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Preprocalcitonin antigen T epitopes

An immunogenic, polynucleotide technology, applied in the field of procalcitonin precursor antigen T epitope

Active Publication Date: 2013-05-01
法国古斯塔夫鲁西研究院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aberrant expression of the gene CALCA in lung cancer cells is poorly understood, but is partly caused by promoter demethylation (BAYLIN et al., Cancer Res, 46, 2917-22, 1986) and loss of transcriptional repression (SYMES et al. ., FEBS Lett, 306, 229-33, 1992) lead to

Method used

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  • Preprocalcitonin antigen T epitopes
  • Preprocalcitonin antigen T epitopes
  • Preprocalcitonin antigen T epitopes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Materials and methods

[0042] Derivatives of NSCLC tumor cell lines and autologous CTL clones

[0043] IGR-Heu NSCLC was derived from LCC samples of patients Heu (HLA-A2, A68, B7, B35, C4, C7) as previously reported (ECCHHAKIR et al., Cancer Res, 61, 4078-83, 2001) cell lines and maintained in culture medium. According to the method described in the literature (ECCHHAKIR et al., Int Immunol, 12, 537-46, 2000), the Heu161 clone was derived from autologous TIL, and in a 96-well V-shaped microtiter plate (Nunc, Roskilde, Denmark) Amplify.

[0044] Cytotoxicity assay and TNFβ release

[0045] Cultures were used in triplicate in 96-well plates with round bottoms and passed through common 4-h 51 Cr release assay to measure cytotoxic activity. Percent specific cytotoxicity is usually calculated; SD (standard deviation) + MTC; ECACC, Salisbury, UK) and DMS53 (HLA-A2 - SCLC; ECACC) cell lines were used as targets.

[0046] TNF[beta] was detected by measuri...

Embodiment 2

[0058] Example 2: CTL clones that can recognize autologous lung cancer cells

[0059] Patient Heu(HLA-A2+ ) is a currently disease-free lung cancer patient 11 years after resection of the primary tumor. The LCC cell line IGR-Heu was derived from the patient's resected tumor in 1996. Primary tumor permeable monocytes were isolated and stimulated with irradiated IGR-Heu tumor cells, irradiated autologous EBV-transformed B cells, and IL-2. Responding lymphocytes were cloned by limiting dilution and stimulated with the same mixture of tumor cells and EBV-B cells. Several tumor-specific CTL clones were obtained and differentiated into three groups based on their TCRVβ sequences (ECHCHAKIR et al., Int Immunol, 12, 537-46, 2000). We previously reported that the first two sets of clones (denoted Heu127 and Heu171) recognized antigenic peptides encoded by mutated sequences in the α-actinin-4 (ACTN4) gene (ECHCHAKIR et al., Cancer Res, 61, 4078-83, 2001; ECCHHAKIR et al., Proc Natl A...

Embodiment 3

[0060] Example 3: Identification of genes encoding antigens recognized by HEU161 clones

[0061] Poly(A) + RNA The cDNA library prepared from IGR-Heu cells was cloned into the expression plasmid pCEP4 (ECHCHAKIR et al., Cancer Res, 61, 4078-83, 2001). The library was divided into 264 pools containing approximately 100 recombinant clones, and DNA was prepared from each pool. 293-EBNA cells were co-transfected with DNA from each pool and the HLA-A*0201 construct. After 24h, the CTL clone Heu161 was added to the transfectants; after another 24h, the supernatant was collected, and their TNFβ content was measured with TNF-sensitive WEHI-164c13 cells (ESPEVIK & NISSEN-MEYER, J Immunol Methods, 95, 99-105, 1986). A large proportion (85 / 264) of the cDNA pool proved positive, suggesting that a surprisingly high frequency of about 0.4% of the cDNA clones encoded Ag. One cDNA pool was subcloned, and a cDNA clone called 150 was isolated ( figure 1 B).

[0062] cDNA 150 is 956 bp lon...

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Abstract

The present invention is related to Preprocalcitonin antigen T epitopes, presented by the Major Histocompatibility Complex I (MHC I). These peptides can be used in cancer immunotherapy.

Description

technical field [0001] The present invention relates to procalcitonin precursor T epitope and its application in cancer immunotherapy. Background technique [0002] Analysis of tumor-reactive CTLs derived from patients with different solid tumors has led to promising new therapeutic approaches for malignant diseases, either by developing T cells in vitro before transferring them into patients with IL-2 ( GATTINONI et al., Nat Rev Immunol, 6, 383-93, 2006), or by identifying their target Ags which can then be applied in therapeutic vaccines. A number of tumor-associated Ags have been identified that are recognized by CTLs from PBLs, or from tumor-infiltrating lymphocytes (TILs). Most of this work has been performed with malignant melanoma tumors. Unfortunately, clinical studies have shown that despite the increased frequency of antitumor CD8 T cells, the efficacy of current therapeutic vaccines is still limited to patients with metastatic melanoma (ROSENBERG et al., Nat Med...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/585A61K38/04C12N15/11
CPCA61K39/0011C07K14/585A61K2039/53A61K39/00A61P35/00A61P37/04A61K39/4644A61K39/4611
Inventor 法腾·埃拉阿热文森特·施特罗班特皮埃尔·G·库利耶法蒂埃·马米-舒艾卜
Owner 法国古斯塔夫鲁西研究院
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