Human interferon alpha-2b recombinant bacillus calmette-guerin, construction method and identification method thereof

Abstract

The invention relates to a recombinant BCG vaccine rBCG-IFN alpha-2b for secreting human IFN alpha-2b and a construction method and an identification method thereof, wherein a BCG vaccine Ag85B signal peptide fragment which has the function of secretion and genes of human IFN alpha-2b are cloned to pMV261 by the genetic engineering technology, and a BCG vaccine shuttle expression vector pMV261-Ag85B-IFN alpha-2b is obtained; and the vector is induced into BCG by the electrotransformation technology, and the recombinant BCG vaccine rBCG-IFN alpha-2b is established; and the human IFN alpha-2b can be highly efficiently secreted in virtue of the secretion function of the pMV261-Ag85B-IFN alpha-2b on BCG replication and signal peptide. The recombinant BCG vaccine rBCG-IFN alpha-2b obtained not only keeps the immunogenicity of the prior BCG but also can continuously secrete the cell factor-the IFN alpha-2b, thereby improving the immunocompetence of the BCG; the IFN alpha-2b can be directly acted on tumor cells to inhibit proliferation and induced differentiation of the tumor cells, has good antitumor action, and can reduce the application dosage and reduce the toxic and side effect caused by the BCG; and the IFN alpha-2b solves the problems of large application dosage, high incidence rate of side effects, short response time and expensive cost caused by exogenous IFN alpha-2b.

Description

Technical field: [0001] The invention relates to an anti-bladder cancer drug in clinical medicine, in particular to a recombinant BCG vaccine of human interferon alpha-2b, and a construction method and identification method thereof. Background technique: [0002] Transitional epithelial carcinoma of the bladder is the most common malignant tumor of the urinary system in my country, and it is easy to recur and progress after surgery. Therefore, mitomycin, doxorubicin, thiotepa, Bacille Calmette-Guerin (BCG) and other drugs should be regularly used for bladder perfusion to prevent tumor recurrence. Among them, BCG is currently considered the most effective drug. However, clinically, about 30% of patients still have unsatisfactory BCG treatment and prevention effects, especially for invasive bladder cancer. In addition, BCG bladder perfusion has a higher incidence of side effects. In recent years, many studies have applied low-dose BCG combined with human interferon α-2b (IFNα-2b) b...

AI Technical Summary

Problems solved by technology

However, clinically, there are still about 30% of patients with unsatisfactory BCG treatment and prevention effects, especially for invasive bladder cancer.

In addition, BCG intravesical infusion has a high incidence of toxic side effects

In recent years, many studies have used low-dose BCG combined with intravesical infusion of human interferon α-2b (IFNα-2b) to improve anticancer efficacy and reduce side effects, but the long-term efficacy of combined application is not good, and exogenous added IFNα-2b has the problems of large application dose, high incidence of side effects, short duration of action, and high cost

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