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Construction method of Beta-alexin-2-tumor vaccine and uses thereof

A technology of tumor vaccines and defensins, applied in the field of tumor vaccines, can solve problems such as unidentified antigens and unclear effects

Inactive Publication Date: 2009-03-11
INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] However, the method of Biragyn et al. is only applicable to tumors for which relevant antigens have been identified. In fact, so far, most tumors have not identified relevant antigens.
Moreover, Biragyn et al. only confirmed the anti-tumor effect of MBD2 in lymphoma, and its role in other malignant tumors such as leukemia is still unclear

Method used

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  • Construction method of Beta-alexin-2-tumor vaccine and uses thereof
  • Construction method of Beta-alexin-2-tumor vaccine and uses thereof
  • Construction method of Beta-alexin-2-tumor vaccine and uses thereof

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Embodiment Construction

[0030] 1. Construction and Characterization of MBD2 Tumor Vaccine

[0031] 1. Materials and methods

[0032] 1.1 Cell lines, main reagents and plasmids The mouse lymphocytic leukemia cell line L1210 was purchased from the Cell Bank of the Chinese Academy of Medical Sciences, and was routinely cultured in RPMI 1640 medium and 10% (V / V) FBS. Lipofectamine TM 2000), OPTI-MEM low serum medium are products of Invitrogen Company. RNase A was purchased from Sangon Company. Pyrobest TM DNA polymerase, E X Taq enzymes, restriction endonucleases, and a small plasmid extraction kit (MiniBEST Plasmid Purification Kit) were purchased from TaKaRa (Dalian) Company. T 4DNA ligase, TRIzol, and M-MLV reverse transcription kit were purchased from Invitrogen. The gel recovery kit was purchased from Huashun Biological Engineering Co., Ltd. Mouse GM-CSF and IL-4 were purchased from Peprotech. Transwell migration plates were purchased from COSTAR, USA (5 μm pore size). Goat anti-mouse β-...

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Abstract

The invention discloses a method for constructing a beta-defensin-2 tumor vaccine and the application thereof, wherein, the beta-defensin-2 is represented by MBD2. The method comprises the following steps: LPS is adopted to stimulate a mammalian skin in order to obtain a skin tissue expressing MBD2; an RT-PCR method is adopted to clone MBD2 matured gene fragment of the obtained skin cell, and an overlap PCR method is adopted to connect a mouse Igk signal peptide with the MBD2 matured fragment, so as to construct a MBD2 secretion expression vector pMBD2; and a lipofectamine method is adopted to separately transfect an empty vector and the pMBD2 into L1210 cells to obtain a stable expression cell line so as to construct a MBD2 tumor vaccine. The invention needs no identification of tumor-associated antigen, and is applicable to the majority of tumors, tumor-associated antigens of which is not determined, and provides a new strategy for the immunotherapy of leukemia.

Description

technical field [0001] The present invention relates to a method of tumor vaccine and its application, especially the method of constructing β-defensin 2 tumor vaccine and its application. Background technique [0002] Hematopoietic malignancy is a serious disease that endangers human beings with a high mortality rate. The current conventional treatment methods are chemotherapy and stem cell transplantation, but the specificity of chemotherapy drugs is low. While killing leukemia cells in the patient, it also kills normal blood cells and their progenitor cells, and inhibits the patient's hematopoietic and immune functions. Although stem cell transplantation can improve the prognosis of leukemia patients, only some patients are suitable for transplantation. Moreover, no matter chemotherapy or transplantation, only about 30%-50% of patients can achieve long-term disease-free survival. The recurrence and drug resistance of a considerable number of patients with residual leuke...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K35/36A61P35/02C12N15/09C12N15/63
Inventor 马小彤徐玢安莉莉林永敏吴克复
Owner INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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