参考文献

Cyclic adenosine monophosphate (cAMP)-dependent phosphodiesterase (PDE) inhibitors such as pentoxifylline (PTX) suppress cAMP degradation and promote cAMP-dependent signal transduction. PDE inhibitors increase bone formation and bone mass in preclinical models and are used clinically to treat psoriatic arthritis by targeting inflammatory mediators including activated T cells. T cell activation requires two signals: antigen-dependent CD3-activation,which stimulates cAMP production; and CD28 co-stimulation,which downregulates cAMP-signaling,through PDE activation. PDE-inhibitors consequently suppress T cell activation by disrupting CD28 co-stimulation. Interestingly,we have reported that when CD8+ T cells are activated in the absence of CD28 co-stimulation,they secrete Wnt-10b,a bone anabolic Wnt ligand that promotes bone formation. In the present study,we investigated whether the bone anabolic activity of the PDE-inhibitor PTX,has an immunocentric basis,involving Wnt-10b production by CD8+ T cells. When wild-type (WT) mice were administered PTX,biochemical markers of both bone resorption and formation were significantly increased,with net bone gain in the axial skeleton,as quantified by micro-computed tomography ($\mu$CT). By contrast,PTX increased only bone resorption in T cell knockout (KO) mice,causing net bone loss. Reconstituting T cell-deficient mice with WT,but not Wnt-10b knockout (KO) CD8+ T cells,rescued bone formation and prevented bone loss. To study the role of cAMP signaling in Wnt-10b expression,reverse-transcription polymerase chain reaction (RT-PCR) and luciferase-reporter assays were performed using primary T cells. PDE inhibitors intensified Wnt-10b promoter activity and messenger RNA (mRNA) accumulation in CD3 and CD28 activated CD8+ T cells. In contrast,inhibiting the cAMP pathway mediators protein kinase A (PKA) and cAMP response element-binding protein (CREB),suppressed Wnt-10b expression by T cells activated in the absence of CD28 co-stimulation. In conclusion,the data demonstrate a key role for Wnt-10b production by CD8+ T cells in the bone anabolic response to PDE-inhibitors and reveal competing T cell-independent pro-resorptive properties of PTX,which dominate under T cell-deficient conditions. Selective targeting of CD8+ T cells by PDE inhibitors may be a beneficial approach for promoting bone regeneration in osteoporotic conditions. {\textcopyright} 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. View Publication

Due to viral envelope glycoprotein D binding to cellular membrane HVEM receptor,HSV-1 can infect certain dendritic cells,which becomes an event in the viral strategy to interfere with the host's immune system. We previously generated the HSV-1 mutant strain M6,which produced an attenuated phenotype in mice and rhesus monkeys. The attenuated M6 strain was used to investigate how HSV-1 infection of dendritic cells interferes with both innate and adaptive immunity. Our study showed that dendritic cells membrane HVEM receptors could mediate infection of the wild-type strain and attenuated M6 strain and that dendritic cells infected by both viruses in local tissues of animals exhibited changes in transcriptional profiles associated with innate immune and inflammatory responses. The infection of pDCs and cDCs by the two strains promoted cell differentiation to the CD103+ phenotype,but varied transcriptional profiles were observed,implying a strategy that the HSV-1 wild-type strain interferes with antiviral immunity,probably due to viral modification of the immunological phenotype of dendritic cells during processing and presentation of antigen to T cells,leading to a series of deviations in immune responses,ultimately generating the deficient immune phenotype observed in infected individuals in the clinical. View Publication

N6-methyladenosine (m6A) modification is involved in diverse immunoregulation,while the relationship between m6A modification and immune tolerance post kidney transplantation remains unclear. Expression of Wilms tumor 1-associating protein (WTAP),an m6A writer,was firstly detected in tolerant kidney transplant recipients (TOL). Then the role of WTAP on regulatory T (Treg) cell differentiation and function in CD4+ T cells from kidney transplant recipients with immune rejection (IR) was investigated. The potential target of WTAP and effect of WTAP on immune tolerance in vivo were subsequently verified. WTAP was upregulated in CD4+ T cells of TOL and positively correlated with Treg cell proportion. In vitro,WTAP overexpression promoted Treg cell differentiation and enhanced Treg cell-mediated suppression toward na?ve T cells. Forkhead box other 1 (Foxo1) was identified as a target of WTAP. WTAP enhanced m6A modification of Foxo1 mRNA in coding sequence (CDS) region,leading to up-regulation of Foxo1. Overexpression of m6A demethylase removed the effect of WTAP overexpression,while Foxo1 overexpression reversed these effects. WTAP overexpression alleviated allograft rejection in model mice,as evidenced by reduced inflammatory response and increased Treg population. Our study suggests that WTAP plays a positive role in induction of immune tolerance post kidney transplant by promoting Treg cell differentiation and function. leading to up-regulation of Foxo1. Overexpression of m6A demethylase removed the effect of WTAP overexpression while Foxo1 overexpression reversed these effects. WTAP overexpression alleviated allograft rejection in model mice as evidenced by reduced inflammatory response and increased Treg population. Our study suggests that WTAP plays a positive role in induction of immune tolerance post kidney transplant by promoting Treg cell differentiation and function." View Publication

The larval stage of the helminthic cestode Echinococcus multilocularis can inflict tumor-like hepatic lesions that cause the parasitic disease alveolar echinococcosis in humans,with high mortality in untreated patients. Opportunistic properties of the disease have been established based on the increased incidence in immunocompromised patients and mouse models,indicating that an appropriate adaptive immune response is required for the control of the disease. However,cellular interactions and the kinetics of the local hepatic immune responses during the different stages of infection with E. multilocularis remain unknown. In a mouse model of oral infection that mimics the normal infection route in human patients,the networks of the hepatic immune response were assessed using single-cell RNA sequencing (scRNA-seq) of isolated hepatic CD3+ T cells at different infection stages. We observed an early and sustained significant increase in natural killer T (NKT) cells and regulatory T cells (Tregs). Early tumor necrosis factor (TNF)- and integrin-dependent interactions between these two cell types promote the formation of hepatic lesions. At late time points,downregulation of programmed cell death protein 1 (PD-1) and ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1)-dependent signaling suppress the resolution of parasite-induced pathology. The obtained data provide fresh insight into the adaptive immune responses and local regulatory pathways at different infection stages of E. multilocularis in mice. View Publication

BACKGROUND This study aimed to investigate the role of circulating tumor cells (CTCs) and circulating cancer stem-like cells (cCSCs) before and after one cycle of chemotherapy and assessed the effects of early changes in CTCs and cCSCs on the outcomes of patients with metastatic breast cancer. METHODS Patients with stage IV invasive ductal carcinoma of the breast who received first-line chemotherapy between April 2014 and January 2016 were enrolled. CTCs and cCSCs were measured before the first cycle of chemotherapy (baseline) and on day 21,before the second cycle of chemotherapy commenced; a negative selection strategy and flow cytometry protocol were employed. RESULTS CTC and cCSC counts declined in 68.8 and 45.5% of patients,respectively. Declines in CTCs and cCSCs following the first chemotherapy cycle were associated with superior chemotherapy responses,longer progression-free survival (PFS),and longer overall survival (OS). An early decline in cCSCs remained an independent prognostic indicator for OS and PFS in multivariate analysis. CONCLUSIONS A cCSC decline after one cycle of chemotherapy for metastatic breast cancer is predictive of a superior chemotherapy response and longer PFS and OS,implying that cCSC dynamic monitoring may be helpful in early prediction of treatment response and prognosis. View Publication

BACKGROUND Osteoclasts play a crucial role in the maintenance,repair,and remodeling of bones of the adult vertebral skeleton due to their bone resorption capability. Rheumatoid arthritis (RA) and psoriatic arthritis (PsA) are associated with increased activity of osteoclasts. OBJECTIVES Our study aimed to investigate the dynamic proteomic changes during osteoclast differentiation in healthy donors,in RA,and PsA. METHODS Blood samples of healthy donors,RA,and PsA patients were collected,and monocytes were isolated and differentiated into osteoclasts in vitro using macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor $\kappa$B ligand (RANK-L). Mass spectrometry-based proteomics was used to analyze proteins from cell lysates. The expression changes were analyzed with Gene Set Enrichment Analysis (GSEA). RESULTS The analysis of the proteomic changes revealed that during the differentiation of the human osteoclasts,expression of the proteins involved in metabolic activity,secretory function,and cell polarity is increased; by contrast,signaling pathways involved in the immune functions are downregulated. Interestingly,the differences between cells of healthy donors and RA/PsA patients are most pronounced after the final steps of differentiation to osteoclasts. In addition,both in RA and PsA the differentiation is characterized by decreased metabolic activity,associated with various immune pathway activities; furthermore by accelerated cytokine production in RA. CONCLUSIONS Our results shed light on the characteristic proteomic changes during human osteoclast differentiation and expression differences in RA and PsA,which reveal important pathophysiological insights in both diseases. View Publication

Neutrophils are the most abundant leukocytes in human blood and are essential components of innate immunity. Until recently,neutrophils were considered homogeneous and transcriptionally inactive cells,but both concepts are being challenged. Single-cell RNA sequencing (scRNA-seq) offers an unbiased view of cells along a continuum of transcriptional states. However,the use of scRNA-seq to characterize neutrophils has proven technically difficult,explaining in part the paucity of published single-cell data on neutrophils. We have found that modifications to the data analysis pipeline,rather than to the existing scRNA-seq chemistries,can significantly increase the detection of human neutrophils in scRNA-seq. We have then applied a modified pipeline to the study of human peripheral blood neutrophils. Our findings indicate that circulating human neutrophils are transcriptionally heterogeneous cells,which can be classified into one of four transcriptional clusters that are reproducible among healthy human subjects. We demonstrate that peripheral blood neutrophils shift from relatively immature (Nh0) cells,through a transitional phenotype (Nh1),into one of two end points defined by either relative transcriptional inactivity (Nh2) or high expression of type I IFN-inducible genes (Nh3). Transitions among states are characterized by the expression of specific transcription factors. By simultaneously measuring surface proteins and intracellular transcripts at the single-cell level,we show that these transcriptional subsets are independent of the canonical surface proteins that are commonly used to define and characterize human neutrophils. These findings provide a new view of human neutrophil heterogeneity,with potential implications for the characterization of neutrophils in health and disease. View Publication

BACKGROUND In recent years,the T-cell therapy and immune checkpoint inhibitors toward CTLA-4 and PD-1/PD-L1 axis antibody therapy have acquired encouraging success. However,most of patients were still not benefited with lots of troubles,such as low penetration of tissues/cells,strong immunogenicity and cytokine release syndrome,and long manufacturing process and expensive costs. By contrast,the immune-modulating small molecules possessed natural advantages to overcome these obstacles and might achieve greater success. PURPOSE Exploring the potent immune-modulating natural small molecules and revealing what kinds of molecules or structures with the immunomodulatory activity against cancers. METHODS A novel non-cytotoxic T-cell immunomodulating screening model was used to identify the cytotoxic/selective/immunomodulatory bioactivity for 148 natural steroidal saponins. The structure-activity relationships (SARs) research was used to reveal the key groups for immunomodulation/cytotoxicity/selectivity. The negative selection was used to isolate and purify the T-cell. The cell viability assay was used to measure the anti-cancer effect in vitro. The ELISA assay was used to detect the cytokines for IL-1$\beta$,IL-6,TNF-$\alpha$,IFN-$\gamma$,IL-12,perforin and granzyme B (GZMB). The western blotting assay was used to research the immunomodulatory mechanism. The siRNA knockdown was used to generate the IFN-$\gamma$ resistant melanoma cells. The NOG immune-deficient mice were used to evaluate the anti-tumor efficacy in vivo. The peripheral blood samples from 10 cancer patients were used to detect the broad population anti-tumor efficacy. RESULTS It was reported that the correlation among structures and immunomodulation/ cytotoxicity/selectivity,in which opening ring-F with 26-O-glucopyranosyl,disaccharide and trisaccharide chains at C-3,steric hindrance and polarity of C-22 were key immunomodulatory groups. Moreover,taccaoside A was identified as the most potent candidate against cancer cells,including non-small cell lung cancer,triple negative breast cancer,and the IFN-$\gamma$ resistant melanoma,partly through enhancing T lymphocyte mTORC1-Blimp-1 signal to secrete GZMB. Besides,10 patients derived T-cell also would be modulated against cancer cells in vitro. Moreover,the overall survival was great extended (>140 days vs 93 days) with nearly 100% tumor burden disappearance (0 mm3vs 1006 ± 79.5 mm3) in mice. CONCLUSION This work demonstrated one possibility for this concerned purpose,and identified a potent immune-modulating natural molecule taccaoside A,which might contribute to cancer immunotherapy in future. View Publication

This study describes the characterization of one induced pluripotent stem cell line (iPSC) from a healthy female. It is crucial to use iPSCs derived from healthy individuals as controls in genetic disease studies. Thus,we established a human iPSC cell line derived from healthy people. The iPSC cell line was generated in our lab from the peripheral blood mononuclear cells (PBMCs) of a 28-year-old girl. The generated hiPSC line is free of episomal vectors,has a normal karyotype,expresses pluripotency markers and can differentiate into three germ layers in vivo. View Publication

Immunogenicity following an additional dose of Coronavirus disease 2019 (COVID-19) vaccine was investigated in an extended primary series among kidney transplant (KT) recipients. Eighty-five KT participants were randomized to receive either an mRNA (M group; n =??43) or viral vector (V group; n =??42) vaccine. Among them,62% were male,with a median (IQR) age of 50 (43-59) years and post-transplantation duration of 46 (26-82) months. At 2??weeks post-additional dose,there was no difference in the seroconversion rate between the M and V groups (70% vs. 65%,p =??.63). A median (IQR) of anti-RBD antibody level was not statistically different between the M group compared with the V group (51.8 [5.1-591] vs. 28.5 [2.9-119.3] BAU/ml,p =??.18). Furthermore,the percentage of participants with positive SARS-CoV-2 surrogate virus neutralization test results was not statistically different between groups (20% vs. 15%,p =??.40). S1-specific T cell and RBD-specific B cell responses were also comparable between the M and V groups (230 [41-420] vs. 268 [118-510],p =??.65 and 2 [0-10] vs. 2 [0-13] spot-forming units/106 peripheral blood mononuclear cells,p =??.60). In conclusion,compared with an additional dose of viral vector COVID-19 vaccine,a dose of mRNA COVID-19 vaccine did not elicit significantly different responses in KT recipients,regarding either humoral or cell-mediated immunity. (TCTR20211102003). View Publication

Monoclonal antibodies are at the vanguard of the most promising cancer treatments. Whereas traditional therapeutic antibodies have been limited to extracellular antigens,T cell receptor mimic (TCRm) antibodies can target intracellular antigens presented by cell surface major histocompatibility complex (MHC) proteins. TCRm antibodies can therefore target a repertoire of otherwise undruggable cancer antigens. However,the consequences of off-target peptide/MHC recognition with engineered T cell therapies are severe,and thus there are significant safety concerns with TCRm antibodies. Here we explored the specificity and safety profile of a new TCRm-based T cell therapy for hepatocellular carcinoma (HCC),a solid tumor for which no effective treatment exists. We targeted an alpha-fetoprotein peptide presented by HLA-A*02 with a highly specific TCRm,which crystallographic structural analysis showed binds directly over the HLA protein and interfaces with the full length of the peptide. We fused the TCRm to the ? and ? subunits of a TCR,producing a signaling AbTCR construct. This was combined with an scFv/CD28 co-stimulatory molecule targeting glypican-3 for increased efficacy towards tumor cells. This AbTC + co-stimulatory T cell therapy showed potent activity against AFP-positive cancer cell lines in vitro and an in an in vivo model and undetectable activity against AFP-negative cells. In an in-human safety assessment,no significant adverse events or cytokine release syndrome were observed and evidence of efficacy was seen. Remarkably,one patient with metastatic HCC achieved a complete remission after nine months and ultimately qualified for a liver transplant. View Publication

The myeloma surface proteome (surfaceome) determines tumor interaction with the microenvironment and serves as an emerging arena for therapeutic development. Here,we use glycoprotein capture proteomics to define the myeloma surfaceome at baseline,in drug resistance,and in response to acute drug treatment. We provide a scoring system for surface antigens and identify CCR10 as a promising target in this disease expressed widely on malignant plasma cells. We engineer proof-of-principle chimeric antigen receptor (CAR) T-cells targeting CCR10 using its natural ligand CCL27. In myeloma models we identify proteins that could serve as markers of resistance to bortezomib and lenalidomide,including CD53,CD10,EVI2B,and CD33. We find that acute lenalidomide treatment increases activity of MUC1-targeting CAR-T cells through antigen upregulation. Finally,we develop a miniaturized surface proteomic protocol for profiling primary plasma cell samples with low inputs. These approaches and datasets may contribute to the biological,therapeutic,and diagnostic understanding of myeloma. View Publication