Multidimensional Single Cell Based STAT Phosphorylation Profiling Identifies a Novel Biosignature for Evaluation of Systemic Lupus Erythematosus Activity
Open Access
- 22 July 2011
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 6 (7), e21671
- https://doi.org/10.1371/journal.pone.0021671
Abstract
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. 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. 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. The basal activation of STATs signaling and reduced response to cytokines may be helpful us to identify the activity and severity of SLE.Keywords
This publication has 35 references indexed in Scilit:
- Phospho Flow Cytometry Methods for the Analysis of Kinase Signaling in Cell Lines and Primary Human Blood SamplesMethods in molecular biology (Clifton, N.J.), 2010
- Single cell phospho-specific flow cytometry can detect dynamic changes of phospho-Stat1 level in lung cancer cellsCytometry Part A, 2010
- Stage Dependent Aberrant Regulation of Cytokine-STAT Signaling in Murine Systemic Lupus ErythematosusPLOS ONE, 2009
- Multiparameter Phospho-Flow Analysis of Lymphocytes in Early Rheumatoid Arthritis: Implications for Diagnosis and Monitoring Drug TherapyPLOS ONE, 2009
- Single-Cell Profiling Identifies Aberrant STAT5 Activation in Myeloid Malignancies with Specific Clinical and Biologic CorrelatesCancer Cell, 2008
- Single-Cell, Phosphoepitope-Specific Analysis Demonstrates Cell Type- and Pathway-Specific Dysregulation of Jak/STAT and MAPK Signaling Associated with In Vivo Human Immunodeficiency Virus Type 1 InfectionJournal of Virology, 2008
- Increased expression of STAT3 in SLE T cells contributes to enhanced chemokine-mediated cell migrationAutoimmunity, 2007
- Phospho‐proteomic immune analysis by flow cytometry: from mechanism to translational medicine at the single‐cell levelImmunological Reviews, 2006
- Prolactin signals via Stat5 and Oct-1 to the proximal cyclin D1 promoterMolecular and Cellular Endocrinology, 2005
- The american rheumatism association 1987 revised criteria for the classification of rheumatoid arthritisArthritis & Rheumatism, 1988