HLA-A2 restrictive epitope polypeptide of LMP2A protein source and purpose thereof

A technology of LMP2A264 and HLA-A2, applied in the direction of peptide/protein components, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problem of patients who are prone to recurrence and metastasis, and cannot completely and effectively remove tumor cells , side effects and other issues

Inactive Publication Date: 2010-02-10
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, NPC cannot be treated with surgery, mainly through radiotherapy and chemotherapy, but this treatment method cannot completel

Method used

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  • HLA-A2 restrictive epitope polypeptide of LMP2A protein source and purpose thereof
  • HLA-A2 restrictive epitope polypeptide of LMP2A protein source and purpose thereof
  • HLA-A2 restrictive epitope polypeptide of LMP2A protein source and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Discovery of EBV-LMP2A-specific CTL epitopes

[0022] In this example, HLA-A2-restricted LMP2A-specific epitope peptides were screened out by combining bioinformatics with in vitro experiments.

[0023] 1) Test method

[0024] The following three bioinformatics prediction software were used to predict the possible epitope peptides of LMP2A protein: SYFPEITHI (http: / / www.uni-tuebingen.de / uni / kxi), MHCPred (http: / / www.jenner.ac. uk / MHCPred / ) and NetMHC (http: / / www.cbs.dtu.dk / services / NetMHC-2.0 / ). Those who met the following conditions at the same time were selected: ① The prediction score was high; ② The polynomial scheme analysis score was greater than the selected threshold (-23 in this study); ③ There were two or more prediction results of the above three methods.

[0025] The inventor selected the following 6 polypeptides for research:

[0026] Table 1. LMP2A-specific CTL candidate epitope peptides

[0027]

[0028] The above peptides were respectivel...

Embodiment 2

[0031] Example 2. Newly discovered peptide QLS induces Th1 cytokine test in vitro test in patients with nasopharyngeal carcinoma

[0032] 1) Test method:

[0033] Peptides were used to stimulate PBMCs of patients with nasopharyngeal carcinoma in vitro, the stimulating concentration of peptides was 20 μg / ml, and cytokine IL-2 was added at the same time, the final concentration was 20 U / ml. After 5 days, the cells were collected to interact with target cells (LMP2A-293T cells), the secretion of intracellular IFN-γ and the killing effect of CTL were detected, and the in vitro immune effect of the polypeptide was evaluated. Such as Figure 2 showed that FACS detected CD8 + / IFN-γ + frequency of T cells.

[0034] 2) Test results (see figure 2)

[0035] The results showed that the number of CD8 cells secreting IFN-γ in HLA-A2 positive NPC patients XLD, XWM, ZHW and CJW PBMCs stimulated in vitro by peptides was significantly higher than that of other non-HLA-A2 positive NPC pa...

Embodiment 3

[0036] Example 3. In vitro test of newly discovered peptide QLS in nasopharyngeal carcinoma patients to induce CTL killing test

[0037] 1) Test method

[0038] The T lymphocytes and LMP2A-293T cells stimulated by the polypeptide were added to the 96-well U-bottom culture plate according to the effect-to-target ratio of 10:1, and the cell killing activity was detected by double-color flow cytometry to detect the death rate of the target cells, and the polypeptide was evaluated in CTL killing activity induced in different patients.

[0039] 2) The results show that,

[0040] CTLs derived from HLA-A2 positive NPC patients can kill target cells, while no obvious killing activity was detected in other non-HLA-A2 NPC patients (patients 1, 2, 3, 5, 7, 10, 11) (see image 3) . Killing activity predicts the ability of in vivo therapy to kill tumor cells, and this result also proves that the newly discovered peptide is mainly used in patients with HLA-A2.1 positive (ie, HLA-restrict...

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Abstract

The invention discloses HLA-A2 restrictive epitope polypeptide of an LMP2A protein source, which is polypeptide LMP2A264 and comprises 9 amino acids in the positions of 264 to 272; the sequence is QLSPLLGAV. Mycobacteria heat shock protein 70 (HSP70) is expressed, identified and purified; a transgene animal model for evaluating EBV relevant polypeptide vaccine in vivo tests is established; the immune research on inducing EBV specific CTL inside and outside healthy volunteers and transgene rats for the new EBV epitope polypeptide LMP2A264 is systemically researched and the tumor prevention andthe tumor treating test of the polypeptide LMP2A264 on LMP2A positive solid tumors are evaluated, thereby proving that the polypeptide LMP2A264 can induce LMP2A specific cell toxic T lymphocyte reaction. Therefore, the polypeptide LMP2A264 can be applied to prepare medicaments for treating or preventing LMP2A positive solid tumors.

Description

Technical field: [0001] The invention relates to an HLA-A2 restricted epitope polypeptide derived from LMP2A protein and its application in the treatment and prevention of solid tumors Background technique [0002] Epstein-Barr virus (EBV) is a member of the γ subfamily of the Herpesviridae family. It was first isolated from African childhood lymphoma by Epstein and Barr in 1964. About 95% of the world's people have been infected with EBV and become carriers. Nasopharyngeal carcinoma (NPC) is a malignant tumor derived from epithelial cells that is highly related to EBV infection, and is one of the common tumors in the head and neck. NPC occurs frequently in Southeast Asia and South China. The current incidence rate in Jiangsu is also increasing, with an average of 10 to 20 new cases per week, and about 960 new cases per year. Clinically, NPC cannot be treated with surgery, mainly through radiotherapy and chemotherapy, but this treatment method cannot completely and effecti...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K38/08A61K47/48A61P35/00
Inventor 姚堃王冰刘根焰周锋谢芳艺陈云冯东举刘英霞
Owner NANJING MEDICAL UNIV
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