Application of agrocybe aegerita galactose agglutinin in preparation of anti-HIV (Human Immunodeficiency Virus) infection medicament
A lectin and application technology, which is applied in the field of pharmacological application of poplar mushroom galectin to achieve the effects of inhibiting infection, improving human immunity and reducing viral load
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experiment example 1
[0025] Experimental Example 1 Activity test of fungal lectin AAL against HIV infection
[0026] Anti-HIV experiments include cytotoxicity experiments and anti-HIV replication experiments.
[0027] Cytotoxicity test (MTT assay): C8166 cells (Wuhan Institute of Virology, Chinese Academy of Sciences) were cultured in 96-well plates (3×10 4 / well), C8166 cells were treated with different concentrations (1.25nM, 6.25nM, 30nM, 150nM, 750nM) of fungal lectin AAL (SEQ ID No. 2), each concentration repeated 3 wells, 37°C, 5% CO 2 After 4 days of incubation, cell viability was detected by MTT assay. The concentration of the drug that causes a 50% reduction in cells is CC50.
[0028]HIV Replication Inhibition Assay: In parallel with the cytotoxicity assay, the HIV replication inhibition assay was detected by a p24 antigen capture ELISA assay. C8166 cells were inoculated with HIV-1 LAI (The virus was from the Institute of Virology, Chinese Academy of Sciences), the virus multiplicity ...
experiment example 2
[0032] Experimental Example 2 Experiment on the effect of fungal lectin AAL on virus replication
[0033] HIV-1 infection in C8166 cells LAI Fungal agglutinin AAL (SEQ ID No. 2) was added at different times (0 hours, 1 hour, 2 hours, 4 hours, 8 hours, 16 hours, 24 hours) after the virus, and virus replication was detected according to the method in Experimental Example 1. The multiplicity of infection (MOI) of the inoculated virus was 0.05, the final drug concentration was 6.25nM, and the culture system was 300ul. Azidothymidine (AZT) was used as a positive control. The experimental results see figure 2 . The experimental results showed that the inhibitory trend of fungal lectin AAL on the virus was similar to that of AZT, but at the same concentration, the inhibitory effect of fungal lectin AAL on HIV replication was better than that of AZT.
experiment example 3
[0034] Experimental Example 3 The inhibition experiment of fungal lectin AAL on the fusion of chronically infected cells and uninfected cells to form syncytia
[0035] To examine the inhibitory effect of the fungal lectin AAL (SEQ ID No.2) on the formation of syncytia between chronically infected C8166 cells and uninfected C8166 cells, the infected cells were treated with different concentrations of AAL (SEQ ID No.2) (0.2nM, 1nM, 5nM, 25nM, 125nM, 625nM) were incubated for 1 h, washed to remove excess AAL, and then uninfected cells were added to the culture system, wherein the ratio of infected cells to uninfected cells was 1:8. Cells were observed after co-culture for 36 h, and syncytia were counted.
[0036] The experimental results see image 3 . It can be seen from the experimental results that the fungal lectin AAL inhibits the formation of syncytia with an EC50 of 1.73 nM.
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