Background Primary Sjogren’s Syndrome (pSS) is a lymphoproliferative disease with autoimmune characteristics, which is characterized by lymphocyte infiltration of exocrine glands and involvement and dysfunction of extraglandular organs. Renal tubular acidosis (RTA) is a common renal involvement in pSS. This study investigated the phenotypic characteristics of peripheral blood lymphocyte subsets and cytokines in pSS patients complicated with RTA (pSS-RTA). Method This retrospective study included 25 pSS patients complicated with RTA and 54 pSS patients without RTA (pSS-no-RTA). To examine the level of peripheral lymphocytes subsets, flow cytometry analysis was used. The level of serum cytokines were detected by flow cytometry bead array(CBA). The influencing factors related to the occurrence of pSS-RTA were identified through logistic regression analyze. Results The absolute number of CD4 + T cells and Th2 cells in peripheral blood were decreased in pSS-RTA patients than pSS-no-RTA patients. Moreover, the absolute number of NK cells and Treg cells were also decreased in pSS-RTA patients than pSS-no-RTA. The level of serum IL-2 was higher in pSS-RTA patients than pSS-no-RTA patients, and is negatively correlated with the number of NK cells, the number and percentage of Th17 cells, and Th17/Treg. Serum IL-2 level is also correlated with various cytokines. Multivariate logistic analysis proved that elevated ESR and ALP were risk factors for pSS complicated with RTA, while Treg was a protective factor. Conclusion The increase of serum IL-2 level and the decrease of peripheral blood NK cells and Treg cells may be the immune mechanism of the development of pSS-RTA disease.

Objective Patients with ankylosing spondylitis (AS) carry an increased burden of cardiovascular diseases (CVD), but features denoting the development of CVD in AS are unclear. This study aimed to evaluate the percentage and absolute number of lymphocytes and CD4+T cells in AS patients complicated with CVD (AS-CVD) and determine whether circulating Th17 cells are associated with the development of CVD in AS. Method A total of 117 AS patients (46 had CVD and 71 had no CVD) were enrolled in this retrospective study. The percentage and absolute number of lymphocytes and CD4+T cells were determined by Flow cytometry. Associations between CVD and clinical markers were analyzed using logistic regression. Results The ratio of Th17/Treg cells (0.30 vs 0.19, p = 0.014) and the absolute number of Th17 cells (7.27 cells/μL vs 4.34 cells/μL, p < 0.001) was significantly elevated in AS-CVD group compared with AS-no-CVD group. Multivariate logistic regression revealed that elevated Th17 cells (OR = 1.20, p = 0.016) were associated with CVD complications in AS. Receiver operating characteristic (ROC) curves showed a contribution of Th17 cell for distinguishing AS patients with CVD, with the areas under the ROC curve (AUCs) of 0.729 (95%CI: 0.632–0.825, p < 0.001). Conclusion Our findings provide evidence for the association between Th17 cells and increased cardiovascular risk in AS. Th17 cells may contribute to accelerated atherogenesis and increased cardiovascular burden in AS and be valuable for early assessment and management of AS-CVD.

BackgroundRheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease that results in the destruction of the bone and cartilage of the joints[1].The imbalance between Th17 and Treg cells has been demonstrated to play a role in RA pathogenesis[2].With the advances in the RA field, curing and even preventing RA has become a new direction of efforts. To explore the immune characteristics in multi-staging RA is helpful for the development of accurate strategies.ObjectivesTo observe the immune characteristics in different staging rheumatoid arthritis patientsMethodsA total of 127 patients with RA were recruited for this study, in cluding 30 early RA(newly diagnosed and treatment-naive,duration of disease ≤3months), 30 midddle RA (newly diagnosed and treatment-naive, duration of disease >3months and≤1 year), 30 advanced RA(newly diagnosed and treatment-naive,duration of disease >1 year) and 37 active treated RA (remissioned after target treatment and this time due to disease recurrence in hospital). All patients were hospitalized in the second hospital of Shanxi medical university and diagnosed with RA fulfilled the ACR 1987 or 2010 criteria.The percentage and absolute numbers of lymphocyte phenotypes and CD4+ T subsets in peripheral blood were examined by flow cytometry.ResultsThere were no significant difference in the percentage and value of T,B,NK,CD4+T,CD8+T andCD4+T subsets. But the Th1/Th2 cell ratios in active treated RA group were increased than that in early RA, middle RA and advanced RA group.The Th17/Treg cell ratios in middle RA and advanced RA group were higher when compared with those of active treated RA group.In addition, the Th1/ Treg cell ratios were decreased in middle and advanced RA group as compared with those in active treated RA.ConclusionTh1/Th2,Th17/Treg and Th1/Treg imbalance varied with disease staging.The different disease staging in RA, the immune function is different. Providing targeted immunotherapy according to disease staging is helpful to improve remission rates and even prevent disease.References[1]Petrovská N, Prajzlerová K, Vencovský J, Šenolt L, Filková M.The pre-clinical phase of rheumatoid arthritis: From risk factors to prevention of arthritis.Autoimmun Rev.2021, 20(5):102797.[2]Alison M Gizinski, David A Fox.T cell subsets and their role in the pathogenesis of rheumatic disease.Curr Opin Rheumatol,2014;26(2):204-10.Figure 1.The comparsion about the lymphocyte subsets and cell ratios in peripheral blood of different stage RA patients.(*p<0.05)Disclosure of InterestsNone declared

Background:The incidence of Ankylosing Spondylitis (AS) complicated with cardiovascular diseases (CVD) has increased in recent years [1]. However, identification of risk factors indicating the development of CAD in AS patients is lacking. Th17 cells are increasingly recognized to be important in atherogenesis [2]. However, the role of these cells in the pathogenesis of ankylosing spondylitis patients complicated with cardiovascular events remains elusive.Objectives:This study aimed to assess the level of circulating Th17 cells as well as other lymphocyte subsets such as Treg, Th, Ts, and NK cells in AS combined with CVD, and further to evaluate whether elevations in special PBMC subpopulations in AS patients indicate an increased risk of CVD.Methods:Samples were assessed from 141 AS patients hospitalized at the Second Hospital of Shanxi Medical University (60 AS patients combined with CVD and 81 AS patients without CVD) and 100 healthy controls. The absolute numbers of lymphocytes and CD4+ T cells in peripheral blood were determined using Flow Cytometer. The association between PBMC subpopulations and CVD development in AS patients were analyzed using multivariable logistic regression.Results:1. Compared with AS group, AS with CVD group exhibited significant increases in the number of Th17 cells (P=0.001) and Treg cells (P=0.046). The ratio of Th17/Treg was also increased (P=0.085).2. Analogous increases in the absolute number (P<0.001) and frequency (P<0.001) of Th1 cells, as well as the ratio of Th1/Th2 (P<0.001) and Th1/Treg (P=0.004) were also present in AS with CVD patients, compared to those without CVD.3. Compared to HCs, 141 AS patients showed significantly decreased Treg cells (P<0.012) and increased Th17/Treg (P=0.001).4. Logistic regression showed age (odds ratio: 1.09; 95% CI: 1.035-1.137), hypertension (odds ratio: 3.31; 95% CI: 1.152-9.528), diabetes (odds ratio: 8.03; 95% CI: 1.251-51.503), and elevated level of Th1 number (odds ratio: 1.01; 95% CI: 1.003-1.016) and DD (D-dimer) (odds ratio: 1.00; 95% CI: 1.000-1.002) were significantly correlated with the onset of CVD in AS patients.5. Smoke, increased Th17 level, and use of NSAIDS were also positively correlated with the onset of CVD although the P-values did not reach significant.Conclusion:Our data indicates aberrant expansion of Th17 cells in AS with CVD patients. Moreover, age, hypertension, diabetes, and increased level of Th1 in PBMC and DD are single independent risk factors for the presence of CVD in AS. The mechanisms of atherogenesis in AS may associate with the elevations in Th1 and Th17 cells. Imbalance of Th1/Th2 and Th17/Treg may be shared etiologic pathways of AS and CVD, providing attractive targets for the prevention and therapy of CVD development in AS patients.References:[1]Kim JH, Choi IA. Cardiovascular morbidity and mortality in patients with spondyloarthritis: A meta-analysis. Int J Rheum Dis (2020). doi: 10.1111/1756-185x.13970.[2]Saigusa R, Winkels H, Ley K. T cell subsets and functions in atherosclerosis. Nat Rev Cardiol. 2020 Jul;17(7):387-401. doi: 10.1038/s41569-020-0352-5.Figure 1.Compared with AS group, AS with CVD group exhibited significant increases in the number of Th17 cells (P=0.001) and Treg cells (P=0.046). The ratio of Th17/Treg was also increased (P=0.085). The absolute number (P<0.001) and frequency (P<0.001) of Th1 cells, as well as the ratio of Th1/Th2 (P<0.001) and Th1/Treg (P=0.004) were also present in AS with CVD patients.Disclosure of Interests:None declared.

BackgroundConnective tissue diseases (CTDs) are a group of diseases with a variety of clinical manifestations. The main drug was glucocorticoids and immunosuppressive drugs, but the results are not satisfactory and the side effects are obvious, increased the incidence of infection, especially opportunistic infections. Infections becomes important causes of morbidity and mortality in CTD patients.ObjectivesTo evaluate the incidence of infection in CTD patients who were clinically considered for co-infection by a combination of metagenomic next-generation sequencing (mNGS) and conventional diagnostic testing methods.MethodsWe analyzed 126 connective tissue diseases (CTD)patients with suspected infections admitted to The Second Hospital of Shanxi Medical University. All patients with CTD were diagnosed according to relevant diagnostic criteria, including 34 systemic lupus erythematosus (SLE), 24 dermatomyositis and polymyositis (DM/PM), 19 rheumatoid arthritis (RA), 10 undifferentiated connective tissue disease(UCTD),16 Sjogren syndrome (SS), 5 mixed connective tissue disease (MCTD), 5 ANCA associated systemtc vasculitis (AAV), 5 adult onset Stillystemtc isease disease(tic criteria, including 34nfections admitted to The Second Hospital of Shanxi (TA), 1 systemic sclerosis (SSC), 1retroperitoneal fibrosis (RPF). All enrolled patients were tested for conventional diagnostic testing methods(CDT) and mNGS.ResultsAmong the 126 patients with CTD who were clinically considered for co-infection, 31 patients were negative for mNGS, and pathogens were detected in 99 of them. In our results, the mNGS and CDT were both positive for pathogens detection in 28 individuals.Of both positive individuals, 2 cases were perfect matches,12 cases were partly matched, 14 cases were totally mismatched. A total of 23 cases were negative for both mNGS and CDT. 70 cases were positive for mNGS only.There were only 5 cases positive for pathogens detection by CDT only. In addition, the results of mNGS showed that 131 patients were virus-positive(54%), 78 patients were prokaryotes-positive (37%) inculding bacteria, mycoplasma and 14 patients were eukaryotes-positive (9%). Of course, someones have mixed infections among these patinets some of these patients, with two or more pathogens. In the mixed infection, 5 cases have no viruses infection, 38 cases with virus infection, including 20 cases of bacteria and viruses infection, 4 cases of bacteria,fungi and viruses infection, 9 cases of viruses mixed infection, 1 case of bacteria,viruses,fungi and mycoplasma infection, 1 case of bacteria,viruses and mycoplasma infection, 1 case of viruses and mycoplasma infection, 1 case of viruses and fungi infection. According to the results, viruses were the most common pathogens identified, followed by prokaryotes and eukaryotes. It is noteworthy that the incidence of Human gammaherpesvirus 4(EBV), Human betaherpesvirus 5(CMV) and Human alphaherpesvirus 1 are more common in virus-positive. The most frequently detected prokaryotes were Acinetobacter baumannii, Mycobacterium tuberculosis complex, followed by Staphylococcus aureus, Prevotella melaninogenica,Staphylococcus homini and Helicobacter pylori. The major pathogens were Pneumocystis jirovecii and Candida albicans among eukaryotes-positive individuals.ConclusionAs a complementary approach to conventional methods, mNGS could help improving the identification of infection in CTD patients.The incidence of viral infection is high in patients with connective tissue disease and close attention should be paid to it in clinical works.Figure 1.A. Comparison of test results between mNGS and conventional diagnostic testing methods(CDT) in CTD patients. B. The classification of mixed infections with or without viruses infection detected by mNGS and conventional diagnostic testing methods(CDT).Figure 2.Distribution of pathogens detected by mNGS. A. Type distribution of pathogens identified by mNGS. Species distribution of viruses of B.viruses, C.Prokayote, D. Eucayon detected by mNGS.Disclosure of InterestsNone declared