Use of B cell-bound HLA-A2 class I monomers to generate high-avidity, allo-restricted CTLs against the leukemia-associated protein Wilms tumor antigen

Savage, Philip; Gao, Liquan; Vento, Kevin; Cowburn, Pam; Man, Stephen; Steven, Neil; Ogg, Graham S.; McMichael, Andrew; Epenetos, Agamemnon A.; Goulmy, Els; Stauss, Hans J.

doi:10.1182/blood-2003-11-3903

Cited by 56 publications

(47 citation statements)

References 21 publications

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“…Thus, the observed cross-reactivity profiles did not correlate with the WT1 specificity of the TCR and therefore could not be easily predicted. Although others have been successful in generating T cells specific for self-antigens presented in the allogeneic setting (8,(38)(39)(40)(41), none of our applied selection strategies resulted in the isolation of T cell clones that exhibited single WT1 specificity in the absence of promiscuous recognition of other peptides, illustrating that cross-reactivity is not a rare phenomenon among such high-avidity clones. In several of our experiments, we coisolated CMV-specific T cells that showed profound peptidespecific functional avidity, illustrating that our in vitro selection strategies did not result in deletion of high-avidity T cells.…”

Section: Discussionmentioning

confidence: 98%

“…However, as the T cell repertoire does not undergo negative selection in response to peptides presented in the context of nonself (allogeneic) MHC (allo-MHC), allo-MHC responses specific for a large repertoire of peptides can still be present in the postselection T cell compartment (36,37). These T cells can exhibit high avidity for Ag, which has prompted efforts to induce tumor Ag-specific T cell responses across MHC barriers (8,(38)(39)(40)(41)(42). Such high-avidity antitumor activity could be transferred to thirdparty T cells using TCR gene transfer strategies, illustrating the intrinsically high affinity of these TCRs.…”

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confidence: 99%

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Allogeneic HLA-A*02–Restricted WT1-Specific T Cells from Mismatched Donors Are Highly Reactive but Show Off-Target Promiscuity

Falkenburg

Melenhorst

Meent

et al. 2011

The Journal of Immunology

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T cells recognizing tumor-associated Ags such as Wilms tumor protein (WT1) are thought to exert potent antitumor reactivity. However, no consistent high-avidity T cell responses have been demonstrated in vaccination studies with WT1 as target in cancer immunotherapy. The aim of this study was to investigate the possible role of negative thymic selection on the avidity and specificity of T cells directed against self-antigens. T cell clones directed against the HLA-A*0201–binding WT1126–134 peptide were generated from both HLA-A*02–positive (self-HLA–restricted) and HLA-A*02–negative [nonself (allogeneic) HLA [allo-HLA]-restricted] individuals by direct ex vivo isolation using tetramers or after in vitro priming and selection. The functional avidity and specificity of these T cell clones was analyzed in-depth. Self-HLA–restricted WT1-specific clones only recognized WT1126–134 with low avidities. In contrast, allo-HLA–restricted WT1 clones exhibited profound functional reactivity against a multitude of HLA-A*02–positive targets, even in the absence of exogenously loaded WT1 peptide, indicative of Ag-binding promiscuity. To characterize this potential promiscuity, reactivity of the T cell clones against 400 randomly selected HLA-A*0201–binding peptides was investigated. The self-HLA–restricted WT1-specific T cell clones only recognized the WT1 peptide. In contrast, the allo-HLA–restricted WT1-reactive clones recognized besides WT1 various other HLA-A*0201–binding peptides. In conclusion, allogeneic HLA-A*02–restricted WT1-specific T cells isolated from mismatched donors may be more tumor-reactive than their autologous counterparts but can show specific off-target promiscuity of potential clinical importance. As a result of this, administration of WT1-specific T cells generated from HLA-mismatched donors should be performed with appropriate precautions against potential off-target effects.

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Section: Discussionmentioning

confidence: 98%

mentioning

confidence: 99%

Allogeneic HLA-A*02–Restricted WT1-Specific T Cells from Mismatched Donors Are Highly Reactive but Show Off-Target Promiscuity

Falkenburg

Melenhorst

Meent

et al. 2011

The Journal of Immunology

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“…The concept that cancer cells can be targeted by T cells recognizing self-peptides presented by foreign HLA was first described by Stauss and coworkers (15)(16)(17). Although these and other studies have shown that certain peptides can be specifically recognized when presented on allogeneic HLA (8,9,12,(18)(19)(20)(21)(22)(23), it seemed possible that they represented exceptions, because there is also evidence that allorestricted cells are more promiscuous with regard to peptide recognition (9,18,(24)(25)(26)(27)(28).…”

Section: Discussionmentioning

confidence: 99%

Alloreactive cytotoxic T cells provide means to decipher the immunopeptidome and reveal a plethora of tumor-associated self-epitopes

Kumari

Wälchli

Fallang

et al. 2013

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

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HLA molecules presenting peptides derived from tumor-associated self-antigens (self-TAA) are attractive targets for T-cell-based immunotherapy of cancer. However, detection of such epitopes is hampered by self-tolerance and limitations in the sensitivity of mass spectrometry. Here, we used T cells from HLA-A2-negative donors as tools to detect HLA-A2-bound peptides from two leukemiaassociated differentiation antigens; CD20 and the previously undescribed cancer target myeloperoxidase. A high-throughput platform for epitope discovery was designed using dendritic cells cotransfected with full-length transcripts of self-TAA and HLA-A2 to allow presentation of all naturally processed peptides from a predefined self-protein on foreign HLA. Antigen-reactive T cells were directly detected using panels of color-coded peptide-HLA multimers containing epitopes predicted by a computer algorithm. Strikingly, cytotoxic T cells were generated against 37 out of 50 peptides predicted to bind HLA-A2. Among these, 36 epitopes were previously undescribed. The allorestricted T cells were exquisitely peptide-and HLA-specific and responded strongly to HLA-A2-positive leukemic cells with endogenous expression of CD20 or myeloperoxidase. These results indicate that the repertoire of self-peptides presented on HLA class I has been underestimated and that a wealth of self-TAA can be targeted by T cells when using nontolerized T-cell repertoires.C ytotoxic T cells (CTLs) selectively kill target cells that express defined peptides in complex with MHC class I molecules on the cell surface. Most tumor-associated antigens (TAA) are wild-type self-proteins, and T cells that recognize peptides from these antigens with high affinity are deleted during thymic development. Thus, the utility of T cells for detection of self-peptides presented on self-HLA is limited by tolerance. This may be one reason why the number of epitopes identified from TAA after 2 decades of intense research amounts to less than 600 (1). The ability to rapidly identify new CTL epitopes would likely facilitate the development of effective immunotherapeutic strategies against cancer.MHC molecules can be isolated from cells and the associated peptides eluted for identification by MS. Ultimately, this approach may provide a description of the entire MHC-bound peptide repertoire: the immunopeptidome (2, 3). This is, however, a daunting task, and it is unclear whether current MS-based protocols already provide the required sensitivity. Indeed, although 100,000-750,000 peptide-MHC class I complexes are expressed for each allelic product on the cell surface (for HLA-A and HLA-B loci) (3, 4), the largest HLA ligandome identified to date contains 14,065 peptides (5). In contrast, the predicted number of different HLA class I ligands would be 352,000 using the well-renowned computer algorithm NetMHCpan, considering that on average, 4.4% of all nonamers bind HLA class I (6) and that a cell contains at least 8 × 10 6 distinct nonamers (7). Thus, there is a very large gap between the numb...

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“…This approach has also been used to treat solid tumors (2). Recently, protocols have been developed to identify, isolate, and expand ex vivo tumor peptide-specific allorestricted T cells (3)(4)(5)(6), anticipating their use for adoptive immunotherapy. However, the plan may be hampered by adventitious generation of peptide-independent alloreactive T cells (3,6), which could induce immunopathology.…”

mentioning

confidence: 99%

“…Recently, protocols have been developed to identify, isolate, and expand ex vivo tumor peptide-specific allorestricted T cells (3)(4)(5)(6), anticipating their use for adoptive immunotherapy. However, the plan may be hampered by adventitious generation of peptide-independent alloreactive T cells (3,6), which could induce immunopathology. Therefore, renewed calls have been made for a more extensive analysis of T cell alloreactivity before clinical application of these strategies (7).…”

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confidence: 99%

Investigation of Peptide Involvement in T Cell Allorecognition Using Recombinant HLA Class I Multimers

Whitelegg

Oosten²,

Jordan³

et al. 2005

The Journal of Immunology

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Alloreactive T cells are involved in injurious graft rejection and graft-vs-host disease. However, they can also evoke beneficial responses to tumor Ags restricted by foreign MHC molecules. Manipulation of these alloreactivities requires information on the basis of T cell allorecognition. The vigorous T cell response to foreign MHC molecules may arise from peptide-independent recognition of polymorphic residues of foreign MHC molecules or peptide-specific recognition of novel peptides presented by foreign MHC molecules. We investigated CD8+ T cell allorecognition using recombinant HLA class I/peptide complexes. Peptide-specific allorecognition was examined using tetramers of HLA-A*0201 representing five peptides derived from ubiquitously expressed self-proteins that are known to bind endogenously to HLA-A*0201. Distinct subsets of CD8+ T cells specific for each HLA-A*0201/peptide combination were detected within four in vitro-stimulated T cell populations specific for foreign HLA-A*0201. Peptide-independent allorecognition was investigated using artificial Ag-presenting constructs (aAPCs) coated with CD54, CD80, and functional densities of a single HLA-A*0201/peptide combination for four different peptides. None of the four T cell populations specific for foreign HLA-A*0201 were stimulated by the aAPCs, whereas they did produce IFN-γ upon stimulation with cells naturally expressing HLA-A*0201. Thus, aAPCs did not stimulate putative peptide-independent allorestricted T cells. The results show that these alloreactive populations comprise subsets of T cells, each specific for a self-peptide presented by foreign class I molecules, with no evidence of peptide-independent components.

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Use of B cell-bound HLA-A2 class I monomers to generate high-avidity, allo-restricted CTLs against the leukemia-associated protein Wilms tumor antigen

Cited by 56 publications

References 21 publications

Allogeneic HLA-A*02–Restricted WT1-Specific T Cells from Mismatched Donors Are Highly Reactive but Show Off-Target Promiscuity

Allogeneic HLA-A*02–Restricted WT1-Specific T Cells from Mismatched Donors Are Highly Reactive but Show Off-Target Promiscuity

Alloreactive cytotoxic T cells provide means to decipher the immunopeptidome and reveal a plethora of tumor-associated self-epitopes

Investigation of Peptide Involvement in T Cell Allorecognition Using Recombinant HLA Class I Multimers

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