Simultaneous monitoring of binding to and activation of tumor-specific T lymphocytes by peptide–MHC

Cohen, Cyril J.; Denkberg, Galit; Schiffenbauer, Yael S.; Segal, Dina; Trubniykov, Ella; Berke, Gideon; Reiter, Yoram

doi:10.1016/s0022-1759(03)00110-8

Cited by 10 publications

(12 citation statements)

References 29 publications

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“…In addition, the fusion of the ␤2m subunit to the CD1 heavy chain through a flexible hydrophilic linker sequence most likely led to enhanced expression and stability of the product. Although we have not directly compared the scCD1 proteins to soluble versions that lack the covalent linkage of ␤2m, this strategy has been successfully used in previous studies for the production of soluble MHC class I and CD1 proteins (42,43). We were able to achieve adequate levels of scCD1 proteins in the supernatants of transfected CHO cells to allow the isolation of sufficient quantities of these proteins for ligand binding studies without requiring a large scale-up of culture volumes.…”

Section: Discussionmentioning

confidence: 99%

Direct Measurement of Antigen Binding Properties of CD1 Proteins Using Fluorescent Lipid Probes

Illarionov

et al. 2004

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

Direct Measurement of Antigen Binding Properties of CD1 Proteins Using Fluorescent Lipid Probes

Illarionov

et al. 2004

Journal of Biological Chemistry

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“…The CellScan is unique in its capability of repetitive measurements of individual cells in a cell population. It has been used to detect activation of lymphocytes both in physiological (Eisenthal et al 1996, Zurgil et al 1996 and pathological situations including: cancer (Ron et al 1995, Merimsky et al 1996, Rahmani et al 1996, Schiffenbauer et al 2002, Cohen et al 2003, autoimmune disorders (Zurgil et al 1997, and atherosclerosis ).…”

Section: Cellscan Testmentioning

confidence: 99%

A Novel System to Diagnose Cutaneous Adverse Drug Reactions Employing the Cellscan—Comparison with Histamine Releasing Test and Inf‐γ Releasing Test

Goldberg

Gilburd

Kravitz

et al. 2005

Journal of Immunology Research

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Background: There are several mechanisms to describe allergic drug reactions yet the methods to diagnose them are limited.Objective: To compare several conventional clinical and laboratory methods to diagnose skin reactions to drugs to a new method of diagnosing drug reactions by the CellScan system.Methods: The study entailed 21 patients who were diagnosed as suffering from drug eruptions, and 105 healthy controls with no history of drug allergy. The drugs were classified into two groups according to suspicion of causing drug allergy: high and low. Most of the patients were on more than one drug, leading to 41 patient -drug interactions (assays). Histamine releasing test (HRT), interferon (INF)-g releasing test and CellScan examination were performed on lymphocytes of the patients and controls.Results: The HRTwas interpreted as positive in 9 out of 18 (50%) patients and in 13 out of 35 (37%) assays. Based on the INF-g releasing test, positive results were observed in 16 out of 21 (76%) patients and in 24 out of 41 (59%) assays. In the CellScan test (CST), positive results were observed in 17 out of 21 (81%) patients and in 29 out of 41 (71%) assays. The rate of identifying the drug for eruption in the high suspicion level drugs was 9 out of 22 (41%) assays in the HRT, 20 out of 24 (83%) assays in the INF-g releasing test, and 21 out of 24 (87%) studies with the CellScan method. The rate of determining of the drug that caused the eruption in the low suspicion level drugs was 4 out of 13 (31%) in the HRT, 4 out of 17 (24%) assays in the INF-g releasing test, and 8 out of 17 (47%) analyses in the CST. When examined in the CellScan, 99 out of 105 (94%) controls were interpreted as negative.Conclusion: This preliminary study indicates that the CellScan seems to be an easy and promising method for the detection of drugs responsible for adverse skin reactions. In contrast to the HRTand to the Interferon-g secretion test, the CellScan method is characterized by its ability to track and monitor the reaction of individual cells. By measuring the kinetic parameters of selected cells before and after adding the suspected drug, we were able to identify the culprit drug. The CellScan method had the highest sensitivity, and the interferon-g secretion test had the highest specificity for detection of the culprit drug. In contrast, the analysis of 105 normal control sera disclosed a high specificity of 94% for the CellScan method.

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“…This observation may result from the fact that a local high concentration of coated MHC-peptide complexes displaying one particular T cell epitope (peptide) is formed on the targeted cells, which greatly exceeds the natural density of such complexes displayed on the surface of cells. Further evidence of this possibility is that MHC tetramers can induce T cell activation by themselves (33); including our recent observation that CTL activation by MHC tetramers without accessorial molecules can be demonstrated at the single-cell level (34). Another possible mode of action of our approach is the induction of Fas͞Fas ligand-mediated apoptosis.…”

Section: Discussionmentioning

confidence: 83%

Tumor-specific Ab-mediated targeting of MHC-peptide complexes induces regression of human tumor xenograftsin vivo

Lev

Noy

Oved

et al. 2004

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

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A cancer immunotherapy strategy is described herein that combines the advantage of the well established tumor targeting capabilities of high-affinity recombinant fragments of Abs with the known efficient, specific, and potent killing ability of CD8 T lymphocytes directed against highly antigenic MHC-peptide complexes. Structurally, it consists of a previously uncharacterized class of recombinant chimerical molecules created by the genetic fusion of single-chain (sc) Fv Ab fragments, specific for tumor cell surface antigens, to monomeric scHLA-A2 complexes containing immunodominant tumor-or viralspecific peptides. The fusion protein can induce very efficiently tumor cell lysis, regardless of the expression of self peptide-MHC complexes. Moreover, these molecules exhibited very potent antitumor activity in vivo in nude mice bearing preestablished human tumor xenografts. These in vitro and in vivo results suggest that recombinant scFv-MHCpeptide fusion molecules could represent an approach to immunotherapy, bridging Ab and T lymphocyte attack on cancer cells. C urrent cancer immunotherapy strategies typically employ two arms of the natural immune system: humoral and cellular. In the first, systemic injection of high-affinity mAbs directed against cell surface tumor-associated antigens has demonstrated statistically significant antitumor activities in clinical trials (1-3). Antitumor Abs that carry effector payloads such as toxins (immunotoxins) or cytokines are also potent molecules currently being tested in various clinical trials (4, 5). The second major approach for specific cancer immunotherapy consists of the potentiation of the cellular arm of the immune system, mainly through CD8 ϩ cytotoxic T lymphocytes (CTLs). Two major strategies are currently being used: (i) active immunization of patients with antigens known to be recognized by T lymphocytes and to activate them (6-8) and (ii) adoptive transfer therapies that enable the selection and activation of highly reactive T cell subpopulations with improved antitumor activities (9). Clinical studies using MHC tetramer staining have demonstrated T lymphocyte responses against the immunizing tumor antigens in the course of vaccination. However, these promising clinical trials using active immunization have suffered from a relatively low percentage of tumor remissions and a lack of correlation between clinical and T lymphocyte responses to the vaccine (9, 10). Furthermore, in using this approach there is the potential risk of selecting tumor cell variants that have undergone HLA loss (11). The adoptive transfer approach has recently demonstrated impressive results (12, 13). Regression of metastatic melanoma was reported in patients undergoing adoptive transfer protocols with highly selected tumor-reactive T cells directed against overexpressed self-derived differentiation antigens after a nonmyeloablative conditioning regimen (12, 13). The efficiency of such T cell-based immunotherapy approaches may be limited by the absence or low expression of either MHC molecul...

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Simultaneous monitoring of binding to and activation of tumor-specific T lymphocytes by peptide–MHC

Cited by 10 publications

References 29 publications

Direct Measurement of Antigen Binding Properties of CD1 Proteins Using Fluorescent Lipid Probes

Direct Measurement of Antigen Binding Properties of CD1 Proteins Using Fluorescent Lipid Probes

A Novel System to Diagnose Cutaneous Adverse Drug Reactions Employing the Cellscan—Comparison with Histamine Releasing Test and Inf‐γ Releasing Test

Tumor-specific Ab-mediated targeting of MHC-peptide complexes induces regression of human tumor xenograftsin vivo

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