Multidimensional Single Cell Based STAT Phosphorylation Profiling Identifies a Novel Biosignature for Evaluation of Systemic Lupus Erythematosus Activity

INTRODUCTION: Dysregulated cytokine action on immune cells plays an important role in the initiation and progress of systemic lupus erythematosus (SLE), a complex autoimmune disease. Comprehensively quantifying basal STATs phosphorylation and their signaling response to cytokines should help us to better understand the etiology of SLE. METHODS: Phospho-specific flow cytometry was used to measure the basal STAT signaling activation in three immune cell types of peripheral-blood mononuclear cells from 20 lupus patients, 9 rheumatoid arthritis (RA) patients and 13 healthy donors (HDs). A panel of 27 cytokines, including inflammatory cytokines, was measured with Bio-Plex™ Human Cytokine Assays. Serum Prolactin levels were measured with an immunoradiometric assay. STAT signaling responses to inflammatory cytokines (interferon α [IFNα], IFNγ, interleukin 2 [IL2], IL6, and IL10) were also monitored. RESULTS: We observed the basal activation of STAT3 in SLE T cells and monocytes, and the basal activation of STAT5 in SLE T cells and B cells. The SLE samples clustered into two main groups, which were associated with the SLE Disease Activity Index 2000, their erythrocyte sedimentation rate, and their hydroxychloroquine use. The phosphorylation of STAT5 in B cells was associated with cytokines IL2, granulocyte colony-stimulating factor (G-CSF), and IFNγ, whereas serum prolactin affected STAT5 activation in T cells. The responses of STAT1, STAT3, and STAT5 to IFNα were greatly reduced in SLE T cells, B cells, and monocytes, except for the STAT1 response to IFNα in monocytes. The response of STAT3 to IL6 was reduced in SLE T cells. CONCLUSIONS: The basal activation of STATs signaling and reduced response to cytokines may be helpful us to identify the activity and severity of SLE

Pregnancy outcomes among Chinese women with and without systemic lupus erythematosus: A retrospective cohort study

Objective To completely and quantifiably determine the effect of systemic lupus erythematosus (SLE) on pregnancy outcomes in a Chinese cohort. Design A retrospective cohort study. Setting Data were collected at a tertiary medical centre located in Shanghai, China, from September 2011 to May 2017. Participants We assigned 338 pregnant women with SLE to the study cohort and 1014 randomly selected pregnant women without SLE (three for every woman with SLE) to a comparison cohort. The relevant medical records of all pregnant women were retrospectively reviewed. Cases of multiple pregnancy and cases in which an artificial abortion was performed for personal reasons were excluded. Primary and secondary outcome measures Maternal and fetal outcomes were primary outcomes, and management of antenatal care was the secondary outcome. Results The risks of pregnancy-induced hypertension (OR 2.68, 95% CI 1.75 to 4.09), pre-eclampsia (OR 3.13, 95% CI 1.95 to 5.03) and premature rupture of membranes (OR 2.53, 95% CI 1.46 to 4.40) were significantly different between women with and without SLE. Gestational diabetes was negatively associated with SLE in pregnant women (OR 0.49, 95% CI 0.28 to 0.85). Pregnant women with SLE displayed significantly higher rates of fetal loss (OR 10.23, 95% CI 5.08 to 20.59), including spontaneous abortion (OR 4.42, 95% CI 1.52 to 12.80), therapeutic abortion (OR 16.57, 95% CI 5.80 to 47.35) and stillbirth (OR 13.25, 95% CI 1.49 to 118.11), and a higher risk of preterm birth (OR 3.15, 95% CI 2.21 to 4.50), intrauterine growth restriction (OR 2.20, 95% CI 1.35 to 3.58), a child who was small for the gestational age (OR 1.86, 95% CI 1.11 to 3.13), a caesarean section (OR 4.73, 95% CI 3.30 to 6.80) or a neonatal intensive care unit admission (OR 3.48, 95% CI 2.21 to 5.48) than women in the non-SLE population after adjusting for confounding factors. Conclusions In this study, SLE significantly increased the risk of adverse pregnancy outcomes. Therefore, a preconception assessment and close antenatal monitoring by both rheumatologists and obstetricians should be performed in pregnant women with SLE.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

The utility of urinary biomarker panel in predicting renal pathology and treatment response in Chinese lupus nephritis patients.

Given the urgent need for non-invasive biomarkers of LN, we aim to identify novel urinary biomarkers that facilitate diagnosis, assessment of disease activity and prediction of treatment response in a retrospective SLE cohort. A total of 154 SLE patients and 55 healthy controls were enrolled, among whom 73 were active LN patients. We measured renal activity by renal SLEDAI. The treatment response of the active LN patients who finished 6-month induction therapy was assessed based on the American College of Rheumatology response criteria. The expression levels of 10 urinary biomarkers (UBMs): β2-MG, calbindin D, cystatin C, IL-18, KIM-1, MCP-1, nephrin, NGAL, VCAM-1, and VDBP were tested using Luminex high-throughput proteomics technology. All but urinary nephrin levels were significantly increased in active LN compared to healthy controls. uCystatinC, uMCP-1, uKIM-1 levels were significantly higher in active LN group compared to inactive LN group. Correlation analysis revealed positive correlation between uCystatinC, uKIM-1, uMCP-1, uNGAL, uVDBP and RSLEDAI score. In renal pathology, uCystatinC, uKIM-1, uVCAM-1, and uVDBP positively correlated with activity index (AI) while uVCAM-1 positively correlated with chronicity index (CI). Moreover, the combination of uVCAM-1, uCystatinC, uKIM-1 discriminated proliferative LN from membranous LN with an AUC of 0.80 (95%CI: 0.69-0.90). Most importantly, baseline uNGAL demonstrated good prediction ability to discriminate responders from non-responders in active LN patients after 6-month induction therapy. Using a multiplex bead technique, we have identified the combination of uVCAM-1, uCystatinC, uKIM-1 as a biomarker panel to reflect renal pathology and NGAL as a promising urinary biomarker to both reflect disease activity and predict treatment response

Prediction of fetal loss in Chinese pregnant patients with systemic lupus erythematosus: a retrospective cohort study.

To develop a predictive model for fetal loss in women with systemic lupus erythematosus (SLE).SCOPUS: ar.jinfo:eu-repo/semantics/publishe

pSTATs in SLE patients after cytokine stimulation.

<p>The MFI for the phosphorylation of STAT1 (5A), STAT3 (5B), and STAT5 (5C) in the T cells of SLE patients decreased significantly after IFNα stimulation for 15 min. The MFI for the phosphorylation of STAT1 (5D), STAT3 (5E), and STAT5 (5F) decreased in the B cells of SLE patients after IFNα stimulation for 15 min. The MFI for the phosphorylation of STAT1 (5G) and STAT3 (5H) in the monocytes of SLE patients decreased after IFNα stimulation for 15 min. The MFI for pSTAT3 (5I) decreased significantly in the T cells of SLE patients after IL6 stimulation for 15 min. MFI, mean fluorescence index; SLE, systemic lupus erythematosus; HD, healthy donors; IFNα, interferon α.</p

Association between the mean fluorescence index (MFI) of pSTAT5 in B cells and cytokine levels in SLE patients.

<p>The concentrations of serum IL2 (A), IFNγ (B) and G-CSF (C),correlated positively with the MFI of STAT5 in the B cells of SLE patients. (D) The serum prolactin concentration correlated positively with the MFI of pSTAT5 in the T cells of SLE patients. MFI, mean fluorescence index; IL, interleukin; IFNγ, interferon γ; G-CSF, granulocyte colony-stimulating factor; PRL, prolactin.</p

Comparison pSTATs expression between SLE patient and healthy donor (HD) after cytokine stimulation.

<p>A representative experiment is shown. The MFI for the phosphorylation of STAT1 (5A), STAT3 (5B), and STAT5 (5C) in the T cells of SLE patient was compared with that of HD. The MFI for the phosphorylation of STAT1 (5D), STAT3 (5E), and STAT5 (5F) in the B cells of SLE patient was compared with that of HD. The MFI for the phosphorylation of STAT3 (5G) and STAT5 (5H) in the monocytes of SLE was compared with that of HD. The MFI for pSTAT3 (5I) in the T cells of SLE patient was compared with that of HD. MFI, mean fluorescence index; SLE, systemic lupus erythematosus; HD, healthy donor; IFNα, interferon α.</p