The Reactivation Effects And Mechanisms Of Selective Class ? Histone Deacetylase Inhibitor CC-4a On Latent HIV-1

Background:Human Immunodeficiency Virus(HIV)is a double-stranded positive-strand RNA virus.AIDS(Acquired Immune Deficiency Syndrome,AIDS)caused by its infection is a highly harmful infectious disease.The main reason why HIV is difficult to be completely controlled or eliminated is that a latent HIV reservoir is formed in the patient's body.This mechanism prompts HIV to evade surveillance of the host immune system and the elimination of antiviral drugs.In 2012,the emergence of the "shock and kill" strategy raised new ideas for the treatment of AIDS and continues to this day.This strategy refers to inducing the replication and expression of latent viral genes through the latent infection activator(LRA),and then using anti-retroviral drugs or the autoimmune system to clear the progeny virus and its host cells to achieve the functional cure of AIDS.There are many types of latent activators,including histone deacetylase inhibitors(HDACi),which can inhibit histone deacetylase,increase the degree of acetylation of histones,and reduce the degree of entanglement of histones and DNA on the nucleosomes,which contains long terminal repeat(LTR)of HIV-1 genomes and facilitates the transcriptional expression of HIV genes.It is considered to be an effective latent activator of HIV-1.Objectives:This topic uses HIV-1 latently infected cell lines to detect whether the self-synthesized type I histone deacetylase inhibitor CC-4a has the activity to activate latent HIV-1,and further explore the combination of HDACi and antiviral drugs in the "shock" and kill "strategy and its mechanism of activating HIV-1,which provides a new drug reference for the elimination and functional cure of HIV-1.Methods:1.Flow cytometry,ELISA and RT-PCR tests were conducted to detect the activation effect of compound CC-4a on HIV-1 latent cell line and the combined effect of CC-4a and other latent reversing agents.2.MTS method was used to detect the toxicity of CC-4a to experimental cells.3.The Luciferase assay was operated to explore the IC50 value of antiviral drugs for different HIV-1 isolates after CC-4a and antiviral drugs are used together.4.Western blot experiments were carried out to explore the mechanisms of CC-4a activation of latent HIV-1.Results:1.CC-4a could promote HIV-1 RNA and protein expression in J-Lat 10.6 and ACH2 cells in a concentration-and time-dependent manner.2.After 48 hours of CC-4a treatment of the experimental cells,the cell viability remained above 90%.In addition,compared with cells that are riot infected with HIV-1,CC-4a could selectively promote the enhanced cleavage of Caspase 3 and PARP proteins in cells infected with HIV-1.3.After CC-4a treatment of human peripheral blood mononuclear cells(PBMCs),the expression of T cell activation surface markers CD25 and CD69 did not increase significantly,and the number of cells co-expressing HIV-1 receptor CD4 and co-receptor CCR5 decreased.4.After CC-4a and Prostratin co-treated J-Lat 10.6 and ACH2 cells,the effect of activating HIV-1 was significantly different from that of the single-use group.5.CC-4a did not change the antiviral activity of antiviral drugs,and antiviral drugs would not affect the activation effect of CC-4a.6.After CC-4a treated the two HIV-1 latent cell models,the level of intracellular histone acetylation increased;intracellular p65 protein phosphorylation increased,nuclear accumulation increased,and the expression of IkBa protein decreased;PDTC could inhibit CC-4a activation effect.Conclusion:1.CC-4a can effectively activate latent HIV-1.2.CC-4a has low cytotoxicity to experimental cells and can induce apoptosis of cells infected with HIV-1.3.CC-4a will not cause extensive T cell activation and high expression of inflammatory factors,and can reduce the expression of HIV-1 receptor CD4 and co-receptor CCR5.4.CC-4a has a synergistic effect with the classic latent reversing agen Prostratin,and reduces T cell activation caused by Prostratin.5.CC-4a and cART drugs do not affect each other's pharmacological effects.6.CC-4a significantly increases histone acetylation,thereby promoting HIV gene transcription;the classic NF-?B pathway is also involved in CC-4a-induced HIV latent activation.