Elevated type I IFN signalling directly affects CD8+ T-cell distribution and autoantigen recognition of the skeletal muscles in active JDM patients.

The link between type I IFN and adaptive immunity, especially T-cell immunity, in JDM still remained largely unclear. This study aimed to understand the effect of elevated type I IFN signaling on CD8+ T cell-associated muscle damage in juvenile dermatomyositis (JDM). This study used flow cytometry (FC) and RT‒PCR were used to examine the circulating cell ratio and type I IFN response. And scRNA-seq was used to examine peripheral immunity in 6 active JDM patients, 3 stable JDM patients, 3 juvenile IMNM patients and 3 age-matched healthy children. In vivo validation experiments were conducted using a mouse model induced by STING agonists and an experimental autoimmune myositis model (EAM). In vitro experiments were conducted using isolated CD8+ T-cells from JDM patients and mice. We found that active JDM patients showed an extensive type I IFN response and a decreased CD8+ T-cell ratio in the periphery (P < 0.05), which was correlated with muscle involvement (P < 0.05). Both new active JDM patients and all active JDM patients showed decreased CD8+ TCM cell ratios compared with age and gender matched stable JDM patients (P < 0.05). Compared with new pediatirc systemic lupus erythematosus (SLE) patients, new active JDM patients displayed decreased CD8+ T-cell and CD8+ TCM cell ratios (P < 0.05). Active JDM patient skeletal muscle biopsies displayed an elevated type I IFN response, upregulated MHC-I expression and CD8+ T-cell infiltration, which was validated in EAM mice. sc-RNAseq demonstrated that type I IFN signalling is the kinetic factor of abnormal differentiation and enhances the cytotoxicity of peripheral CD8+ T cells in active JDM patients, which was confirmed by in vivo and in vitro validation experiments. In summary, the elevated type I IFN signalling affected the differentiation and function of CD8+ T cells in active JDM patients. Skeletal muscle-infiltrating CD8+ T cells might migrate from the periphery under the drive of type I IFN and increased MHC I signals. Therapies targeting autoantigen-specific CD8+ T cells may represent a potential new treatment direction.

The Association between Perceived Family Financial Stress and Adolescent Suicide Ideation: A Moderated Mediation Model.

Nowadays, suicide (especially adolescents' suicide) has been an increasingly prominent social problem worldwide; suicide ideation, as an important predictor, has been the focus of relevant studies and practices. Against this background, the present study aimed to examine the association between perceived family financial stress and adolescents' suicidal ideation, as well as the potential roles of depression and parent-child attachment. A sample of 526 junior middle school students was recruited voluntarily to participate in this cross-sectional study, and the results indicated that the prevalence of suicidal ideation among junior high school students was 15.45%; perceived family financial stress was positively associated with suicidal ideation, and depression could significantly mediate this relation; parent-child attachment significantly moderated the mediating effect of depression (in particular, the relation between depression and suicidal ideation); specifically, this relation was stronger among adolescents with lower values of parent-child attachment. These findings could deepen our understanding of the influences of perceived family financial condition and the risky factors of adolescents' suicidal ideation, which could provide guidance for the prevention and intervention of adolescents' depression and suicidal ideation.

Identification and Characterization of CD8+ CD27+ CXCR3- T Cell Dysregulation and Progression-Associated Biomarkers in Systemic Lupus Erythematosus.

Systemic Lupus Erythematosus (SLE) etiopathogenesis highlights the contributions of overproduction of CD4+ T cells and loss of immune tolerance. However, the involvement of CD8+ T cells in SLE pathology and disease progression remains unclear. Here, the comprehensive immune cell dysregulation in total 263 clinical peripheral blood samples composed of active SLE (aSLE), remission SLE (rSLE) and healthy controls (HCs) is investigated via mass cytometry, flow cytometry and single-cell RNA sequencing. This is observed that CD8+ CD27+ CXCR3- T cells are increased in rSLE compare to aSLE. Meanwhile, the effector function of CD8+ CD27+ CXCR3- T cells are overactive in aSLE compare to HCs and rSLE, and are positively associated with clinical SLE activity. In addition, the response of peripheral blood mononuclear cells (PBMCs) is monitored to interleukin-2 stimulation in aSLE and rSLE to construct dynamic network biomarker (DNB) model. It is demonstrated that DNB score-related parameters can faithfully predict the remission of aSLE and the flares of rSLE. The abundance and functional dysregulation of CD8+ CD27+ CXCR3- T cells can be potential biomarkers for SLE prognosis and concomitant diagnosis. The DNB score with accurate prediction to SLE disease progression can provide clinical treatment suggestions especially for drug dosage determination.

Low-Dose Staphylococcal Enterotoxin C2 Mutant Maintains Bone Homeostasis via Regulating Crosstalk between Bone Formation and Host T-Cell Effector Immunity.

Studies in recent years have highlighted an elaborate crosstalk between T cells and bone cells, suggesting that T cells may be alternative therapeutic targets for the maintenance of bone homeostasis. Here, it is reported that systemic administration of low-dose staphylococcal enterotoxin C2 (SEC2) 2M-118, a form of mutant superantigen, dramatically alleviates ovariectomy (OVX)-induced bone loss via modulating T cells. Specially, SEC2 2M-118 treatment increases trabecular bone mass significantly via promoting bone formation in OVX mice. These beneficial effects are largely diminished in T-cell-deficient nude mice and can be rescued by T-cell reconstruction. Neutralizing assays determine interferon gamma (IFN-γ) as the key factor that mediates the beneficial effects of SEC2 2M-118 on bone. Mechanistic studies demonstrate that IFN-γ stimulates Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling, leading to enhanced production of nitric oxide, which further activates p38 mitogen-activated protein kinase (MAPK) and Runt-related transcription factor 2 (Runx2) signaling and promotes osteogenic differentiation. IFN-γ also directly inhibits osteoclast differentiation, but this effect is counteracted by proabsorptive factors tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1ß) secreted from IFN-γ-stimulated macrophages. Taken together, this work provides clues for developing innovative approaches which target T cells for the prevention and treatment of osteoporosis.

O-alg-THAM/gel hydrogels functionalized with engineered microspheres based on mesenchymal stem cell secretion recruit endogenous stem cells for cartilage repair.

Lacking self-repair abilities, injuries to articular cartilage can lead to cartilage degeneration and ultimately result in osteoarthritis. Tissue engineering based on functional bioactive scaffolds are emerging as promising approaches for articular cartilage regeneration and repair. Although the use of cell-laden scaffolds prior to implantation can regenerate and repair cartilage lesions to some extent, these approaches are still restricted by limited cell sources, excessive costs, risks of disease transmission and complex manufacturing practices. Acellular approaches through the recruitment of endogenous cells offer great promise for in situ articular cartilage regeneration. In this study, we propose an endogenous stem cell recruitment strategy for cartilage repair. Based on an injectable, adhesive and self-healable o-alg-THAM/gel hydrogel system as scaffolds and a biophysio-enhanced bioactive microspheres engineered based on hBMSCs secretion during chondrogenic differentiation as bioactive supplement, the as proposed functional material effectively and specifically recruit endogenous stem cells for cartilage repair, providing new insights into in situ articular cartilage regeneration.

Long-term follow-up of Janus-kinase inhibitor and novel active disease biomarker in juvenile dermatomyositis.

OBJECTIVE: To evaluate the use of Janus kinase inhibitor (JAKi) in treating JDM and develop cytokine biomarkers of active disease. METHODS: This study involved a retrospective cohort study that evaluated JAKi in 101 JDM patients as well as a cross-sectional study of cytokines in 128 JDM patients and 30 controls between November 2017 and December 2021 in the Affiliated Children's Hospital of Capital Institute of Pediatrics (China). RESULTS: During the median follow-up period of 19 months, 65.5% of the patients had improved rashes, and CAT-BM scores decreased. Overall, 39.6% of JDM patients eliminated glucocorticoids. Muscle strength was improved in all patients who had abnormal muscle strength before JAKi use. Patients and parents provided positive subjective reviews of JAKi, and no serious adverse events were reported. Potential side effects of JAKi included abnormal leukopoenia (14/95) and cough (16/83), which affected over 10% of the JDM patients. In the cytokine analysis, 12/34 cytokines were significantly elevated in active JDM patients. Compared with active JDM patients with multiple phenotypes, active JDM patients with only rashes demonstrated lower cytokine levels. Anti-NXP2-positive active patients had lower cytokine levels compared with those without positive anti-NXP2 antibodies. Among all increased cytokines, IL-1RA changed most dramatically, reaching over 793 times the mean of normal values. We developed a panel composed of six cytokines to differentiate active or stable status in our cohort (area under the curve = 0.8486, P < 0.05). CONCLUSION: The preliminary evidence suggested that JAKi is a relatively safe and effective alternative for JDM patients. Cytokine profiles could well reflect the inflammatory status of JDM patients.

Interferon response and profiling of interferon response genes in peripheral blood of vaccine-naive COVID-19 patients.

Introduction: There is insufficient understanding on systemic interferon (IFN) responses during COVID-19 infection. Early reports indicated that interferon responses were suppressed by the coronavirus (SARS-CoV-2) and clinical trials of administration of various kinds of interferons had been disappointing. Expression of interferon-stimulated genes (ISGs) in peripheral blood (better known as interferon score) has been a well-established bioassay marker of systemic IFN responses in autoimmune diseases. Therefore, with archival samples of a cohort of COVID-19 patients collected before the availability of vaccination, we aimed to better understand this innate immune response by studying the IFN score and related ISGs expression in bulk and single cell RNAs sequencing expression datasets. Methods: In this study, we recruited 105 patients with COVID-19 and 30 healthy controls in Hong Kong. Clinical risk factors, disease course, and blood sampling times were recovered. Based on a set of five commonly used ISGs (IFIT1, IFIT2, IFI27, SIGLEC1, IFI44L), the IFN score was determined in blood leukocytes collected within 10 days after onset. The analysis was confined to those blood samples collected within 10 days after disease onset. Additional public datasets of bulk gene and single cell RNA sequencing of blood samples were used for the validation of IFN score results. Results: Compared to the healthy controls, we showed that ISGs expression and IFN score were significantly increased during the first 10 days after COVID infection in majority of patients (71%). Among those low IFN responders, they were more commonly asymptomatic patients (71% vs 25%). 22 patients did not mount an overall significant IFN response and were classified as low IFN responders (IFN score < 1). However, early IFN score or ISGs level was not a prognostic biomarker and could not predict subsequent disease severity. Both IFI27 and SIGLEC1 were monocyte-predominant expressing ISGs and IFI27 were activated even among those low IFN responders as defined by IFN score. In conclusion, a substantial IFN response was documented in this cohort of COVID-19 patients who experience a natural infection before the vaccination era. Like innate immunity towards other virus, the ISGs activation was observed largely during the early course of infection (before day 10). Single-cell RNA sequencing data suggested monocytes were the cell-type that primarily accounted for the activation of two highly responsive ISGs (IFI44L and IFI27). Discussion: As sampling time and age were two major confounders of ISG expression, they may account for contradicting observations among previous studies. On the other hand, the IFN score was not associated with the severity of the disease.

SCANCell reveals diverse inter-cluster interaction patterns in systemic lupus erythematosus across the disease spectrum.

MOTIVATION: High-dimensional mass cytometry (CyTOF), which provides both cellular signatures and inter-cluster interactions like the antagonism between immune activation and suppression, and the pro-inflammatory synergy, sheds light on the cellular and molecular basis of disease pathogenesis. However, revealing the aberrance of inter-cluster communication networks in CyTOF datasets remains a significant challenge. RESULTS: Here, we developed Sample Classification and direct Association Network among Cell clusters (SCANCell) that quantifies the direct association (DA) network of cell clusters. SCANCell was applied to profile inter-cluster interaction patterns of a well-recruited systemic lupus erythematosus (SLE) cohort, including 8 healthy controls, 10 active SLE patients (APs) and 8 remission SLE patients (RPs). SCANCell identified decreased inter-cluster interactions of CD8+ T cells in APs compared with RPs, and enhanced DA of CD8+ T cells after stimulation with immunostimulatory cytokine interleukin-2 in vitro. These discoveries prove that SCANCell can uncover pathology- and drug stimulation-associated inter-cluster interactions, which potentially benefits understanding of pathogenesis and novel therapeutic strategies. AVAILABILITY AND IMPLEMENTATION: The main processing scripts of SCNACell are available at https://github.com/Lxc417/SCANCell. Other codes for the following data statistics are available from the corresponding author upon request. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Reduced Sleep in the Week Prior to Diagnosis of COVID-19 is Associated with the Severity of COVID-19.

BACKGROUND: The rapid outbreak of coronavirus disease 2019 (COVID-19) is a major health concern, in response to which widespread risk factor research is being carried out. OBJECTIVE: To discover how physical activity and lifestyle affect the epidemic as well as the disease severity and prognosis of COVID-19 patients. METHODS: This multicenter, retrospective cohort study included 203 adults infected with COVID-19 and 228 uninfected adults in three Chinese provinces, with 164 (80.7%) of the infected participants and 188 (82.5%) of the uninfected participants answering a doctor-administered telephone questionnaire on lifestyle. The binary logistic regression model and the ordinal logit model were used to observe relevance. RESULTS: Comparing sick and non-sick patients, we found that irregular exercise (P=0.004), sedentary lifestyle (P=0.010), and overexertion (P<0.001) may be associated with the susceptibility to COVID-19. In symptomatic patients, using the recommended status as a reference, risk of severe infection increased with decreased sleep status, being 6.729 (95% CI=2.138-21.181) times higher for potentially appropriate sleep (P=0.001) and peaking at 8.612 (95% CI=1.913-38.760) times higher for lack of sleep (P=0.005). Reduction in average daily sleep time significantly increased the likely severity (P=0.002). DISCUSSION: Through further examination of damage of external lung organs, we found that lack of sleep affected not only disease severity but also prognosis. Based on these findings, the public should prioritize a healthy lifestyle and get adequate sleep in response to the outbreak. The study of life habits may bring new ideas for the prevention and treatment of COVID-19.

Electrochemical impedance spectroscopy study on polymerization of L-lysine on electrode surface and its application for immobilization and detection of suspension cells.

Poly-L-lysine (PLL), which has been employed as a conductive polymer in the construction of some electrochemical sensors, can be prepared using L-lysine by cyclic voltammetry (CV) with a wide potential range. However, the presented explanation and description about its polymerization mechanism seems oversimplified because the self-reaction of electrode and the electrolysis of solvent at high potential are ignored. This work presents an intensive investigation on the relevant reactions during the process of PLL-polymerization using CV, X-ray photoelectron spectroscopy, Fourier transform-infrared spectroscopy, and electrochemical impedance spectroscopy. At a higher positive potential, the transfer from lysine molecules to cation radicals and the polymerization reaction on the glassy carbon electrode (GCE) could be achieved, accompanied by the activation of GCE, the formation of oxygen-containing functional groups, and the generation of oxygen derived from the oxidation of water. The adsorbed oxygen had a seriously negative effect on the formation of PLL unless it suffered reduction at a lower negative potential. The charge transfer through the electrochemical polymerized PLL film was seriously hindered by the immobilization of suspension cells due to the electrostatic interaction. The charge-transfer resistance difference (ΔR(ct)) was increased with the enhancement of the cell number (N(cells)) and the 1/ΔR(ct) value displayed a linear response with 1/N(cells) in the range of 5.0 × 10(2)-1.0 × 10(5) cells with a detection limit of 180 cells estimated at a signal-to-noise ratio of 3. A sensitive electrochemical sensor for the quantitative detection of suspension cells was developed.

Electrochemical immunoassay for carcinoembryonic antigen using gold nanoparticle-graphene composite modified glassy carbon electrode.

Carcinoembryonic antigen (CEA), which is typically associated with certain tumors and developing fetus, is widely used as a clinical tumor marker for some familiar cancers. In this work, a simple and sensitive electrochemical CEA sensor was developed by employing immunoreaction. Gold nanoparticle-decorated graphene composites (Au-GN) were successfully synthesized based on the reduction of HAuCl4 in the presence of graphene. Horseradish peroxidase-labeled anti-CEA antibody (HRP-anti-CEA) and HRP were successively adsorbed on the Au-GN modified glassy carbon electrode. The stepwise assembly process of the immunosensor was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The introduction of CEA antigens on the electrode surface reduced the electrochemical response of the electron transfer mediator due to the strong steric effect. Under the optimized conditions, the peak current change derived from the differential pulse voltammetry (DPV) measurements (ΔIDPV) was proportional to the CEA concentration from 0.10 to 80 ng mL(-1) with a detection limit of 0.04 ng mL(-1) (S/N=3). In addition, this new protocol shows good selectivity, stability and reproducibility. The determination of CEA in human serum samples was performed and received in excellent accordance with the results determined by the enzyme-linked immunosorbent assay (ELISA).

Interaction between maternal passive smoking during pregnancy and CYP1A1 and GSTs polymorphisms on spontaneous preterm delivery.

OBJECTIVE: The present study aimed to examine the association between maternal passive smoking during pregnancy and the risk of spontaneous PTD and to explore the potential interaction of the single or joint gene polymorphism of CYP1A1 and GSTs with maternal passive smoking on the risk of spontaneous PTD. METHOD: We investigated whether the association between maternal passive smoking and PTD can be modified by 2 metabolic genes, i.e. cytochrome P4501A1 (CYP1A1) and glutathione S-transferases (GSTs), in a case-control study with 198 spontaneous preterm and 524 term deliveries in Shenzhen and Foshan, China. We used logistic regression to test gene-passive smoking interaction, adjusting for maternal socio-demographics and prepregnancy body mass index. RESULTS: Overall, maternal passive smoking during pregnancy was associated with higher risk of PTD (adjusted odds ratio = 2.20 [95% confidence interval: 1.56-3.12]). This association was modified by CYP1A1 and GSTs together, but not by any single genotype. For cross-categories of CYP1A1 Msp I and GSTs, maternal passive smoking was associated with higher risk of PTD among those women with CYP1A1 "TC/CC"+ GSTs "null", but not among women with other genotypes; and this interaction was significant (OR = 2.66 [95% CI: 1.19-5.97]; P-value: 0.017). For cross-categories of CYP1A1 BsrD I and GSTs, maternal passive smoking was associated with higher risk of PTD only among those women with CYP1A1"AG/GG"+ GSTs "null", but not among women with other genotypes; and this interaction was significant (OR = 3.00 [95% CI: 1.17-7.74]; P-value: 0.023). CONCLUSIONS: Our findings suggest that the combined genotypes of CYP1A1 and GSTs can help to identify vulnerable pregnant women who are subject to high risk of spontaneous PTD due to passive smoking.

Enhancing the efficiency of Epstein-Barr viral serologic test in the diagnosis of nasopharyngeal carcinoma.

OBJECTIVE: To investigate the efficiency of concurrent application of VCA-IgA, EA-IgA and EA-IgG serological tests in diagnosing nasopharyngeal carcinoma (NPC). METHODS: The sera of 266 untreated NPC patients and 347 healthy adults were collected. In addition to the conventional immunoenzymatic method of VCA-IgA test, enzyme-linked immunosorbent assay (ELISA) was adopted as an alternative to test the antibody level of EA-IgG and EA-IgA. A new statistical formula was used to evaluate the odds ratio of different combinations of these three tests. RESULTS: The sensitivity and specificity of VCA-IgA, EA-IgG and EA-IgA concurrently were as high as 95.11% and 97.41%, respectively, which were higher than those of single test (90.60% and 94.52% for VCA-IgA, 93.98% and 93.66% for EA-IgG, 89.84% and 88.18% for EA-IgA). Furthermore, the odds ratio of 3-test positivity (1 912.5) was higher than those of 2-test positivity (27.903 2 for VCA-IgA and EA-IgG, 11.169 0 for EA-IgG and EA-IgA, 8.032 8 for VCA-IgA and EA-IgA), which were even higher than those of 1-test positivity (0.121 4 for VCA-IgA, 0.170 5 for EA-IgG and 0.048 8 for EA-IgA). CONCLUSION: ELISA is more accurate in reflecting the antibody level of EA-IgG and EA-IgA than the conventional immunoenzymatic method. The concurrent application of VCA-IgA, EA-IgG and EA-IgA test can markedly improve the sensitivity, specificity and odds ratio as well, thus resulting in enhancing the efficiency of diagnosing nasopharyngeal carcinoma serologically.