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Application of USP11 in inhibition of degradation of cytokine IL6

A technology for inhibiting cells and factors, applied in the application, introduction of foreign genetic material and modified cells, genetic engineering, etc., can solve problems such as abnormal expression and no IL6 degradation

Active Publication Date: 2022-07-12
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

USP11 is a gene involved in the ubiquitination process of the body. Studies have found that it is abnormally expressed in a variety of tumors, and there is no research on its involvement in IL6 degradation.

Method used

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  • Application of USP11 in inhibition of degradation of cytokine IL6
  • Application of USP11 in inhibition of degradation of cytokine IL6
  • Application of USP11 in inhibition of degradation of cytokine IL6

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Mice were divided into groups and the sepsis model was induced by intraperitoneal injection of LPS. Experimental mice were grouped: 10 WT mice (USP11 wild type) and 10 KO mice (USP11 knockout), respectively, received intraperitoneal injection of LPS at a dose of 5 mg / kg.

[0027] 1. Take USP11 knockout C57 mice as experimental group and control group as USP11 wild-type C57 mice, and inject 5mg / kg LPS subcutaneously.

[0028] 2. The mouse serum was taken at a specific time point, and the IL-6 content was detected by ELISA.

[0029] 3. After USP11 gene knockout, the content of IL-6 in the blood of mice was significantly lower than that of non-knockout mice after 4 hours of LPS stimulation. Note that USP11 effectively reduces the degradation rate of IL-6, see figure 1 .

Embodiment 2

[0031] 293 cells were transfected with recombinant adenovirus Ad-USP11. The 293 cells were observed under the microscope after the cells were confluent to 70%, then the cells were digested and centrifuged. Divided into the experimental group (Ad-USP11) and the control group (Nc-USP11), discard the supernatant, add virus diluent and cell culture medium to each well at one time, mix well, and replace the normal medium after culturing for 24 hours. cell state. After 48 h, cells were digested with trypsin, centrifuged, and the cells were resuspended to observe the state for subsequent experiments. Take the above cultured cells, add LPS (1ug / ml) to stimulate 293 cells to produce IL-6.

Embodiment 3

[0033] 293 cells were transfected with lentiviral sh-USP11. When the confluence of 293 cells was 70%, the cells were slid down with trypsin, counted, and the cell concentration was adjusted. Resuspend cells in medium and add lentiviral reagent dropwise. The experimental group is (sh-USP11) and the control group is (sh-NC). Lentivirus dosage: virus volume=(MOI×cell number) / virus titer. After 12 h, the medium was replaced with normal medium, and the culture was continued to observe the state of the cells. 72 hours after infection, the infection efficiency was detected by qRT-PCR for subsequent experiments. Take the above cultured cells, add LPS (1ug / ml) to stimulate 293 cells to produce IL-6.

[0034] The changes in the expression level of IL-6 after LPS (1ug / ml) treatment of the cells of Example 2 and Example 3 were observed, and the correlation between USP11 and IL-6 was verified at the cellular level. The 293 cells overexpressing USP11 and the control group were stimulat...

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Abstract

IL6 is an important cytokine and plays an important role in the processes of infection resistance, tumor immunity, immune cell development and the like, so that maintenance of a certain level of IL6 is very important in the processes of preventing the IL6 from being degraded too early. The USP11 is a gene participating in the ubiquitination process of an organism, and the research finds that the USP11 is abnormally expressed in various tumors and does not participate in the research of IL6 degradation. The invention discloses a new function of USP11, namely an effect of protecting IL6 from degradation, and provides a new target spot and idea for anti-infection and tumor treatment.

Description

technical field [0001] The invention belongs to biotechnology, and specifically discloses the application of USP11 in inhibiting the degradation of cytokine IL6. Background technique [0002] IL6 is an important cytokine that plays an important role in anti-infection, tumor immunity, immune cell development, etc. Therefore, maintaining a certain level of IL6 to prevent premature degradation is very important in the above process. . USP11 is a gene involved in the body's ubiquitination process. Studies have found that it is abnormally expressed in a variety of tumors, and there is no research on its involvement in IL6 degradation. SUMMARY OF THE INVENTION [0003] There are few studies on the degradation of IL6 in the prior art. The present invention discloses a new function of USP11-protecting IL6 from degradation, and provides new targets and ideas for anti-infection and tumor treatment. [0004] The present invention adopts following technical scheme: [0005] Use of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57C12N15/861C12N15/867C12N5/10A61K38/48A61K45/00A61P35/00A61P37/04
CPCC12N9/485C12Y304/19012C12N15/86C12N5/0686A61K38/4813A61K45/00A61P35/00A61P37/04C12N2710/10043C12N2740/15043C12N2510/00Y02A50/30
Inventor 祁小飞
Owner SUZHOU UNIV
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