HIV-1 Mediated Inflammatory Response And Its Mechanism

BACKGROUND:Acquired immune deficiency syndrome(AIDS),also known as AIDS,was first discovered in the world in 1981.The main cause of this problem is that the human infected by the Human Immunodeficiency Virus(HIV-1),which is essentially a retrovirus that attacks the human immune system and damages many tissues and organs of the human body,leading to increased opportunistic infections and malignant tumors.With the promotion of cocktail therapy and antiretroviral treatment and the worldwide attention to AIDS and the investment of various medical resources,AIDS has become a controllable disease for patients.With the longevity of HIV-infected people,the number of HIV-infected people has also shown an increasing trend.The infected people are showing an aging trend.Only 20%of patients can continue to use drug treatment depending on their condition.Therefore,the infective and diseased have to face various complications,which leads to a serious decline in the quality of life of the patient.At present,no researchers have studied how HIV-1 causes ocular inflammatory lesions.After HIV-1 infecting the body,it induces cells to produce a large number of cytokines such as interferon and inflammatory factors.In the existing research articles,only HIV-1 trans-regulatory protein(Tat)can stimulate macrophage to produce neurotoxicity inflammatory factors(TNF-α,IL-1β).However,in our curreint study,we found that HIV-1,in addition to its own trans-regulatory protein(Tat)which can cause cellular inflammatory factors,the dsDNA produced by the reverse transcription of viral RNA during the replication of host cells by HIV-1 could cause the expression of intracellular inflammasome AIM2 which result in production of neurotoxic inflammatory factors.Hardly any researches have found the relationship between HIV-1 and imflammasome.In our research,we selected Muller cell which is the most important glial cells of the retina of and run through the whole layer of retina.Muller cells play a very important role in the retinal system and participate in the whole process of retinopathy.Our research revealed the role of-inflammatory bodies in the ocular complications of HIV-1 infection and elucidated the mechanisms of inflammation occurrence through vitro experiments.Objectives:1.Culture human retinal glial cells(Millercells)in vitro to establish a retinal glial cell model in vitro.2.Study the pathway of HIV-1 entry into human retinal glial cells and the molecular events involved.3.Elucidate the mechanism of HIV-1-induced retinal diseases.4.Provide new strategies for the search of potential drug treatment targets.Method:1.Muller cell surface receptor expression assay2.Packaging of different tropic clone viruses and detection of virus titer3.Detection of whether Muller cells produce HIV-1 progeny virus4.Western blotting was used to detect the expression changes of’AIM2,GSDMD and Caspase-1.5.Western blotting and confocal microscopy for detection of ASC oligomerization in Muller cells.6.Western blotting assay dected anti-HIV-1 drugs effect on the expression of AIM2.7.ELISA detection of LDH release in cell supernatant after HIV-1 infection of Muller cells8.Detection of changes in IL-β and IL-18 in cell supernatants after UV-inactivation and heat inactivation of HIV-1 infected Muller cells.RESULTS:1.Muller cells express CD4 receptor and CCR5r CXCR4 receptors.HIV-1 with different tropism can infect Muller cells and HIV-lcan produce progeny virus in Muller cells.2.The expression levels of inflammatory factors IL-1β,IL-6 and IL-8 mRNA were up-regulated after different tropic HIV-1 infected Muller cells,and the inflammatory factors in the supernatant also increased.3.Different tropic HIV-1 infections of Muller cells stimulate intracellular AIM2 expression.4.Different tropic HIV-1 infection of Muller cells can induce Caspase-1 cleavage in cells,and the specific inhibitor VX-765 of Caspase-1,it can inhibit the release of inflammatory factors IL-1β and IL-18.5.Different tropic HIV-1 infections of Muller cells can induce cleavage of GSDMD in Muller cells and release LDH in cell supernatants.6.Different tropic HIV-1 infections of Muller cells can induce oligomerization of intracellular ASC.7.Knockdown of ASC,GSDMD,Caspase-1,ASC can significantly reduce the release of factors caused by HIV-1 infection of Muller cells.8.Compared with other different mechanisms anti-HIV-1 drugs,antiretroviral drugs can irnhibit the production of AIM2 in Muller cells induced by HIV-1,and inhibit the production of neurotoxic inflammatory factors IL-1β and IL-18.CONCLUSION:When HIV-1 infects Muller cells,it causes a sharp increase in various inflammatory factors(such as IL-1β,IL-6,IL-18,etc.),and HIV-1 stimulates pattern recognition receptors in Muller cells(AIM2).The expression of AIM2 increased sharply.At the same time,the activation of AIM2 will recruit the cohesive protein ASC,At the same time,the ASC will be oligomerized.The subsequent recruitment of Caspase-1 and protein GSDMD will cause the two proteins to be cleaved after activation.Inhibition of IL-1β,IL-18 inflammatory factors into mature body secreting cells,based on the important role of Muller cells in the retinal system,inflammation caused by HIV-1 infection of Muller cells may be the cause of retinal damage,this topic can be HIV-1 provides new strategies for prevention and treatment of retinal complications.