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Primary Sjögren's syndrome (pSS) is a chronic systemic autoimmune disease characterized by B cell hyperactivation and hypergrammaglobulinemia. Currently, the role of metabolic pathways in the B cells of pSS patients is poorly defined. Here, we showed that upon cytosine phosphate-guanine (CpG)/sCD40L/IL-4 stimulation, B cells proportionally increased glycolysis and oxygen consumption, and compared with B cells from healthy controls (HCs), B cells from pSS patients exhibited higher glycolysis capacity and maximal oxidative respiration (OXPHOS). We also found that glucose transporter 1 (GLUT1) expression in B cells from pSS patients was significantly higher than that in B cells from HCs. Treatment with 2-deoxy-d-glucose (2-DG) inhibited the activation of B cells in pSS patients. Both 2-DG and Metformin inhibited the proliferation, formation of plasma/plasmablasts and decreased the IgG and IgM levels in the supernatants of B cells from pSS patients. Furthermore, inhibition of mTORC1 by rapamycin had an effect similar to that of 2-DG, suppressing B cell activation, proliferation and antibody production. Taken together, we demonstrated that B cells from pSS patients are more metabolically active than those from HCs and suggested that the mTORC1-GLUT1 glycolysis pathways were the major drivers of B cell hyperactivation and autoantibody production in pSS patients.

Objectives. Immunoglobulin-G4-related disease (IgG4-RD) is a distinct systemic autoimmune-mediated disease that characterized by tissue fibrosis. The purpose of this study is to investigate the epigenetic modifications in IgG4-RD. Methods. A genome-wide DNA methylation study was performed in B cells, CD4+ T cells and salivary gland tissue from IgG4-RD patients and matched controls using the Illumina HumanMethylation 850K BeadChip. Results. We identified differentially methylated CpG sites including 44 hypomethylated and 166 hypermethylated differentially methylated probes (DMPs) in B cells, 260 hypomethylated and 260 hypermethylated DMPs in CD4 + T cells from 10 IgG4-RD patients compared with 10 healthy controls. We also identified 36945 hypomethylated and 78380 hypermethylated DMPs in salivary gland tissue of 4 IgG4-RD patients compared with 4 controls. We found that DPM2 (cg21181453), IQCK (cg10266221), ABCC13 (cg05699681, cg04985582) were hypermethylated and MBP (cg18455083) was hypomethylated in B cells, CD4+ T cells and tissue of IgG4-RD patients compared with controls. Also, we observed hypomethylated HLA class in both B cells and CD4+ T cells from IgG4-RD patients compared with controls. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DMPs in salivary gland of IgG4-RD patients demonstrated enrichment of pathways in regulation of immune cells responses and fibrosis. Conclusion. Our study is the first DNA methylation study in peripheral B cells, CD4+ T cells as well as salivary gland tissues from patients with IgG4-related disease. Our findings highlighted the role of epigenetic modification of DNA methylation and identified several genes and pathways that might involve in the pathogenesis of IgG4-RD.

Immunoglobulin-G4-related disease (IgG4-RD) is a distinct systemic autoimmune-mediated disease manifesting as chronic inflammation and tissue fibrosis. Since the role of DNA methylation in the pathogenesis of IgG4-RD is still unclear, we conduct this study to investigate epigenetic modifications in IgG4-RD.A genome-wide DNA methylation study was conducted with B cells, CD4We identified differentially methylated CpG sites including 44 hypomethylated and 166 hypermethylated differentially methylated probes (DMPs) in B cells and 260 hypomethylated and 112 hypermethylated DMPs in CD4This is the first DNA methylation study in peripheral B cells, CD4

Juvenile-onset recurrent respiratory papillomatosis (JORRP) is the most common benign laryngeal neoplasm in children and is considered to be primarily caused by human papillomavirus (HPV) types 6 and 11. In the present study, we performed RNA sequencing (RNA-seq) of 8 tumors and 4 adjacent nontumor tissues to explore the transcriptional profiles of JORRP tumors. A total of 1,151 upregulated genes involved in the interleukin-17 (IL-17) signaling pathway and 1,620 downregulated genes involved in dysregulated inflammatory responses were reported. Immunohistochemistry (IHC) assays confirmed the upregulation of IL-17C in JORRP tumors compared with paired adjacent nontumor tissues. Real-time PCR (RT-PCR) assays showed positive correlations between CXCL1 (CXC chemokine ligands 1) and CXCL8 and the Derkay Clinic Score of JORRP patients. We further overexpressed the HPV6 or HPV11 E6 and E7 oncogenes in SNU-1076 head and neck squamous cell carcinoma (HNSCC) cell lines and carried out RNA-seq. We found that HPV6-E6-E7 gene overexpression resulted in only 16 upregulated genes and 1 downregulated gene; however, HPV11-E6-E7 gene overexpression resulted in 1,776 upregulated genes and 461 downregulated genes compared with the control cell lines. The differentially expressed genes (DEGs) of HPV11-E6-E7 gene overexpression were positively enriched in the DNA replication-related terms by Gene Ontology (GO) analysis and the IL-17 signaling pathway by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Taken together, our present findings revealed IL-17 signaling pathway-related gene profiles that might contribute to disease pathogenesis and that the HPV11 E6 and E7 oncogenes promote disease progression by enhancing tumor growth and activating the IL-17 signaling pathway in JORRP patients. IMPORTANCE Juvenile-onset recurrent respiratory papillomatosis (JORRP) is primarily caused by human papillomavirus 6 (HPV6) and HPV11 infection; however, the gene signatures of tumors are currently less understood. In the present study, we performed RNA sequencing and found upregulated genes associated with the IL-17 signaling pathway and downregulated genes associated with inflammatory-related pathways. Further RNA sequencing was performed in HPV6-E6-E7- or HPV11-E6-E7-overexpressing SNU-1076 HNSCC cells lines to explore the potential pathogenic molecular mechanisms of HPV virus. We found that HPV11-E6-E7 overexpression resulted in gene expression related to DNA replication and the IL-17 signaling pathway. Our results suggested enriched that the IL-17 signaling pathway resulting from HPV11 infection might contribute to JORRP pathogenesis.

The presence or absence of anti-citrullinated peptide antibodies (ACPA) and associated disparities in patients with rheumatoid arthritis (RA) implies disease heterogeneity with unknown diverse immunopathological mechanisms. Here we profile CD45+ hematopoietic cells from peripheral blood or synovial tissues from both ACPA+ and ACPA- RA patients by single-cell RNA sequencing and identify subsets of immune cells that contribute to the pathogenesis of RA subtypes. We find several synovial immune cell abnormalities, including up-regulation of CCL13, CCL18 and MMP3 in myeloid cell subsets of ACPA- RA compared with ACPA+ RA. Also evident is a lack of HLA-DRB5 expression and lower expression of cytotoxic and exhaustion related genes in the synovial tissues of patients with ACPA- RA. Furthermore, the HLA-DR15 haplotype (DRB1/DRB5) conveys an increased risk of developing active disease in ACPA+ RA in a large cohort of patients with treatment-naive RA. Immunohistochemical staining shows increased infiltration of CCL13 and CCL18-expressing immune cells in synovial tissues of ACPA- RA. Collectively, our data provide evidence of the differential involvement of cellular and molecular pathways involved in the pathogenesis of seropositive and seronegative RA subtypes and reveal the importance of precision therapy based on ACPA status., Patients with rheumatoid arthritis are commonly stratified by ACPA serology, with positivity being associated with more severe disease and joint destruction. Here the authors present a single cell RNA sequencing resource comparing peripheral blood and synovial tissue cells from patients with ACPA+ versus ACPA- rheumatoid arthritis.

ObjectivesLack of effective biomarkers in anti-citrullinated protein antibody (ACPA)-negative rheumatoid arthritis (RA) impedes early diagnosis and treatment. This study aimed to identify novel autoantibodies in RA and verify their diagnostic values in ACPA-negative RA based on protein microarray technology.MethodsA total of 1011 sera from 559 RA (276 ACPA-positive and 283 ACPA-negative), 239 disease controls (DCs) and 213 healthy controls (HCs) were collected and sampled on two sequential microarrays and ELISA and western blot (WB) detection, for novel autoantibodies discovery, validation and verification, respectively. The high-density protein microarray printed with a broad spectrum of recombinant human proteins was first employed to screen candidate autoantibodies, then focused microarrays composed of candidate autoantigens were used for validation, followed by ELISA and WB to verify the presence of the most promising candidates in ACPA-negative RA.ResultsNine novel autoantibodies were identified by two sequential microarrays with positivity 17.93%–27.59% and specificities >90% in ACPA-negative RA. Among these, anti-PTX3 and anti-DUSP11 autoantibodies presented with the highest sensitivity and were consistently verified by ELISA and WB. Pooling samples of all cohorts, the positivities of anti-PTX3 and anti-DUSP11 in ACPA-negative RA were 27.56% and 31.80%, respectively, similar to those in ACPA-positive RA, and significantly higher than in HCs (4.69% and 2.35%) and DCs (10.04% and 8.49%) (pConclusionAnti-PTX3 and anti-DUSP11 autoantibodies are newly identified biomarkers for diagnosis of ACPA-negative RA.

The pathogenesis of rheumatoid arthritis (RA), a systemic autoimmune disease characterized by autoreactive T-cell accumulation and pro-inflammatory cytokine overproduction, is unclear. Systematically addressing T-cell receptor (TCR) repertoires of different CD4

Gut microbiota play an essential role in shaping immune cell responses. The liver was continuously exposed to metabolic products of intestinal commensal bacterial through portal vein and alteration of gut commensal bateria was always associated with increased risk of liver inflammation and autoimmune disease. Considered as a unique immunological organ, the liver is enriched with a large number of innate immune cells. Herein, we summarize the available literature of gut microbiota in shaping the response of hepatic innate immune cells including NKT cells, NK cells, γδ T cells and Kupffer cells during health and disease. Such knowledge might help to develop novel and innovative strategies for the prevention and therapy of innate immune cell-related liver disease.

Background Representing the first line of host defense against virus infections and an essential link between innate and adaptive immune response, the role of dendritic cells (DCs) in peripheral blood of juvenile-onset recurrent respiratory papillomatosis (JORRP) patients and association with disease activity were still not established. Materials and Methods In our present study, 28 JORRP patients and 28 age and sex matched healthy controls were enrolled. The percentage, phenotype and cytokine secretion of DC and was measured by flow cytometry. Plasma cytokine were detected by the enzyme-linked immunosorbent assay (ELISA). Results We found that the percentage of myeloid DC (mDC) was significantly lower in JORRP patients compared to healthy controls and was negatively correlated with interval times, but not surgical times or disease onset. Moreover, the activation marker, CD40 and CD86 was significantly up-regulated on the surfaces of mDC in JORRP patients compared with healthy controls. Neither the percentage nor activation of plasmacytoid DC (pDC) showed statistical difference between JORRP patients and healthy controls. HLA-DR expression on both mDC and pDC was down-regulated in JORRP group and negatively correlated with surgical times. Antigen presenting ability of DC was greatly impaired in JORRP patients of higher number of operations and shorter interval time. Plasma IL-10 as well as IL-10 secreted by mDC was higher in JORRP patients compared with healthy control. Finally, we detected an up-regulated TLR2 and TLR4 expression on mDCs and TLR4 expression was positively correlated with HLA-DR expression on mDC of JORRP patients. Conclusion Our results demonstrate an abnormal TLR2 and TLR4 expression in mDCs may contribute to suppressive immune response to HPV6 or HPV11 infection and associated with disease activity in JORRP patients.

Severe combined immunodeficiency (SCID) is the most serious disorder among primary immunodeficiency diseases threatening children's life. Atypical SCID variant, presenting with mild reduced T cells subsets, is often associated with infection susceptibility but poor clinical diagnosis. The atypical X-SCID patient in the present study showed a mild clinical presentation with a TlowNK+B+ immunophenotype. The patient has reduced T- cell subpopulations with a subdued thymic output measured by sjTRECs. Further analysis showed that T cells maintained a normal proliferation and a broad Vβ repertoire. NK cells, however, exhibited a skewed development toward immature CD3-CD16+CD56- cells. Genetic analysis revealed a novel deletion at nucleotide 52 in exon 1 of IL2RG gene. Sequence alignment predicted a truncated IL2RG protein missing signal peptide derived from a possible alternative reading frame. The novel mutation in IL2RG gene identified in our study may help the early diagnosis of atypical X-SCID.

Background:Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease. Epidemiological studies in SLE have been reported in the literature in many countries and ethnic groups. Although SLE in China has been described in the past, there has not been a detailed evaluation of SLE patients in Xinjiang of China, a largely Uygur population.Objectives:To describe the clinical featuresand immunological features of 484 SLE subjects.Methods:484 adult patients followed in the The People’s Hospital of the Xinjiang Uygur Autonomous Region, 211 patients with Uygur SLE as welI as 273patients with Han SLE.Results:Of the 211 Uygur SLE patients, 195 (92.4%) were female and 16 (7.6%) were male (female:male=12.2:1),the mean age at SLE onset was 34.67±11.57 years, mean disease duration was 20.77±35.16 months.Of the 273 Han SLE patients,247(90.5%)were female and 26(9.5%) were male,the mean age at SLE onset was 36.68±14.44 years,mean disease duration was 41.64±61.89 months.(2)between the Uygur and Han SLE patients,the Raynaud phenomenon(χ2=6.066 P=0.014), Chest pain(χ2=7.906 P=0.005), headache(χ2=4.572 P=0.029)has obvious differences(table 1).(3)The anti-nuclear (χ2=8.108 P=0.004), anti-AHA (χ2=4.952 P=0.026)were higer in Uygur SLE patients than those in Han SLE patients;the Uygur SLE patients has been anemia(χ2=6.904,P=0.009), high level of immunoglobulin (χ2=8.939,P=0.003),decrease of complement(χ2=6.330 P=0.012).(table 2)figure 1. Clinical manifestationsClinical manifestationsUygur SLE patients(n=211)Han SLE (n=273)χ2Prash106(50.2%)157(57.5%)2.5370.111Photosensitivity40(19%)56(20.5%)0.1810.670Alopecia73(34.6%)101(37.0%)0.2980.585Oral ulcers49(23.2%)64(23.5%)0.0060.937Raynaud phenomenon28(13.3%)60(22%)6.0660.014livedo reticularis5(2.4%)7(2.6%)0.0200.866arthralgia/arthritis105(49.8%)159(58.5%)3.6230.057abnormal liver-function8(3.8%)7(2.6%)0.5970.440Chest pain28(13.3%)16(5.9%)7.9060.005Suffocation49(23.2%)52(19%)1.2560.262palpitation27(12.8%)41(15%)0.4870.485Shortness of breath24(11.4%)35(12.8%)0.2330.630Ophthalmia2(0.9%)6(2.2%)1.1540.283Visual impairment1(0.5%)3(1.1%)0.5670.457hemiplegia1(0.5%)0(0.0%)1.2970.436Mental disorder4(1.9%)11(4.0%)1.8040.179headache14(6.6%)7(2.6%)4.5720.029Lower Limb Edem36(17.1%)34(12.5%)2.0420.153pleurisy37(17.5%)43(15.8%)0.2560.613pericarditis38(18%)33(12.1%)3.2730.070pulmonary fibrosis7(3.3%)9(3.3%)0.0000.982figure 2. immunological manifestationsmanifestationsUygur SLE patients(n=211)Han SLE (n=273)χ2Pantinuclear antibodies185(87.7%)212(77.7%)8.1080.004anti-dsDNA115(54.5%)144(52.7%)0.1470.701anti-SSA101(47.9%)149(54.6%)2.1470.143anti-SSB45(21.3%)63(23.1%)0.2100.647anti-Sm44(21%)68(25%)1.0880.297ACL29(13.8%)26(9.5%)2.1610.142antiU1-RNP74(35.1%)100(36.6%)0.1260.723anti-AHA60(28.4%)54(19.8%)4.9520.026Low white blood cell46(22.1%)79(29%)2.9380.087anemia90(42.9%)85(36.3%)6.9040.009Thrombocytopenia36(17.1%)51(18.7%)0.2120.645Urine protein positive84(39.8%)114(41.9%)0.2170.641Rise of urine RBC29(13.7%)35(12.8%)0.0880.766Increased immunoglobulin82(38.9%)71(26.1%)8.9390.003Complement decline120(57.1%)124(45.6%)6.3300.012Conclusion:The Uygur SLE patients have their own clinical and immunological characteristics, which has guiding significance in the diagnosis, treatment and prognosis of SLE.References:[1]Martyna TS,Hanna SS, Marek F. Clinical and immunological characteristics of Polish patients with systemic lupus erythematosus. Adv Clin Exp Med. 2018;27(1):57–61[2]Maloney K C, Ferguson T S, Stewart H D, et al. Clinical and immunological characteristics of 150 systemic lupus erythematosus patients in Jamaica: A comparative analysis[J]. Lupus, 2017, 26(13):961203317707828.[3]Ching K H, Burbelo P D, Christopher T, et al. Two Major Autoantibody Clusters in Systemic Lupus Erythematosus[J]. PLoS ONE, 2012, 7(2):e32001.Disclosure of Interests:None declared

Background:AIDS is a deadly infectious disease caused by the HIV. When HIV infects a host, it may induce production of autoantibodies due to the structural antigen similarity between viral proteins and selfantigens.The molecular mimicry between HIV protein and self-antigens could cause antibody cross-reactions and lead to development of autoimmune disease.Objectives:To explore the clinical value of serum autoantibodies and human leukocyte antigen (HLA-B27) molecular testing in Uygur patients with human immunodeficiency virus (HIV) infection.Methods:A total of 727 HIV-infected Uygur patients who visited Kuche Infectious Diseases Hospital during May 2016 to March 2017 were include in this study. The other 390 healthy people were enrolled as controls. Serum antinuclear antibodies (ANA) and ANA Profile, anti-cyclic citrullinated peptide (CCP) antibody, and HLA-B27 molecule were tested.Results:Among 727 HIV-infected Uygur patients, 317 were males and 410 were females with mean age (35.52±13.44) years old. The mean duration of disease was (6.34±3.05)years. There were 697(95.87%) patients receiving Highly active antiretroviral therapy (HAART) with mean duration of treatment (6.34±3.05)years. Rheumatic manifestations were recorded in 238 (32.74%) HIV-infected Uygur patients, mainly with dry mouth and dry eye (15.41%), alopecia (9.90%), arthralgia (8.94%), ect. Compared with the health controls, positive ANA was more common in HIV infected Uygur patients (33.42%vs.17.43%,P< 0.001) with low titers (ANA titer:1:100). HIV-infected Uygur patients had higher positive anti-u1-RNP antibodies positive rate (1.10%), but lower anti-SSA antibodies positive rate (0.14%) and anti-CCP antibodies positive rate (0.28%). Patients with positive ANA in HAART group were significantly less than that in non-treatment group (38.72%vs.50.00%,P=0.049).Only one female patient was HLA-B27 positive (0.14%), which was significantly lower than that in healthy controls (3.08%) (PConclusion:Rheumatic manifestations are common in HIV-infected Uygur patients. Several autoantibodies are positive, but the coincidence of rheumatic diseases is rare. It’s noted that patients with Rheumatic manifestations and low titre positive ANA should be considered as a differential diagnosis of HIV infection.Disclosure of Interests:None declared

Background:Connective tissue diseases are a group of inflammatory, immune mediated disorders.Interstitial lung disease (ILD) is associated with significant morbidity and mortality.Currently,, scientists are still looking for serum markers to diagnose interstitial lung disease.Althought serum KL-6 level has been studied in ILD of various aetiologies and revealed to be an important serum marker for ILD,but differences in KL-6 expression related to ethnic and/or genetic variants may exist.Objectives:To evaluate the diagnosis of the serum Krebs von den Lungen-6 (KL-6) for CTD-ILD in the Uygur population of China.Methods:117 Patients with CTD-ILD (CTD-ILD group) and 182 patients with CTD (CTD group) who visited the department of rheumatology and immunology of People’s Hospital of Xinjiang Uygur Autonomous Region between January, 2015 and December, 2019 were included. Serum KL-6 levels were measured by chemiluminescent enzyme immunoassay kit.Results:The significantly higher levels of KL-6 were determined in the RA-ILD group than RA group [569(287.5,984)U/ml vs 194(152,266.5)U/ml](PFigure 1.Comparison of serum KL-6 concentrations in CTD-ILD group and CTD group.Fig 2.Receiver-operating characteristic curve(ROC) of KL-6 for the diagnosis of CTD-ILDConclusion:The serum KL-6 is a important biomarker for the diagnosis of CTD-ILD and Serum KL-6 could be a clinically useful biomarker in screening CTD-ILD in the Uygur population of China.References:[1]Woodhead F, Wells A U, Desai S R. Pulmonary Complications of Connective Tissue Diseases[J]. Clinics in Chest Medicine, 2008, 29(29):149–164.Tanaka S, Hattori N, Ishikawa N, et al. Krebs von den Lungen-6 (KL-6) is a progn -ostic biomarker in patients with surgically resected nonsmall cell lung cancer. Int J Cancer 2012; 130:377–87.[2]Ogz E O, Kucuksahin O, Turgay M, et al. Association of serum KL-6 levels with interstitial lung disease in patients with connective tissue disease: a cross-sectional study. Clinical Rheumatology, 2016, 35(3):663-666.Disclosure of Interests:None declared

Increasing evidence supports the concept that dysregulated B cell differentiation and activation are pivotal in the initiation and progression of autoimmune disorders, in both antibody (Ab)-dependent and Ab-independent mechanisms. Antimitochondrial antibodies (AMA), the serologic hallmark of PBC, are present in greater than 95% of patients with PBC, with a long latency period between the detection of autoantibodies and the onset of clinical disease. However, the mechanisms involved in the role of B cells in the pathogenesis of PBC have remained enigmatic. Herein, we have applied scRNA sequencing to simultaneously characterize B cell repertoire and transcriptome status in blood-liver paired samples from patients with PBC. Our data reveal that there is a dramatic increase in the heterogeneity of the blood CD27− B-cell population in PBC as compared to control. Strikingly, this overwhelmingly expanded naïve-like population were IgM+/IgD+, and encoded by germline sequences. In contrast with blood B cells, the dominant liver CD27− B-cell population is comprised of a high frequency of class-switched clonal B cells, which strongly express MHC class II genes. Upon stimulation with CpG (a surrogate TI-antigen), PBC CD27− B-cell populations markedly expanded and were accompanied by a significantly reduced expression of MARCKS, a newly identified molecule that suppresses active BCR signaling, as compared to healthy controls. Our data indicate that PBC CD27− B-cells exhibit abnormalities and heterogeneity, which may arise from a consistent exposure to TI-antigens. Our findings underscore the significance of B cells in PBC and suggest that more efforts should be placed on PBC B cell immunobiology and as potential therapeutic targets.

The pathogenesis of rheumatoid arthritis (RA), a systemic autoimmune disease characterized by autoreactive T-cell accumulation and pro-inflammatory cytokine overproduction, is unclear. Systematically addressing T-cell receptor (TCR) repertoires of different CD4+ T-cell subsets could help understand RA pathogenesis. Here, peripheral CD4+ T cells from treatment-naive RA patients and healthy controls were sorted into seven subsets including naive, effector, central memory, effector memory (EMT), Th1, Th17, and regulatory T cells. T-cell receptor β chain repertoires were then analyzed by next-generation sequencing. We identified T-cell clonal expansion in EMT and Th17 cells of RA patients, with highly similar TCR repertoires. Ex vivo experiments demonstrated the preferred differentiation from EMT to Th17 cells in RA. Notably, we showed that TCR diversity and abundance of differentiated T cells of Th17 were significantly correlated with RA disease activity. Based on these observations, we propose that abnormal differentiation from EMT to Th17 and expansion of Th17 play pivotal role in RA pathogenesis.

Excessive inflammatory cytokines play crucial roles in the pathogenesis of rheumatoid arthritis (RA), however, the underlying mechanism remains unclear. In this study, we demonstrated that pentaxin 3 (PTX3), an essential component of innate immunity, was elevated in RA and preferentially bound to CD14+ monocytes. C1q promoted the binding and resulted in increased cell proliferation, activation and caspase-1-related late apoptotic cells (7-AAD+annexin V+), as well as enhanced release of inflammatory cytokines including TNF-α, IL-1β and IL-6. Serum from RA patients, compared with healthy controls, induced gasdermin D (GSDMD)-dependent pyroptosis in monocytes, and this ability was associated with disease activity. Moreover, PTX3 synergized with C1q to promote pyroptosis in RA-serum pre-incubated monocytes by coordinately enhancing NLRP3 inflammasome over-activation and inducing GSDMD cleavage, cell swelling with large bubbles, caspase-1-dependent cell death and inflammatory cytokine release including IL-6. On the other hand, IL-6 promoted PTX3 plus C1q-induced pyroptosis in both normal and RA serum pre-incubated monocytes. These findings collectively implicated an important role of IL-6 in driving PTX3 plus C1q-mediated pyroptosis in RA and shed lights on a potential new treatment strategy targeting pyroptosis-mediated persistent inflammatory cytokine release.

Excessive inflammatory cytokines play crucial roles in the pathogenesis of rheumatoid arthritis (RA), however, the underlying mechanism remains unclear. In this study, we demonstrated that pentaxin 3 (PTX3), an essential component of innate immunity, was elevated in RA and preferentially bound to CD14+ monocytes. C1q promoted the binding and resulted in increased cell proliferation, activation and caspase-1-related late apoptotic cells (7-AAD+annexin V+), as well as enhanced release of inflammatory cytokines including TNF-α, IL-1β and IL-6. Serum from RA patients, compared with healthy controls, induced gasdermin D (GSDMD)-dependent pyroptosis in monocytes, and this ability was associated with disease activity. Moreover, PTX3 synergized with C1q to promote pyroptosis in RA-serum pre-incubated monocytes by coordinately enhancing NLRP3 inflammasome over-activation and inducing GSDMD cleavage, cell swelling with large bubbles, caspase-1-dependent cell death and inflammatory cytokine release including IL-6. On the other hand, IL-6 promoted PTX3 plus C1q-induced pyroptosis in both normal and RA serum pre-incubated monocytes. These findings collectively implicated an important role of IL-6 in driving PTX3 plus C1q-mediated pyroptosis in RA and shed lights on a potential new treatment strategy targeting pyroptosis-mediated persistent inflammatory cytokine release.

Background The connective tissue diseases is a group of inflammatory, immune-mediated diseases.CTD often leads to autoimmunity and subsequent tissue injury. It is an important contribute to thoracic changes, particularly interstitial lung disease, is are the main causes of mortality and morbidity among patients with connective tissue diseases. Prognosis and response to therapy are the most pressing challenges for connective tissue disease-associated ILD (CTD-ILD). At present, the basic methods for the diagnosis of various types of ILD includes high-resolution computed tomography, bronchoscopy, and surgical lung biopsy. In addition, continuous lung function tests are commonly used to monitor disease activity and predict the outcome of patients with ILDs, But these tests require specific inspection machines and Repeatability is not good. At present, many biomarkers have been developed to detect ILDs, and the most important biomarkers is KL-6 and lung surface active protein A (SP- A) and surfactant protein D (SP – D) witch secreted by alveolar epithelial type II cells. But Relevant studies have shown that the sensitivity, specificity and accuracy of KL-6 are higher than SP-A and SP-D. Objectives To evaluate the diagnosis of the serum Krebs von den Lungen-6 (KL-6) for the interstitial lung disease(ILD) associated with connective tissue diseases(CTD). Methods We retrospectively analysed the medical records of 50 patients with CTD associated ILD,46 CTD patients without ILD. Measurement of serum KL-6 levels and pulmonary function tests performed in parallel were reviewed.T test, X2 test, non-parametric test, SPSS and correlation analysis were used for data analysis. Results The significantly higher levels of KL-6 were determined in the CTD-ILD group than in either the CTD without pulmonary involvement group(P Conclusions The serum KL-6 is a valuable biomarker for CTD-ILD diagnosis and even as a predictive factor could be used to identify the clinical development of ILD. Disclosure of Interest None declared

CD4+CD25+ regulatory T cells (Tregs) are involved in the regulation of physiological and pathological hepatic immune responses, but the roles are not well explored in natural killer (NK) cell-mediated liver diseases. In this study, using the NK cell-mediated oversensitive liver injury model of hepatitis B virus transgenic (HBs-Tg) mice triggered by a low dose of concanavalin A, it was observed that an increased number of CD4+CD25+Foxp3+ Tregs were accumulated in the liver, along with the recovery of liver injury. Adoptive transfer of hepatic Tregs from HBs-Tg mice but not wild B6 mice could significantly attenuate the oversensitive liver injury via inhibiting liver accumulation and decreasing NK cell group 2D-mediated activation of NK cells in the recipient HBs-Tg mice. Furthermore, upregulated expression of membrane-bound TGF-β (mTGF-β) and OX40 on hepatic Tregs were demonstrated to account for inhibiting the NK cell-mediated hepatic injury in HBs-Tg mice through cell-cell contact, confirmed by antibody blockade and cell Transwell experiments in vivo and in vitro. Our findings for the first time indicated that CD4+CD25+ Tregs directly suppressed NK cell-mediated hepatocytotoxicity through mTGF-β and OX40/OX40L interaction in a cell-cell contact manner in HBV-associated liver disease.

Whether humoral immunity plays a role in HPV type 6 or 11 virus-mediated Juvenile-onset Recurrent Respiratory Papillomatosis (JORRP) remains unknown. In the present study, serum total IgG level in 44 JORRP patients was significantly decreased compared with that in 40 healthy controls. Moreover, expanded CD3−CD19+ B cells with down-regulation of CD23, CD40, HLA-DR and up-regulation of CD86 expression were found in the peripheral blood of JORRP patients. Flow cytometry analysis of B-cell compartment showed that the frequency of both CD19+CD27hi plasma cells and CD19+CD27+ memory B cells were decreased in JORRP patients. Importantly, although the proportion of circulating CXCR5+PD1hi Tfh cells was not changed, the function of Tfh cells were greatly impaired with reduced ability of IL-21 secretion to promote B cell maturation. Association analysis by the Kaplan-Meier method revealed that IL-21 secreting Tfh cell was positively correlated to the CD27+ B cell subset frequency, the serum IgG level and the frequency of recurrence in JORRP patients, but negatively correlated to the percentage of IgD+CD27− B cell. We concluded that a reduced IL-21 secretion by Tfh cells may limit B cell maturation and antibody production in JORRP patients and Tfh cell-derived IL-21 might be associated with JORRP outcome in clinic.

Defects in the adaptive immune response to human papillomavirus-6 and -11 are among the most important mechanisms for Juvenile-onset Recurrent Respiratory Papillomatosis (JORRP) development. However, the percentage of CD8+ T cells and peripheral TH1/TH2 immune responses in Juvenile-onset Recurrent Respiratory Papillomatosis is still not well addressed due to limited sample sizes.Twenty-three patients who were diagnosed with JORRP and underwent surgical intervention at the Beijing TongRen Hospital from October 2015 to March 2016 were enrolled in our study. The CD8We found that the proportions of CD4We conclude that only T