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Herpes simplex virus and recombinant virus as well as host cell and medicinal combination thereof

A herpes simplex virus, host cell technology, applied in the direction of viral peptide, virus/phage, recombinant DNA technology, etc., can solve the problems of slow proliferation rate, low virus infection rate, etc., to achieve strong lysis ability and high specific infection rate , the effect of copying fast

Active Publication Date: 2008-07-30
北京奥源和力生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genomes of herpes simplex virus type I that infect different races have significantly different restriction endonuclease decomposition patterns, that is, there are restriction fragment length polymorphisms (restriction fragment length polymorphism, RFLP) (see "Quantitative analysis of genomic polymorphism of herpes simplex virus type 1 strains from six countries: studies of molecular evolution and molecular epidemiology of the virus", H. Sakaoka et al., Journal of General Virology, Vol 75, 513-527, 1994), herpes simplex virus 17+ strains from white Therefore, when herpes simplex virus type 17+ strains that infect white people are used for gene therapy on yellow people, they have the disadvantages of low virus infection rate and slow proliferation in host cells

Method used

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  • Herpes simplex virus and recombinant virus as well as host cell and medicinal combination thereof
  • Herpes simplex virus and recombinant virus as well as host cell and medicinal combination thereof
  • Herpes simplex virus and recombinant virus as well as host cell and medicinal combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0194] This example illustrates the isolation of herpes simplex virus according to the invention.

[0195] (1) Collection of virus

[0196] An adult Chinese male, 37 years old, Han nationality, with a history of multiple recurrent oral herpes, without other diseases, showed the typical clinical manifestations of recurrent infection of herpes simplex virus type I-herpetic vesicles around the corner of the mouth were sampled two days later.

[0197] Take a 15 ml sterile centrifuge tube, which contains 3 ml of sterile phosphate buffered saline (PBS) containing 100 μg / ml kanamycin, which contains 9 g / L of NaCl, 0.21 g / ml of liters of KH 2 PO 4 and 0.97 g / L of Na 2 HPO 4 .12H 2 O: Squeeze the herpetic vesicle with a sterile cotton swab until the light yellow secretion flows out, then dip the secretion with a cotton swab, and rinse the end of the obtained cotton swab with the secretion in the above-mentioned PBS buffer for 10 seconds. Take out the cotton swab, and use a steril...

Embodiment 2

[0205] This example demonstrates the lytic ability of the herpes simplex virus of the present invention in human nasopharyngeal carcinoma cells (CNE-1).

[0206] (1) Determination of herpes simplex virus titer

[0207] Using DMEM medium containing 10% fetal bovine serum, in six-well cell culture plate at 5 × 10 5 Inoculate Vero cells (Vero cells) at a density of 2 cells / well and store at 37°C in 5% CO 2 Cultivate in the incubator for 24 hours until each well of Vero cells is 80% full and ready for use.

[0208] Under sterile conditions, collect the supernatant culture solution of the Vero cell culture cultured with herpes simplex virus of the present invention in Example 1, and use serum-free DMEM medium to gradiently dilute to 1 × 10 respectively. 2 Double solution, 1×10 3 Double solution, 1×10 4 Double solution, 1×10 5 Double solution, 1×10 6 Double solution and 1×10 7 100 microliters of each dilution.

[0209] Under sterile conditions, discard the DMEM medium contai...

Embodiment 3

[0223] This example demonstrates the lytic ability of the herpes simplex virus of the present invention to normal tumor cells.

[0224] According to the method of Example 2, human breast cancer cells MCF-7, human colon cancer cells HT-29 and Human lung adenocarcinoma cell A549 (Second Research Institute of the Academy of Military Medical Sciences), and the survival number of tumor cells at different time points after transfection were tested, and the results are shown in Table 3.

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Abstract

The invention provides a herpes simplex virus which is separated from yellow race. The virus has high infection rate on the specificity of the host cells of yellow race, has high reproduction speed; so that can be directly used for infecting and dissolve the tumour cells of yellow race, and can also be used as a carrier to carry gene medicine into the cells of yellow race to conduct gene treatment.

Description

technical field [0001] The present invention relates to a virus, a recombinant virus using the virus as a carrier, a host cell including the virus and / or the recombinant virus, and a pharmaceutical composition including the virus and / or the recombinant virus. Specifically, the present invention relates to a herpes simplex virus, a recombinant virus using the virus as a vector, a host cell comprising the virus and / or the recombinant virus, and a host cell comprising the virus and / or the recombinant virus pharmaceutical composition. Background technique [0002] Herpes simplex virus (HSV) has a spherical shape, and the complete virus consists of a core, a capsid, a tegument and an envelope. The viral core contains double-stranded DNA, wound into a filamentous scroll. The capsid is icosahedral, with a diameter of about 100 nanometers and consists of 162 shell particles. The capsid is covered by a membrane of uneven thickness. The outermost layer of the virus is a typical li...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/869C07K14/08C12N15/38C12N15/63A61K39/245
Inventor 李小鹏李晓波
Owner 北京奥源和力生物技术有限公司
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