Application of Lambda interferon in resisting human immunodeficiency virus

Abstract

The invention discloses an application of Lambda interferon in resisting human immunodeficiency virus. IFN-Lambda resisting HIV infection can inhibit the infection and replication of HIV-1 in human macrophage, which is presented by the following processes: (1) IFN-Lambda1 / Lambda2 differentiates mature human macrophage through pretreatment, then HIV-1R5 type strain respectively infects the cells, a DMEM culture medium is used for washing and then a DMEM culture medium containing 10 percent of fetal calf serum is added for culture; and (2) after infection, the formed multinucleate giant cells of IFN-Lambda treatment group are rather less than those of a virus contrast group, and the HIV-1 reverse transcriptase and envelope protein P24 in culture supernatant fluid of the IFN-Lambda treatment group are detected to be rather less than that of the virus contrast group. After infection, the HIV-1RT in the culture supernatant fluid is respectively detected. The effect of inhibiting the HIV-1 is obvious after infection. The IFN-Lambda treatment group has obvious time and dose-effect relationship in inhibiting the activity of HIV-1RT after infection. The function of IFN-Lambda in resisting the HIV-1 is similar to that of other interferons and has the antivirus effect. The IFN-Lambda shows obvious effect of inhibiting the replication of virus as regards to the human macrophage before, in and after the infection of the HIV-1 virus.

Description

technical field

[0001] The invention belongs to the field of antiviral drugs, in particular, the invention relates to the application of a new type of interferon-λ interferon (interferon lambda, IFN-λ) in anti-human immunodeficiency virus (HIV-1). Background technique

[0002] Human immunodeficiency virus (HIV) is the cause of acquired immunodeficiency syndrome (AIDS). Patients are characterized by CD4+T cell reduction, accompanied by repeated opportunistic infections, malignant tumors, and degeneration of the central nervous system. During the twenty-seven years since the HIV / AIDS epidemic broke out, about 65 million people worldwide were infected with HIV, and 25 million of them died. If nothing is done to combat the HIV / AIDS epidemic, it will turn into a historic catastrophe with an estimated millions of new infections by the end of this decade.

[0003] Because HIV mainly invades CD4+ T cells and mononuclear phagocytes, and relies on a variety of active molecules in inf...

Examples

Embodiment 1

[0021] Example 1: Toxic effect of IFN-λ on human macrophages

[0022] Differentiated mature human macrophages were added with different concentrations of IFN-λ1 / λ2 and cultured for 72 hours. The cytotoxic effect of IFN-λ1 / λ2 was detected by MTS method. The results showed that the concentration of IFN-λ1 / λ2 ranged from 1ng / ml to 1000ng / ml is less than 10% for human macrophage toxicity, and there is no significant difference in statistical analysis, so it can be considered that IFN-λ1 / λ2 has no toxicity to human macrophages ( figure 1) .

Embodiment 2

[0023] Embodiment 2: IFN-λ anti-human immunodeficiency virus (HIV-1) infection in vitro

[0024] IFN-λ1 / λ2 (100ng / ml) pretreated differentiated mature human macrophages for 24 hours, HIV-1R5 strains Bal, Jago and JRFL (P24 protein content were all 30ng / 10 6 cells) were infected for 2 hours, washed three times with DMEM medium, then added DMEM (V / V) medium containing 10% fetal bovine serum, 37 ° C, 5% CO 2 (V / V) culture; on the 8th day after infection, observed under the light microscope, the macrophages in the HIV-1 infection group presented a typical multinucleated giant cell morphology, and the multinucleated giant cells in the IFN-λ treatment group were significantly less than the virus control group ( figure 2) .

[0025] Cell supernatants were collected on day 8 post-infection and isotopic 32 P-labeled HIV reverse transcriptase method and ELISA method were used to detect HIV-1 viral reverse transcriptase (RT) and P24 protein. The content of P24 was significantly lower...

Embodiment 3

[0026] Example 3: IFN-λ anti-HIV-1 infection in vitro has a time effect:

[0027] IFN-λ1 / λ2 (100ng / ml) pretreated differentiated mature human macrophages for 24 hours, infected with HIV-1 Bal strain for 2 hours, washed three times with DMEM medium, and then added DMEM containing 10% fetal bovine serum (V / V) Medium, 37°C, 5% CO 2 (V / V) culture; cell supernatants were collected on the 4th day, 8th day and 12th day of infection respectively, and isotope was applied 32 P marker HIV reverse transcriptase method to detect RT. The results showed that on the 8th day after infection, the effect of inhibiting HIV-1 infection was obvious ( Figure 4, P Figure 4, P<0.05). It shows that IFN-λ inhibits the replication and infection of HIV-1 in human macrophages and presents a time relationship.

technical field

[0001] The invention belongs to the field of antiviral drugs, in particular, the invention relates to the application of a new type of interferon-λ interferon (interferon lambda, IFN-λ) in anti-human immunodeficiency virus (HIV-1). Background technique

[0002] Human immunodeficiency virus (HIV) is the cause of acquired immunodeficiency syndrome (AIDS). Patients are characterized by CD4+T cell reduction, accompanied by repeated opportunistic infections, malignant tumors, and degeneration of the central nervous system. During the twenty-seven years since the HIV / AIDS epidemic broke out, about 65 million people worldwide were infected with HIV, and 25 million of them died. If nothing is done to combat the HIV / AIDS epidemic, it will turn into a historic catastrophe with an estimated millions of new infections by the end of this decade.

[0003] Because HIV mainly invades CD4+ T cells and mononuclear phagocytes, and relies on a variety of active molecules in inf...

Examples

Embodiment 1

[0021] Example 1: Toxic effect of IFN-λ on human macrophages

[0022] Differentiated mature human macrophages were added with different concentrations of IFN-λ1 / λ2 and cultured for 72 hours. The cytotoxic effect of IFN-λ1 / λ2 was detected by MTS method. The results showed that the concentration of IFN-λ1 / λ2 ranged from 1ng / ml to 1000ng / ml is less than 10% for human macrophage toxicity, and there is no significant difference in statistical analysis, so it can be considered that IFN-λ1 / λ2 has no toxicity to human macrophages ( figure 1) .

Embodiment 2

[0023] Embodiment 2: IFN-λ anti-human immunodeficiency virus (HIV-1) infection in vitro

[0024] IFN-λ1 / λ2 (100ng / ml) pretreated differentiated mature human macrophages for 24 hours, HIV-1R5 strains Bal, Jago and JRFL (P24 protein content were all 30ng / 10 6 cells) were infected for 2 hours, washed three times with DMEM medium, then added DMEM (V / V) medium containing 10% fetal bovine serum, 37 ° C, 5% CO 2 (V / V) culture; on the 8th day after infection, observed under the light microscope, the macrophages in the HIV-1 infection group presented a typical multinucleated giant cell morphology, and the multinucleated giant cells in the IFN-λ treatment group were significantly less than the virus control group ( figure 2) .

[0025] Cell supernatants were collected on day 8 post-infection and isotopic 32 P-labeled HIV reverse transcriptase method and ELISA method were used to detect HIV-1 viral reverse transcriptase (RT) and P24 protein. The content of P24 was significantly lower...

Embodiment 3

[0026] Example 3: IFN-λ anti-HIV-1 infection in vitro has a time effect:

[0027] IFN-λ1 / λ2 (100ng / ml) pretreated differentiated mature human macrophages for 24 hours, infected with HIV-1 Bal strain for 2 hours, washed three times with DMEM medium, and then added DMEM containing 10% fetal bovine serum (V / V) Medium, 37°C, 5% CO 2 (V / V) culture; cell supernatants were collected on the 4th day, 8th day and 12th day of infection respectively, and isotope was applied 32 P marker HIV reverse transcriptase method to detect RT. The results showed that on the 8th day after infection, the effect of inhibiting HIV-1 infection was obvious ( Figure 4, P Figure 4, P<0.05). It shows that IFN-λ inhibits the replication and infection of HIV-1 in human macrophages and presents a time relationship.

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