gene associations and dominant autoantigen targets are well suited for application of this technology, whereas other autoimmune disorders with more heterogeneous genetics or antigen targets are less optimal.…”

Section: Discussionmentioning

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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Section: Discussionmentioning

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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“…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%

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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“…The reasons for the discrepancy between MHC class I and MHC class II tetramers are still unclear. It is probably the convergence of many parameters: 1) low frequency of CD4 ϩ T cells (11,12,16,17), 2) low affinity of MHC-TCR interactions, 3) difficulties in the preparation of soluble class II-peptide complexes, 4) lower affinity of MHC class II-binding peptides as compared with class I-binding peptides, 5) sliding of the peptide within the groove (28,40,41), 6) importance of the flanking residues for binding and recognition, 7) nonequivalent functions of CD4 and CD8 accessory molecules, and 8) need to have active metabolic processes for staining (38). More importantly, it points out the fundamental differences between CD4 ϩ and CD8 ϩ T cells in term of physiology.…”

Section: Discussionmentioning

confidence: 99%

Detection of Low-Avidity CD4+ T Cells Using Recombinant Artificial APC: Following the Antiovalbumin Immune Response

Mallet-Designe¹,

Stratmann²,

Homann

et al. 2003

The Journal of Immunology

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Subtle differences oppose CD4+ to CD8+ T cell physiologies that lead to different arrays of effector functions. Interestingly, this dichotomy has also unexpected practical consequences such as the inefficacy of many MHC class II tetramers in detecting specific CD4+ T cells. As a mean to study the CD4+ anti-OVA response in H-2d and H-2b genetic backgrounds, we developed I-Ad- and I-Ab-OVA recombinant MHC monomers and tetramers. We were able to show that in this particular system, despite normal biological activity, MHC class II tetramers failed to stain specific T cells. This failure was shown to be associated with a lack of cooperation between binding sites within the tetramer as measured by surface plasmon resonance. This limited cooperativeness translated into a low “functional avidity” and very transient binding of the tetramers to T cells. To overcome this biophysical barrier, recombinant artificial APC that display MHC molecules in a lipid bilayer were developed. The plasticity and size of the MHC-bearing fluorescent liposomes allowed binding to Ag-specific T cells and the detection of low numbers of anti-OVA T cells following immunization. The same liposomes were able, at 37°C, to induce the full reorganization of the T cell signaling molecules and the formation of an immunological synapse. Artificial APC will allow T cell detection and the dissection of the molecular events of T cell activation and will help us understand the fundamental differences between CD4+ and CD8+ T cells.

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HLA class II tetramers: Tools for direct analysis of antigen-specific CD4+ T cells

Cited by 67 publications

References 36 publications

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

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

Detection of Low-Avidity CD4+ T Cells Using Recombinant Artificial APC: Following the Antiovalbumin Immune Response

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