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Application of TMED2 as treatment target spot of Ebola viral disease

An Ebola virus and target technology, applied in the field of biomedicine, can solve the problems of lack of vaccination options for treatment measures, increase of natural or man-made, infection of Ebola virus, etc.

Active Publication Date: 2020-06-02
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Ebola virus infection has geographical limitations, with the expansion of human activities and the increasing frequency of cross-border exchanges, the probability of natural or artificial Ebola virus infection has been greatly increased, coupled with the lack of treatment measures and vaccination options, making Ebola Bora virus emerges as deadly biothreat pathogen

Method used

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  • Application of TMED2 as treatment target spot of Ebola viral disease
  • Application of TMED2 as treatment target spot of Ebola viral disease
  • Application of TMED2 as treatment target spot of Ebola viral disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1, immunoprecipitation and immunoblotting detection of the interaction between VP35 and TMED2

[0076] Co-transfect pcDNA3.0-Flag-VP35 and pEGFP-TMED2 plasmids in HEK293 cells, passage the cells into cell culture dishes as needed, and transfer the culture dishes into CO 2 Cultivate in an incubator, and transfect when the inoculated cells grow to 70-90% confluence, using Lipofectamine from Thermo Company TM 3000 transfection reagent, carry out plasmid transfection according to the manual: mix 1 μg pcDNA3.0-Flag-VP35 plasmid and 1 μg pEGFP-TMED2 plasmid with 4 μl P3000 at the ratio of plasmid:P3000=1:2 TM Add 100 μl Opti-MEM TM Dilute in culture medium to prepare DNA premix; mix 6 μl Lipofectamine according to plasmid: Lipo3000=1:3 TM Add 3000 to 100 μl Opti-MEM TM Diluted in the culture medium, in the diluted Lipofectamine TM Add the diluted DNA premix solution to the 3000 reagent, mix well and incubate at room temperature for 10-15 minutes, then add the DNA-...

Embodiment 2

[0080] Example 2, live cell imaging detection of co-localization of VP35 and TMED2

[0081] Inoculate HepG cells into the chamber cover glass culture system one day in advance, and when the cells grow to a suitable density, transfect pcDNA3.0-mcherry-VP35 plasmid and pEGFP-TMED2 plasmid (for the operation method, see cell transfection), transfect After 24 hours of transfection, the chamber coverslip culture system was placed under a live cell imager to observe and take pictures.

[0082] The result is as figure 2 As shown, live-cell imaging technique found that VP35 co-localized with TMED2 in the cytoplasm.

Embodiment 3

[0083] Example 3. Immunofluorescent staining detection of co-localization of VP35 and TMED2 in inclusion bodies

[0084] Inoculate HepG cells into a 6-well plate containing sterile coverslips one day in advance, and when the cells grow to a suitable density, transfect Ebola virus minimal genome plasmids: pCAGGS-NP (125ng), pCAGGS-VP35 (125ng) , pCAGGS-VP30 (75ng), pCAGGS-L (1000ng), p4cis-vRNA-Rluc (250ng) and pCAGGS-T7 (250ng) (see cell transfection for the operation method), after 48 hours of transfection, suck out the medium in the well , wash the cells 3 times with PBS at room temperature, and absorb the residual PBS for the last time; add 2ml 4% paraformaldehyde to each well and fix it at room temperature for 20-30min, absorb the paraformaldehyde, and wash the cells 3 times with PBS; add 2ml 0.3% Triton X-100 (prepared with 1×PBS) was perforated at room temperature for 15-20min, the Triton X-100 was aspirated, and the cells were washed 3 times with PBS; 2ml of blocking so...

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PUM

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Abstract

The invention discloses application of TMED2 as a treatment target spot of an Ebola viral disease. The invention provides the application of the TMED2 as a target spot in any of the following: treatment of Ebola viral infection, treatment of the Ebola viral disease, inhibition of replication of Ebola virus, inhibition of propagation of the Ebola virus in cells and inhibition of formation of Ebolavirus inclusion bodies. The TMED2 can serve as a target spot; a TMED2 suppressant can serve as a candidate drug for preventing and treating the Ebola viral disease and is applied to the treatment of the Ebola viral infection. The TMED2 has a great application value in the treatment of the Ebola viral infection.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of TMED2 as a therapeutic target for Ebola virus disease. Background technique [0002] Ebola virus disease (formerly Ebola hemorrhagic fever) is a severe and often fatal disease caused by Ebola virus infection, with a case fatality rate ranging from 25% to 90%. Ebola virus is considered to be the prototype pathogen of viral hemorrhagic fever. The virus was transmitted to humans through wild animals and spread among humans. The Ebola outbreak in West Africa from 2014 to 2016 was the largest and most complex outbreak ever. In 2018, the Ebola outbreak broke out again in Congo. As of November 26, 2019, a total of 3,304 cases of Ebola had been reported. Bora cases, including 3186 confirmed cases and 118 probable cases, of which 2199 died (overall case fatality rate about 67%). Although Ebola virus infection has geographical limitations, with the expansion of human activit...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/7105A61K48/00A61P31/14
CPCA61K45/00A61K31/7105A61K48/005A61P31/14
Inventor 曹诚杨伟洪高婷靳彦文朱林刘曜宁
Owner ACADEMY OF MILITARY MEDICAL SCI
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