cgas-sting pathway activators and their uses
A pathway, cell technology, applied in the field of activators, which can solve the problem of unclear identification and clearance
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Embodiment 1
[0118] Example 1.Mn 2+ Treatment confers resistance to viral infection in host cells
[0119] Experiment (a)
[0120] with 100 μM MnCl 2THP1 cells cultured in RPMI-1640 medium (Gibco) supplemented with 10% FBS were pretreated for 12 hours, and then treated with HSV-1-GFP (obtained from Chinese Academy of Medical Sciences) at MOI of 0.5, VSV-GFP (obtained from Cells were infected from Rongbin zhou, University of Science and Technology of China) or NDV-GFP (obtained from Chen Wang, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences). After 24 hours, cells were photographed and measured using flow cytometry (FACSCalibur) and analyzed using FlowJo (eg figure 1 shown). figure 1 The use of Mn is clearly shown in 2+ Pretreatment confers resistance to viral infection in host cells.
[0121] As mentioned above, with figure 2 or image 3 MnCl at the indicated concentrations 2 THP1 cells were treated with either DMXAA (DMX) for 24 hours, then cells were inf...
Embodiment 2
[0128] Example 2. Intravenous injection of Mn 2+ confer virus resistance in mice
[0129] Experiment (a)
[0130] Take eight-week-old SPF grade C57BL / 6 (purchased from Beijing Weitong Lihua Co., Ltd.), mix liquid A and liquid B (as described above) according to the ratio in the table, and inject through tail vein. 16-24 hours later, herpes simplex virus type I (HSV-1), vesicular stomatitis virus (VSV) type of New Jersey and vaccinia virus (VacV) were injected, respectively. Virus titers in mice were detected using the plaque assay after 48 hours, and virus lethality was monitored.
[0131] Table 2. Intravenous injection of Mn 2+ Virus titers in mice
[0132] A liquid liquid B Mncl 2 Usage amount
HSV-1 titer VSV titer VacV titer 200μL 0μL 0mg / kg 198μL 2μL 1mg / kg 196μL 4μL 2mg / kg 190μL 10μL 5mg / kg drop hundreds of times 180μL 20μL 10mg / kg drop hundreds of times
[0133] Ta...
Embodiment 3
[0139] Example 3.Mn 2+ Effects of treatment on downstream gene expression
[0140] Experiment (a)
[0141] with 100 μM MnCl 2 THP1 cells were treated for 12 hours with , VACV, IFNβ or medium (control), RNA was extracted and sequenced. By calculating log 2 ((treated RPKM) / (control RPKM)) to obtain a heat map of gene expression (eg Image 6 shown in A).
[0142] Experiment (b)
[0143] with 200 μM MnCl 2 THP1 cells were treated and discarded after 2, 12, 24 hours containing MnCl 2 The medium and replaced with fresh medium, NW means that the MnCl-containing medium is not discarded 2 culture medium, Image 6 B shows the production of I-IFN.
[0144] Experiment (c)
[0145] With the indicated concentrations of MnCl 2 , Mn(OAc) 2 or Mn(OAc) 3 THP1 cells were treated for 24 hours, then supernatants were analyzed using an I-IFN bioassay, Image 6 C shows that treatment with different manganese salts induces I-IFN production, indicating that both divalent manganese salts...
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