scholarly journals Generation of Recombinant Monoclonal Antibodies from Single B Cells Isolated from Synovial Tissue of Rheumatoid Arthritis Patients

2018 ◽  
pp. 159-187 ◽  
Author(s):  
Elisa Corsiero ◽  
Lucas Jagemann ◽  
Michele Bombardieri ◽  
Costantino Pitzalis
Keyword(s):  
Rheumatoid Arthritis ◽  
Monoclonal Antibodies ◽  
B Cells ◽  
Synovial Tissue

scholarly journals Mast cells in early rheumatoid arthritis associate with disease severity and support B cell autoantibody production

2018 ◽  
Vol 77 (12) ◽  
pp. 1773-1781 ◽  
Author(s):  
Felice Rivellese ◽  
Daniele Mauro ◽  
Alessandra Nerviani ◽  
Sara Pagani ◽  
Liliane Fossati-Jimack ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
Mast Cells ◽  
Disease Activity ◽  
B Cell ◽  
Synovial Tissue ◽  
Autoantibody Production ◽  
Early Ra ◽  
Treatment Naïve

ObjectivesMast cells (MCs) are involved in the pathogenesis of rheumatoid arthritis (RA). However, their contribution remains controversial. To establish their role in RA, we analysed their presence in the synovium of treatment-naïve patients with early RA and their association and functional relationship with histological features of synovitis.MethodsSynovial tissue was obtained by ultrasound-guided biopsy from treatment-naïve patients with early RA (n=99). Immune cells (CD3/CD20/CD138/CD68) and their relationship with CD117+MCs in synovial tissue were analysed by immunohistochemistry (IHC) and immunofluorescence (IF). The functional involvement of MCs in ectopic lymphoid structures (ELS) was investigated in vitro, by coculturing MCs with naïve B cells and anticitrullinated protein antibodies (ACPA)-producing B cell clones, and in vivo in interleukin-27 receptor alpha (IL27ra)-deficient and control mice during antigen-induced arthritis (AIA).ResultsHigh synovial MC counts are associated with local and systemic inflammation, autoantibody positivity and high disease activity. IHC/IF showed that MCs reside at the outer border of lymphoid aggregates. Furthermore, human MCs promote the activation and differentiation of naïve B cells and induce the production of ACPA, mainly via contact-dependent interactions. In AIA, synovial MC numbers increase in IL27ra deficient mice, in association with ELS and worse disease activity.ConclusionsSynovial MCs identify early RA patients with a severe clinical form of synovitis characterised by the presence of ELS.

Selective Infiltration of B Cells Committed to the Production of Monoreactive Rheumatoid Factor in Synovial Tissue of Patients with Rheumatoid Arthritis

1993 ◽  
Vol 69 (1) ◽  
pp. 16-22 ◽  
Author(s):  
Masayuki Hakoda ◽  
Tsuyoshi Ishimoto ◽  
Sakura Hayashimoto ◽  
Kazuhiko Inoue ◽  
Atsuo Taniguchi ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
Rheumatoid Factor ◽  
Synovial Tissue

scholarly journals FRI0005 DIVERSITY OF ANTI-CITRULLINATED PROTEIN ANTIBODY COMPOSITIONS INFLUENCE SYNOVIAL FIBROBLAST REACTIVITY

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 573.2-574
Author(s):  
M. Sun ◽  
B. Réthi ◽  
A. Krishnamurthy ◽  
V. Joshua ◽  
A. Circiumaru ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Monoclonal Antibodies ◽  
Synovial Tissue ◽  
Trichostatin A ◽  
Synovial Fibroblasts ◽  
Cross Reactivity ◽  
Anacardic Acid ◽  
Cellular Targets ◽  
Fibroblast Migration ◽  
Citrullinated Protein

Background:Anti-citrullinated protein antibodies (ACPAs) play an important role in rheumatoid arthritis (RA) pathogenesis. We hypothesized that the effect of these antibodies is mediated by their binding to synovial fibroblasts and inducing an increased mobility of fibroblasts1.Objectives:In our study, we analyzed and compared fibroblast modulation by ACPA pools obtained from different patients or by a set of monoclonal ACPAs with different fine specificity that were obtained from different tissue sites.Methods:Synovial fibroblasts were isolated from RA patients synovial tissue biopsies. Individual polyclonal ACPA and control IgGs were purified from sera of four ACPA-positive RA patients by affinity purification on protein G and CCP-2 columns. Monoclonal antibodies were derived from memory B cell isolated from blood2, synovial fluid or bronchoalveolar lavage of RA patients. Whole antibodies and F(ab’)2 fragments were tested in fibroblast migration using IncuCyte live-cell analysis. Blocking experiments were performed with soluble citrullinated proteins in SF migration. Cross-reactivity of the antibodies to citrullinated and acetylated epitopes was tested using PAD inhibitors (Cl-amidine and GSK199), histone acetyltransferases (anacardic acid) and deacetylases (trichostatin A). Binding patterns of monoclonal ACPAs, both whole and F(ab’)2 fragments were analyzed in synovial biopsies obtained from both healthy donors and RA patients.Results:Three out of four tested individual ACPA were able to promote fibroblast migration. Five out of nine tested monoclonal ACPAs stimulated fibroblast migration. One of these antibodies, clone 1325:01B09 is characterized by cross-reactivity to citrullinated, homocitrullinated and acetylated targets. The effect of 1325:01B09 on fibroblast migration was completely abolished by Cl-amidine or by pre-incubating the antibody with citrullinated fibrinogen or histone but not citrullinated enolase or vimentin. Despite the cross-reactivity to acetylated epitopes, neither anacardic acid nor trichostatin A could modulate the 1325:01B09 effect on fibroblast migration. F(ab’)2 fragments of this antibody stimulated fibroblast migration and labelled podoplanin-positive fibroblasts in inflamed RA synovium similarly to the intact antibody, indicating an Fc-independent effect.Conclusion:The effect on fibroblast mobility was likely to be mediated by binding to citrullinated epitopes but not through Fc receptors. Detection of fibroblast modulating ACPAs in majority of RA patients indicated that fibroblasts might be key cellular targets in disease pathogenesis, although individual variability might exist in the composition of ACPA cellular targets.References:[1]Sun M, Rethi B, Krishnamurthy A, et al. Anticitrullinated protein antibodies facilitate migration of synovial tissue-derived fibroblasts. Ann Rheum Dis 2019;78(12):1621-31. doi: 10.1136/annrheumdis-2018-214967 [published Online First: 2019/09/05][2]Amara K, Lena Israelsson, Ragnhild Stålesen, et al. A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material. Bio-protocol 2019;9(16)Disclosure of Interests:Meng Sun: None declared, Bence Réthi: None declared, Akilan Krishnamurthy: None declared, Vijay Joshua: None declared, Alexandra Circiumaru: None declared, Marianne Engström: None declared, Caroline Grönwall: None declared, Vivianne Malmström Grant/research support from: VM has had research grants from Janssen Pharmaceutica, Khaled Amara: None declared, Lars Klareskog: None declared, Heidi Wähämaa: None declared, Anca Catrina: None declared

scholarly journals Potential Biological Molecules and Immune Cell for Rheumatoid Arthritis Synovial Lesions

2021 ◽  
Author(s):  
Mingyi Yang ◽  
Yani Su ◽  
Yao Ma ◽  
Ke Xu ◽  
Aihaiti Yirixiati ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Gene Ontology ◽  
Dendritic Cells ◽  
B Cells ◽  
Signaling Pathway ◽  
Synovial Tissue ◽  
Immune Cell ◽  
Hub Genes ◽  
Data Set ◽  
Synovial Lesions

Abstract Objective: To study the potential biomarkers and related pathways in rheumatoid arthritis (RA) synovial lesions, as well as immune cell, to provide theoretical basis and research directions for the mechanism and treatment of RA. Methods: Download the RA synovial tissue microarray data set (GSE77298, GSE55457 and GSE55235) from Gene Expression Omnibus (GEO), The“limma” package of R to identify differentially expressed genes (DEGs), DAVIA Perform GO (Gene Ontology, gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes, Kyoto Gene Encyclopedia) enrichment analysis. STRING (Search Tool for the Retrieval of Interacting Genes) constructs a Protein Protein Interaction Network (PPI), R screens Hub genes, and mines targeted miRNAs of Hub genes based on multiple databases. In R,the immune cell of RA synovial tissue samples are obtained through the three packages of "e1071", "parallel" and "preprocessCore" with "CIBERSORT" software. Results: Ten Hub genes (KIAA0101, FOXM1, EGFR, CDC20, BUB1B, TYMS, TOP2A, RRM2, JUN and CCNA2) and 2 key miRNAs (miR-520d-5p and miR) related to RA synovial lesions were finally identified -139-5p), significantly enriched in epithelial cell signaling, ECM-receptor interaction, estrogen signaling pathway, cell cycle, ErbB signaling pathway and GnRH signaling pathway in Helicobacter pylori infection. Immune cell analysis found that resting dendritic cells, B cells memory, dendritic cells activated, plasma cells, macrophages M1, mast cells resting and T cells regulatory have high expression in RA, while neutrophils, B cells naive and natural killer cells activated have low expression in RA. Conclusion:The Hub genes, key miRNAs, related pathways, and immune cell obtained in this study provide a certain basis for the etiology and treatment of RA synovial lesions.

THU0043 The Alternative CD20 Transcript Variant is not Expressed in B Cells and Synovial Tissue from Patients with Rheumatoid Arthritis

2013 ◽  
Vol 72 (Suppl 3) ◽  
pp. A178.2-A178
Author(s):  
C. Gamonet ◽  
M. Deschamps ◽  
B. Gaugler ◽  
P. Saas ◽  
I. Auger ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
Synovial Tissue ◽  
Transcript Variant

scholarly journals A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material

BIO-PROTOCOL ◽  
2019 ◽  
Vol 9 (17) ◽  
Author(s):  
khaled Amara ◽  
Lena Israelsson ◽  
Ragnhild Stålesen ◽  
Peter Sahlström ◽  
Johanna Steen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Monoclonal Antibodies ◽  
Rheumatoid Arthritis Patient ◽  
B Cells ◽  
Human B Cells

scholarly journals POS0388 INTERLEUKIN 40 (IL-40) IS UP-REGULATED IN RHEUMATOID ARTHRITIS (RA) AND ASSOCIATED WITH DISEASE ACTIVITY, LEVELS OF AUTOANTIBODIES AND CHEMOKINES

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 424.1-424
Author(s):  
A. Navrátilová ◽  
L. Andres Cerezo ◽  
H. Hulejova ◽  
V. Becvar ◽  
D. Tegzová ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
Synovial Fluid ◽  
Disease Activity ◽  
B Cell ◽  
Synovial Tissue ◽  
Lupus Erythematosus ◽  
Synovial Fibroblasts ◽  
Clinical Activity

Background:Interleukin 40 (IL-40) is newly identified B cell - associated cytokine implicated in humoral immune responses and in B cell development. As B cells play a pivotal role in autoimmunity, we aimed to investigate the function of IL-40 in rheumatoid arthritis (RA).Objectives:The aim of our study was to determine the function of IL-40 in RA.Methods:IL-40 expression in the synovial tissue was determined by immunohistochemistry and immunofluorescence (n=4-5). IL-40 was analysed in the serum/synovial fluid of patients with RA (n=69), systemic lupus erythematosus (SLE; n=69), osteoarthritis (OA; n=44), and in healthy controls (HC; n=25). Given the association of IL-40 with B cells, we analysed the effect of rituximab therapy on the serum IL-40 in 19 patients with RA after 16 and 24 weeks of the therapy. The clinical activity of patients with RA was assessed according to the 28 joint count Disease Activity Score (DAS28). Levels of C-reactive protein (CRP) and autoantibodies were measured by routine laboratory techniques. In vitro experiments were performed in RA synovial fibroblasts (n=9). Levels of cytokines and inflammatory mediators were determined in serum, synovial fluid and supernatants using ELISA or multiplex immunoassay.Results:IL-40 was overexpressed in RA synovial tissue compared to OA, particularly by synovial fibroblasts and immune cells such as B and T lymphocytes, macrophages and neutrophils. The levels of IL-40 were significantly higher in the synovial fluid of RA patients compared to OA (33.2 (6.6-68.9) vs. 0.7 (0.1-2.4) ng/ml; p<0.0001). In addition, IL-40 was increased in the serum of RA patients compared to SLE, OA or HC (4.8 (1.7-24.9) vs. 1.4 (1.0-1.9), 1.6 (0.6-3.1) or 1.5 (0.7-2.7) ng/ml; p<0.0001 for all) and decreased after 16 (p<0.01) and 24 weeks (p<0.001) in a subgroup of rituximab treated patients with RA. IL-40 levels in RA patients correlated with autoantibodies rheumatoid factor (IgM) and anti-citrullinated protein antibody (ACPA) in the serum (p<0.0001 and p<0.01) as well as in the synovial fluid (p<0.0001 and p<0.001). IL-40 in RA synovial fluid was also significantly associated with DAS28 (p<0.05), synovial fluid leukocyte count (p<0.01), number of swollen joints (p<0.05) and neutrophil attractants IL-8 (p<0.01) and MIP-1α (p<0.01). RA synovial fibroblasts exposed to recombinant IL-40 increased secretion of IL-8 (p<0.01), MCP-1 (p<0.05) and MMP-13 (p<0.01) compared to unstimulated cells in in vitro conditions.Conclusion:Our results show for the first time that IL-40 is elevated in RA and decreases following B-cell depletion therapy. The association of IL-40 with autoantibodies and chemokines may imply its potential involvement in RA development. Moreover, IL-40 up-regulates the secretion of chemokines and MMP-13 by synovial fibroblasts, indicating its role in the regulation of inflammation and tissue destruction in RA.Acknowledgements:Supported by MHCR 023728 a SVV 260 523Disclosure of Interests:None declared

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