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We applied AMS 14C dates, Mg/Ca estimated sea surface temperature (SST), planktonic foraminiferal abundance, coarse component and X-ray diffraction mineral composition analyses for an International Ocean Discovery Program (IODP) sediment Hole U1467C in the Maldives inner sea, to reveal factors that affected the depositional process in the Maldives inner sea over the past 30 kyr. We found that the linear sedimentation rate (LSR) in Holocene (6.8 cm/kyr) was obviously higher than that in glacial period (3.45 cm/kyr); the average SST during the Holocene was 2–3°C higher than that in glacial stage. High abundance of planktonic foraminifera occurred during the glacial period, which was consistent with the variation of pelagic biogenic component. Reef-sourced material showed apparent high percentage during the Holocene (43.5%), in contrast to the low values during the glacial stage (20.7%). Terrigenous matter, only accounting for a small percentage in carbonate platform, was relatively high during the glacial period than that in the Holocene. We therefore conclude that reef-sourced material, dominated in the glacial-interglacial sediment sources, was mainly affected by sea level and climate changes. During glacial stage, sea level low-stand and weakened weathering in Maldives limited the input of the eroded material into the inner sea, resulting in low LSR; while the Holocene high sea level accompanied with rapid growth of the reef atoll and enhanced weathering, brought more reef-sourced material to the inner sea, leading to increased LSR and lowered abundance of planktonic foraminifera. The sea level and climate-controlled reef-sourced material changes are the key to understand the sedimentary process of the Maldives inner sea. Our study will shed some light on the evolution of glacial-interglacial sedimentary process of carbonate platform.

New planktonic and benthic foraminiferal stable isotope records from core YDY05 (northeastern Indian Ocean) provide new insights into paleoceanographic changes in the northeastern Indian Ocean since the last glacial period. The distinct δ18O decrease was observed since the beginning of the deglaciation to the mid-Holocene (∼8–6 kyr BP), possibly reflecting a reduction in surface salinity in the central Bay of Bengal (BoB) water, which probably resulted from strengthened precipitation, concurrent enhanced river discharge and rising sea-level, related to the intensification of Indian Summer Monsoon (ISM). Variations in benthic δ13C and δ13CPlanktonic-Benthic in our core site reflect significant variations in source water characteristics over the LGM-Holocene. The large δ13CPlanktonic-Benthic offset during the glacial period suggests a more sluggish deep water circulation, and lower δ13CPlanktonic-Benthic from the deglaciation to the Holocene suggests an enhanced deep water circulation in the central BoB. The drastic depletion in benthic δ13C during the glacial period suggests a significant reduction of North Atlantic Deep Water (NADW) intrusion and a progressive influx of Antarctic Bottom Water (AABW) and 12C-rich Circumpolar Deep Water (CDW) into the central BoB. In contrast, since the deglaciation, the central BoB experienced a drastically increased intrusion of better ventilated and 13C-rich NADW. The differences in benthic δ18O between the LGM section and the Holocene exceeds the ice volume effect by ∼0.5‰, providing further evidence that the deep water mass of the central BoB was influenced by the less dense NADW, instead of the AABW, since the last deglaciation.

High-risk B-cell acute lymphoblastic leukemia (B-ALL) is an aggressive disease, often characterized by resistance to chemotherapy. A frequent feature of high-risk B-ALL is loss of function of the IKAROS (encoded by the IKZF1 gene) tumor suppressor. Here, we report that IKAROS regulates expression of the BCL2L1 gene (encodes the BCL-XL protein) in human B-ALL. Gain-of-function and loss-of-function experiments demonstrate that IKAROS binds to the BCL2L1 promoter, recruits histone deacetylase HDAC1, and represses BCL2L1 expression via chromatin remodeling. In leukemia, IKAROS’ function is impaired by oncogenic casein kinase II (CK2), which is overexpressed in B-ALL. Phosphorylation by CK2 reduces IKAROS binding and recruitment of HDAC1 to the BCL2L1 promoter. This results in a loss of IKAROS-mediated repression of BCL2L1 and increased expression of BCL-XL. Increased expression of BCL-XL and/or CK2, as well as reduced IKAROS expression, are associated with resistance to doxorubicin treatment. Molecular and pharmacological inhibition of CK2 with a specific inhibitor CX-4945, increases binding of IKAROS to the BCL2L1 promoter and enhances IKAROS-mediated repression of BCL2L1 in B-ALL. Treatment with CX-4945 increases sensitivity to doxorubicin in B-ALL, and reverses resistance to doxorubicin in multidrug-resistant B-ALL. Combination treatment with CX-4945 and doxorubicin show synergistic therapeutic effects in vitro and in preclinical models of high-risk B-ALL. Results reveal a novel signaling network that regulates chemoresistance in leukemia. These data lay the groundwork for clinical testing of a rationally designed, targeted therapy that combines the CK2 inhibitor, CX-4945, with doxorubicin for the treatment of hematopoietic malignancies.

Vaccinia virus (VV) is the most studied member of the poxvirus family, is responsible for the successful elimination of smallpox worldwide, and has been developed as a vaccine vehicle for infectious diseases and cancer immunotherapy. We have previously shown that the unique potency of VV in the activation of CD8+T cell response is dependent on efficient activation of the innate immune system through Toll-like receptor (TLR)-dependent and -independent pathways. However, it remains incompletely defined what regulate CD8+T cell response to VV infection. In this study, we showed that γδT cells play an important role in promoting CD8+T cell response to VV infection. We found that γδT cells can directly present viral antigens in the context of MHC-I for CD8+T cell activation to VVin vivo, and we further demonstrated that cell-intrinsic MyD88 signaling in γδT cells is required for activation of γδT cells and CD8+T cells. These results illustrate a critical role for γδT cells in the regulation of adaptive T cell response to viral infection and may shed light on the design of more effective vaccine strategies based on manipulation of γδT cells.

Excessive exposure to manganese (Mn) can cause cognitive impairment, a common feature of Alzheimer's disease (AD), but the mechanisms remain unclear. Amyloid precursor protein (APP) is key to AD pathogenesis, and whether APP and its secretase processing are involved in Mn-induced cognitive impairment remains unknown. In the present study, we established a model of Mn-induced neurotoxicity in vivo (male C57BL/6, 0–100 mg/kg Mn, 90 days, gastric gavage) and in vitro (Neuro-2a (N2a) cells, 0–800 μM Mn for 24 h; APP overexpression and APP shRNA N2a cells, 0 and 800 μM Mn for 24 h). We found impaired cognition of Mn-treated mice. Both in vivo and in vitro results consistently showed that Mn exposure inhibited the expression of APP, α-secretase, soluble APP alpha protein (sAPPα), and synapse proteins as well as the activity of α-secretase. However, Mn exposure showed no effect on the protein levels of β-secretase, Aβ40, and Aβ42 or the activity of β-secretase. Collectively, these findings demonstrate key roles of APP and its α-secretase processing in the regulation of Mn-induced cognitive impairment, which may act as a target for ameliorating Mn-induced neurotoxicity.

The etiology of multiple myeloma (MM) remains incompletely understood; however, epidemiologic studies have suggested a possible link between exposure to environmental aromatic hydrocarbons-which serve as exogenous ligands for the aryl hydrocarbon receptor (AHR), which has been implicated in cancer biology-and development of monoclonal gammopathy of undetermined significance (MGUS) and MM. Herein, we demonstrate the functional expression of AHR in MM cell lines and primary human MM samples. AHR is expressed in putative MM 'stem cells' and advanced clinical stages of MM, and functionally contributes to MM tumor cell phenotype and proliferation. Antagonism of AHR directly impairs MM cell viability and increases MM cell susceptibility to immune-mediated clearance. Furthermore, our findings indicate that AHR antagonism may represent an effective means to enhance the function of other drugs, such as anti-CD38 antibodies, in future clinical studies. Taken together, these data identify AHR as a novel target for MM therapy.

Vaccinia virus (VV) is the most studied member of the poxvirus family, is responsible for the successful elimination of smallpox worldwide, and has been developed as a vaccine vehicle for infectious diseases and cancer immunotherapy. We have previously shown that the unique potency of VV in the activation of CD8+ T cell response is dependent on efficient activation of the innate immune system through Toll-like receptor (TLR)-dependent and -independent pathways. However, it remains incompletely defined what regulate CD8+ T cell response to VV infection. In this study, we showed that γδT cells play an important role in promoting CD8+ T cell response to VV infection. We found that γδT cells can directly present viral antigens in the context MHC-I for CD8+ T cell activation to VV in vivo, and we further demonstrated that cell-intrinsic MyD88 signaling in γδT cells is required for activation of γδT cells and CD8+ T cells. These results illustrate a critical role for γδT cells in the regulation of adaptive T cell response to viral infection and may shed light on the design of more effective vaccine strategies based on manipulation of γδT cells.ImportanceTargeting the immune systems has powerful potentials to treat many disorders, such as some cancers and viral infections. By understanding how the immune system responds to model infections, we can better determine strategies to manipulate our immune systems. Vaccinia virus is responsible for the worldwide elimination of smallpox and produces one of the longest immune responses known in humans. We know from previous findings that NK cells are required for initial immune response and CD8+ T cells are required for the elimination of the virus. How CD8+ T cells are activated in response to Vaccinia virus is not fully understood. This manuscript found that γδT cells activate CD8+ T cells in response to Vaccinia virus infection through MyD88 pathway

The South China Sea (SCS) is an important moisture and heat source in the Asian monsoon system, and its environmental changes influence economic and social stability in Southeast Asia and East Asian. Previous records on sea surface temperature (SST) and salinity (SSS) were mostly from one sediment core, and/or only showed fragmentary information. It is hard to provide insights into the mechanisms of the overall paleoceanographical environmental variability in the northern SCS. In this study, paired δ18O and Mg/Ca records of planktonic foraminifer Globigerinoides ruber from Core S0204B nearby the Luzon Strait (21°13.818′N, 118°3.077′, water depth 1822 m), and Core GHE 27L (19.85°N, 115.34°E, water depth of 1533 m) in the central northern SCS, were analyzed, so as to better understand the physical process of the SST and SSS changes in the northern SCS and its relationship with the tropical Pacific. The results demonstrate that the SST is relatively stable and seawater δ18O (δ18Osw) values, a salinity proxy, shows a decrease trend since 7.0 ka BP in Core S0204B. By contrast, SST and δ18Osw in Core GHE 27L exhibit an overall increasing trend at the same time, indicating apparent regional discrepancy on the changing trend of the SST and SSS in the northern SCS. We suggest that the thermal and hydrographic conditions in Core S0204B were primarily influenced by the outer western Pacific water. However, the thermal and hydrographic conditions in Core GHE 27L may be predominantly controlled by the weakened Asian monsoon during the mid-late Holocene, which was induced by the southern migration of Intertropical Convergence Zone (ITCZ) and stronger EI Nino Southern Oscillation.

The programmed death-1 (PD-1) and the PD ligand 1 (PD-L1) interaction represents a key immune checkpoint within the tumor microenvironment (TME), and PD-1 blockade has led to exciting therapeutic advances in clinical oncology. Although IFN-γ–dependent PD-L1 induction on tumor cells was initially thought to mediate the suppression on effector cells, recent studies have shown that PD-L1 is also expressed at high level on tumor-associated macrophages (TAMs) in certain types of tumors. However, the precise role of PD-L1 expression on TAMs in suppressing antitumor immunity within the TME remains to be defined. Using a myeloid-specific Pdl1-knockout mouse model, here we showed definitive evidence that PD-L1 expression on TAMs is critical for suppression of intratumor CD8+ T cell function. We further demonstrated that tumor-derived Sonic hedgehog (Shh) drives PD-L1 expression in TAMs to suppress tumor-infiltrating CD8+ T cell function, leading to tumor progression. Mechanistically, Shh-dependent upregulation of PD-L1 in TAMs is mediated by signal transducer and activator of transcription 3, a cascade that has not been previously reported to our knowledge. Last, single-cell RNA sequencing analysis of human hepatocellular carcinoma revealed that PD-L1 is mainly expressed on M2 TAMs, supporting the clinical relevance of our findings. Collectively, our data revealed an essential role for Shh-dependent PD-L1 upregulation in TAMs in suppressing antitumor immunity within the TME, which could lead to the development of new immunotherapeutic strategies for treating cancer.

The epithelial splicing regulatory proteins, ESRP1 and ESRP2 are essential for mammalian development through regulation of a global program of alternative splicing of genes involved in maintenance of epithelial cell function. To further inform our understanding of the molecular functions of ESRP1 we performed enhanced crosslinking immunoprecipitation coupled with high throughput sequencing (eCLIP) in epithelial cells of mouse epidermis. The genome-wide binding sites of ESRP1 were integrated with RNA-Seq analysis of alterations in splicing and total gene expression that result from epidermal ablation of Esrp1 and Esrp2. These studies demonstrated that ESRP1 functions in splicing regulation occur primarily through direct binding in a position-dependent manner to either promote exon inclusion or skipping. In addition, we also identified widespread binding of ESRP1 in 3′ and 5′ untranslated regions (UTRs) of genes involved in epithelial cell function suggesting that its post-transcriptional functions extend beyond splicing regulation.

Learning pollen transport mechanisms is the basis for correct interpretation of fossil pollen data. Firstly, 33 seabed surface sediment samples from the northeastern Indian Ocean were selected for pollen analysis. Utilizing the 90°E Indian Ridge as a dividing line, more Trilete spores (mainly Microlepia)and herbaceous pollen were present to the west or northwest of the line, possibly originating in Sri Lanka and India. More Polypodiaceae spores and tree pollen (Pinus excluded) were present to the east or southeast of the boundary, possibly originating in Sumatra Island. Based on the PCA analysis, the ocean current from the southeast to the northwest and the distance from land masses around the study area affect the percentage of pollen. Then, based on a fossil pollen analysis on the YDY05 core from the northeastern Indian Ocean, we determined that since 16 ka BP, the pollen terrestrial area might change from Sumatra Island to Sri Lanka Island and India, probably due to the climate and sea level change.

The Southern Yellow Sea Cold Water Mass (YSCWM) is closely related to the modern circulation system of the east China shelf seas, which has significantly influenced regional marine environmental changes. The study of the Holocene evolution of the YSCWM will greatly improve our understanding of the mechanisms of regional environmental change. Benthic foraminifera are sensitive to bottom water environmental changes and can serve as useful indicators in bottom water environmental reconstruction. In this study, benthic foraminifera were analyzed in core N02 from the northwestern margin of the southern Yellow Sea Mud to decipher the phase evolution of the YSCWM during the last 7 kyr. Benthic foraminifera census counts and Q-mode factor analysis indicate that the Holocene sedimentary environment can be divided into three stages: From 6.9–5.0 ka, the fauna was dominated by Ammonia ketienziensis , indicating that the YSCWM was at its strongest during the last 7 kyr, while the Yellow Sea Coastal Current (YSCC) had a weak influence on the bottom water of the study area. From 5.0–2.9 ka, the relative abundance of Hanzawaia nipponica remarkably increased while the abundance of A. ketienziensis decreased significantly, reflecting that the strength of the YSCWM was relatively weak and the range of the YSCWM might have contracted. The influence of the YSCC on the bottom water might have slightly increased, although its influence was still weak during this time. A notable increase in low-temperature and low-salinity species, such as Protelphidium tuberculatum and Buccella spp. has occurred since 2.9 ka, indicating that the YSCC has had a strong influence on bottom water during this period, while the strength of the YSCWM has been at its weakest during the last 7 kyr. Generally, the influence of the YSCWM and the YSCC on the bottom water properties of the study area show an obvious seesaw pattern, with one’s influence increasing while the other’s influence decreases and vice versa. The fluctuations in the strength of YSCWM during the Holocene may be caused by the different effect allocations of regional climatic factors (i.e. El Nino Southern Oscillation, East Asian Winter Monsoon, summer insolation in the northern hemisphere, etc.) acting on the circulation system during different periods.

The Holocene evolution of the Indian Ocean Monsoon (IOM), especially the spatiotemporal nature of its maximum precipitation period, remains controversial. Here we reconstruct high-resolution local δ18Osw and sea surface temperature (SST) values by pairing Mg/Ca values and the δ18O data from planktonic foraminifera Globigerinoides ruber (white) in the southern Andaman Sea in order to understand IOM rainfall variability during the Holocene. Local δ18Osw values show that local maximum rainfall occurred between 10.6 ka and 8.6 ka in the southern Andaman Sea, before declining markedly during the 8.6–5.0 ka period, and then decreasing slightly after 5.0 ka. The Mg/Ca-SST was relatively low during the 10.3–8.6 ka period, before increasing slightly between 8.6 ka and 4.0 ka; it declined thereafter. But the sedimentation rate at the site of Core ADM-C1 relating to monsoonal precipitation of the source region of sediments exhibited its highest value during the 8.6–5.5 ka period. After making further comparisons with other published records for this region, we would propose that maximum IOM precipitation during the early Holocene has exhibited a clearly meridional migration: it was located in the tropics around 7°N during the 10.6–8.6 ka period, migrated northward to the subtropics during the 8.6–5.5 ka period, before migrating southward again after 5.0 ka. We would suggest that the northward shift of maximum precipitation during the Early Holocene in the IOM region was in response to the progressively northward migration of the Intertropical Convergence Zone (ITCZ) and the change of El Nino Southern Oscillation (ENSO) and Indian Ocean Dipole (IOD) activities.

We compared the characteristics of cases of highly pathogenic avian influenza (HPAI) and low pathogenic avian influenza (LPAI) A(H7N9) virus infections in China. HPAI A(H7N9) case-patients were more likely to have had exposure to sick and dead poultry in rural areas and were hospitalized earlier than were LPAI A(H7N9) case-patients.

Vaccinia virus (VV) can potently activate NK- and T-cell responses, leading to efficient viral control and generation of long-lasting protective immunity. However, immune responses against viral infections are often tightly controlled to avoid collateral damage and systemic inflammation. We have previously shown that granulocytic myeloid-derived suppressor cells (g-MDSCs) can suppress the NK-cell response to VV infection. It remains unknown what regulates T-cell responses to VV infection in vivo. In this study, we first showed that monocytic MDSCs (m-MDSCs), but not g-MDSCs, from VV-infected mice could directly suppress CD4+ and CD8+ T-cell activation in vitro. We then demonstrated that defective recruitment of m-MDSCs to the site of VV infection in CCR2-/- mice enhanced VV-specific CD8+ T-cell response and that adoptive transfer of m-MDSCs into VV-infected mice suppressed VV-specific CD8+ T-cell activation, leading to a delay in viral clearance. Mechanistically, we further showed that T-cell suppression by m-MDSCs is mediated by indication of iNOS and production of NO upon VV infection, and that IFN-γ is required for activation of m-MDSCs. Collectively, our results highlight a critical role for m-MDSCs in regulating T-cell responses against VV infection and may suggest potential strategies using m-MDSCs to modulate T-cell responses during viral infections.

Allogeneic hematopoietic cell transplantation (HCT) represents a potentially curative treatment for a variety of hematologic malignancies due to the well-recognized graft-versus-leukemia/lymphoma (GVL) effect that is mediated by donor-derived alloreactive T cells. However, graft-versus-host disease (GVHD) is mediated by the same T cells and remains a significant clinical problem associated with substantial morbidity and mortality. In this issue of the JCI, Ni and colleagues used several murine models of GVHD to evaluate the effect of CD4+ T cell depletion on GVL versus GVHD and revealed that depletion of CD4+ T cells leads to the upregulation of PD-L1 by recipient tissues and donor CD8+ T cells. Interaction of PD-L1 with PD-1 in GVHD-targeted tissues resulted in CD8+ T cell exhaustion and apoptosis, thereby preventing GVHD, whereas PD-L1 interactions with CD80 in lymphoid tissue promoted CD8+ T cell survival and expansion, thereby enhancing the GVL response. By separating these seemingly similar alloreactive T cell responses based on the context of interaction, the results of this study may lay the groundwork for the development of effective clinical strategies to enhance GVL while minimizing GVHD following allogeneic HCT.

NK cells are critical for the control of viral infections. Studies have shown that efficient NK cell activation in response to infection with VV in vivo requires multiple pathways, including the NKG2D pathway. We have recently shown that IL-18 is necessary for the activation of NK cells through upregulation of the NKG2D ligand Rae-1 on DCs upon VV infection. However, how IL-18R signaling on the accessory cells contributes to Rae-1 up-regulation remains to be defined. In this study, we found IL-18-mediated Rae-1 up-regulation in accessory cells, including macrophages and DCs, to be dependent on the MyD88-PI3K pathway. We further found that IL-18 signaling through PI3K led to inhibition of GSK-3, which we found to be a negative regulator of Rae-1. Finally, we demonstrated that in vivo inhibition of GSK-3 could restore Rae-1 up-regulation on IL18R−/− DCs and partially rescue NK-cell activation against VV, leading to improved viral control in IL-18R−/− mice. Our results showed that IL18-dependent Rae-1 up-regulation on accessory cells is mediated by the MyD88-PI3K-GSK3 pathway. These observations may provide important insights into the design of effective NK cell-based immunotherapies.

Heparanase is an endo-β-D-glucuronidase capable of cleaving heparan sulfate (HS) side chains contributing to break down of the extracellular matrix. Increased expression of heparanase has been found in numerous malignancies, and is associated with a poor prognosis. It has generated significant interest as a potential anti-neoplastic target because of the multiple roles it plays in tumor growth and metastasis. The pro-tumorigenic effects of heparanase are enhanced by the release of HS side chains, with subsequent increase in bioactive fragments and increased cytokine levels; both promoting tumor invasion, angiogenesis and metastasis. Preclinical experiments have shown heparanase inhibitors to substantially reduce tumor growth and metastasis leading to clinical trials with heparan sulfate mimetics. In this review we will examine heparanase’s role in tumor biology, its interaction with heparan surface proteoglycans, specifically syndecan-1; as well as the mechanism of action for heparanase inhibitors developed as anti-neoplastic therapeutics.

A palynological investigation of a 514 cm long continuous core section from Qinghai Crater Lake, western Yunnan Province, southwestern China, was performed in order to reconstruct regional vegetation and climate from 15.8 cal kyr BP to the present. The results show that the area was covered with deciduous broadleaved forest dominated by deciduous Quercus between 15.8 and 12.8 cal kyr BP, suggesting slightly cold and dry conditions. From 12.8 to 8.3 cal kyr BP, the vegetation continued to be dominated by deciduous broadleaved forest, but there was also a sudden increase in Castanopsis/Lithocarpus and a slight drop in deciduous Quercus and Alnus, reflecting a gradual increase in both temperature and humidity. During the period 8.3–4.6 cal kyr BP, the mixed deciduous forests became more diverse and dense, with thermophilous and hygrophilous species expanding to their maximum extents, implying a warm and rather humid climate. From 4.6 to 0.5 cal kyr BP, the vegetation was succeeded by open mixed deciduous forest dominated by Alnus, with a retreating evergreen broadleaved forest, indicating a cool and slightly dry climate. After 0.5 cal kyr BP, the vegetation became less diverse and more open, but evergreen broadleaved trees increased their spread markedly, suggesting that the climate had become warmer and drier. In recent decades, human activity resulted in a large increase in Alnus and Pinus around this lake basin. Specifically, we concluded that the warm and moist climate detected between 8.3 and 4.6 cal kyr BP may have occurred in response to the high sea surface temperatures and high sea levels in the Bay of Bengal existent at that time.

Diesel exhaust particles (DEPs) are common airborne ultrafine particles (UFPs); however, few studies have examined their effects on the gastrointestinal tract. To investigate the interaction of gut microbiota and DEPs‐induced colonic injury, adult C57BL/6 mice are kept in whole‐body inhalation chambers and exposed to filtered room air (FRA) or DEPs (300 µg m−3) 1 h per day for 28 consecutive days. DEPs exposure results in colon epithelial injury with inflammatory cell infiltration and mucus depletion. Abundance of Lactobacillus in murine feces is transiently increased following 7‐day DEPs exposure and then decreased until the end of 28‐day exposure. A reduction of the colonic mucus layer thickness is observed in mice receiving gut microbiota from DEPs‐exposed mice. Mechanistically, RNA‐sequencing suggests disruption of the nitrogen metabolism pathway in DEPs‐exposed NCM460 cells. Upregulation of carbonic anhydrase 9 (CA9) expression levels is observed in epithelia following DEPs exposure both in vivo and in vitro. Oral administration of probiotics protects the mice against DEPS‐induced colon epithelial injury. The results strongly suggest the involvement of gut microbiota in response to DEPs exposure and subsequently epithelial injury in vivo. Supplementation with probiotic may be a potential way to protect against UFPs‐induced colon epithelial injury.

The activation and recruitment of NK cells to the site of viral infection are crucial for virus control. However, it remains largely unknown what controls the recruitment of the activated NK cells to the infection site. In a model of intraperitoneal infection with vaccinia virus (VV), we showed that poly(ADP-ribose) polymerase-1 (PARP-1), a sensor of DNA damage, is critical for NK cell recruitment to the site of infection and viral control in vivo. We further demonstrated that PARP-1 promotes the production of CCL2 and that the CCL2-CCR2 axis is essential for NK cell recruitment to the infection site. In addition, we demonstrated that peritoneal macrophages are the main producer of PARP-1–dependent CCL2 secretion. Mechanistically, PARP-1 functions as a regulator of NF-κB by promoting its nuclear translocation and binding to its response sequences in macrophages upon VV infection. Taken together, our results reveal a potentially previously unknown role for PARP-1–dependent CCL2 production in NK cell migration and viral control and may provide important insights into the design of effective NK cell–based therapies for viral infections and cancer.

Tumor-associated macrophages (TAMs) usually display an antiinflammatory M2-like phenotype to facilitate tumor growth. However, what drives M2 polarization of TAMs and how TAMs suppress antitumor immunity within the tumor microenvironment (TME) remain largely undefined. Using several murine tumor models, we showed that hedgehog (Hh) signaling in myeloid cells is critical for TAM M2 polarization and tumor growth. We also found that tumor cells secrete sonic hedgehog (SHH), an Hh ligand, and that tumor-derived SHH drives TAM M2 polarization. Furthermore, Hh-induced functional polarization in TAMs suppresses CD8(+) T cell recruitment to the TME through the inhibition of CXCL9 and CXCL10 production by TAMs. Last, we demonstrated that Krüppel-like factor 4 (Klf4) mediates Hh-dependent TAM M2 polarization and the immunosuppressive function. Collectively, these findings highlight a critical role for tumor-derived SHH in promoting TAM M2 polarization, a mechanism for TAM-mediated immunosuppression, and may provide insights into the design of new cancer immunotherapeutic strategies.

Manganese (Mn), an essential micronutrient, has the potential to induce apoptosis. The NT3/TrkC ligand/receptor pair known as part of the classic neurotrophic theory plays a critical role in neuronal survival. However, whether the NT3/TrkC-mediated signaling pathways are involved in Mn-induced apoptosis of cortical neurons remains unknown. The present study was designed to investigate the interactions between NT3/TrkC-mediated signaling pathways and Mn-induced apoptosis in cortical neurons. This study showed that subacute Mn exposure significantly increased the levels of pro-apoptotic Bax while decreasing the levels of anti-apoptotic Bcl 2 in the cortex compared with the corresponding control. Markedly reduced NT3 and TrkC levels along with decreased Ras/MAPK and PI3/Akt signaling in the cortex were observed following subacute Mn exposure. We further found increased levels of Bax, cleaved caspase-3, and the total apoptosis rate, and decreased levels of Bcl 2, NT3, TrkC, and Ras/MAPK and PI3/Akt signaling in Mn-treated primary cortical neurons. Pretreatment with hNT3 or Z-VAD-FAM ameliorated Mn-induced apoptosis by increasing the levels of NT3 and TrkC and its Ras/MAPK and PI3/Akt signaling pathways. Taken together, our findings clearly indicate that NT3/TrkC and mediated Ras/MAPK and PI3/Akt signaling pathways play a crucial role in Mn-induced neurotoxicity.

Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track tumor development and quantify tumor burden in vivo. For this purpose, the introduction of genes that confer tumors with bioluminescent properties has been a critical advance for oncologic studies in rodents. Methods of introducing bioluminescent genes, such as firefly luciferase, by viral transduction has allowed for the production of tumor cell lines that can be followed in vivo longitudinally over long periods of time. Here we describe methods for the production of stable luciferase expressing tumor cell lines by lentiviral transduction.

Vildagliptin promotes beta cell survival by inhibiting cell apoptosis. It has been suggested that chronic ER (endoplasmic reticulum) stress triggers beta cell apoptosis. The objective of the study is to explore whether the pro-survival effect of vildagliptin is associated with attenuation of endoplasmic reticulum stress in islets of db/db mice.Vildagliptin was orally administered to db/db mice for 6 weeks, followed by evaluation of beta cell apoptosis by caspase3 activity and TUNEL staining method. Endoplasmic reticulum stress markers were determined with quantitative RT-PCR, immunohistochemistry and immunoblot analysis.After 6 weeks of treatment, vildagliptin treatment increased plasma active GLP-1 levels (22.63±1.19 vs. 11.69±0.44, P0.001), inhibited beta cell apoptosis as demonstrated by lower amounts of TUNEL staining nuclei (0.37±0.03 vs. 0.55±0.03, P0.01) as well as decreased caspase3 activity (1.48±0.11 vs. 2.67±0.13, P0.01) in islets of diabetic mice compared with untreated diabetic group. Further, vildagliptin treatment down-regulated several genes related to endoplasmic reticulum stress including TRIB3 (tribbles homolog 3) (15.9±0.4 vs. 33.3±1.7, ×10⁻³, P0.001), ATF-4(activating transcription factor 4) (0.83±0.06 vs. 1.42±0.02, P0.001) and CHOP(C/EBP homologous protein) (0.07±0.01 vs. 0.16±0.01, P0.001).Vildagliptin promoted beta cell survival in db/db mice in association with down-regulating markers of endoplasmic reticulum stress including TRIB3, ATF-4 as well as CHOP.

IL-21 has been shown to play an important role in the CD8 T cell response during acute and chronic viral infections. However, the role of IL-21 signaling in the CD4 T cell response to viral infection remains incompletely defined. In a model of infection with vaccinia virus, we show that intrinsic IL-21 signaling on CD4 T cells was critical for the formation of memory CD4 T cells in vivo. We further reveal that IL-21 promoted CD4 T cell survival in a mechanism dependent on activation of the STAT1 and STAT3 signaling pathways. In addition, the activation of Akt is also required for IL-21-dependent survival of CD4 T cells in vivo. These results identify a critical role for intrinsic IL-21 signaling in CD4 T cell survival and memory formation in response to viral infection in vivo and may provide insights into the design of effective vaccine strategies.

Tumor-associated macrophages (TAMs), representing most of the leukocyte population in solid tumors, demonstrate great phenotypic heterogeneity and diverse functional capabilities under the influence of the local tumor microenvironment. These anti-inflammatory and protumorigenic macrophages modulate the local microenvironment to facilitate tumor growth and metastasis. In this review, we examine the origin of TAMs and the complex regulatory networks within the tumor microenvironment that facilitate the polarization of TAMs toward a protumoral phenotype. More extensively, we evaluate the mechanisms by which TAMs mediate angiogenesis, metastasis, chemotherapeutic resistance and immune evasion. Lastly, we will highlight novel interventional strategies targeting TAMs in preclinical studies and in early clinical trials that have significant potential in improving efficacy of current chemotherapeutic and/or immunotherapeutic approaches.

NK cells are important for the control of vaccinia virus (VV) in vivo. Recent studies have shown that multiple pathways are required for effective activation of NK cells. These include both TLR-dependent and -independent pathways, as well as the NKG2D activating receptor that recognizes host stress-induced NKG2D ligands. However, it remains largely unknown what controls the upregulation of NKG2D ligands in response to VV infection. In this study using C57BL/6 mice, we first showed that IL-18 is critical for NK-cell activation and viral clearance. We then demonstrated that IL-18 signaling on both NK cells and DCs is required for efficient NK-cell activation upon VV infection in vitro. We further showed in vivo that efficient NK-cell activation in response to VV is dependent on DCs and IL-18 signaling in non-NK cells, suggesting an essential role for NK cell-extrinsic IL-18 signaling in NK-cell activation. Mechanistically, IL-18 signaling in DCs promotes expression of Rae-1, an NKG2D ligand. Collectively, our data reveal a previously unrecognized role for NK cell-extrinsic IL-18 signaling in NK-cell activation through upregulation of NKG2D ligands. These observations may provide insights into the design of effective NK-cell-based therapies for viral infections and cancer.

Viral hepatitis remains a global health challenge despite recent progress in the development of more effective therapies. Although virus-specific CD8+ and CD4+ T cell responses are essential for viral clearance, it remains largely unknown what regulates T cell-mediated viral clearance. Thus, a better understanding of the regulation of anti-viral T cell immunity would be critical for the design of more effective therapies for viral hepatitis. Using a model of adenovirus-induced hepatitis, here we showed that adenoviral infection induced recruitment of Ly6Chi monocytes to the liver in a CCR2-dependent manner. These recruited Ly6Chi monocytes suppressed CD8+ and CD4+ T cell responses to adenoviral infection, leading to a delay in viral clearance. In vivo depletion of Ly6Chi monocytes markedly enhanced anti-viral T cell responses and promoted viral clearance. Mechanistically, we showed that induction of iNOS and the production of NO by Ly6Chi monocytes are critical for the suppression of T cell responses. In addition, a contact-dependent mechanism mediated by PD-1 and PD-L1 interaction is also required for T cell suppression by Ly6Chi monocytes. These findings suggest a critical role for Ly6Chi monocytes in the regulation of T cell immunity in viral hepatitis and may provide new insights into development of more effective therapies for treating viral hepatitis based on targeting the immunosuppressing monocytes.

Here we present the first detailed planktonic and benthic δ13C records and benthic foraminiferal assemblage records from the north-eastern Indian Ocean to decipher the palaeoceanographic changes during the last 56 kyr. We identified three different palaeoceanographic stages, clearly differentiated by significant variations in the benthic foraminiferal assemblages and δ13C records. The results of this study indicate that productivity was generally higher during the glacial periods than during the Holocene. Comparison of the benthic foraminiferal assemblage distributions and planktonic and benthic δ13C records show a significant correlation between productivity and the bottom water oxygenation on glacial-interglacial timescales. Productivity gradually increased during the period between 56 and 27.5 kyr. During this period, the dominance of Melonis spp. and Oridosalis umbonatus were correlated with conditions of intermediate to high surface productivity and moderate bottom water oxygenation. Increased higher equatorial productivity and low bottom water oxygenation during the period between 27.5 and 15 kyr are supported by planktonic δ13C and faunal records. During this period, the dominance of benthic foraminifera assemblages characterised by Uvigerina peregrina indicates sustained continuous phytodetritus flux to the seafloor from enhanced surface water productivity and relatively low bottom water oxygenation. The absence or minimal occurrences of high productivity indicating U. peregrina, the dominance of intermediate to low productivity indicating fauna, and relatively low planktonic δ13C records suggest low productivity and active deep-water oxygenation after 15 kyr. Concurrent river discharge and rising sea levels during this period are indicated by negative δ18O of G. ruber at the site.

Graft-versus-host disease (GVHD) remains the most common cause of nonrelapse-related morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although T-cell depletion and intensive immunosuppression are effective in the control of GVHD, they are often associated with higher rates of infection and tumor recurrence. In this study, we showed that heparan sulfate (HS), an extracellular matrix component, can activate Toll-like receptor 4 on dendritic cells in vitro, leading to the enhancement of dendritic cell maturation and alloreactive T-cell responses. We further demonstrated in vivo that serum HS levels were acutely elevated at the onset of clinical GVHD in mice after allo-HSCT. Treatment with the serine protease inhibitor α1-antitrypsin decreased serum levels of HS, leading to a reduction in alloreactive T-cell responses and GVHD severity. Conversely, an HS mimetic that increased serum HS levels accelerated GVHD. In addition, in patients undergoing allo-HSCT for hematologic malignancies, serum HS levels were elevated and correlated with the severity of GVHD. These results identify a critical role for HS in promoting acute GVHD after allo-HSCT, and they suggest that modulation of HS release may have therapeutic potential for the control of clinical GVHD.

Natural CD4+CD25+Foxp3+ regulatory T cells (Tregs) are critical for the control of immune responses to pathogens. However, most studies have focused on chronic infections, in which pathogen-specific Tregs contribute to pathogen persistence and, in some cases, concomitant immunity. How Tregs behave and function following acute infections remains largely unknown. In this article, we show that pathogen-specific Tregs can be activated and expand upon acute viral infections in vivo. The activated Tregs then contract to form a memory pool after resolution of the infection. These memory Tregs expand rapidly upon a secondary challenge, secrete large amounts of IL-10, and suppress excessive immunopathological conditions elicited by recall expansion of non-Tregs via an IL-10–dependent mechanism. Our work reveals a memory Treg population that develops after acute viral infections and may help in the design of effective strategies to circumvent excessive immunopathological effects.

CD4 T cell help plays an important role in promoting CD8 T cell immunity to pathogens. In models of infection with vaccinia virus (VV) and Listeria monocytogenes, CD4 T cell help is critical for the survival of activated CD8 T cells during both the primary and memory recall responses. Still unclear, however, is how CD4 T cell help promotes CD8 T cell survival. In this study, we first showed that CD4 T cell help for the CD8 T cell response to VV infection was mediated by IL-21, a cytokine produced predominantly by activated CD4 T cells, and that direct action of IL-21 on CD8 T cells was critical for the VV-specific CD8 T cell response in vivo. We next demonstrated that this intrinsic IL-21 signaling was essential for the survival of activated CD8 T cells and the generation of long-lived memory cells. We further revealed that IL-21 promoted CD8 T cell survival in a mechanism dependent on activation of the STAT1 and STAT3 pathways and subsequent upregulation of the prosurvival molecules Bcl-2 and Bcl-xL. These results identify a critical role for intrinsic IL-21 signaling in CD8 T cell responses to an acute viral infection in vivo and may help design effective vaccine strategies.

Natural killer (NK) cells, as part of the innate immune system, play a key role in host defense against viral infections. Recent advances have indicated that NK cell activation and function are regulated by the interplay between inhibitory and activating signals. Thus, a better understanding of mechanisms responsible for NK cell activation and function in the control of viral infections will help develop NK cell-based therapies. In this review, we will first discuss how NK cells are activated in response to viral infections. We will then focus on the recruitment of activated NK cells to the site of infection as well as on NK cell effector mechanisms against virally infected cells.

Despite high transduction efficiency in vivo, the application of recombinant E1-deleted adenoviral vectors for in vivo gene therapy has been limited by the attendant innate and adaptive immune responses to adenoviral vectors. NK cells have been shown to play an important role in innate immune elimination of adenoviral vectors in vivo. However, the mechanisms underlying NK cell activation and function in response to adenoviral vectors remain largely undefined. In this study, we showed that NK cell activation upon adenoviral infection was dependent on accessory cells such as dendritic cells and macrophages and that cell contact-dependent signals from the accessory cells are necessary for NK cell activation. We further demonstrated that ligands of the NK activating receptor NKG2D were upregulated in accessory cells upon adenoviral infection and that blockade of NKG2D inhibited NK cell activation upon adenoviral infection, leading to a delay in adenoviral clearance in vivo. In addition, NKG2D was required for NK cell-mediated cytolysis on adenovirus-infected targets. Taken together, these results suggest that efficient NK cell activation and function in response to adenoviral infection is critically dependent on the NKG2D pathway, which understanding may assist in the design of effective strategies to improve the outcome of adenovirus-mediated gene therapy.

Naturally occurring regulatory T cells (T(Reg)) suppress multiple cell types of the immune system to maintain dominant tolerance to protect from autoimmunity, down-modulate anti-tumor immunity and restrain allergic diseases. In addition to these functions, T(Reg) can alter effector responses to invading pathogens, leading to a variety of outcomes affecting both the host and infecting microorganisms. Here, we review how T(Reg) can influence the immune responses to chronic infections where pathogen-specific T(Reg) can contribute to pathogen persistence and, in some cases, concomitant immunity, as well as control immunopathology associated with robust immune responses. We also review the data on T(Reg) during acute infection, focusing on the questions these studies raise regarding the most appropriate model(s) to examine T(Reg) during infection. Finally, we discuss the ways in which the T(Reg) function can be altered by invading pathogens and how these can be exploited to develop methods therapeutically to influence disease and vaccine outcomes.

After hematopoietic allogeneic stem cell transplantation (HCT), B cells reconstitute in the presence of alloantigen in a nucleic acid (NA)-rich environment. Ongoing tissue and cellular damage results in release of endogenous NA that may serve as ligand for endosomal Toll-Like Receptors (TLRs). Alloantibodies and autoantibodies, including NA-binding antibodies, have been associated with the presence of cGVHD manifestations. In patients after HCT, B cells are constitutively activated via key proximal B Cell Receptor (BCR) signaling molecules, Syk and BLNK (Allen JL et al Blood 2014, Flynn R et al Blood 2015) . In mouse models of autoimmunity, TLR7 or TLR9 can operate in conjunction with the BCR to mediate autoantibody production (Suthers A et al. Front Immunol. 2017). Whether TLR7/9-BCR signaling contributes to cGVHD remains unknown. We studied responses by TLR9 and TLR7 in B cells from patients >12 months post-HCT who had active, inactive or no cGVHD at the time of sample acquisition and were not receiving high dose steroid. In contrast to a previous study (She K et al. BBMT 2007) and consistent with findings by others (de Masson A et al Blood 2015), we did not find a consistent increase in B cell response to TLR9 agonist. Instead we found significantly higher TLR7 transcript expression in B cells from patients with active cGVHD (n=7) compared to no/inactive cGVHD patient B cells (n=11, p=0.042). TLR7 over-expression in mice leads to anti-RNA antibody production by immature B cells (Giltiay NV et al JEM 2013). We employed ELISA to measure antibodies to a known RNA-containing autoantigen, Ro-52 in a group of 66 plasma samples. We found that significantly more patients with active cGVHD (n=32) had anti-Ro-52 antibody compared to inactive cGVHD patients (n=22) or patients without cGVHD (n=12) (Fisher exact test for both comparisons = p To determine if TLR7 activation was increased in cGVHD B cells, we measured IL-6 production. Without ex vivo stimulation, cGVHD patient B cells produced significantly more IL-6 (p=0.039) (Fig.1A). After TLR7 stimulation with R848, B cells from patients with active cGVHD had significantly increased IL-6 production compared those from patients without (p= 0.008) or those with inactive cGVHD (p=0.017) (Fig.1A). We next investigated the influence of the BCR-activated state in cGVHD on TLR7 responses and signaling. Using flow cytometry, we measured Ki-67 expression as a marker of activation after ex vivo stimulation with both low level surrogate BCR antigen and R848. We found that B cells from active cGVHD patients were significantly more responsive compared to no cGVHD patients (p=0.015) and healthy donors (p=0.045), revealing increased synergistic BCR-TLR7 signaling in cGVHD B cells (Fig.1B). To further investigate this, we employed qPCR to measure levels of the proximal BCR signaling molecule, Lyn, known to negatively regulate anti-NA antibody production (Lamagna et al. J. Immunol. 2014). We found that Lyn expression was significantly decreased in B cells from active cGVHD patients compared to healthy donor (p=0.001), inactive (p=0.042) or no (p=0.007) cGVHD patients (Fig.1C). Lyn can also negatively regulate TLR7/9 activation by directly associating with Interferon Regulatory Factor 5 (IRF5), which is important for pro-inflammatory cytokine production, including IL-6 (Ban et al, J.Immunol. 2016). Consistent with a role for Lyn in TLR7 activation, we found that IRF5 was significantly increased in active cGVHD patient B cells compared to all other groups (healthy donor, p=0.035; inactive cGVHD, p=0.011; no cGVHD, p=0.006) (Fig.1D). Notably, no difference in expression was found in downstream TLR7 transcription factors, IRF3 and IRF7 . Using a mouse model (Zhang C et al Blood 2006), we found significantly higher IRF5 expression in splenic B cells in animals with cGVHD manifestations (p=0.005). Ongoing mouse work will ascertain mechanistic roles for Lyn and IRF5 in TLR7 signaling by cGVHD B cells. Together data support a potentially pathogenic role of TLR7 signaling in cGVHD B cells. Development of agents that block this newly elucidated TLR7-BCR signaling axis in cGVHD is warranted. This work was supported by National Institutes of Health grant R01 HL129061 Figure 1 Figure 1. Disclosures Rizzieri: Erytech: Research Funding; Shire: Research Funding.

Recombinant adeno-associated viruses (AAVs) have been used widely for in vivo gene therapy. However, adaptive immune responses to AAV have posed a significant hurdle in clinical application of AAV vectors. Recent advances have suggested a crucial role for innate immunity in shaping adaptive immune responses. How AAV activates innate immunity, and thereby promotes AAV-targeted adaptive immune responses, remains unknown. Here we show that AAV activates mouse plasmacytoid DCs (pDCs) via TLR9 to produce type I IFNs. In vivo, the TLR9-MyD88 pathway was crucial to the activation of CD8+ T cell responses to both the transgene product and the AAV capsid, leading to loss of transgene expression and the generation of transgene product-specific and AAV-neutralizing antibodies. We further demonstrate that TLR9-dependent activation of adaptive immunity targeting AAV was mediated by type I IFNs and that human pDCs could be activated in vitro to induce type I IFN production via TLR9. These results reveal an essential role for the TLR9-MyD88-type I IFN pathway in induction of adaptive immune responses to AAV and suggest that strategies that interfere with this pathway may improve the outcome of AAV-mediated gene therapy in humans.

Tumor-specific T-cell tolerance represents one major mechanism of tumor-induced immune evasion. Myeloablative chemotherapy with stem cell transplantation may offer the best chance of achieving a state of minimal residual disease and, thus, minimize tumor-induced immune evasion. However, studies have shown that tumor-specific T-cell tolerance persists after transplantation. Here, we showed that CD4+CD25+ regulatory T (TReg) cells play a critical role in tumor-specific CD8+ T-cell tolerance after transplantation. Removal of TReg cells from the donor lymphocyte graft did not overcome this tolerance because of rapid conversion of donor CD4+CD25− T cells into CD4+CD25+Foxp3+ TReg cells in recipients after transplantation, and depletion of TReg cells in recipients was necessary for the reversal of tumor-specific tolerance. These results suggest that strategies capable of overcoming T-cell tolerance in recipients are required to promote antitumor immunity after transplantation. Toward this goal, we showed that dendritic cell (DC) vaccines coadministered with the TLR9 ligand, CpG could effectively overcome tumor-specific tolerance, leading to significant prolongation of tumor-free survival after transplantation. We further showed that CpG-induced type I interferon was critical for the reversal of tumor-specific tolerance in vivo. Collectively, these results may suggest effective immunotherapeutic strategies for treating cancer after stem cell transplantation.

Recombinant viral vectors such as adenovirus and adenovirus-associated virus have been used widely as vehicles for gene therapy applications because of the high efficiency with which they transfer genes into a wide spectrum of cells in vivo. However, enthusiasm for the use of viral vectors in gene therapy has been tempered by significant problems of attendant host cellular and humoral immune responses that limit their safety and efficacy in vivo. Advances in immunology have suggested a crucial role for the innate immune system in the induction of immune responses to viruses. Thus, a better understanding of the mechanisms by which the host's innate immune system recognizes viruses and viral vectors will help in the design of effective strategies to improve the outcome of viral vector-mediated gene therapy. In this review we first discuss our current understanding of innate immune recognition of viruses in general, and then focus on the innate immune responses to viral vectors for gene therapy.

Recent advances have suggested a crucial role of the innate immunity in shaping adaptive immune responses. How activation of innate immunity promotes adaptive T-cell responses to pathogens in vivo is not fully understood. It has been thought that Toll-like receptor (TLR)–mediated control of adaptive T-cell responses is mainly achieved by the engagement of TLRs on antigen-presenting cells to promote their maturation and function. In this study, we showed that direct TLR2–myeloid differentiating factor 88 (MyD88) signaling in CD8 T cells was also required for their efficient clonal expansion by promoting the survival of activated T cells on vaccinia viral infection in vivo. Effector CD8 T cells that lacked direct TLR2-MyD88 signaling did not survive the contraction phase to differentiate into long-lived memory cells. Furthermore, we observed that direct TLR2 ligation on CD8 T cells promoted CD8 T-cell proliferation and survival in vitro in a manner dependent on the phosphatidylinositol 3-kinase (PI3K)–Akt pathway activation and that activation of Akt controlled memory cell formation in vivo. These results identify a critical role for intrinsic TLR2-MyD88 signaling and PI3K-Akt pathway activation in CD8 T-cell clonal expansion and memory formation in vivo and could lead to the development of new vaccine approaches.

Recombinant adenoviruses have been used widely for gene therapy due to their high transduction efficiency in vivo. However, the attendant innate immune response to adenoviral vectors has limited their applications for in vivo gene therapy. Recent studies have shown that adenoviruses activate the innate immunity through both Toll-like receptor-dependent (TLR-dependent) and TLR-independent pathways, leading to the production of type I interferons (IFNs) and other inflammatory cytokines. Furthermore, type I IFNs play a pivotal role in innate immune elimination of adenoviral vectors in vivo. It remains to be defined how type I IFNs regulate innate immune clearance of adenoviral vectors. In this study, we showed in vivo that natural killer (NK) cells were activated and accumulated in the liver upon intravenous administration of adenoviral vectors, leading to the loss of adenoviral genome and the reduction of transgene expression. We further demonstrated that type I IFNs were critical for the activation of NK cells. This was achieved by direct action of type I IFNs on NK cells. Overall, our observations reveal a critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo and may help design effective strategies to improve the outcome of adenovirus-mediated gene therapy.

Recombinant adenoviral vectors have been widely used for gene therapy applications and as vaccine vehicles for treating infectious diseases such as human immunodeficiency virus disease. The innate immune response to adenoviruses represents the most significant hurdle in clinical application of adenoviral vectors for gene therapy, but it is an attractive feature for vaccine development. How adenovirus activates innate immunity remains largely unknown. Here we showed that adenovirus elicited innate immune response through the induction of high levels of type I interferons (IFNs) by both plasmacytoid dendritic cells (pDCs) and non-pDCs such as conventional DCs and macrophages. The innate immune recognition of adenovirus by pDCs was mediated by Toll-like receptor 9 (TLR9) and was dependent on MyD88, whereas that by non-pDCs was TLR independent through cytosolic sensing of adenoviral DNA. Furthermore, type I IFNs were pivotal in innate and adaptive immune responses to adenovirus in vivo, and type I IFN blockade diminished immune responses, resulting in more stable transgene expression and reduction of inflammation. These findings indicate that adenovirus activates innate immunity by its DNA through TLR-dependent and -independent pathways in a cell type-specific fashion, and they highlight a critical role for type I IFNs in innate and adaptive immune responses to adenoviral vectors. Our results that suggest strategies to interfere with type I IFN pathway may improve the outcome of adenovirus-mediated gene therapy, whereas approaches to activate the type I IFN pathway may enhance vaccine potency.

Owing to drug synergy effects, drug combinations have become a new trend in combating complex diseases like cancer, HIV and cardiovascular diseases. However, conventional synergy quantification methods often depend on experimental dose-response data which are quite resource-demanding. In addition, these methods are unable to interpret the explicit synergy mechanism. In this review, we give representative examples of how systems biology modeling offers strategies toward better understanding of drug synergy, including the protein-protein interaction (PPI) network-based methods, pathway dynamic simulations, synergy network motif recognitions, integrative drug feature calculations, and "omic"-supported analyses. Although partially successful in drug synergy exploration and interpretation, more efforts should be put on a holistic understanding of drug-disease interactions, considering integrative pharmacology and toxicology factors. With a comprehensive and deep insight into the mechanism of drug synergy, systems biology opens a novel avenue for rational design of effective drug combinations.

Heparan sulfate (HS) is an essential component of the extracellular matrix (ECM), which serves as a barrier to tumor invasion and metastasis. Heparanase promotes tumor growth by cleaving HS chains of proteoglycan and releasing HS-bound angiogenic growth factors and facilitates tumor invasion and metastasis by degrading the ECM. HS mimetics, such as PG545, have been developed as antitumor agents and are designed to suppress angiogenesis and metastasis by inhibiting heparanase and competing for the HS-binding domain of angiogenic growth factors. However, how PG545 exerts its antitumor effect remains incompletely defined. Here, using murine models of lymphoma, we determined that the antitumor effects of PG545 are critically dependent on NK cell activation and that NK cell activation by PG545 requires TLR9. We demonstrate that PG545 does not activate TLR9 directly but instead enhances TLR9 activation through the elevation of the TLR9 ligand CpG in DCs. Specifically, PG545 treatment resulted in CpG accumulation in the lysosomal compartment of DCs, leading to enhanced production of IL-12, which is essential for PG545-mediated NK cell activation. Overall, these results reveal that PG545 activates NK cells and that this activation is critical for the antitumor effect of PG545. Moreover, our findings may have important implications for improving NK cell–based antitumor therapies.

There has been rising interest in the discovery of novel drug indications because of high costs in introducing new drugs. Many computational techniques have been proposed to detect potential drug-disease associations based on the creation of explicit profiles of drugs and diseases, while seldom research takes advantage of the immense accumulation of interaction data. In this work, we propose a matrix factorization model based on known drug-disease associations to predict novel drug indications. In addition, genomic space is also integrated into our framework. The introduction of genomic space, which includes drug-gene interactions, disease-gene interactions, and gene-gene interactions, is aimed at providing molecular biological information for prediction of drug-disease associations. The rationality lies in our belief that association between drug and disease has its evidence in the interactome network of genes. Experiments show that the integration of genomic space is indeed effective. Drugs, diseases, and genes are described with feature vectors of the same dimension, which are retrieved from the interaction data. Then a matrix factorization model is set up to quantify the association between drugs and diseases. Finally, we use the matrix factorization model to predict novel indications for drugs.

Vaccinia virus (VV) has been used extensively as a vaccine vehicle in the clinical application for infectious diseases and cancer. Previous studies have suggested that the unique potency of VV-based vaccine lies in its effective activation of the innate immune system. However, how VV activates innate immune pathways remains largely unknown. In this study, we showed that VV elicited innate immune response through both Toll-like receptor (TLR)–dependent and –independent pathways. The TLR pathway was mediated by TLR2 and MyD88, leading to the production of proinflammatory cytokines, whereas activation of the TLR-independent pathway resulted in the secretion of IFN-β. More importantly, both TLR-dependent and -independent pathways were required for activating innate and adaptive immunity to VV in vivo. These findings represent the first evidence that innate immune recognition of VV is mediated by TLR2, demonstrate that one pathogen can target both TLR and non-TLR innate immune pathways to work together in achieving efficient activation of host defense, and suggest potential new strategies for the design of effective vaccines.

What controls the fate of the T-cell response remains incompletely defined. Gain of effector function facilitated by costimulation has been thought to be a crucial factor in determining the outcome of the T-cell response, i.e. long-term memory in the presence of costimulation versus tolerance induction in the absence of costimulation. In this study, we show that while costimulation or cognate CD4 helps to promote the acquisition of effector function during the initial phase of the CD8 T-cell response, the fate of effector CD8 T cells is controlled by the duration of subsequent antigenic stimulation. Effector CD8 T cells differentiate into memory cells only after clearance of antigen, whereas in the presence of persistent antigen, effector CD8 T cells are tolerized. Furthermore, protective immunity against tumour cannot develop in the persisting antigen environment. These results suggest that removal of persisting antigen by other means might be a prerequisite for effective immunotherapy in cancer.

Induction of peripheral T cell tolerance is mediated by bone marrow-derived dendritic cells that cross-present self-antigen to self-reactive T cells. The current model for peripheral CD8(+) T cell tolerance is that TCR engagement by self-antigen in the absence of costimulation results in abortive activation without development of effector function. Here we demonstrate in vivo that high-dose self-antigen ("signal 1") can compensate for lack of costimulation ("signal 2"), leading to full activation of and development of effector function by self-reactive T cells. In the setting of low-dose self-antigen, acquisition of effector function by self-reactive T cells is dependent on costimulation via CD40 ligation in vivo. However, gain of effector function in either setting does not prevent eventual tolerance of self-reactive CD8(+) T cells. These results suggest that the mechanisms for peripheral CD8(+) T cell tolerance are more complex than the proposed "signal 1 in the absence of signal 2" hypothesis. Further exploration of these mechanisms will have direct impact on the design of effective immunotherapy for autoimmune diseases, chronic infections and cancers.