Mechanisms of allergy and clinical immunology
Cytometry by time-of-flight immunophenotyping identifies a blood Sjögren's signature correlating with disease activity and glandular inflammation

https://doi.org/10.1016/j.jaci.2016.01.024Get rights and content

Background

Mass cytometry has recently emerged as a promising tool for clinical research. However, few studies have demonstrated its benefit for patient stratification and biomarker identification. Primary Sjögren's syndrome (pSS) is a prototype of chronic autoimmune disease, the pathogenesis of which remains unclear and for which treatment does not exist.

Objective

This observational case-control study was designed to discover new cellular biomarkers and therapeutic targets in patients with pSS.

Methods

Forty-nine patients with pSS and 45 control subjects were enrolled for clinical evaluation and mass cytometry quantification of 34 protein markers in blood. For a third of these subjects, matched labial salivary gland biopsy specimens were also analyzed by mass cytometry and immunohistochemistry.

Results

In salivary gland biopsy specimens from patients with pSS, we identified a high number of activated CD8+ T cells, terminally differentiated plasma cells, and activated epithelial cells, pointing to new pathogenic mechanisms for future clinical intervention. In blood, we identified a 6-cell disease signature defined by decreased numbers of CD4 and memory B lymphocytes, decreased plasmacytoid dendritic cell numbers, and increased representation of activated CD4 and CD8 T cells and plasmablasts. These blood cellular components correlated with clinical parameters and, when taken together, clustered patients into subsets with distinct disease activity and glandular inflammation.

Conclusion

This first application of mass cytometry to a well-stratified clinical cohort and small biopsy tissues establishes the benefits of such an approach for the discovery of new biomarkers and therapeutic targets. Similar high-dimensional immunophenotyping strategies could be implemented in longitudinal and interventional clinical settings in this and other disease areas.

Section snippets

Study participants

All patients with pSS (n = 49) met the American European Consensus criteria (AECG) diagnostic criteria.20 Patients were referred to the Department of Rheumatology of Hôpitaux Universitaires Paris-Sud (France) between December 2013 and September 2014. Control subjects (n = 45) were either healthy volunteers or patients with sicca syndrome with a negative diagnostic result for pSS. Clinical evaluation confirmed that the latter had neither autoantibodies nor a positive biopsy result. The study was

Identification of a cellular signature in peripheral blood from patients with pSS

The correlation between flow and mass cytometry was very good in PBMCs from 9 healthy donors (average R2 = 0.92 ± 0.09), which confirmed the validity of mass cytometry for clinical immunophenotyping (see Fig E1 in this article's Online Repository at www.jacionline.org). Characteristics of the 49 patients with pSS and 45 control subjects are summarized in Table E3 in this article's Online Repository at www.jacionline.org. Using the viSNE algorithm,21 we identified 40 peripheral blood cell types

Discussion

This study is the first large-scale clinical application of mass cytometry to stratify a patient population with a chronic disease. In addition, this is also the first study that performs immunophenotyping of paired blood and biopsy samples to correlate cellular changes in the blood and target tissue of the disease. The strength of this high-dimensional and unbiased immunophenotyping approach relies on its ability to identify new cellular variations and, importantly, to enable the simultaneous

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    Supported by an unrestricted grant (UPSud/SAIC N 97731) from Biogen to Université Paris-Sud.

    Disclosure of potential conflict of interest: M. Mingueneau is an employee of Biogen. S. Boudaoud received a grant from Biogen and is an employee of INSERM. S. Haskett is an employee of Biogen. T. L. Reynolds is an employee of Biogen. G. Nocturne has received a grant from Biogen. E. Norton is an employee of Biogen. X. Zhang is an employee of Biogen. M. Constant is an employee of Biogen. D. Park is an employee of Biogen. W. Wang is an employee and holds stock in Biogen. T. Lazure is an employee of Assitance Publique Hopitaux de Paris. A. Ergun is an employee of Biogen. X. Mariette received a grant from Biogen and provided consultancy for Biogen. The rest of the authors declare that they have no relevant conflicts of interest.

    These authors contributed equally to this work.

    These authors contributed equally to this work.

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