HLA-DQ Tetramers Identify Epitope-Specific T Cells in Peripheral Blood of Herpes Simplex Virus Type 2-Infected Individuals: Direct Detection of Immunodominant Antigen-Responsive Cells

Kwok, William W.; Liu, Andrew W.; Novak, Erik J.; Gebe, John A.; Ettinger, Ruth A.; Nepom, Gerald T.; Reymond, Sigrid N.; Koelle, David M.

doi:10.4049/jimmunol.164.8.4244

Cited by 116 publications

(97 citation statements)

References 23 publications

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“…Class II molecules, however, are noncovalent dimers of α and β chains that display variable stability in solution (20). To reliably generate stable class II molecules in soluble form, Kwok et al have constructed class II molecules incorporating leucine zipper motifs that replace the transmembrane and cytoplasmic portions of the molecules (10,21). The advantage of this approach, with respect to others involving the synthesis of covalent single-chain class II/peptide molecules (22), is that empty class II molecules can be loaded with any selected peptide, increasing tremendously the number of epitopes that can be studied.…”

Section: Discussionmentioning

confidence: 99%

Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Ayyoub

Dojcinovic

Pignon

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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MHC-peptide tetramers have become essential tools for T-cell analysis, but few MHC class II tetramers incorporating peptides from human tumor and self-antigens have been developed. Among limiting factors are the high polymorphism of class II molecules and the low binding capacity of the peptides. Here, we report the generation of molecularly defined tetramers using His-tagged peptides and isolation of folded MHC/peptide monomers by affinity purification. Using this strategy we generated tetramers of DR52b (DRB3*0202), an allele expressed by approximately half of Caucasians, incorporating an epitope from the tumor antigen NY-ESO-1. Molecularly defined tetramers avidly and stably bound to specific CD4 + T cells with negligible background on nonspecific cells. Using molecularly defined DR52b/ NY-ESO-1 tetramers, we could demonstrate that in DR52b + cancer patients immunized with a recombinant NY-ESO-1 vaccine, vaccineinduced tetramer-positive cells represent ex vivo in average 1:5,000 circulating CD4 + T cells, include central and transitional memory polyfunctional populations, and do not include CD4 + CD25 + CD127 − regulatory T cells. This approach may significantly accelerate the development of reliable MHC class II tetramers to monitor immune responses to tumor and self-antigens.S oluble MHC-peptide tetramers, allowing the direct visualization, characterization, and isolation of antigen-specific T cells, have become essential tools for T-cell analysis. MHC class I tetramers incorporating short CTL peptide epitopes, originally developed by Altman and Davis (1) have been generated for a large number of murine and human alleles, incorporating a variety of peptides of microbial, tumor, and self-antigen origin (2). The development of MHC class II tetramers, however, and particularly of those incorporating peptides from tumor and self-antigens, has been far less successful (3-5). One limiting factor is the high polymorphism of the human MHC class II molecules, especially those encoded by the DRB1 locus, the most frequently studied. Another limiting factor is the binding affinity of antigenic peptides derived from tumor and self-antigens, which is generally lower than that of peptides from pathogens.NY-ESO-1 (ESO), a tumor-specific antigen of the cancer/testis group frequently expressed in tumors of different histological types (6), is an important candidate for the development of generic cancer vaccines (7). In a recent vaccination trial using a recombinant ESO protein (rESO) administered with Montanide ISA 51 and CpG ODN 7909, we have observed induction of CD4 + T cell responses in all vaccinated patients (8). By assessing vaccineinduced CD4 + T cells, we have identified an immunodominant epitope , core region ESO 123-137 ) restricted by HLADR52b (DRB3*0202), an allele expressed by half of Caucasians (9). DRB3-, DRB4-, and DRB5-encoded molecules are less polymorphic than those encoded by DRB1, and are therefore attractive candidates for the development of generic MHC class II tetramers.Our initial attempts to const...

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Section: Discussionmentioning

confidence: 99%

Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Ayyoub

Dojcinovic

Pignon

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…For assay of viral antigenic epitopes, we have previously shown that stimulation with UV-inactivated virus is effective a Underlined residue is the proposed first anchor for binding to the respective allele as determined by peptide truncation studies (data not shown). (19). Similarly, peptide pools can be used for initial stimulation in the event that whole Ag is not available.…”

Section: Discussionmentioning

confidence: 99%

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Novak

Liu²,

Gebe³

et al. 2001

The Journal of Immunology

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129

107

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T cell responses to Ags involve recognition of selected peptide epitopes contained within the antigenic protein. In this report, we describe a new approach for direct identification of CD4+ T cell epitopes of complex Ags that uses human class II tetramers to identify reactive cells. With a panel of 60 overlapping peptides covering the entire sequence of the VP16 protein, a major Ag for HSV-2, we generated a panel of class II MHC tetramers loaded with peptide pools that were used to stain peripheral lymphocytes of an HSV-2 infected individual. With this approach, we identified four new DRA1*0101/DRB1*0401- and two DRA1*0101/DRB1*0404-restricted, VP16-specific epitopes. By using tetramers to sort individual cells, we easily obtained a large number of clones specific to these epitopes. Although DRA1*0101/DRB1*0401 and DRA1*0101/DRB1*0404 are structurally very similar, nonoverlapping VP16 epitopes were identified, illustrating high selectivity of individual allele polymorphisms within common MHC variants. This rapid approach to detecting CD4+ T cell epitopes from complex Ags can be applied to any known Ag that gives a T cell response.

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“…Peptide-loaded tetramers were produced as described previously (15). Briefly, recombinant DRB1*0401, in which the transmembrane domain was replaced by a leucine zipper, were produced in Drosophila S2 cells.…”

Section: Tetramer Stainingmentioning

confidence: 99%

Differential Antigen Sensitivity and Costimulatory Requirements in Human Th1 and Th2 Antigen-Specific CD4+ Cells with Similar TCR Avidity

Holzer

Kwok

Nepom

et al. 2003

The Journal of Immunology

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The differentiation of naive CD4+ Th cells into Th1 and Th2 phenotypes is influenced by cytokines, concentration of Ag, accessory molecules, and the affinity of the MHC-TCR interaction. To study these factors in human memory T cells, T cell lines with Th1 or Th2 phenotypes specific for the peptide hemagglutinin (HA)307–319 in the context of DRB1*0401 were established from the peripheral blood of an individual previously vaccinated for influenza virus. Flow cytometric analysis with fluorescent-labeled MHC class II tetramers was used to analyze TCR avidity: the Th2 line bound the HLA-DR*0401-HA307–319 tetramers with higher mean avidity, although the range of binding avidity largely overlapped with the Th1 line. High-affinity Th1 and Th2 lines were established for further study by FACS sorting. When activated with plate-bound HLA-DR*0401-HA307–319 monomers, the Th1 line proliferated and produced IFN-γ without additional costimulation whereas the Th2 line required the addition of soluble anti-CD28 Ab to induce proliferation and IL-5 production, but this requirement could be overcome with high concentrations of plate-bound monomer alone. IL-2 production was dependent on costimulation in both cell lines. These findings demonstrate that upon antigenic rechallenge, Th1 and Th2 cells differ in their response to Ag-specific stimulation. Th2 cells were sensitive to the strength of signal to a greater degree than Th1 cells and required costimulation through CD28 for maximal proliferation. These distinctions between Th1 and Th2 activation are not consistent with a simple avidity model of Ag recognition and indicate both qualitative and quantitative differences in determining cell lineage commitment.

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HLA-DQ Tetramers Identify Epitope-Specific T Cells in Peripheral Blood of Herpes Simplex Virus Type 2-Infected Individuals: Direct Detection of Immunodominant Antigen-Responsive Cells

Cited by 116 publications

References 23 publications

Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Differential Antigen Sensitivity and Costimulatory Requirements in Human Th1 and Th2 Antigen-Specific CD4+ Cells with Similar TCR Avidity

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HLA-DQ Tetramers Identify Epitope-Specific T Cells in Peripheral Blood of Herpes Simplex Virus Type 2-Infected Individuals: Direct Detection of Immunodominant Antigen-Responsive Cells

Cited by 116 publications

References 23 publications

Monitoring of NY-ESO-1 specific CD4 + T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Monitoring of NY-ESO-1 specific CD4 + T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Differential Antigen Sensitivity and Costimulatory Requirements in Human Th1 and Th2 Antigen-Specific CD4+ Cells with Similar TCR Avidity

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Product

Resources

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Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers

Monitoring of NY-ESO-1 specific CD4 ⁺ T cells using molecularly defined MHC class II/His-tag-peptide tetramers