Aberrant phenotypes of circulating γδ-T cells may be involved in the onset of systemic lupus erythematosus.

OBJECTIVE: Human gamma-delta T cells (γδ-T cells) play crucial roles in both innate and adaptive immune responses. However, much less is known about the immune status of γδT cells in systemic lupus erythematosus (SLE) patients. The objective of this study was to explore potential relationships between the frequency of γδ-T-cell subpopulations and disease activity, autoantibody titres and renal involvement in patients with SLE. METHODS: Circulating γδ-T cells and their subsets (Vδ1+ T cells, Vδ2+ T cells and γδ-T-cell subpopulations defined by expression of surface receptors, including NKG2D, NKp30, NKp46 and PD-1), were identified via flow cytometry. Sixty active SLE patients were selected, including 41 new-onset and 19 relapsing cases. One hundred healthy controls (HCs) were enrolled as the control group. Percentages of these cell subsets in SLE patients and HCs and their relationships with disease activity were analysed. Twenty-two of the 41 new-onset SLE patients were assessed before and after treatment. Changes in the frequencies of these cell subsets and their relationships with renal involvement were also analysed. RESULTS: Compared with that in HCs, the percentage of total γδ-T cells among CD3+ T cells in SLE patients was significantly lower. An imbalance in the proportions of Vδ1+ and Vδ2+ T cells among γδ-T cells was observed. The proportion of Vδ1+ T cells among γδ-T cells was significantly greater in SLE patients than in HCs, while the proportion of Vδ2+ T cells was significantly lower. Expression levels of PD-1, NKG2D, NKp30 and NKp46 in Vδ1+ T cells and Vδ2+ T cells from SLE patients were generally significantly increased, except for expression of NKG2D in Vδ2+ T cells. Moreover, Vδ2+ T cells, Vδ1+ T cells and Vδ1+PD-1+ T cells were associated with disease activity, and an increase in Vδ2+ T-cell frequency and a decrease in PD-1 expression by γδ-T cells might be associated with effective treatment. Interestingly, our results indicated that Vδ2+ T cells and their Vδ2+NKp30+ T-cell subpopulation might be associated with renal involvement in SLE. CONCLUSION: A broad range of anomalies in the proportions of γδ-T-cell subsets and γδ-T cells in SLE patients may be involved in the pathogenesis of SLE. There is a strong association between Vδ2+ T cells and their Vδ2+NKp30+ T-cell subpopulation and LN occurrence. Our results indicate that γδ-T cells and their subpopulations might be key players in disease immunopathology and renal involvement in SLE.

Time-Frequency Aliased Signal Identification Based on Multimodal Feature Fusion.

The identification of multi-source signals with time-frequency aliasing is a complex problem in wideband signal reception. The traditional method of first separation and identification especially fails due to the significant separation error under underdetermined conditions when the degree of time-frequency aliasing is high. The single-mode recognition method does not need to be separated first. However, the single-mode features contain less signal information, making it challenging to identify time-frequency aliasing signals accurately. To solve the above problems, this article proposes a time-frequency aliasing signal recognition method based on multi-mode fusion (TRMM). This method uses the U-Net network to extract pixel-by-pixel features of the time-frequency and wave-frequency images and then performs weighted fusion. The multimodal feature scores are used as the classification basis to realize the recognition of the time-frequency aliasing signals. When the SNR is 0 dB, the recognition rate of the four-signal aliasing model can reach more than 97.3%.

[Expression of regulatory B cells in peripheral blood of patients with systemic lupus erythematosus].

OBJECTIVE: To explore the expression of regulatory B cells (Bregs) in peripheral blood of patients with systemic lupus erythematosus (SLE) and elucidate the function of Bregs in the pathogenesis of SLE. METHODS: A total of 38 active SLE patients, 22 inactive SLE subjects and 20 healthy controls were enrolled. The expressions of IL-10+ CD19+ Breg and CD19+ CD24hi CD38hi Breg in peripheral blood mononuclear cells (PBMC) were evaluated by flow cytometry. And IL-10 was measured in the culture supermatants by enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression of IL-10+ CD19+ Breg in PBMCs of active SLE patients was (1.54±0.64)%. And it was lower than those in inactive SLE patients (2.42±0.75)% (P<0.01) and healthy controls (4.35±1.00)% (P<0.01). And the expression of CD19+ CD24hi CD38hi Breg was (1.26±0.45)%. Also it was lower than those in inactive SLE subjects (2.01±0.61)% (P<0.01) and healthy controls (3.14±0.87)% (P<0.01).The level of IL-10 significantly decreased in culture supermatants of active SLE patients. And it was lower than those in inactive SLE subjects (P<0.05) and healthy controls (P<0.01). The percentage of IL-10+ CD19+ Breg and CD19+ CD24hi CD38hi Breg in PBMCs of SLE patients was positively correlated with the level of IL-10 in culture supernatants (r=0.652, P<0.01 and r=0.574, P<0.01). CONCLUSION: The expression of IL-10-related Bregs significantly decreases in PBMCs of active SLE patients. And Bregs may play some role in the pathogenesis of SLE.

Prolactin increases tumor necrosis factor alpha expression in peripheral CD14 monocytes of patients with rheumatoid arthritis.

Tumor necrosis factor (TNF)-α is one of the major proinflammatory mediators of rheumatic arthritis (RA); the regulatory factors for TNF-α release is not fully understood. This study aims to investigate the role of prolactin receptor (PRLR) activation in regulating the expression and release of TNF-α from CD14(+) monocytes. The results showed that the expression of PRLR was detectable in CD14(+) monocytes of healthy subjects, which was markedly increased in RA patients. Exposure to PRL in the culture increased the expression and release of TNF-α from CD14(+) monocytes, which was abolished by the PRLR gene silencing or blocking the mitogen activated protein (MAPK) pathway. We conclude that exposure to PRL increases TNF-α release from CD14(+) monocytes of RA patients, which can be abolished by PRLR gene silencing or treating with MAPK inhibitor.

Hypomethylation of interleukin 6 correlates with renal involvement in systemic lupus erythematosus.

OBJECTIVE: To analyze peripheral blood interleukin 6 (IL-6) methylation status and its clinical significance in patients with systemic lupus erythematosus (SLE). MATERIAL AND METHODS: Blood samples from 41 adult patients with SLE, and 20 healthy controls were collected. The methylation status of IL-6 was determined by methylation specific polymerase chain reaction (MSP). The IL-6 expression was detected by real-time PCR. Correlations between IL-6 methylation status and clinical features or laboratory findings in patients with SLE were investigated. RESULTS: The methylation status and expression of IL-6 in peripheral blood could reflect the level in peripheral blood mononucleated cells (PBMCs) of SLE. Significantly positive correlation was found between IL-6 hypomethylation and renal disorder. Interleukin 6 hypomethylation was found negatively correlated with serum C3. CONCLUSIONS: The detection of IL-6 methylation status in peripheral blood could reflect the status in PBMC with SLE. Interleukin 6 may play a role in renal disorder with SLE patients. Interleukin 6 could be considered as a new biomarker for predicting SLE flare.

[Detection significance of Th17 cells in peripheral blood of patients with rheumatoid arthritis in clinical remission].

OBJECTIVE: To explore the significance and evaluate the early structural erosion through the expressions of Th17 cells in peripheral blood of patients with rheumatoid arthritis (RA) in clinical remission. METHODS: A total of 41 active RA patients without structural erosion were selected. Intracelluar flow cytometric detection of Th17 cells in peripheral blood was performed. And the supernatant level of interleukin (IL)-17A was determined simultaneously in RA patients and control groups at baseline and endpoint of 24-month therapy. The correlations were analyzed between Th17 cells and RA disease activity index DAS28. They were classified into radiographic progression (P, n = 10) and radiographic non-progression groups (NP, n = 26) by the Sharp/van der Heijde score (SHS) at the endpoint. The differences of Th17 cells and IL-17A levels were analyzed between P (SHS > 0.5) and NP groups (SHS ≤ 0.5). RESULTS: The expression of Th17 cells in active RA patients was significantly higher than that of controls [(1.63 ± 0.45)% vs (0.91 ± 0.26)%, P < 0.01]. And the results of IL-17A level were similar [1510 ± 280) vs (320 ± 31) ng/L, P < 0.05]. The expression of Th17 cells was positively correlated with DAS28 score (r = 0.87, P < 0.01). Thirty-six RA patients were followed up at the endpoint and all of them stayed in clinical remission (DAS28 < 2.6). The peripheral blood expressions of Th17 cells of P group were significantly higher than those of NP group . At the same time, no differences of IL-17A levels existed between two groups. CONCLUSION: Structural erosion still progresses in some RA patients despite an apparent clinic remission. And a high-level peripheral expression of Th17 hints at structural erosion.

Diagnostic Values of METTL1-Related Genes and Immune Characteristics in Systemic Lupus Erythematosus.

Purpose: Methyltransferase like 1 (METTL1) regulates epitranscriptomes via the m7G modification in mammalian mRNA and microRNA. Systemic lupus erythematosus (SLE) is caused by abnormal immune reactivity and has diverse clinical manifestations. RNA methylation as a mechanism to regulate gene expression is widely implicated in immune regulation. However, the role of m7G in immune response of SLE has not been extensively studied. Patients and Methods: Expression of METTL1 was identified in the public dataset GSE122459 and validated in an independent cohort of SLE patients. We investigated the association between METTL1-expression and clinical manifestations of SLE. Subsequently, differentially expressed genes (DEG) that were correlated with METTL1-expression in GSE122459 were used for functional enrichment analysis. The correlation between infiltrating immune cells and METTL1, as well as candidate biomarkers identified to be correlated with either METTL1 or immune cell infiltration were assessed by single-sample GSEA. Potential mechanisms were explored with Gene ontology and KEGG pathway enrichment. Diagnostic performances of candidate biomarkers in SLE were analyzed. Results: The mRNA and protein expression of METTL1 in SLE patients were significantly decreased in both datasets. METTL1-coexpressed DEGs were enriched in several key immune-related pathways. Activated CD8 T cells, activated CD4 T cells, memory B cells and type 2 helper T cells were different between patients with high and low METTL1 expression. Further, activated CD8 T-cells, activated CD4 T-cells, memory B-cells were correlated with METTL1. The genes of LAMP3, CD83, PDCD1LG2, IGKVD3D-20, IGKV5-2, IGKV2D-30, IGLV3-19 and IGLV4-60 were identified as candidate targets that were correlated with immune cell proportion. Moreover, LAMP3, CD83, and PDCD1LG2 expression were of diagnostic value in SLE as indicated by ROC analysis. Conclusion: Our findings suggested that METTL1 and its candidate targets LAMP3, CD83, PDCD1LG2 may be used for diagnosing SLE and could be explored for developing targeted molecular therapy for SLE.

[Expression and significance of Th17 and Treg cells in peripheral blood of patients with systemic lupus erythematosus].

OBJECTIVE: To explore the expression and significance of Th17 and Treg cells in peripheral blood of patients with systemic lupus erythematosus (SLE). METHODS: Thirty active SLE patients (including 17 SLE patients with lupus nephritis), 20 inactive SLE patients and 20 healthy controls were enrolled. The expressions of Th17 cells and CD4+CD25+Foxp3+Treg cells in peripheral blood mononuclear cells (PBMC) were evaluated by flow cytometry. The correlations between the expression of Th17 cells, CD4+CD25+Foxp3+Treg cells and disease activity (SLEDAI), as well as the ratios of Th17 cells and Treg cells (Th17/Treg) in SLE patients and healthy controls were analyzed respectively. RESULTS: The expression of Th17 cells in PBMC of SLE patients was higher than that in healthy controls [(1.39 ± 0.60)% vs (0.80 ± 0.33)%, P < 0.01] while the expression of CD4+CD25+Foxp3+Treg cells decreased in SLE patients [(3.09 ± 1.54)% vs (6.04 ± 1.49)%, P < 0.01]. The increased expression of Th17 cells and reduced CD4+CD25+Foxp3+Treg cells in PBMC were positively correlated with SLEDAI and negatively correlated with complements C3 and C4. There were increased expression of Th17 cells and reduced CD4+CD25+Foxp3+Treg cells in PBMC of lupus nephritis versus SLE patients without nephritis. CONCLUSION: There is an abnormal elevation of Th17 cells and decrease of CD4+CD25+Foxp3+Treg cells in PBMC of SLE patients. The imbalance between Th17 and Treg cells may play a critical role in the pathogenesis of SLE.

Inflammatory markers and risk factors of RA patients with depression and application of different scales in judging depression.

To evaluate the association of inflammatory markers and depression in RA patients and the risk factors in RA with depression, a cross-sectional study was conducted in a cohort of RA patients from southern China.Two hundred-fifteen RA patients were enrolled. The demographic and disease-related characteristics were recorded and inflammatory markers in sera were measured. RA patients were guided to fill out PHQ-9 scale by themselves, the psychological state was evaluated by psychiatry experts and graded according to the HAMD-17 scale. The consistency of the two scales in judging depression was evaluated. RA with depression group had HAMD-17 scores greater than 7. The levels of CRP, ESR, fibrinogen, SAA, IL-2, IL-6, TNF-α, IFN-γ, IL-4, and IL-10 were measured and compared. Logistic regression analysis was performed to find the risk factors of RA with different depression levels. One hundred-five (48.84%) RA patients had HAMD-17 scores greater than 7. High consistency was found between HAMD-17 and PHQ-9 in predicting depression. RA patients with depression were more likely to have tender joints, lower income, no employment, higher disease activity, joint deformities and glucocorticoid treatment. The depressed RA patients had higher serum levels of IL-6, CRP, fibrinogen, and SAA. IL-6, CRP, fibrinogen, and SAA were positive correlated with depression in RA patients. PHQ-9 can replace HAMD-17 in clinical application to judge depression.

Phenotypical changes and clinical significance of CD4+/CD8+ T cells in SLE.

OBJECTIVE: T cells display significant phenotypical changes and play multiple roles in promoting the immune response in SLE. The frequencies of T cell subpopulations in SLE are still not well understood. To better understanding the phenotypic abnormalities of T cells in SLE will help us to clarify disease immunopathology and to find promising biomarkers for disease monitoring and control. METHODS: Peripheral blood CD4+ and CD8+ T cells and their subsets were determined by flow cytometry. Forty-one active SLE patients were selected, including 28 new-onset patients and 13 relapsing patients. One hundred healthy controls (HCs) were enrolled as the control group. The percentages of these cell subsets between patients with SLE and HCs and their relationships with disease activity and autoantibody titers were analysed. Thirteen of 28 new-onset SLE patients were assessed before and after treatment. The changes in the frequencies of these cell subsets and their relationships with renal response were analysed. RESULTS: There was a broad range of anomalies in the proportion of T cell subsets in patients with SLE compared with that of the HCs. Compared with the HCs, a higher frequency of memory T cells and a lower frequency of naïve T cells were noted in patients with SLE. In addition, an imbalance of CD28+ and CD28- cells in CD4+ T cells was observed in patients with SLE. We found that the expanded CD4+CD28- T cells did not decrease after treatment in patients who had impaired renal responses. It was very interesting to exhibit a negative correlation in the frequency between the CD4+CD28- T cells and T regulatory (Treg) cells and a positive correlation between the frequency of CD4+CD28+ T cells and Treg cells in this study. Increased CD8+HLADR+ T cell and CD8+CD38+HLADR+ T cell counts were observed in patients with SLE, suggesting an impaired cytotoxic capacity of CD8+ T cells in SLE. Additionally, we found that CD8+CD38+HLADR+ T cells were closely associated with disease activity, autoantibody titres and renal prognosis. CD4+ CXCR5-PD1+ T cells were expanded in patients with SLE in this study and were associated with disease activity in SLE. Th1 (T helper type 1) cells and Treg cells were decreased, but frequencies of T follicular helper (Tfh) cells, Th2 cells, Th17 cells and Tfh17 cells were increased. A strong correlation between Th17 cells and Tregs with renal involvement was observed in this study. CONCLUSION: The proportions of CD4+CD28- T cells, CD4+CXCR5-PD1+ T cells, CD8+HLADR+ T cells and CD8+CD38+HLADR+ T cells increased in patients with SLE and could be associated with disease activity and renal prognosis.

miR-98 Modulates Cytokine Production from Human PBMCs in Systemic Lupus Erythematosus by Targeting IL-6 mRNA.

OBJECTIVE: There is evidence that interleukin-6 (IL-6) upregulation plays a critical role in immunopathology of systemic lupus erythematosus (SLE). MicroRNA- (miRNA-) 98 was predicted to bind with the 3'-untranslated region (3'-UTR) of IL-6 gene. We hypothesized miR-98 through its regulation of IL-6 gene expression to influence cytokine production from peripheral blood mononuclear cells (PBMCs) in SLE. METHODS: The expression of miR-98 and IL-6 mRNA in the PBMCs of 41 SLE patients and 20 healthy controls (HC) was detected by quantitative reverse transcription PCR (qRT-PCR). The correlations between miR-98 expression and clinical features were evaluated. Luciferase reporter assay was performed to identify miR-98 targets. miR-98 mimics, miR-98 inhibitor, and IL-6 overexpression vector were generated. Cell viability of PBMCs was assessed using MTT assay. Gene expression and protein level were determined by qRT-PCR and Western blotting. TNF-α, IL-8, IL-1ß, and IL-10 levels in cultured supernatants were quantified using ELISA. RESULTS: The expression of miR-98 was downregulated in PBMCs of SLE patients, and its expression is negatively associated with IL-6 levels. miR-98 expression was correlated with disease activity, lupus nephritis, and anti-dsDNA antibody. IL-6 mRNA was a target gene of miR-98. IL-6 overexpression promoted the proliferation of PBMCs and increased the levels of TNF-α, IL-8, IL-1ß, and IL-10. Those effects were further enhanced by miR-98 inhibitor, while were suppressed by miR-98 mimics. miR-98 regulated the levels of STAT3 phosphorylation via its target gene IL-6. CONCLUSION: The current study revealed that miR-98 could ameliorate STAT3-mediated cell proliferation and inflammatory cytokine production via its target gene IL-6 in patients with SLE. These results suggest that miR-98 might serve as a potential target for SLE treatment and other IL-6-mediated diseases.

Fetal and Maternal Outcomes of Planned Pregnancy in Patients with Systemic Lupus Erythematosus: A Retrospective Multicenter Study.

OBJECTIVE: To investigate the fetal and maternal outcomes as well as predictors of APOs in women with SLE who conceived when the disease was stable, the so-called "planned pregnancy." Methods. A retrospective multicenter study of 243 patients with SLE who underwent a planned pregnancy was performed. APOs in fetus and mothers were recorded. RESULTS: The average age at conception was 28.9 ± 3.9 years. Duration of SLE prior to pregnancy was 4.4 ± 4.3 years. Fetal APOs occurred in 86 (86/243, 35.4%) patients. Preterm births, intrauterine growth retardation (IUGR), fetal distress, and fetal loss accounted for 22.2%, 14.8%, 11.1%, and 4.9%, respectively. Forty-two preterm infants (42/54, 77.8%) were delivered after the 34th week of gestation. All the preterm infants were viable. Fifty-two patients (52/243, 21.4%) had disease flares, among which 45 cases (45/52, 86.5%) were mild, 6 (6/52, 11.5%) were moderate, and 1 (1/52, 1.9%) was severe. Disease flares were mainly presented as active lupus nephritis (41/52, 78.8%), thrombocytopenia (10/52, 19.2%), and skin/mucosa lesions (9/52, 17.3%). Pregnancy-induced hypertension (PIH) occurred in 29 patients, among which 3 were gestational hypertension and 26 were preeclampsia. Multiple analysis showed that disease flares (OR, 8.1; CI, 3.8-17.2) and anticardiolipin antibody positivity (OR, 7.4; CI, 2.5-21.8) were associated with composite fetal APOs. CONCLUSION: Planned pregnancy improved fetal and maternal outcomes, presenting as a lower rate of fetal loss, more favorable outcomes for preterm infants, and less severe disease flares during pregnancy.