An improved assembly assay for peptide binding to HLA-B*2705 and H-2Kk class I MHC molecules

Tan, Linda; Andersen, Mads Hald; Elliott, Tim; Haurum, John S.

doi:10.1016/s0022-1759(97)00142-7

Cited by 15 publications

(11 citation statements)

References 49 publications

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“…All peptides were purchased from KJ Ross-Petersen ApS (RJ Ross-Petersen ApS, Holte, Denmark) and provided at Ͼ80% purity as verified by HPLC and MS analysis. The assembly assay was carried out as described previously (12). It uses a HLA-B*2705-transfected T2 cell line (T2-B27) (a kind gift from P. Cresswell) that are defective in their ability to load endogenous peptides into MHC class I because of a defect in the transport of peptides within the cell.…”

Section: Methodsmentioning

confidence: 99%

Immunogenicity of Constitutively Active V599EBRaf

et al. 2004

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Activating BRAF somatic missense mutations within the kinase domain are present in 60 -66% of melanomas. The vast majority of these represent a single substitution of glutamate for valine (V599E). Here, we demonstrate spontaneous HLA-B*2705-restricted cytotoxic T-cell responses against an epitope derived from V599E BRaf. These T-cell responses were mutation specific as the corresponding epitope derived from wildtype BRaf was not recognized. The loss of the V599E BRAF genotype during progression from primary to metastatic melanoma in patients with V599E BRaf specific T-cell responses suggests an active immune selection of nonmutated melanoma clones by the tumor-bearing host.

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Section: Methodsmentioning

confidence: 99%

Immunogenicity of Constitutively Active V599EBRaf

et al. 2004

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“…35,36 Briefly, 5 × 10 6 T2 cells per experimental point were washed in warm PBS (37°C). Cells were resuspended in warm (37°C) methionine/cysteine free RPMI supplemented with 10% FCS (the FCS was dialysed against PBS prior to use) and 10 mM HEPES (Met/Cys free R10) at 10 7 cells/ml and incubated at 37°C, 5% CO 2 for 30-45 min.…”

Section: Assembly Assay For Peptide Binding To Hla Class I Moleculesmentioning

confidence: 99%

Peptides spanning the junctional region of both the abl/bcr and the bcr/abl fusion proteins bind common HLA class I molecules

et al. 2000

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The Philadelphia (Ph) chromosome, resulting from the t(9;22) translocation, is characteristic of chronic myeloid leukemia (CML). As a result of this translocation, two novel chimeric genes are generated and the bcr/abl and abl/bcr fusion proteins expressed. The bcr/abl fusion mRNA is present in all CML patients, whereas the reciprocal abl/bcr fusion mRNA is detectable in about 80% of the Ph + CML patients. These fusion proteins may undergo enzymatic degradation in the cytosol and give rise to MHC class I restricted peptide epitopes originating from the junctional regions of the translocation products, which thus may serve as novel tumor specific antigens. Previously, other groups have tested peptides corresponding to the junctional region of the bcr/abl protein for their binding capacity to HLA class I molecules and have identified a few candidate epitopes. Peptides originating from the abl/bcr fusion protein have on the other hand so far been neglected, for no apparent reason. We have now extended these studies to include also the reciprocal abl/bcr translocation product by testing a large panel of synthetic peptides corresponding to the junctional regions of both the abl/bcr and the bcr/abl fusion proteins for their ability to stabilize HLA class I molecules. We find that the abl/bcr translocation product may be an even more important source of CML specific peptide antigens and together the junctional sequences of both these proteins contain peptide sequences which bind efficiently to a number of HLA molecules (HLA-A1, -A2, -A3, -A11, -B7, -B27, -B35) and thus may serve as candidate CML specific tumor antigens. Leukemia (2000) 14, 419-426.

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“…The binding affinity of synthetic peptides (Invitrogen, Carlsbad, CA, USA) to HLA-A1, -A2, -A3, or -A11 molecules metabolically labeled with [ 35 S]-methionine was measured in the assembly assay, as described previously. 26,27 The assay is based on peptide-mediated stabilization of empty HLA molecules released upon cell lysis, from the TAP-deficient cell lines. Stably folded HLA-molecules were immune-precipitated using the HLA class I-specific, conformation-dependent mAb W6/32, and separated by isoelectric focusing (IEF) gel electrophoresis.…”

Section: Introductionmentioning

confidence: 99%

Identification of Novel Survivin-Derived CTL Epitopes with Different HLA-A-Restriction Profiles

Reker

Meier

Holten-Andersen

et al. 2004

Cancer Biology & Therapy

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The identification of tumor antigens, which are essential for the survival of tumor cells is a new avenue to prevent antigen loss variants emerging due to immunoselection, particularly during immune therapy. In the search for such immunogenic tumor antigens, we recently identified spontaneous cytotoxic lymphocyte (CTL) responses against the inhibitor of apoptosis protein survivin. Thus, we identified two HLA-A2-restricted, survivin-derived CTL epitopes, which both were targets for spontaneous CTL responses in melanoma, breast cancer, and CLL. Here, we extend these data and describe the characterization of novel HLA-A1-, HLA-A2-, HLA-A3-, and HLA-A11-restricted survivin epitopes on the basis of spontaneous CTL responses in cancer patients. These epitopes significantly increase the number of patients eligible for immunotherapy based on survivin derived peptides. Additionally, the collective targeting of several restriction elements is likely to decrease the risk of immune escape by HLA-allele loss.

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An improved assembly assay for peptide binding to HLA-B*2705 and H-2Kk class I MHC molecules

Cited by 15 publications

References 49 publications

Immunogenicity of Constitutively Active V599EBRaf

Immunogenicity of Constitutively Active V599EBRaf

Peptides spanning the junctional region of both the abl/bcr and the bcr/abl fusion proteins bind common HLA class I molecules

Identification of Novel Survivin-Derived CTL Epitopes with Different HLA-A-Restriction Profiles

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