Cutting Edge: Predetermined Avidity of Human CD8 T Cells Expanded on Calibrated MHC/Anti-CD28-Coated Microspheres

Walter, Steffen; Herrgen, Leah; Schoor, Oliver; Jung, Gundram; Wernet, Dorothee; Bühring, Hans‐Jörg; Rammensee, Hans‐Georg; Stevanović, Stefan

doi:10.4049/jimmunol.171.10.4974

Cited by 53 publications

(43 citation statements)

References 18 publications

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“…16,17 In contrast, in vitro priming of T cells results in a T-cell repertoire containing a large variety of avidities, 18,19 comparable with the repertoire of naive T cells, indicating that during in vitro activation and expansion no selection for high avidity occurs. Because under normal circumstances allo-HLA molecules are not encountered, alloreactive T cells which can be present within the naive as well as the memory T-cell pool, 20,21 will not have undergone selection for T cells with high avidity for allo-HLA molecules.…”

Section: Introductionmentioning

confidence: 99%

Allo-HLA–reactive T cells inducing graft-versus-host disease are single peptide specific

Amir

Steen

Hagedoorn

et al. 2011

Blood

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T-cell alloreactivity directed against nonself-HLA molecules has been assumed to be less peptide specific than conventional T-cell reactivity. A large variation in degree of peptide specificity has previously been reported, including single peptide specificity, polyspecificity, and peptide degeneracy. Peptide polyspecificity was illustrated using synthetic peptide-loaded target cells, but in the absence of confirmation against endogenously processed peptides this may represent low-avidity T-cell reactivity. Peptide degeneracy was concluded based on recognition of Ag-processing defective cells. In addition, because most investigated alloreactive T cells were in vitro activated and expanded, the previously determined specificities may have not been representative for alloreactivity in vivo. To study the biologically relevant peptide specificity and avidity of alloreactivity, we investigated the degree of peptide specificity of 50 different allo-HLA-reactive T-cell clones which were activated and expanded in vivo during GVHD. All but one of the alloreactive T-cell clones, including those reactive against Ag-processing defective T2 cells, recognized a single peptide allo-HLA complex, unique for each clone. Downregulation of the expression of the recognized Ags using silencing shRNAs confirmed single peptide specificity. Based on these results, we conclude that biologically relevant alloreactivity selected during in vivo immune response is peptide specific. (Blood. 2011;118(26):6733-6742)

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Section: Introductionmentioning

confidence: 99%

Allo-HLA–reactive T cells inducing graft-versus-host disease are single peptide specific

Amir

Steen

Hagedoorn

et al. 2011

Blood

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“…Such primed T cells might enhance tumor specificity and limit Graft versus Host Disease complications of current DLI strategies [63]. For this purpose, a promising alternative is represented by the quite fast generation of epitope-specific T cells through artificial APC [119]. This strategy, in fact, allowed us to obtain an epitope-specific CTL population ( Figure 20) in a shorter time if compared to traditional protocols, and to successfully generate a large number of epitope-positive clones (Figure 21), which could be in turn expanded for DLI intentions, even if we still need to confirm their functionality.…”

Section: Discussionmentioning

confidence: 99%

“…MHC class-I/peptide monomers were refolded and biotinylated using standards protocols [103,119] ), and 2.5-7.5mg of the selected peptide diluted 10mg/ml in DMSO. The reaction was then incubated overnight at 10°C while gently shaking.…”

Section: Monomers Refolding and Multimers Productionmentioning

confidence: 99%

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1086 IGHV1-69 as a Promising Candidate for the Development of a Shared Immunotherapy to B-cell Lymphomas

Muraro¹,

Martorelli²,

Bomben³

et al. 2012

European Journal of Cancer

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B-cell Non-Hodgkin Lymphomas (B-NHL) are a heterogeneous group of cancers, broadly diffused worldwide and often relapsing after standard treatment and rituximab. Therapeutic vaccines targeting B-NHL idiotype (Id) represent a promising approach to maintain the complete response induced by standard treatments. However, customized idiotypic vaccination still remains a non-approved, experimental therapeutic option, mostly due to the personalized use and penalized by the lack of reliable clinical or biological markers of patient eligibility and responsiveness. Nevertheless, the molecular characterization of different lymphoid tumor histotypes revealed a set of stereotyped immunoglobulins among distinct B-cell lymphoma types. On this ground, we focused our attention on the IGHV1-69 protein, frequently expressed in HCV-associated lymphomas, chronic lymphocytic leukaemia, and auto-immunity related lymphoproliferations, and we characterized the ex vivo immunogenicity of this protein.Seventy IGHV1-69 sequences obtained from patients affected by different B-NHLs or pre-malignant lymphoproliferations were compared to design an optimized sequence characterized by the highest degree of similarity among studied cancers, and thus CDR3 hypervariable region free. Within this "immunogenically" optimized sequence, we identified 13 potential HLA class-I cytotoxic T lymphocyte (CTL) epitopes and synthesized the corresponding pentamers (Pent). We assessed by flow cytometry the presence and extent of epitope-specific T-cell responses in peripheral blood of patients with IGHV1-69 + B-cell lymphoproliferative disorders and healthy donors, and validated these data in IFN-γ ELISPOT (Enzyme-linked immunosorbent spot) assays. Finally, we boosted in vitro IGHV1-69-specific responses by stimulating peripheral blood lymphocytes (PBL) from donors and patients with different protocols for the generation of epitope-specific CTL cultures.Interestingly, the IGHV1-69 Pent + population observed in patients' samples was generally larger than in donors, supporting the existence of spontaneous memory T-cell responses against IGHV1-69, at least for some HLA-restrictions. Surprisingly, in patients' samples, IGHV1-69-recognizing T cells displayed higher IFN-γ release in ELISPOT assays compared to viral-specific T cells. Moreover, we obtained peptide-specific CTL lines, which showed a weak but specific lysis against peptide-pulsed targets, especially when derived from patients' PBLs. In addition, we were able to generate IGHV1-69-epitope specific CTL clones from healthy donors CD8 + T cells, employing synthetic artificial APC, developed to elicit and expand low-avidity tumor-directed human CTL lines. Finally, IGHV1-69-induced CTL lines showed specific lysis also towards an IGHV1-69 naturally expressing cell line, suggesting that IGHV1-69 memory T-cell responses could be boosted for therapeutic purposes.These results show that IGHV1-69 constitutes a potential target for the development of a subset-specific Id vaccine. Furthermore, multimer (tetramers...

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“…Correlation of mRNA with protein levels (36) and HLA ligand levels (27) is weak. Thus, protein levels of VEGF were assessed by a bead-based sandwich immunoassay (32) in RCC099 and RCC110 (Fig.…”

Section: Quantitative Analysis Of Vegf Protein Levels In Rccsmentioning

confidence: 99%

A Cryptic Vascular Endothelial Growth Factor T-Cell Epitope: Identification and Characterization by Mass Spectrometry and T-Cell Assays

et al. 2008

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Cutting Edge: Predetermined Avidity of Human CD8 T Cells Expanded on Calibrated MHC/Anti-CD28-Coated Microspheres

Cited by 53 publications

References 18 publications

Allo-HLA–reactive T cells inducing graft-versus-host disease are single peptide specific

Allo-HLA–reactive T cells inducing graft-versus-host disease are single peptide specific

1086 IGHV1-69 as a Promising Candidate for the Development of a Shared Immunotherapy to B-cell Lymphomas

A Cryptic Vascular Endothelial Growth Factor T-Cell Epitope: Identification and Characterization by Mass Spectrometry and T-Cell Assays

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