Deep Immunophenotyping and Clustering Identifies Biomarkers Predictive of Lymphoma in Primary Sjögren's Syndrome

Abstract

Objective: Patients with primary Sjögren’s Syndrome (pSS) are prone to develop non-Hodgkin’s lymphoma (NHL-pSS) but relevant biomarkers are lacking. We aimed to determine new biomarkers predictive of NHL in patients with pSS. 

Methods: 206 pSS patients fulfilling ACR/EULAR 2016 criteria were included and divided into three groups: pSS, lymphoproliferative pSS (L-pSS), and NHL-pSS. Deep flow cytometry immunophenotyping of B and T cell compartments as well as serum IFNα quantification were coupled to clinical, biological and histopathological data analysis. 

Results: We identified CD11c+FcRL5+ tissue-like memory B cells and IFNγ+TNFα+ conventional T cells as significantly associated with NHL in pSS. These clusters showed progressive enrichment in L-pSS and NHL-pSS as compared to pSS. The combination of these two populations abundances discriminates NHL-pSS patients with a sensibility of 78.9% and a specificity of 76.8%, thus overcoming alone the performance of the sum of conventional clinical and biological markers such as cryoglobulinemia vasculitis, parotid enlargement, RF antibodies, low C4 and elevated serum IFNα (73.7% and 76.8% respectively). CD11c+ FcRL5+ tissue-like memory B cells were associated with occurrence of mucosa-associated lymphoid tissue marginal-zone lymphoma (MALT NHL), while IFNγ+TNFα+ conventional T cells were more indicative of non-MALT B-cell NHLs. 

Conclusion: We unveil novel biomarkers of NHL in pSS based on an integrative analysis coupling deep immunophenotyping and clinical, biological and histopathological data. Furthermore, these markers allow distinguishing B-cell NHL subtypes in pSS.

Funding: This PhD was funded by a research grant issued by the French Ministry of Health (research year grant). Furthermore, the authors did not receive any specific funding for carrying out the work underlying this manuscript.

Declaration of Interest: The authors declare no competing financial interests, and no personal conflicts of interest. The work presented here is patented under the international patent application No. PCT/EP2022/060498.

Ethical Approval: The study was approved by the local ethics committee of Pitié-Salpêtrière Hospital – CPP-IV Ile de France and was conducted in accordance with the Declaration of Helsinki. All participants provided written informed consent.

Keywords: Primary Sjögren's syndrome, lymphomagenesis, non-Hodgkin's lymphoma, biomarkers, B cells, T cells, flow cytometry, immunophenotyping, IFNα.