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Potent antitumor activity of CCDC134 in vivo.

CCDC134 is a poorly characterized secreted protein that may act as an immune cytokine. Here, we show that CCDC134 is differentially expressed on resting and activated immune cells and that it promotes CD8+ T-cell activation, proliferation, and cytotoxicity by augmenting expression of the T-cell effector molecules IFNγ, TNFα, granzyme B, and perforin. CCDC134 facilitated infiltration of CD8+ T cells with enhanced cytolytic activity into tumors, demonstrating strong antitumor effects in a CD8+ T-cell–dependent manner. Mechanistically, in CD8+ T cells, exposure to CCDC134 promoted cell proliferation through the JAK3–STAT5 pathway, a classic feature of many cytokines of the common γ-chain (γc) cytokine receptor family. Overall, our results provide evidence that CCDC134 may serve as a member of the γc cytokine family and illustrate its potent antitumor effects by augmenting CD8+ T-cell–mediated immunity. Cancer Res; 74(20); 5734–45. ©2014 AACR.

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As the adeno-associated virus (AAV) vectors hold unique advantages over other viral vectors, AAV gene therapy has accumulated rapid progress and development. Liver-targeted gene therapy by AAV vectors has been successfully applied in clinical trials for many diseases. Low transduction efficiency and high prevalence of neutralizing antibodies (Nabs), however, are the major obstacles to further translate this therapeutic strategy into clinical trials. Pre-clinical evaluation on hepatocytes could help to elucidate the tropism of AAV serotypes for liver-targeted gene therapy, and could also provide a test model to develop novel AAV mutants with Nabs evasion and high liver tropism. Here, we described the basic laboratory procedure to apply the AAV vector to transduce human hepatocytes in vitro and in vivo with some tips gained from our own experience.

It is generally believed that the main source of antibodies is B lymphocytes. In this study, our results first revealed that B cell-deficient mice could not produce antibodies specific for TI-Ags; however, mice with B cell deficiency could produce TD-Ag-specific antibodies, although antibody production was delayed compared with that in BALB/c mice after primary TD-Ag challenge. Subsequently, we identified that mouse lung epithelial cells could produce and secrete Ig, including IgM, IgA or IgG, which could display TD-Ag-specific antibody activity. Notably, the production of TD-Ag-specific antibodies by lung epithelial cells was found to be dependent on CD4+ T cells but not CD8+ T cells. Our findings indicate for the first time that B cells are not the only source of TD-Ag-specific antibodies but are essential for rapid TD-Ag-specific antibody production by non-B cells. This discovery may reveal a new mechanism for the production of specific antibodies.

Adeno-associated virus (AAV) vectors have been successfully used in patients with bleeding disorders and blindness. For human liver targeting, two major factors restrict effective AAV transduction after systemic administration of AAV vectors: human hepatocyte tropism and neutralizing antibodies (Nabs). In this study, we attempted to isolate AAV variants with the ability to transduce human hepatocytes and escape Nabs using a directed evolution approach in vivo. After four cycles of selection, 14 AAV capsid mutants were identified from a capsid shuffling library selected in the presence of human Intravenous Immunoglobulin (IVIG) and isolated from human hepatocytes xenografted into chimeric mice. AAV neutralization assays using IVIG showed that most of the mutants showed the Nab escape pattern in a manner similar to that of AAV8 or AAV9 and better than that of other AAV serotypes. Different mutants displayed varying capacities to escape Nab activity from individual serum samples collected from healthy subjects or hemophilia patients. The mutant AAV LP2-10 was found in 12 colonies out of 25, which was composed of capsids from AAV serotypes 2, 6, 8, and 9, with VP3 subunits derived from AAV8 swapped with AAV6 from residues 261 to 272. The mutant AAV LP2-10 manifested a higher ability than that of other serotypes to escape Nabs in IVIG and most human serum samples. After injection of AAV vectors encoding a self-complementary GFP cassette into chimeric mice, LP2-10 transduced human hepatocytes with efficiency similar to that of AAV8. In summary, AAV mutants can be isolated in humanized mice with both human hepatocyte tropism and the ability to evade Nab activity., Graphical Abstract, AAV mutant vectors were identified from a capsid shuffling library selected in the presence of human IVIG and isolated from human hepatocytes xenografted into chimeric mice. LP2-10, whose VP3 subunits were derived from AAV8 and AAV6, manifested the highest ability to escape Nabs in IVIG and most human serum samples.

The rapid spread of COVID-19 results in a pandemic throughout the world, however, there are currently no specific treatments available. We report the first case of ozonated autohemotherapy for a critically ill patient with COVID-19. The patient was diagnosed with severe acute respiratory distress syndrome (ARDS) and life-threatening refractory hypoxemia within 72 hours of the intensive-care unit (ICU) admission. To improve the oxygen delivery, the ozonated autohemotherapy was performed with 40 µg/ml of ozone in 100 ml of blood for 5 days on this patient, who then recovered from ARDS uneventfully and discharged from hospital after viral clearance. This case suggests ozonated autohemotherapy might be an alternative non-invasive medical treatment for critically ill COVID-19 patients.

Adeno-associated virus (AAV) vectors have been successfully applied in hemophilia clinical trials. However, this approach is limited to patients without AAV-neutralizing antibodies (NAbs). In this study, we explored the feasibility of AAV re-administration in hemophilia A dogs treated initially 8 years ago with AAV8.canine FVIII. After the re-administration in two NAb-negative dogs with AAV8 vectors carrying human factor VIII (hFVIII), along with the proteasome inhibitor bortezomib, we observed a phenotypic improvement in both dogs that persisted in one dog. Phenotypic improvement disappeared at 59 days after re-administration in the other dog, and specific cytotoxic T lymphocytes (CTLs) to the capsid were detected at day 17, but not to hFVIII. hFVIII inhibitors were observed at day 59 and gradually increased. Mechanistic studies demonstrated an increase in pro-inflammatory cytokines, a decrease in immunomodulatory cytokines, as well as lower Tregs after re-administration. These results suggest that hFVIII inhibitor development may contribute to the therapeutic failure via immune response activation. Interestingly, it takes about 30–50 days for AAV NAb titers to decrease by half. Collectively, this study suggests that re-administration of the same AAV serotype after long-term follow-up is feasible and that the study of AAV NAb kinetics will provide important information for predicating the efficacy of re-administration. Keywords: AAV, hemophilia, neutralizing antibodies, NAbs, re-administration, human factor VIII, hFVIII, hFVIII inhibitor

To date, IgG in the tumor microenvironment (TME) has been considered a product of B cells and serves as an antitumor antibody. However, in this study, using a monoclonal antibody against cancer-derived IgG (Cancer-IgG), we found that cancer cells could secrete IgG into the TME. Furthermore, Cancer-IgG, which carries an abnormal sialic acid modification in the CH1 domain, directly inhibited effector T-cell proliferation and significantly promoted tumor growth by reducing CD4(+) and CD8(+) T-cell infiltration into tumor tissues. Mechanistic studies showed that the immunosuppressive effect of sialylated Cancer-IgG is dependent on its sialylation and binding to sialic acid-binding immunoglobulin-type lectins (Siglecs) on effector CD4(+) and CD8(+) T cells. Importantly, we show that several Siglecs are overexpressed on effector T cells from cancer patients, but not those from healthy donors. These findings suggest that sialylated Cancer-IgG may be a ligand for Siglecs, which may serve as potential checkpoint proteins and mediate tumor immune evasion.

Immunoglobulin (Ig&kappa, ) has been reported to be expressed in sorted liver epithelial cells of &mu, MT mice, and the sequence characteristics of hepatocyte-derived Ig&kappa, were different from those of classical B-cell-derived Ig&kappa, However, the physiological function of hepatocyte-derived Ig&kappa, is still unclear. The expression of Ig&kappa, was firstly identified in primary hepatocytes and normal liver cell line (NCTC1469), and hepatocyte-derived Ig&kappa, expression was elevated and displayed unique localization in hepatocytes of concanavalin A (ConA)-induced hepatitis model. Moreover, Ig&kappa, knockout mice were more sensitive to ConA-induced hepatitis and had higher serum aspartate aminotransferase (AST) levels, more severe histological injury and a greater number of terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL)-positive cells as compared with littermate controls. Furthermore, knockdown of Ig&kappa, in primary hepatocytes and NCTC1469 cells led to accelerated activation of the mitochondrial death pathway and caspase-3 cleavage in vitro, which might be related to inhibition of NF-&kappa, B signaling pathway and activation of JNK via the cytoskeleton dynamics. Taken together, these results indicate that hepatocyte-derived Ig&kappa, mediates cellular resistance to ConA-induced liver injury by inhibiting activation of caspase-3 and the mitochondrial death pathway, suggesting that Ig&kappa, plays an important role in hepatocyte survival and exerts a protective effect against ConA-induced liver injury in mice.

We report a case series of ozonated autohemotherapy of four COVID-19 patients, classified as critically ill (1 patient), severe (1 patient), and moderate (2 patients). Each ozonated autohemotherapy treatment was performed at a concentration of 40 μg/ml of ozone per 100 ml of blood. The number of treatments varied from 1 to 9 depending on the disease severity. All 4 patients, including 1 critically ill patient with severe acute respiratory distress syndrome (ARDS) and life-threatening refractory hypoxemia, recovered uneventfully and were discharged from the hospital after viral clearance. The younger sibling of the critically ill patient was also diagnosed with COVID-19 and developed ARDS with hypoxemia, who received mechanical ventilation through an endotracheal tube and extracorporeal membrane oxygenation (ECMO) support. The overall medical cost for 18 days spent in the intensive care unit (ICU) and 56 days of hospitalization was $139,935 USD. On the other hand, our critically ill patient underwent 9 ozonated autohemotherapy treatments and spent 10 days in the ICU and was discharged on hospital day 30; his hospitalization cost amounted to $15,466.50 USD. This case series suggests that ozonated autohemotherapy may be an alternative noninvasive medical treatment for COVID-19 patients.

Data from clinical trials for hemophilia B using adeno-associated virus (AAV) vectors have demonstrated decreased transgenic coagulation factor IX (hFIX) expression 6-10 weeks after administration of a high vector dose. While it is likely that capsid-specific cytotoxic T lymphocytes eliminate vector-transduced hepatocytes, thereby resulting in decreased hFIX, this observation is not intuitively consistent with restored hFIX levels following prednisone application. Although the innate immune response is immediately activated following AAV vector infection via TLR pathways, no studies exist regarding the role of the innate immune response at later time points after AAV vector transduction. Herein, activation of the innate immune response in cell lines, primary human hepatocytes, and hepatocytes in a human chimeric mouse model was observed at later time points following AAV vector transduction. Mechanistic analysis demonstrated that the double-stranded RNA (dsRNA) sensor MDA5 was necessary for innate immune response activation and that transient knockdown of MDA5, or MAVS, decreased IFN-�� expression while increasing transgene production in AAV-transduced cells. These results both highlight the role of the dsRNA-triggered innate immune response in therapeutic transgene expression at later time points following AAV transduction and facilitate the execution of effective strategies to block the dsRNA innate immune response in future clinical trials.

// Qinyuan Liao 1, 2, * , Wei Liu 1, 2, * , Yang Liu 1, 2, * , Fulin Wang 4 , Chong Wang 1, 2 , Jingxuan Zhang 3 , Ming Chu 1, 2 , Dongyang Jiang 1, 2 , Lin Xiao 1, 2 , Wenwei Shao 1, 2 , Zhengzuo Sheng 1 , Xia Tao 5 , Lei Huo 6 , C. Cameron Yin 7 , Youhui Zhang 8 , Gregory Lee 9 , Jing Huang 1, 2 , Zihai Li 10 , Xiaoyan Qiu 1, 2, 3 1 Department of Immunology, School of Basic Medical Sciences, Peking University, Beijing, 100191, China 2 Peking University Center for Human Disease Genomics, Beijing, 100191, China 3 Key Laboratory of Medical Immunology, Ministry of Health, Beijing, 100191, China 4 Department of Pathology, Chinese PLA General Hospital, Beijing, 100853, China 5 Department of Gynecology, Peking University First Hospital, Beijing, 100034, China 6 Division of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, 77030, USA 7 Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, 77030, USA 8 Department of Immunology, Cancer Institute & Hospital, Chinese Academy of Medical Science, Beijing, 100021, China 9 Andrology Lab, University of British Columbia Centre for Reproductive Health, Vancouver, BC V5Z 4H4, Canada 10 Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, SC 29425, USA * These authors have contributed equally to this work Correspondence to: Xiaoyan Qiu, e-mail: qiuxy@bjmu.edu.cn Keywords: IgG, RP215, epithelial stem/progenitor-like cells, tumor metastasis Received: July 05, 2015 Accepted: October 02, 2015 Published: October 12, 2015 ABSTRACT High expression of immunoglobulin G (IgG) in many non-B cell malignancies and its non-conventional roles in promoting proliferation and survival of cancer cells have been demonstrated. However, the precise function of non-B IgG remains incompletely understood. Here we define the antigen specificity of RP215, a monoclonal antibody that specifically recognizes the IgG in cancer cells. Using RP215, our study shows that IgG is overexpressed in cancer cells of epithelial lineage, especially cells with cancer stem/progenitor cell-like features. The RP215-recognized IgG is primarily localized on the cell surface, particularly lamellipodia-like structures. Cells with high IgG display higher migration, increased invasiveness and metastasis, and enhanced self-renewal and tumorgenecity ability in vitro and in vivo . Importantly, depletion of IgG in breast cancer leads to reduced adhesion, invasion and self-renewal and increased apoptosis of cancer cells. We conclude that high expression of IgG is a novel biomarker of tumor progression, metastasis and cancer stem cell maintenance and demonstrate the potential therapeutic benefits of RP215-recognized IgG targeted strategy.

The impact of reservoirs on downstream river channel change has been a scientific issue in fluvial geomorphology during the last few decades. However, it is still a difficult issue as to how to express quantitatively the channel adjustment in the Inner Mongolian reach of the Yellow River induced by the joint operation of upstream reservoirs. Based on the shape parameters of channel cross-sections at four gauging stations in this river reach over a flooding season in two periods, 1978–1982 and 2008–2012, the present work investigated the channel changes in terms of shape parameter change rate under the same controlling water level in each flooding season at the channel cross-sections. Results showed that most of the change rates of the parameters evidently increased over a flooding season in both periods. However, the change rate of each parameter at the cross-sections decreased evidently in the latter period, compared with the former period. At the same time, the distribution pattern of the change rate of the shape parameters along the cross-sections thus changed from a convex curve in the former period to an S-shaped curve in the latter period. The obvious decrease of the change rates is related to the joint operation of the Liujiaxia and Longyangxia reservoirs. The reservoirs stored a large volume of water and decreased the peak discharge and maximum velocity in the flooding season; as a result, the erosion ability of the flood decreased accordingly. With the joint operation of the large reservoirs, the Inner Mongolian channel shrunk markedly. Therefore, the channel will present the possibility of an extreme flood in the future. Consequently, it is reasonable to adjust the function of the reservoirs in future. The total water and sediment discharges and the peak discharge in flooding seasons should be effectively controlled. Continuous shrinkage of the channel can thus be avoided and it can be ready for a potential extreme flood.

Aims Cancer cell-derived immunoglobulin (Ig)G (cancer-IgG) has been found to be involved in the pathogenesis and progression of many cancers, including lung cancer. The aim of the present study was to investigate the relationship between cancer-IgG expression in lung adenocarcinoma (ADC) and clinicopathological characteristics and clinical outcome. Methods and results Immunohistochemical analysis was performed using an RP215 monoclonal antibody to determine cancer-IgG expression in 140 lung ADC patients. Cell migration and invasion were analysed in A549 cell line after short interfering RNA (siRNA) knockdown of IgG and cell sorting by flow cytometry. Our results show that RP215 immunostaining score is correlated significantly with local invasion (P

CCDC134 is a poorly characterized secreted protein that may act as an immune cytokine. Here, we show that CCDC134 is differentially expressed on resting and activated immune cells and that it promotes CD8+ T-cell activation, proliferation, and cytotoxicity by augmenting expression of the T-cell effector molecules IFNγ, TNFα, granzyme B, and perforin. CCDC134 facilitated infiltration of CD8+ T cells with enhanced cytolytic activity into tumors, demonstrating strong antitumor effects in a CD8+ T-cell–dependent manner. Mechanistically, in CD8+ T cells, exposure to CCDC134 promoted cell proliferation through the JAK3–STAT5 pathway, a classic feature of many cytokines of the common γ-chain (γc) cytokine receptor family. Overall, our results provide evidence that CCDC134 may serve as a member of the γc cytokine family and illustrate its potent antitumor effects by augmenting CD8+ T-cell–mediated immunity. Cancer Res; 74(20); 5734–45. ©2014 AACR.

Using DEM and topographic and geological maps alongside field surveys, this study investigated the characteristics and retreat processes of the knickpoints along the mainstream and major tributaries of the Suoshui watershed. Seven disequilibrium transient knickpoints are found along the mainstream, and 0-6 are on the trunk of each of the major tributaries. The knickpoints are labeled from KP0 to KP7 from the lowest to the highest according to their spatial position. The average drop across each knickpoint is in the range of 9-19 m, and the cumulative fall from KP7 to KP0 is about 107 m. Using a brute force two-parameter search, the coefficient m in the stream power incision model for the Suoshui watershed is found to be 0.525, and the coefficient K is variable with an inverse relationship with channel steepness. Using the stream power incision model with these parameters, the time for the highest knickpoint (KP7) to retreat from the mouth of the Suoshui River to its present location is found to be ca. 0.454 Ma. Based on the relationship between the cumulative fall and migration time of knickpoints, the average downcutting rate of the Suoshui River is estimated to be 0.224 mm/a in the past 0.454 Ma. By comparing the downcutting rates with the height of typical sandstone pillars in the Suoshui watershed, the formation of these pillars might commence in the early Pleistocene. (C) 2014 Elsevier B.V. All rights reserved.

In recent years, climate change and anthropogenic activities have threatened the water supply in the middle reaches of the Yellow River, prompting this study into the variation of water resources and the influencing factors, taking the Huangfuchuan basin as an example. Firstly, changes in climatic aridity and annual runoff in the Huangfuchuan basin from 1956 to 2009 were analysed. Then, the influence of changes in climatic aridity, water use for irrigation and soil conservation measures were calculated using an analysis of principal components regression. The results show that climatic aridity has increased in the recent three decades with two abrupt changes around 1961 and 1998, and that annual runoff has decreased continually with two abrupt changes around 1979 and 1999. The rapid development of sediment check dams in the 1970s could be the reason for the abrupt change around 1979. The abrupt change around 1999 could be the result of both the intensification of changes in climatic aridity and the large-scale construction of water and soil conservation measures after 1983, the further improvement of these measures after 1993 and ecological restoration measures of converting cropland to forest implemented since 1997. By quantifying the effects of those factors that influence runoff variation, it was found that anthropogenic activities were more important than climate change in the two periods between 1979–1998 and 1999–2006, but the influence of changes in climatic aridity increased from the first to the second period. For the runoff reduction related to anthropogenic activities, the primary cause was water diversion for irrigation in the first period, and it was soil conservation measures in the second period.

article i nfo Suspended sediment dynamics are still imperfectly understood, especially in the upper Yellow River where few studies on this issue have been conducted. This study analyzed the variability in suspended sediment concentration at different temporal scales (within-flood, monthly-seasonal and annual). Using a dataset up to 49 years long (1954-2002) at Toudaoguai station, this paper provided insight into the characteristics of suspended sediment transport in the Ningxia-Inner Mongolia reaches of the upper Yellow River. The results show that, in a single flood event, changes in SSC are not simultaneous with changes in discharge, resulting in counterclockwise hysteretic loops, clockwise loops or figure-eight loops. The C-Q loops and sediment rating curves for different seasons had different characters. The diversiform C-Q loops in the Inner Mongolia reaches of the upper Yellow River may be mainly caused by the sediment inputs from tributaries, the differ- ent propagation velocity of sediment and water discharge, spatial and temporal storage-depletion processes of available sediment. At annual scale, the variations of annual mean discharge and SSC displayed a similar gradually deceasing trend, and the relation between water discharge and SSC during 1954-2002 could be de- scribed by a cubic polynomial function. Since the joint operation of Liujiaxia and Longyangxia reservoirs after 1986, the transport capacity of the Upper Yellow River has been much reduced.

Objectives Hyperplasia of synovial fibroblasts, infiltration with lymphocytes and tissue hypoxia are major characteristics of rheumatoid arthritis (RA). Extensive data support a key role for toll-like receptors (TLRs) in RA. Little is known regarding the impact of hypoxia on TLR-induced inflammation in RA. The aim of this study was to reveal the effects of hypoxia and its regulator, hypoxia-inducible factor-1α (HIF-1α), on the inflammatory response of RA synovial fibroblasts (RASF) to TLR ligands. Methods Hypoxia was induced in RASF by incubation with Na 2 S 2 O 4 . TLR3 ligand polyIC, TLR2 ligand peptidoglycan, TLR4 ligand LPS and TLR9 ligand CpG were used to stimulate the cells. Effects of hypoxia on TLR-induced inflammatory mediators were determined by RT-PCR, qPCR and ELISA. Overexpression of HIF-1α as well as knocking-down its expression was used to reveal its fundamental role. RASF-induced inflammatory T cell expansion was determined by flow cytometry analysis of T helper (Th)1/Th17 cells, and IFN-γ/IL-17 production by ELISA after RASF/T cell coculture. Results Hypoxia potentiated the expression of inflammatory cytokines, metalloproteinases and VEGF in RASF stimulated by different TLR ligands, especially polyIC, a synthetic mimic of dsRNA from viruses or apoptotic cells. HIF-1α played a fundamental role in this synergy. Moreover, HIF-1α overexpression enhanced RASF-mediated expansion of inflammatory Th1 and Th17 cells, leading to proinflammatory IFN-γ and IL-17 production. Conclusions Our findings suggest that hypoxia and HIF-1α may function in conjunction with TLR-stimulated innate immune responses to drive inflammation in RA. This pathway may serve as a therapeutic target for the disease.

Based on the long-term hydrological and meteorological series. DEM, soil database and documents of soil conservation measures of the middle Yellow River, investigations are made on the spatial and temporal changes of runoff and the impact of 11 factors, which include rainfall, temperature, terrain slope, drainage density, gravel, sand, silt, and organic carbon content in soil, water consumption, and soil conservation measures. The results show that the total runoff generated from the middle Yellow River had a decreasing trend in the past 60 years, with two abrupt falls around the years 1971 and 1991. In the spatial dimension, runoff modulus grew from the north to the south and from the west to the east, and the largest gradient of spatial change of runoff modulus happened during the period of 1956-1970 before the first falls of the total runoff. In the period 1954-2009. the average annual rate of temporal change of runoff modulus increased acceleratedly outward from the northwest of the middle Yellow River with a concentric ring pattern. Correlation analysis of runoff modulus with potential influencing factors in the spatial perspective reveals that the spatial distribution of the runoff modulus is principally the results of the regional variation of natural conditions. The results of the correlation analysis of the temporal series of runoff coefficient and influencing factors suggest that climate change, hydraulic engineering and soil conservation measures, were all the major causes of runoff reducing in the second half of the last century in the middle Yellow River. For the whole middle Yellow River, climate change contributed over 40% of the runoff deviations in the 1960s, 1970s and 1990s. while water consumption induced also over 40% runoff reduction in the 1960s. 1970s and 1980s. The runoff reduction due to hydraulic engineering was about 2-3 times of that caused by soil conservation measures, and both of them increased continuously from the 1960s to the 1980s, and kept at a higher level in the 1990s. The contribution of different causes to runoff deviation was variable in different drainage areas. Generally, in the semi-arid areas climate change played a decreasing role in runoff reduction comparing with other causes, while in the semi-humid areas it induced a higher and more variable proportion of runoff deviation according to the decade means. (C) 2012 Elsevier B.V. All rights reserved.

In this study, the relation between water discharge and suspended-sediment concentration (SSC) was analyzed based on daily water discharge and SSC recorded at six stations in the Ningxia-Inner Mongolia reaches of the upper Yellow River in flood seasons from 1952 to 1986. The Pettitt statistical method was used to test the abrupt changes in the time series of discharge and sediment concentration. The results indicate that the time series of discharge in Ningxia-Inner Mongolia reaches of the upper Yellow River can be divided into two periods, i.e., before 1969 and between 1969 and 1986. In these two periods, the third-order polynomial function could be used to describe the relationship between water discharge and SSC during the flood periods, indicating that sediment transport in the Ningxia-Inner Mongolia reaches was influenced by the complex processes of water and sediment inputs from the upstream and tributaries and also by the channel boundary conditions. The relation between SSC and the water discharge of low and moderate floods ( Q 3 /s) was fitted by sediment rating curves of a power function form. The sediment rating parameter b is higher in the upper section of the Ningxia-Inner Mongolia reaches due to a higher stream power there, and both the cross-section shape and river bed material determine the value of log( a ). Although a 40% decrease in sediment transport during the flood periods due to the impoundment of the Liujiaxia Reservoir and sediment-checking dams and the implementation of soil and water conservation programmes in the Ningxia-Inner Mongolia reaches of the upper Yellow River, the sediment transport regimes during 1952–1986 did not change.

Background Immunoglobulins(Igs)play an important role in host defence and were initially thought to be expressed solely by B cells. However, recent data suggest that Igs are also expressed in other lineages. Recently, Ig transcripts were detected in cardiomyocytes, but whether the functional Ig protein is expressed by cardiomyocytes has not been thoroughly elucidated. Methods Gene Expression Omnibus (GEO) microarray database analysis was used to analyse IgM heavy chain expression in the myocardium of mice. Immunohistochemistry (IHC), ELISA and Western blot were used to identify IgM in cardiomyocytes of both Balb/c mice and μMT mice (B cell-deficient mice), as well as in cultured cardiomyocytes of neonatal mice and in the myocardial cell line HL-1. Moreover, RT-PCR and cDNA sequencing were used to determine the VDJ rearrangement of the IgM heavy chain. Results In this study, we first analysed transcription of the IgM heavy chain in heart tissue in mice by mining the GEO database, and we observed that IgM heavy chain transcripts were expressed in heart tissues. Subsequently, IgM was found to be expressed in cardiomyocytes in mice; the IgM was primarily localized on the cell membranes and intercalated discs of murine heart cells and in the cytoplasm and cell membranes of isolated cardiomyocytes and HL-1. Importantly, the functional IgM heavy chain transcripts exhibit a unique VDJ rearrangement pattern. Furthermore, IgM can be secreted and deposited in the extracellular space of the myocardium under ischaemic/hypoxic conditions. Conclusions Our data indicate for the first time that IgM is expressed by cardiomyocytes in mice and suggest that its physiological function warrants further investigation.

Numerous studies have demonstrated that free Ig light chain (FLC), a novel inflammation mediator, participates in many inflammatory diseases by activating mast cells and extending the survival of neutrophils. However, it remains unclear whether FLC is involved in colitis and colitis-associated colon carcinogenesis (CAC). In this study, we found a significant increase in FLC in murine models of DSS (Dextran Sulfate Sodium Salt)-induced colitis and CAC compared to controls. Peptide F991, a functional blocker of FLC, significantly attenuated colitis progression, which included abrogating the development of diarrhea and tumor burden, elevating survival rate, greatly reducing the infiltration of inflammatory cells (such as ROS+ active neutrophils), especially reducing tumorigenesis in CAC. Furthermore, we demonstrated that F991 inhibited the activation of the inflammasome by reducing the expression of cleaved caspase-1 and the maturation of IL-1β and IL-18. Altogether, our findings demonstrate that FLC can promote the pathogenesis of colitis and CAC and may be used as novel biomarker for the diagnosis of inflammatory bowel disease. Additionally, F991 may become a potential therapeutic option for colitis or colorectal cancer.

Growing evidence indicates that B cells are not the only source of immunoglobulin (Ig). To investigate this discovery further, we used μMT mice, which have a disruption of the first transmembrane exon of the μ heavy chain and do not express the membrane form of IgM. These mice lack mature B cells and thus serve as a good model to explore Ig expression by liver epithelial cells. We found that Ig heavy chains (μ, δ, γ and α) and light chains (κ and λ) were expressed in sorted liver epithelial cells of μMT mice. Surprisingly, each heavy chain class showed its respective variable region sequence characteristics in their variable region, instead of sharing the same VDJ usage, which suggests that class switching does not occur in liver epithelial cells. Moreover, the γ and α chains, but not the μ and δ chains, showed mutations in the variable region, thus indicating that different classes of Ig have different activities. Our findings support the concept that non-B cells, liver epithelial cells here, can produce different classes of Ig.

Currently, natural IgM antibodies are considered to be the constitutively secreted products of B-1 cells in mice and humans. In this study, we found that mouse epithelial cells, including liver epithelial cells and small intestinal epithelial cells (IECs), could express IgM that also showed natural antibody activity. Moreover, similar to the B-1 cell-derived natural IgM that can be upregulated by TLR9 agonists (mimicking bacterial infection), the expression of epithelial cell-derived natural IgM could also be significantly increased by TLR9 signaling. More importantly, the epithelial cell-derived IgM was polyreactive, and it could recognize single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), lipopolysaccharide (LPS), and insulin with low affinity; additionally, TLR9 agonists could enhance it in a MyD88-dependent manner. Furthermore, epithelial cell-derived IgM could bind various bacteria; therefore, it could be involved in anti-infection responses. Together, these results highlight the fact that epithelial cells are an important source of natural IgM, in addition to that produced by B-1 cells, and IgM contributes to the innate immune responses in local tissues, further demonstrating that the epithelium is a first line of defense in the protection against invading microbes.

The innate immune system of the skin is thought to depend largely on a multi-layered mechanical barrier supplemented by epidermis-derived antimicrobial peptides. To date, there are no reports of antimicrobial antibody secretion by the epidermis. In this study, we report the expression of functional immunoglobulin G (IgG) and immunoglobulin A (IgA), previously thought to be only produced by B cells, in normal human epidermal cells and the human keratinocyte line HaCaT. While B cells express a fully diverse Ig, epidermal cell-expressed IgG or IgA showed one or two conservative VHDJH rearrangements in each individual. These unique VDJ rearrangements in epidermal cells were found neither in the B cell-derived Ig VDJ databases published by others nor in our positive controls. IgG and IgA from epidermal cells of the same individual had different VDJ rearrangement patterns. IgG was found primarily in prickle cells, and IgA was mainly detected in basal cells. Both epidermal cell-derived IgG and IgA showed potential antibody activity by binding pathogens like Staphylococcus aureus, the most common pathogenic skin bacteria, but the microbial-binding profile was different. Our data indicates that normal human epidermal cells spontaneously express IgG and IgA, and we speculate that these Igs participate in skin innate immunity.

Mammalian IgG and IgE are thought to have evolved from IgY of nonmammalian tetrapods; however, no diversification of IgY subclasses has been reported in reptiles or birds, which are phylogenetically close to mammals. To our knowledge, we report the first evidence of the presence of multiple IgY-encoding (υ) genes in snakes. Two υ genes were identified in the snake Elaphe taeniura, and three υ genes were identified in the Burmese python (Python molurus bivittatus). Although four of the υ genes displayed a conventional four-H chain C region exon structure, one of the υ genes in the Burmese python lacked the H chain C region 2 exon, thus exhibiting a structure similar to that of the mammalian γ genes. We developed mouse mAbs specific for the IgY1 and IgY2 of E. taeniura and showed that both were expressed in serum; each had two isoforms: one full-length and one truncated at the C terminus. The truncation was not caused by alternative splicing or transcriptional termination. We also identified the μ and δ genes, but no α gene, in both snakes. This study provides valuable clues for our understanding of Ig gene evolution in tetrapods.

It is well known that B-1 B cells are the main cell type that is responsible for the production of natural immunoglobulin M (IgM) and can respond to infection by increasing IgM secretion. However, we unexpectedly found that some epithelial cells also can express rearranged IgM transcript that has natural IgM characteristics, such as germline-encoded and restricted rearrangement patterns. Here we studied IgM expression in human non-B cells and found that IgM was frequently expressed by many human epithelial cancer cells as well as non-cancer epithelial cells. Moreover, CD79A and CD79B, two molecules that are physically linked to membranous IgM on the surface of B cells to form the B cell antigen receptor complex, were also expressed on the cell surface of epithelial cancer cells and co-located with IgM. Like the natural IgM, the epithelial cancer cell-derived IgM recognized a series of microbial antigens, such as single-stranded DNA, double-stranded DNA, lipopolysaccharide, and the HEp-2 cell antigen. More important, stimulation of the toll-like receptor 9 (TLR9), which mimics bacterial infection, substantially increased the secretion of IgM in human epithelial cancer cells. These findings indicate that human epithelial cancer cells as well as non-cancer epithelial cells can spontaneously produce IgM with natural antibody activity.

CCDC134 (coiled-coil domain containing 134) was firstly identified to be a novel classical secretory protein by our lab. Although we previously identified CCDC134 as a novel MAPK-regulator which down-regulated the Raf-1/MEK/ERK and JNK/ SAPK pathways in some cancer cell lines. However, the critical function of CCDC134 still remains unclear. In view of CCDC134 characteristic, which was widely expressed in lymphocytes, especially up-regulated expression in activated CD4+, CD8+ T lymphocytes and maturated DC, we proposed the hypothesis if CCDC134 was involved in immune response as a potential cytokine. Here, we first found that CCDC134 greatly promoted activation, proliferation and cytotoxicity of CD8+ T cells via augmenting the expression of activation molecules (CD25, CD69) and effector molecules, such as IFN-γ, TNF-α, granzyme B and perforin in vitro. Moreover, by a series of experimental evidences, we have found that CCDC134 displayed strong anti-tumor activity in a CD8+ T cells, but not CD4+ T cells, and IFN-γ-dependent manner in vivo. In addition, CCDC134 markedly promoted phosphorylation of STAT5, but not STAT1, STAT3 in CD8+ T cells and CD8+ T cells proliferation augmented by CCDC134 was dependent on STAT5. Our data collectively suggested that CCDC134, perhaps was served as a so far unidentified cytokine-like protein, which showed strong anti-tumor effects via augmenting CD8+ T cells-mediated tumor immunity.

As we known that IgG have served as prototypical effector molecules of immune defense. However, monomeric IgG, such as the IVIG purified from the serum of thousands of healthy donors, has been demonstrated as an immune inhibitor at high doses as a commonly administered for the treatment of a number of autoimmune diseases. Igs has long been considered to be only produced by B cells, however, by a series of experimental evidences from ours and others, it has been found that other types of non-hematopoietic cells, especially cancer cells could also produce IgG, and the IgG is involved in the cell survival and carcinogenesis. In this study, we unexpectedly found that like the IVIG, epithelial cancer-derived IgG, but not the normal IgG, also showed strong immune inhibitory effect to CD4+ or CD8+ T lymphocytes at low dose. More importantly, the inhibitory effect of cancer-derived IgG was stronger than IVIG. Subsequently, we have analyzed the mechanism of the immune inhibition. Our results suggested that cancer-derived IgG can not directly inhibit the activation and proliferation of CD4+ or CD8+ T cells, their immune inhibitory effect should be dependent on myeloid cells, at least partly, dependent on the monocytes. In addition, we found that the immune inhibitory effect of cancer-derived IgG was not dependent on the Treg. Although the mechanism is remain unclear, we believe that the IgG is so far unknown arms of tumor cells to against the immune system for their immune escape.

The tumor suppressor protein p53 is a well-known transcription factor that functions as a critical component of the genotoxic stress response via regulating the expression of effector proteins that control cellular fate following DNA damage. The human p300/CBP-associated factor (PCAF)-containing histone acetyltransferase (HAT) complex is important for the stability and activity of p53 via its acetylation. The human homolog of yeast alteration/deficiency in activation 2a (ADA2a) is a stable component of the human PCAF-containing HAT complex. In this study, we demonstrated that p53 and hADA2a physically interact with each other in human HEK 293T cells. Using overexpression and small interfering RNA-mediated knockdown, we demonstrated that hADA2a stabilizes p53 via promoting its acetylation at lysine 320 — a PCAF-dependent acetylation site. Furthermore, hADA2a can potentiate the transcriptional activity of p53 at the BAX and p21 promoters to induce cell apoptosis and cell cycle arrest. Overall, our results establish that hADA2a, a component of the PCAF-containing histone acetyltransferase co-activator complex, is a mediator of acetylation-dependent stabilization and activation of p53 in mammalian cells.