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Reproducible kidney cancer therapeutic DNA vaccine

A DNA vaccine and replication-type technology, applied in gene therapy, pharmaceutical formulations, anti-tumor drugs, etc., can solve problems such as reproducible DNA vaccines that have not yet appeared, achieve enhanced induction of efficient specific cellular immunity and humoral immunity, and improve Effects of immune potency and large storage space

Inactive Publication Date: 2014-07-30
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the replicable DNA vaccine has a very broad prospect in the development and production of vaccines, and there is no replicable DNA vaccine available for the treatment of kidney cancer.

Method used

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  • Reproducible kidney cancer therapeutic DNA vaccine
  • Reproducible kidney cancer therapeutic DNA vaccine
  • Reproducible kidney cancer therapeutic DNA vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1. Construction and expression of replicable renal cancer therapeutic DNA vaccine pSVK-G250FGB

[0059] 1. Construction of a replicable DNA vaccine vector

[0060] 1. Construction of the recombinant vector pUC19-PS-KANA carrying PS and KANA fusion gene (PS-KANA)

[0061] The construction method of the recombinant vector pUC19-PS-KANA is as follows:

[0062] 1) Amplify the PS gene: use the replicon DNA vaccine vector pSCA1 containing the ampicillin resistance gene (see the literature Yu YZ, Sun ZW, Yu W Y. Chinese Journal of Biotechnology, 2005, 21(5): 33- for the construction method 38) as template, in primer PSCA-F

[0063] (5’-CCGTCTAGAGATCATAATCAGCCAT-3’) and PSCA-R

[0064] (5'-CCGGCATGCCTCGAGACTAGTCTGTCAGACCAAG-3') guide PCR amplification of the flanking sequence of the ampicillin resistance gene, the 5'flanking sequence contains the restriction endonuclease Xba I recognition site, the 3'end The flanking sequence contains the restriction enzyme Sph I recognition sit...

Embodiment 2

[0122] Example 2. Construction of a renca mouse cell line stably expressing human G250 and a human G250 mouse kidney cancer tumor-bearing model

[0123] Because it is necessary to stably express human G250 mouse kidney cancer model in the research process of kidney cancer vaccine or immunotherapy targeting G250, the research work of this part of the experiment is to establish a renca mouse cell line stably expressing human G250 and human G250 mouse kidney cancer tumor-bearing model. The human G250 gene was amplified by PCR, and inserted into the pIRES-neo eukaryotic expression vector according to the DNA recombination technology to obtain the recombinant expression plasmid pIRES-neo-G250. The plasmid was stably transfected into renca mouse cells by cationic liposome-mediated method. Positive clones were selected by adjusting the concentration of G418. Western blotting and immunofluorescence tests verified that the human G250 gene was transfected into mouse renca cell lines. Usin...

Embodiment 3

[0178] Example 3. Detection of tumor suppressor activity and immunological mechanism of the replicable renal cancer therapeutic DNA vaccine pSVK-G250FGB

[0179] In Example 3, Figure 13-22 The plasmid used for the immunization of mice in the middle E group is marked as pSVK-sig-tG250-Fc-GPI-IRES-GM / B7 and the replicable renal cancer therapeutic DNA vaccine pSVK-G250FGB is the same plasmid, but the name is simple and complicated. The differences are hereby explained.

[0180] The renal cancer therapeutic DNA vaccine pSVK-G250FGB constructed in Example 1 was delivered by intramuscular injection plus electroporation to immunize the tumor-bearing mice obtained in Example 2. By observing and recording the tumor growth of mice in each group, the tumor suppressor activity of the replicable DNA vaccine pSVK-G250FGB was studied. The ELISA method was used to detect the specific antibody titers in the serum of immunized mice, the Elispot method was used to detect the activation of lymphocy...

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Abstract

The invention discloses a reproducible kidney cancer therapeutic DNA vaccine. The reproducible kidney cancer therapeutic DNA vaccine is obtained by inserting a compound gene sig-tG250-Fc-GPI-IRES-GM / B7 into a reproducible DNA vaccine carrier pSVK. The reproducible kidney cancer therapeutic DNA vaccine is a DNA vaccine capable of inducing self anti-tumor immune response, has good safety and broad application prospect and provides a new method and thinking for prevention and treatment of malignant tumors.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a replicable renal cancer therapeutic DNA vaccine. Background technique [0002] Renal cell carcinoma (RCC) is one of the most malignant tumors of the kidney and a common tumor in the urinary system. The incidence of renal cell carcinoma has increased year after year in the past 30 years (Rohrmann K, Staehler M, Haseke N, et al. Immunotherapy in metastatic renal cell carcinoma. World J Urol (2005) 23: 196-201). When the patient is diagnosed, it has reached an advanced stage. At the same time, 20%-30% of renal cancers after radical resection will have metastasis. There is no satisfactory treatment for RCC that has lost the opportunity of surgery (Schrader AJ, Varga Z , Hegele A, et al. Second-line strategies for metastatic renal cell carcinoma: classics and novel approaches. J Cancer Res Clin Oncol (2006) 132:137–149). Therefore, the treatment of advanced kidney cancer urgentl...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/395A61P35/00
Inventor 于继云阎瑾琦田仁礼李云奇肖毅张巍张亮王宇刘宁
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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