Alcohol and Cannabinoids Differentially Affect HIV Infection and Function of Human Monocyte-Derived Dendritic Cells(MDDC)

During human immunodeficiency virus (HIV) infection, alcohol has been known to induce inflammation while cannabinoids have been shown to have an anti-inflammatory role. For instance cannabinoids have been shown to reduce susceptibility to HIV-1 infection and attenuate HIV replication in macrophages. Recently, we demonstrated that alcohol induces cannabinoid receptors and regulates cytokine production by monocyte-derived dendritic cells (MDDC). However, the ability of alcohol and cannabinoids to alter MDDC function during HIV infection has not been clearly elucidated yet. In order to study the potential impact of alcohol and cannabinoids on differentiated MDDC infected with HIV, monocytes were cultured for 7 days with GM-CSF and IL-4, differentiated MDDC were infected with HIV-1Ba-L and treated with EtOH (0.1 and 0.2%), THC (5 and 10 μM), or JWH-015 (5 and 10 μM) for 4-7 days. HIV infection of MDDC was confirmed by p24 and Long Terminal Repeats (LTR) estimation. MDDC endocytosis assay and cytokine array profiles were measured to investigate the effects of HIV and substances of abuse on MDDC function. Our results show the HIV + EtOH treated MDDC had the highest levels of p24 production and expression when compared with the HIV positive controls and the cannabinoid treated cells. Although both cannabinoids, THC and JWH-015 had lower levels of p24 production and expression, the HIV + JWH-015 treated MDDC had the lowest levels of p24 when compared to the HIV + THC treated cells. In addition, MDDC endocytic function and cytokine production were also differentially altered after alcohol and cannabinoid treatments. Our results show a differential effect of alcohol and cannabinoids, which may provide insights into the divergent inflammatory role of alcohol and cannabinoids to modulate MDDC function in the context of HIV infection

Withaferin A Suppresses Beta Amyloid in APP Expressing Cells: Studies for Tat and Cocaine Associated Neurological Dysfunctions

Neurological disorders are the biggest concern globally. Out of ~36 million human immunodeficiency virus (HIV) positive people, about 30%–60% exhibit neurological disorders, including dementia and Alzheimer’s disease (AD) like pathology. In AD or AD like neurological disorders, the pathogenesis is mainly due to the abnormal accumulation of extracellular amyloid beta (Aβ). In this era of antiretroviral therapy, the life span of the HIV-infected individuals has increased leading towards increased neurocognitive dysfunction in nearly 30% of HIV-infected individuals, specifically older people. Deposition of the Aβ plaques in the CNS is one the major phenomenon happening in aging HIV patients. ART suppresses the viral replication, but the neurotoxic protein (Tat) is still produced and results in increased levels of Aβ. Furthermore, drugs of abuse like cocaine (coc) is known to induce the HIV associated neurocognitive disorders as well as the Aβ secretion. To target the Tat and coc induced Aβ secretion, we propose a potent bifunctional molecule Withaferin A (WA) which may act as a neuro-protectant against Aβ neurotoxicity. In this study, we show that WA reduces secreted Aβ and induced neurotoxicity in amyloid precursor protein (APP)-plasmid transfected SH-SY5Y cells (SH-APP). In this study, we show that in SH-APP cells, Aβ secretion is induced in the presence of HIV-1 Tat (neurotoxic) and drug of abuse coc. Our fluorescent microscopy studies show the increased concentration of Aβ40 in Tat (50 ng/ml) and coc (0.1 μM) treated SH-APP cells as compared to control. Our dose optimization study show, lower concentrations (0.5–2 μM) of WA significantly reduce the Aβ40 levels, without inducing cytotoxicity in the SH-APP cells. Additionally, WA reduces the Tat and cocaine induced Aβ levels. Therefore, we propose that Aβ aggregation is induced by the presence of Tat and coc and WA is potent in reducing the secreted Aβ and induced neurotoxicity. Our study provides new opportunities for exploring the pathophysiology and targeting the neurological disorders

Anti-inflammatory effects of CBD in human microglial cell line infected with HIV-1

Abstract Human immunodeficiency virus (HIV) infection is associated with a chronic inflammatory stage and continuous activation of inflammasome pathway. We studied the anti-inflammatory effects of the compound cannabidiol (CBD) in comparison with Δ (9)-tetrahydrocannabinol [Δ(9)-THC] in human microglial cells (HC69.5) infected with HIV. Our results showed that CBD reduced the production of various inflammatory cytokines and chemokines such as MIF, SERPIN E1, IL-6, IL-8, GM-CSF, MCP-1, CXCL1, CXCL10, and IL-1 β compared to Δ(9)-THC treatment. In addition, CBD led to the deactivation of caspase 1, reduced NLRP3 gene expression which play a crucial role in the inflammasome cascade. Furthermore, CBD significantly reduced the expression of HIV. Our study demonstrated that CBD has anti-inflammatory properties and exhibits significant therapeutic potential against HIV-1 infections and neuroinflammation

Recent advances, status, and opportunities of magneto-electric nanocarriers for biomedical applications

Magneto-electric (ME) materials with core-shell architecture where the core is made of magnetic materials have emerged as an attractive nanomaterial due to the coupling of magnetic and electric properties in the same material and the fact that both fields can be controlled which allows an on-demand, transport and release of loaded cargo. Over the last decade, biomedical engineers and researchers from various interdisciplinary fields have successfully demonstrated promising properties ranging from therapeutic delivery to sensing, and neuromodulation using ME materials. In this review, we systematically summarize developments in various biomedical fields using the nanoforms of these materials. Herein, we also highlight various promising biomedical applications where the ME nanocarriers are encapsulated in other materials such as gels and liposomes and their potential for promising therapeutics and diagnostic applications

Cocaine and HIV synergistically down-regulate miR-155 and 20a.

<p>MDDCs (1×10⁶ cells/ml) from normal donors were cultured with 3 different concentrations (0.1, 1 and 10 µM) of cocaine for 15 and 24 hrs.Total cellular microRNA was extracted and amplified by specific RT-PCR for miR-155 and 20a gene expression. Cocaine down regulates <b>(A)</b> miR-155 <b>(B)</b> and 20a gene expression. (<b>C)</b> Cocaine treatment does not induce cytotoxicity. Cell suspension (200 µl) was subjected to XTT-PMS (100 µl) followed by a 4 hr incubation period. Cytotoxicity was measured by ELISA reader and cell suppression was calculated by: suppression index (%)  =  ((Abs cocaine – Abs blank)/(Abs control - Abs blank))×100%. (<b>D</b>) MDDCs (1×10 cells/ml) from normal donors were infected with HIV-1_BAL at a concentration of 20 ng for 2 hr, followed by treatment ± cocaine (1 µM) for 7 days. Total cellular microRNA was extracted and amplified by specific RT-PCR for miR-155 and 20a gene expression. Results shown are representative of mean ± SEM of three experiments. Graphs are represented as % of TAI compared to the non-treated control. p-value≤0.05 were considered to be significant.</p

HIV replication increased in MDDCs treated with cocaine.

<p><b>(A)</b> MDDCs (1×10⁶ cells/ml) from normal donors were infected with HIV-1_BAL at a concentration of 20 ng for 2 hr, followed by treatment ± cocaine (1 µM) for 7 days. The culture supernatants were quantitated for p24 Ag. Significance is calculated with respect to HIV-1 treated control cells. (<b>B</b>) Effect of cocaine on HIV-1 transcription was determined using an LTR-CAT reporter data normalized to total protein concentration. (<b>C</b>) HIV-1 transcription was determined in MDDCs from normal donors infected with HIV-luciferase viruses competent for a single round of infection (∼20 ng per (1×10⁶ cells) in the presence/absence of cocaine (1 µM) for 72 hours. Cultures were harvested 72 hours post infection and luciferase assayed. Results shown are representative of mean ± SEM of three experiments. p-value≤0.05 were considered to be significant.</p

miR-155 inhibition enhances HIV-1 replication.

<p>MDDCs (1×10⁶ cells/ml) from normal donors were transfected with anti-miR-155, at a final concentration of 75 nM, followed by infection with HIV-1 (20 ng) and treatment ± cocaine (1 µM). Total RNA was extracted and amplified by RT-PCR for HIV-1 LTR/RU-5 gene, to detect early steps in HIV reverse transcription. Significance is calculated with respect to non targeting HIV-1 control cells. Graphs are represented as % difference from control. Results shown are representative of mean ± SEM of three experiments. p-value≤0.05 were considered to be significant.</p

Overview of Cocaine-enhancement of HIV infectivity in MDC via the down-regulation of miR-155.

<p>(<b>A</b>) In absence of cocaine or HIV miR155 targets DC-SIGN and PU.1, reducing the expression post-transcriptionally. Lowered PU.1 and DC-SIGN expression is observed in mature DCs. (<b>B</b>) In the presence of Cocaine or HIV alone miR-155 levels are reduced resulting in elevated PU.1 and DC-SIGN expression. This phenotype is evident in immature DCs (iDCs). Together HIV and cocaine, synergize to dramatically lower miR-155 resulting in elevated levels of the transcription factor Pu.1. In turn, Pu.1 binds sites within the DC-SIGN promoter as well as the HIV LTR and promotes transcription. The resulting elevated DC-SIGN expression in DCs lead to maintenance an immature DC phenotype and thus increased HIV infectivity. By binding NF-κB sites within the LTR, PU.1 increases viral transcription and pathogenesis.</p

miR-155 modulates PU.1 levels.

<p>MDDCs (1×10⁶ cells/ml) from normal donors were transfected with anti-miR-155, at a final concentration of 75 nM, followed by infection with HIV-1 (20 ng) and treatment ± cocaine (1 µM). Total RNA was extracted and amplified by RT-PCR for PU.1 gene expression. Values were normalized against GAPDH and significance is calculated with respect to control. Graphs are represented as % difference from control. Results shown are representative of mean ± SEM of three experiments. p-value≤0.05 were considered to be significant.</p