Wildfire-induced pollution and its short-term impact on COVID-19 cases and mortality in California.

Globally, wildfires have seen remarkable increase in duration and size and have become a health hazard. In addition to vegetation and habitat destruction, rapid release of smoke, dust and gaseous pollutants in the atmosphere contributes to its short and long-term detrimental effects. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has emerged as a public health concern worldwide that primarily target lungs and respiratory tract, akin to air pollutants. Studies from our lab and others have demonstrated association between air pollution and COVID-19 infection and mortality rates. However, current knowledge on the impact of wildfire-mediated sudden outburst of air pollutants on COVID-19 is limited. In this study, we examined the association of air pollutants and COVID-19 during wildfires burned during August-October 2020 in California, United States. We observed an increase in the tropospheric pollutants including aerosols (particulate matter [PM]), carbon monoxide (CO) and nitrogen dioxide (NO2) by approximately 150%, 100% and 20%, respectively, in 2020 compared to the 2019. Except ozone (O3), similar proportion of increment was noticed during the peak wildfire period (August 16 - September 15, 2020) in the ground PM2.5, CO, and NO2 levels at Fresno, Los Angeles, Sacramento, San Diego and San Francisco, cities with largest active wildfire area. We identified three different spikes in the concentrations of PM2.5, and CO for the cities examined clearly suggesting wildfire-induced surge in air pollution. Fresno and Sacramento showed increment in the ground PM2.5, CO and NO2 levels, while San Diego recorded highest change rate in NO2 levels. Interestingly, we observed a similar pattern of higher COVID-19 cases and mortalities in the cities with adverse air pollution caused by wildfires. These findings provide a logical rationale to strategize public health policies for future impact of COVID-19 on humans residing in geographic locations susceptible to sudden increase in local air pollution.

microRNAs: Emerging players in oral cancers and inflammatory disorders.

Association of oral diseases and disorders with altered microRNA profiles is firmly recognized. These evidences support the potential use of microRNAs as therapeutic tools for diagnosis, prognosis, and treatment of various diseases. In this review, we highlight the association of altered microRNA signatures in oral cancers and oral inflammatory diseases. Advances in our ability to detect microRNAs in human sera and saliva further highlight their clinical value as potential biomarkers. We have discussed key mechanisms underlying microRNA dysregulation in pathological conditions. The use of microRNAs in diagnostics and their potential therapeutic value in the treatment of oral diseases are reviewed.

miR-24, miR-30b, and miR-142-3p regulate phagocytosis in myeloid inflammatory cells.

Micro-RNAs (miRNAs) are small noncoding RNAs that regulate various biological pathways. As their role in phagocytosis remains poorly understood, we investigated their impact on phagocytosis in myeloid inflammatory cells. Seven miRNAs (miR-24, -30b, -101, 142-3p, -652-3p, -652-5p, and -1275) that were differentially expressed during monocyte to macrophage (Mφ) and monocyte to dendritic cell (DC) differentiation were screened for their potential role in phagocytosis. Among these, overexpression of miR-24, miR-30b, and miR-142-3p in human monocyte-derived Mφ, DC, monocytes, and PBMCs significantly attenuate phagocytosis of Escherichia coli and Staphylococcus aureus, as well as the secretion of inflammatory mediators, including TNF-α, IL-6, and IL-12p40. miRNA-mediated changes in cytokine profiles were observed at transcriptional and/or posttranscriptional levels and importantly exhibit miRNA-specific impact. To examine the underlying mechanism, we monitored the expression of phagocytosis pathway-associated genes and identified several genes that were altered in Mφ and DC transfected with miR-24, miR-30b, and miR-142-3p mimics. Some of these genes with altered expression also harbor putative miRNA binding sites. We show that miR-142-3p directly regulates protein kinase Cα (PKCα), a key gene involved in phagocytosis. Interestingly, miR-142-3p and PKCα exhibit antagonistic expression during Mφ and DC differentiation. Short interfering RNA-mediated knockdown of PKCα in Mφ leads to reduced bacterial uptake, further highlighting the role of the gene in phagocytosis. Overall, these results demonstrate that miR-24, miR-30b, and miR-142-3p regulate phagocytosis and associated cytokine production in myeloid inflammatory cells through modulation of various genes involved in the pathway.

MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines.

MicroRNAs are 18-22 nucleotides long, non-coding RNAs that bind transcripts with complementary sequences leading to either mRNA degradation or translational suppression. However, the inherent differences in preferred mode of miRNA regulation among cells of different origin have not been examined. In our previous transcriptome profiling studies, we observed that post-transcriptional regulation can differ substantially depending on the cell in context. Here we examined mechanistic differences in the regulation of a let-7a targeted (wild type) or resistant (mutant) engineered renilla transcript across various mammalian cell lines of diverse origin. Dual luciferase assays show that compared to mutant (mut), the reporter gene containing wild type (wt) let-7a binding sites was efficiently suppressed upon transfection in various cell lines. Importantly, the strength of miRNA regulation varied across the cell lines. Total RNA analysis demonstrates that wt renilla mRNA was expressed to similar or higher levels compared to mut suggesting that translation repression is a predominant mode of miRNA regulation. Nonetheless, transcript degradation was observed in some cell lines. Ago-2 immunoprecipitation show that miRNA repressed renilla mRNA are associated with functional mi-RISC (miRNA-RNA induced silencing complex). Given the immense potential of miRNA as a therapeutic option, these findings highlight the necessity to thoroughly examine the mode of mRNA regulation in order to achieve the beneficial effects in targeting cells.

Leukocyte production of inflammatory mediators is inhibited by the antioxidants phloretin, silymarin, hesperetin, and resveratrol.

Antioxidants possess significant therapeutic potential for the treatment of inflammatory disorders. One such disorder is periodontitis characterised by an antimicrobial immune response, inflammation, and irreversible changes to the supporting structures of the teeth. Recognition of conserved pathogen-associated molecular patterns is a crucial component of innate immunity to Gram-negative bacteria such as Escherichia coli, as well as the periodontal pathogen Aggregatibacter actinomycetemcomitans. In this study, we investigated the antioxidants Phloretin, Silymarin, Hesperetin, and Resveratrol to ascertain whether they altered the production of inflammatory mediators by innately-activated leukocytes. Peripheral blood mononuclear cells were stimulated with lipopolysaccharide purified from Aggregatibacter actinomycetemcomitans, and the production of cytokines, chemokines, and differentiation factors was assayed by enzyme-linked immunosorbent assay, cytometric bead array, and RT-PCR. Significant inhibition of these factors was achieved upon treatment with Phloretin, Silymarin, Hesperetin, and Resveratrol. These data further characterise the potent anti-inflammatory properties of antioxidants. Their ability to inhibit the production of inflammatory cytokines, chemokines, and differentiation factors by a heterogeneous population of leukocytes has clear implications for their therapeutic potential in vivo.

Histological changes in pulp-dentin complex in tooth subjected to traumatic occlusion and subluxation.

Background: This study evaluated the influence of traumatic occlusion in the dentin-pulp complex a molar teeth submitted to subluxation. Material and methods: Ninety Wistar rats were divided into groups Naïve (N), Subluxation (S) and Subluxation with traumatic occlusion (STO) and submitted to histological analysis after 7 and 21 days. A quantitative analysis was submitted to one-way ANOVA and Tukey's post-hoc test, and Chi-square and Bonferronís post-hoc test. Results: S and STO showed a significant increase in blood vessels area (p < 0.0005), amorphous fundamental substance (p < 0.0005) and reactionary dentin formation (p < 0.0005), as well as a decrease in the nuclear profile (p < 0.0005), odontoblast layer (p = 0.013 and p < 0.0005) by day 7 when compared with N. These changes normalized by day 21, except for the reactionary dentin (p < 0.0005) in both S and STO groups. Interestingly, the STO group exhibited significant changes in the increase of pulp calcification (p < 0.0005), presence of tubules with nuclei (p < 0.0005), and inflammatory infiltrate (p < 0.0005), as well reduction of nuclear profile (p < 0.0005), odontoblast layer (p < 0.0005) compared with N and S at day 21. Conclusions: STO impaired the defence response and decreased pulp regeneration capacity by increasing the inflammatory infiltrate and pulp calcification, and decreasing the nucleated cell number in the odontoblast layer and central pulp.

Heterologous expression of rice calnexin (OsCNX) confers drought tolerance in Nicotiana tabacum.

Calnexin (CNX) is an integral membrane protein of endoplasmic reticulum (ER) and is a critical component of ER quality control machinery. It acts as a chaperone and ensures proper folding of newly synthesised glycoproteins. CNX shares a considerable homology with its luminal counterpart calreticulin (CRT). Together, they constitute CNX/CRT cycle which is imperative for proper folding of nascent proteins. CNX deficient organisms develop severe complications because of improper folding of proteins and consequently ER stress. CNX maintains calcium homeostasis by binding to the Ca(2+) which is a central node in various signaling pathways. Phosphorylation of cytoplasmic tail of CNX controls the sarco endoplasmic reticulum calcium ATPase and thus the movement of Ca(2+) in and out of its store-house, i.e. ER. Our studies on Oryza sativa CNX (OsCNX) reveal constitutive expression at various developmental stages and various tissues, thereby proving its requirement throughout the plant development. Further, its expression under various stress conditions gives an insight of the crosstalk existing between ER stress and abiotic stress signaling. This was confirmed by heterologous expression of OsCNX (OsCNX-HE) in tobacco and the OsCNX-HE lines were observed to exhibit better germination under mannitol stress and survival under dehydration stress conditions. The dehydration tolerance conferred by OsCNX appears to be ABA-dependent pathway.

Assessment of Detoxification Strategies for Used Dental Implant Healing Abutments: Macroscopic and Biological Implications.

PURPOSE: Dental implant manufacturers recommend healing abutments (HA) be used for single-patient use; however, reuse on multiple patients following decontamination and sterilization is common. This study aims to evaluate four decontamination strategies utilizing enzymatic agents, available in most clinical settings, to determine the level to which biomaterial can be removed in a group of previously used HA (uHA). Secondly, to determine the degree to which the decontaminated HA are capable of inducing an inflammatory response in-vitro compared to new, never used HA. MATERIALS AND METHODS: Fifty HA were collected following 2-4 weeks of intraoral use and distributed randomly into 5 test groups (Group A-E; n = 10/group). Group A: Enzymatic cleaner foam + Autoclave; Group B: Ultrasonic bath with enzymatic cleaner + Autoclave; Group C: Prophy jet + Enzymatic cleaner foam + Autoclave; Group D: Prophy jet + ultrasonic bath with enzymatic cleaner + Autoclave; Group E: Prophy jet + Autoclave. Ten new, sterile HA served as controls (Group "Control"). Residual protein concentration was determined by a Micro BCA protein assay while HA from each group were stained with Phloxine B and macroscopically examined for the presence of debris. To examine the inflammatory potential, human primary macrophages were exposed to HA and supernatant levels of 9 cytokines/chemokines profiles were analyzed using a multiplex bead assay. RESULTS: All test groups presented with differences in the degree of visual decontamination compared to Controls, with Groups D and E displaying the most effective surface debris removaland reduced protein concentration. Of the detoxification strategies, Groups D and E removed the greatest biomaterial while least effective was Group A. However, compared to Controls, multiplex assays revealed high levels of inflammatory cytokine secretion up to 5 days from all Test Groups (A-E) irrespective of the decontamination method used. CONCLUSION: Our study found that compared to new, never used HA, decontamination of uHA utilizing enzymatic cleaners failed to reestablish inert HA surfaces and prevent an inflammatory immune response in-vitro. Clinicians should not reuse HA even after attempts to decontaminate and sterilize HA surfaces.

Biogenesis, functions and fate of plant microRNAs.

microRNAs (miRNAs), a recently discovered class of small RNAs, are endogenously transcribed non-coding RNAs that are known to control diverse developmental processes and defense responses. They regulate these pathways by fine-tuning the levels of transcripts to which they bind and cause their cleavage or translation repression. Several studies on the processing of miRNA precursors have shed light on the essential structural features for precise release of miRNA duplexes. The identification of a protein that degrade single stranded small RNA has provided us with some understanding of how miRNA flux is maintained in plants. This review focuses on the genome organization, biogenesis, miRNA activity, and the fate of miRNAs.

The future treatment for type 1 diabetes: Pig islet- or stem cell-derived ß cells?

Replacement of ß cells is only a curative approach for type 1 diabetes (T1D) patients to avoid the threat of iatrogenic hypoglycemia. In this pursuit, islet allotransplantation under Edmonton's protocol emerged as a medical miracle to attain hypoglycemia-free insulin independence in T1D. Shortage of allo-islet donors and post-transplantation (post-tx) islet loss are still unmet hurdles for the widespread application of this therapeutic regimen. The long-term survival and effective insulin independence in preclinical studies have strongly suggested pig islets to cure overt hyperglycemia. Importantly, CRISPR-Cas9 technology is pursuing to develop "humanized" pig islets that could overcome the lifelong immunosuppression drug regimen. Lately, induced pluripotent stem cell (iPSC)-derived ß cell approaches are also gaining momentum and may hold promise to yield a significant supply of insulin-producing cells. Theoretically, personalized ß cells derived from a patient's iPSCs is one exciting approach, but ß cell-specific immunity in T1D recipients would still be a challenge. In this context, encapsulation studies on both pig islet as well as iPSC-ß cells were found promising and rendered long-term survival in mice. Oxygen tension and blood vessel growth within the capsules are a few of the hurdles that need to be addressed. In conclusion, challenges associated with both procedures, xenotransplantation (of pig-derived islets) and stem cell transplantation, are required to be cautiously resolved before their clinical application.

Human herpesvirus-encoded MicroRNA in host-pathogen interaction.

Human herpesviruses (HHV) are ubiquitous, linear dsDNA viruses that establish lifelong latency, disrupted by sporadic reactivation. HHV have evolved diverse ingenious mechanisms to evade robust host defenses. Incorporation of unique stem loop sequences that generate viral microRNAs (v-miRs) exemplifies one such evolutionary adaptation in HHV. These noncoding RNAs can control cellular and viral transcriptomes highlighting their ability in shaping host-HHV interactions. We summarize recent developments in functional characterization of HHV-encoded miRNAs in shaping the outcome of host-pathogen interaction. Non-immunogenic dissemination of v-miRs through exosomes confer added advantage to HHV in incessant modulation of host microenvironment. This review delineates the mechanistic role of v-miRs in facilitating viral persistence and tropism by targeting genes associated with cellular (apoptosis, angiogenesis, cell migration, etc.) and viral life cycle (latency, lytic and reactivation). Burgeoning evidences indicate plausible association of v-miRs in various immune-mediated diseases (nasopharyngeal carcinoma, neurological disorders, periodontal diseases, etc.) and herpesvirus-related malignancies indicating their broad-spectrum impact on host cellular pathways. We propose to exploit tisssue and systemic levels of v-miRs as diagnostic and prognostic markers for cancers and immune-mediated diseases. Therapeutic targeting of v-miRs will advance the promising outcomes of preclinical discoveries to bedside application.

Improved air quality and associated mortalities in India under COVID-19 lockdown.

India enforced stringent lockdown measures on March 24, 2020 to mitigate the spread of the Severe Acute Respiratory Syndrome Coronovirus-2 (SARS-CoV-2). Here, we examined the impact of lockdown on the air quality index (AQI) [including ambient particulate matter (PM10 and PM2.5), nitrogen dioxide (NO2), sulfur dioxide (SO2), carbon monoxide (CO), ozone (O3), and ammonia (NH3)] and tropospheric NO2 and O3 densities through Sentinel-5 satellite data approximately 1 d post-lockdown and one month pre-lockdown and post-lockdown. Our findings revealed a marked reduction in the ambient AQI (estimated mean reduction of 17.75% and 20.70%, respectively), tropospheric NO2 density, and land surface temperature (LST) during post-lockdown compared with the pre-lockdown period or corresponding months in 2019, except for a few sites with substantial coal mining and active power plants. We observed a modest increase in the O3 density post-lockdown, thereby indicating improved tropospheric air quality. As a favorable outcome of the COVID-19 lockdown, road accident-related mortalities declined by 72-folds. Cities with poor air quality correlate with higher COVID-19 cases and deaths (r = 0.504 and r = 0.590 for NO2; r = 0.744 and r = 0.435 for AQI). Conversely, low mortality was reported in cities with better air quality. These results show a correlation between the COVID-19 vulnerable regions and AQI hotspots, thereby suggesting that air pollution may exacerbate clinical manifestations of the disease. However, a prolonged lockdown may nullify the beneficial environmental outcomes by adversely affecting socioeconomic and health aspects.

Impaired cell migration and structural defects in myeloid cells overexpressing miR-30b and miR-142-3p.

Macrophages (MΦ) and dendritic cells (DC) play a fundamental role in shaping immune responses by sensing a plethora of Pathogen Associated Molecular Patterns (PAMPs), phagocytosis and antigen presentation to T lymphocytes. These important biological processes require efficient cell movement and an intact cellular morphology for dynamic interaction. The role of microRNAs (miRs) in this regard, however, is not well understood. In the present study, we show that miR-30b and miR-142-3p regulate migration and morphology of MΦ and DC. Transient overexpression of miR-30b and miR-142-3p attenuates migration and these cells display unique morphological deformities observed under electron microscopy. In addition, miR-142-3p overexpression in MΦ impaired phagocytosis of FITC-conjugated latex beads using live microscopy imaging. Interestingly, live cell imaging and F-actin staining revealed marked changes in the cell polarity and actin polymerization status, respectively. To identify miR-142-3p regulated pathways, we profiled global transcriptome changes in miR-142-3p or control mimic transfected DC. Expression of several genes were differentially altered by miR-142-3p and were associated with pathways related to cell movement, cell adhesion, and cytoskeletal rearrangement. Bioinformatics analysis identified a significant subset of downregulated genes with one or more predicted miR-142-3p binding sites in their 3'UTR strongly suggesting direct post-transcriptional impact of these miRNAs on multiple transcripts. Using dual luciferase assays, novel miR-142-3p binding sites were validated for three genes (Vinculin, Dab2 and Skap2) directly associated with cytoskeletal rearrangement and cell movement. In summary, our results show that miR-30b and miR-142-3p are regulators of myeloid cell cytoskeletal homeostasis and morphology.

Long Noncoding RNA in Myeloid and Lymphoid Cell Differentiation, Polarization and Function.

Long noncoding RNA (lncRNA) are a class of endogenous, non-protein coding RNAs that are increasingly being associated with various cellular functions and diseases. Yet, despite their ubiquity and abundance, only a minute fraction of these molecules has an assigned function. LncRNAs show tissue-, cell-, and developmental stage-specific expression, and are differentially expressed under physiological or pathological conditions. The role of lncRNAs in the lineage commitment of immune cells and shaping immune responses is becoming evident. Myeloid cells and lymphoid cells are two major classes of immune systems that work in concert to initiate and amplify innate and adaptive immunity in vertebrates. In this review, we provide mechanistic roles of lncRNA through which these noncoding RNAs can directly participate in the differentiation, polarization, and activation of myeloid (monocyte, macrophage, and dendritic cells) and lymphoid cells (T cells, B cells, and NK cells). While our knowledge on the role of lncRNA in immune cell differentiation and function has improved in the past decade, further studies are required to unravel the biological role of lncRNAs and identify novel mechanisms of lncRNA functions in immune cells. Harnessing the regulatory potential of lncRNAs can provide novel diagnostic and therapeutic targets in treating immune cell related diseases.

Pea Gß subunit of G proteins has a role in nitric oxide-induced stomatal closure in response to heat and drought stress.

Heterotrimeric G proteins consisting of Gα, Gß and Gγ subunits act as downstream effectors to regulate multiple functions including abiotic stress tolerance. However, the mechanism of Gß-mediated heat and drought tolerance is yet to be established. To explore the role of Pisum sativum Gß subunit (PsGß) in heat and drought stress, transgenic tobacco plants overexpressing (OEs) PsGß were raised. Transgenic plants showing ectopic expression of PsGß performed better under heat and drought stress in comparison with vector control plants. The seed germination, relative water content (RWC) and nitric oxide (NO) induction in the guard cells of transgenic plants were significantly higher in contrast to control plants. PsGß promoter was isolated and several stress-responsive elements were identified. The change in Gß expression in response to heat, methyl jasmonate (MeJA), abscisic acid (ABA), drought and salt confirms the presence of heat, low temperature and drought-responsive elements in the PsGß promoter. Also, heat and drought stress caused the release of NO-induced stomatal closure in the leaves of transgenic tobacco plants OEs PsGß. The better performance of transgenic plant OEs PsGß is also attributed to the improved photosynthetic parameters as compared with control plants. These findings suggest a role of PsGß in the signalling pathway leading to NO-induced stomatal closure during heat and drought stress.

MicroRNA: Dynamic Regulators of Macrophage Polarization and Plasticity.

The ability of a healthy immune system to clear the plethora of antigens it encounters incessantly relies on the enormous plasticity displayed by the comprising cell types. Macrophages (MΦs) are crucial member of the mononuclear phagocyte system (MPS) that constantly patrol the peripheral tissues and are actively recruited to the sites of injury and infection. In tissues, infiltrating monocytes replenish MΦ. Under the guidance of the local micro-milieu, MΦ can be activated to acquire specialized functional phenotypes. Similar to T cells, functional polarization of macrophage phenotype viz., inflammatory (M1) and reparative (M2) is proposed. Equipped with diverse toll-like receptors (TLRs), these cells of the innate arm of immunity recognize and phagocytize antigens and secrete cytokines that activate the adaptive arm of the immune system and perform key roles in wound repair. Dysregulation of MΦ plasticity has been associated with various diseases and infection. MicroRNAs (miRNAs) have emerged as critical regulators of transcriptome output. Their importance in maintaining health, and their contribution toward disease, encompasses virtually all aspects of human biology. Our understanding of miRNA-mediated regulation of MΦ plasticity and polarization can be utilized to modulate functional phenotypes to counter their role in the pathogenesis of numerous disease, including cancer, autoimmunity, periodontitis, etc. Here, we provide an overview of current knowledge regarding the role of miRNA in shaping MΦ polarization and plasticity through targeting of various pathways and genes. Identification of miRNA biomarkers of diagnostic/prognostic value and their therapeutic potential by delivery of miRNA mimics or inhibitors to dynamically alter gene expression profiles in vivo is highlighted.

MicroRNA target Fc receptors to regulate Ab-dependent Ag uptake in primary macrophages and dendritic cells.

Phagocytosis commences with particle internalization and culminates with the activation of innate and adaptive immune responses. However, the role of miRNAs in phagocytosis remains largely unknown. In this study, we examined the role of miR-24, miR-30b and miR-142-3p in Ab Fc receptor (FcR)-mediated phagocytosis by macrophages (MΦ) and dendritic cells (DC). The expression of these miRNAs was reduced following phagocytosis of both IgG-opsonized beads and Escherichia coli, indicating their regulatory role in the process. Further, overexpression of these miRNAs impaired the uptake of IgG-coated latex beads, which corroborated the reduced secretion of the pro-inflammatory cytokines TNF-α and IL-8 and down-regulation of PKC-α, as well as superoxide-generating enzyme NADPH oxidase 2 expression level. Mechanistically, MΦ and DC transfected with miRNA mimics show marked reduction in expression of FcRs including FCGR2A, FcɛR1G and FCER2. We show that FcɛR1G expression is not affected at the transcription level, rather it is post-transcriptionally regulated by miR-30b. Finally, we demonstrate that siRNA-mediated knockdown of FcɛR1G leads to reduced uptake of IgG-opsonized beads, indicating its involvement on Ab-mediated phagocytosis. These results uncover miR-24, miR-30b and miR-142-3p as an essential component of FcR-mediated phagocytosis and associated innate immune responses.

MiR-142-3p is a RANKL-dependent inducer of cell death in osteoclasts.

MicroRNA are small, non-coding, single-stranded RNAs that are estimated to regulate ~60% of the human genome. MiRNA profiling of monocyte-to-osteoclast differentiation identified miR-142-3p as a miRNA that is significantly, differentially expressed throughout osteoclastogenesis. Enforced expression of miR-142-3p via transient transfection with miR-142-3p mimic inhibited cell-to-cell contact and fusion, decreased protein kinase C alpha expression, and ultimately reduced cell viability. miR-142-3p was also identified as significantly differentially expressed during monocyte-to-macrophage differentiation and overexpression of miR-142-3p prevented their conversion to osteoclasts. Furthermore, the inhibitory effect of miR-142-3p on osteoclastogenesis extended to the conversion of a third osteoclast precursor cell type- dendritic cells. These results demonstrate miR-142-3p to be a negative regulator of osteoclastogenesis from the 3 main precursor cell types: monocytes, macrophages and dendritic cells. Importantly, decreased survival was dependent upon both miR-142-3p expression and RANK-signaling, with no harmful effects detected in the absence of this combination. As such, miR-142-3p represents a novel target for the selective removal of osteoclasts by targeting of osteoclastogenic pathways.

miR-24, miR-30b and miR-142-3p interfere with antigen processing and presentation by primary macrophages and dendritic cells.

Antigen uptake, processing and presentation by antigen presenting cells (APCs) are tightly coupled processes which consequently lead to the activation of innate and adaptive immune responses. However, the regulatory role of microRNA (miRNAs) in these critical pathways is poorly understood. In this study, we show that overexpression of miR-24, miR-30b and miR-142-3p attenuates uptake and processing of soluble antigen ovalbumin (Ova) in primary human macrophages and dendritic cells. MiRNA mimic transfected APCs exhibit defects in antigen presentation (Ova and CMV antigen) to CD4+ T-cells leading to reduced cell proliferation. Using transgenic OT-II mice we demonstrated that this impairment in T-cell proliferation is specific to antigen provided i.e., Ova. Further, human T-cells co-cultured with miRNA transfected dendritic cells secrete low levels of T helper (Th)-1 polarization associated cytokines. Analysis of molecules regulating APC and T-cell receptor interaction shows miRNA-mediated induced expression of Programmed Death-Ligand 1 (PD-L1) which inhibits T-cell proliferation. Blocking PD-L1 with antibodies rescues miRNA-mediated inhibition of T cell priming by DCs. These results uncover regulatory functions of miR-24, miR-30b and miR-142-3p in pairing innate and adaptive components of immunity.