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Article ; Online: Immune checkpoints in rheumatoid arthritis: progress and promise.

Small, Annabelle / Lowe, Katie / Wechalekar, Mihir D

2023 Volume 14, Page(s) 1285554

Abstract: Rheumatoid arthritis (RA) is one of the most prevalent autoimmune inflammatory conditions, and while the mechanisms driving pathogenesis are yet to be completely elucidated, self-reactive T cells and immune checkpoint pathways have a clear role. In this ... ...

Abstract	Rheumatoid arthritis (RA) is one of the most prevalent autoimmune inflammatory conditions, and while the mechanisms driving pathogenesis are yet to be completely elucidated, self-reactive T cells and immune checkpoint pathways have a clear role. In this review, we provide an overview of the importance of checkpoint pathways in the T cell response and describe the involvement of these in RA development and progression. We discuss the relationship between immune checkpoint therapy in cancer and autoimmune adverse events, draw parallels with the involvement of immune checkpoints in RA pathobiology, summarise emerging research into some of the lesser-known pathways, and the potential of targeting checkpoint-related pathways in future treatment approaches to RA management.
MeSH term(s)	Humans ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Neoplasms/therapy ; T-Lymphocytes
Language	English
Publishing date	2023-11-24
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1285554
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Editorial: Synovial pathobiology and pathogenesis of inflammatory arthritis.

Small, Annabelle / Humby, Frances / Wechalekar, Mihir D

Frontiers in medicine

2022 Volume 9, Page(s) 1008562

Language	English
Publishing date	2022-08-16
Publishing country	Switzerland
Document type	Editorial
ZDB-ID	2775999-4
ISSN	2296-858X
ISSN	2296-858X
DOI	10.3389/fmed.2022.1008562
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Article ; Online: Synovial biopsies in inflammatory arthritis: precision medicine in rheumatoid arthritis.

Small, Annabelle / Wechalekar, Mihir D

Expert review of molecular diagnostics

2020 Volume 20, Issue 3, Page(s) 315–325

Abstract: ... ...

Abstract	Introduction
MeSH term(s)	Animals ; Arthritis, Rheumatoid/diagnosis ; Arthritis, Rheumatoid/etiology ; Biomarkers ; Biopsy ; Disease Management ; Disease Susceptibility ; Gene Expression Profiling ; Humans ; Immunohistochemistry/methods ; Precision Medicine/methods ; Synovial Membrane/pathology ; Transcriptome
Chemical Substances	Biomarkers
Language	English
Publishing date	2020-01-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2112530-2
ISSN	1744-8352 ; 1473-7159
ISSN (online)	1744-8352
ISSN	1473-7159
DOI	10.1080/14737159.2020.1707671
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Article: Attenuation of the BTLA/HVEM Regulatory Network in the Circulation in Primary Sjögren's Syndrome.

Small, Annabelle / Cole, Suzanne / Wang, Jing J / Nagpal, Sunil / Hao, Ling-Yang / Wechalekar, Mihir D

Journal of clinical medicine

2022 Volume 11, Issue 3

Abstract: Primary Sjögren's syndrome (SjS) is an inflammatory autoimmune disorder which targets the lacrimal and salivary glands, resulting in glandular dysfunction. Currently, the immune drivers of SjS remain poorly understood and peripheral biomarkers of disease ...

Abstract	Primary Sjögren's syndrome (SjS) is an inflammatory autoimmune disorder which targets the lacrimal and salivary glands, resulting in glandular dysfunction. Currently, the immune drivers of SjS remain poorly understood and peripheral biomarkers of disease are lacking. The present study therefore sought to investigate the immune cell constituents of the SjS peripheral blood, and to assess the role of the BTLA/HVEM/CD160 co-stimulatory network by characterizing expression within the periphery. Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood of
Language	English
Publishing date	2022-01-21
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662592-1
ISSN	2077-0383
ISSN	2077-0383
DOI	10.3390/jcm11030535
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Article ; Online: Neutrophils Require Activation to Express Functional Cell-Surface Complement Receptor Immunoglobulin.

Small, Annabelle G / Perveen, Khalida / Putty, Trishni / Patel, Nikita / Quinn, Patrick / Wechalekar, Mihir D / Hii, Charles S / Quach, Alex / Ferrante, Antonio

Frontiers in immunology

2022 Volume 13, Page(s) 840510

Abstract: The phagocytosis-promoting complement receptor, Complement Receptor Immunoglobulin (CRIg), is exclusively expressed on macrophages. It has been demonstrated that expression in macrophages could be modulated by inflammatory mediators, including cytokines. ...

Abstract	The phagocytosis-promoting complement receptor, Complement Receptor Immunoglobulin (CRIg), is exclusively expressed on macrophages. It has been demonstrated that expression in macrophages could be modulated by inflammatory mediators, including cytokines. This raised the possibility that a major phagocyte, the neutrophil, may also express CRIg following activation with inflammatory mediators. Here we show that resting peripheral blood neutrophil lysates subjected to protein analysis by Western blot revealed a 35 kDa CRIg isoform, consistent with the expression of CRIg mRNA by RT-PCR. By flow cytometry, CRIg was detected intracellularly and in very minor amounts on the cell surface. Interestingly, expression on the cell surface was significantly increased to functional levels after activation with inflammatory mediators/neutrophil activators; N-Formylmethionine-leucyl-phenylalanine, tumor necrosis factor (TNF), Granulocyte-Macrophage Colony stimulating Factor (GM-CSF), bacterial lipopolysaccharide, leukotriene B4 and phorbol myristate acetate. The increase in expression required p38 MAP kinase and protein kinase C activation, as well as intracellular calcium. Neutrophils which were defective in actin microfilament reorganization due to a mutation in ARPC1B or inhibition of its upstream regulator, Rac2 lose their ability to upregulate CRIg expression. Inhibition of another small GTPase, Rab27a, with pharmacological inhibitors prevented the increase in CRIg expression, suggesting a requirement for the actin cytoskeleton and exocytosis. Engagement of CRIg on TNF-primed neutrophils with an anti-CRIg monoclonal antibody increased the release of superoxide and promoted the activation of p38 but not ERK1/ERK2 or JNK MAP kinases. The TNF-induced increase in killing of
MeSH term(s)	Cytokines/metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor/metabolism ; Immunoglobulins/metabolism ; Inflammation Mediators/metabolism ; Neutrophils/metabolism ; Receptors, Complement/genetics ; Receptors, Complement/metabolism ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	Cytokines ; Immunoglobulins ; Inflammation Mediators ; Receptors, Complement ; Tumor Necrosis Factor-alpha ; Granulocyte-Macrophage Colony-Stimulating Factor (83869-56-1)
Language	English
Publishing date	2022-03-04
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2022.840510
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Article ; Online: Transcriptomic profiling of programmed cell death 1 (PD-1) expressing T cells in early rheumatoid arthritis identifies a decreased CD4 + PD-1 + signature post-treatment.

Lowe, Katie / Small, Annabelle / Song, Qingxuan / Hao, Ling-Yang / Murray-Brown, William / Proudman, Susanna / Smith, Malcolm D / Nagpal, Sunil / Wechalekar, Mihir D

Scientific reports

2023 Volume 13, Issue 1, Page(s) 2847

Abstract: Programmed cell death protein 1 (PD-1)-expressing T cells are expanded in individuals with established rheumatoid arthritis (RA). However, little is known about their functional role in the pathogenesis of early RA. To address this, we investigated the ... ...

Abstract	Programmed cell death protein 1 (PD-1)-expressing T cells are expanded in individuals with established rheumatoid arthritis (RA). However, little is known about their functional role in the pathogenesis of early RA. To address this, we investigated the transcriptomic profiles of circulating CD4
MeSH term(s)	Humans ; CD4-Positive T-Lymphocytes ; Transcriptome ; Programmed Cell Death 1 Receptor/genetics ; Programmed Cell Death 1 Receptor/metabolism ; Arthritis, Rheumatoid/drug therapy ; Arthritis, Rheumatoid/genetics ; Antirheumatic Agents/pharmacology ; Antirheumatic Agents/therapeutic use ; Antirheumatic Agents/metabolism ; Apoptosis
Chemical Substances	Programmed Cell Death 1 Receptor ; Antirheumatic Agents
Language	English
Publishing date	2023-02-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-29971-5
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Article: Facilitating THP-1 macrophage studies by differentiating and investigating cell functions in polystyrene test tubes

Small, Annabelle / Alex Quach / Antonio Ferrante / Marwah Al-Baghdadi / Nikki Lansdown

Journal of immunological methods. 2018,

2018

Abstract: Macrophage cell lines are a useful model to explore the properties of primary macrophages. However, a major limitation in the use of these cells is that when they are differentiated, they become adherent and hence present with the same limitation as ... ...

Abstract	Macrophage cell lines are a useful model to explore the properties of primary macrophages. However, a major limitation in the use of these cells is that when they are differentiated, they become adherent and hence present with the same limitation as natural macrophages. The cells need to be detached and are often subjected to detachment techniques such as detachment buffers containing proteolytic enzymes or scraping with a rubber ‘policeman’. These steps are time-consuming, reduce cell yields as well as cell viability and function. We have therefore investigated the possibility of differentiating the human macrophage THP-1 cell line in polystyrene FACS tubes to enable cells to be directly used for investigations by flow cytometry. Here we demonstrate that when the human macrophage cell line THP-1 are cultured in FACS tubes with phorbol myristate acetate added, they undergo differentiation into macrophages, assessed morphologically and by autofluorescence expression, in a similar manner to those cultured in tissue culture dishes. The cells can be readily washed and adjusted in concentration by centrifugation in the same tubes and can be directly tested for expression of cell surface markers and function by flow cytometry. This avoids the use of either detachment reagents or physical cell scraping. Consequently, we showed that the tube culture method results in increased cell yield and viability compared to those subjected to detachment procedures. The tube method generated functional macrophages which expressed the complement receptors, CR3 and CR4, and effectively phagocytosed complement opsonised Staphylococcus aureus via these receptors.
Keywords	acetates ; buffers ; cell viability ; centrifugation ; complement ; culture dishes ; flow cytometry ; human cell lines ; humans ; macrophages ; models ; polystyrenes ; proteinases ; receptors ; rubber ; Staphylococcus aureus
Language	English
Size	p. .
Publishing place	Elsevier B.V.
Document type	Article
Note	Pre-press version
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2018.06.019
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Differential expansion of T peripheral helper cells in early rheumatoid arthritis and osteoarthritis synovium.

Murray-Brown, William / Guo, Yanxia / Small, Annabelle / Lowe, Katie / Weedon, Helen / Smith, Malcolm D / Lester, Susan E / Proudman, Susanna M / Rao, Navin L / Hao, Ling-Yang / Nagpal, Sunil / Wechalekar, Mihir D

RMD open

2022 Volume 8, Issue 2

Abstract: Objectives: Programmed cell death protein 1 (PD-1)-expressing T cells are implicated in the pathogenesis of autoimmune inflammatory diseases such as rheumatoid arthritis. A subset of CXCR5 ... - ... T cells, termed T peripheral helper (Tph) cells, ... ...

Abstract	Objectives: Programmed cell death protein 1 (PD-1)-expressing T cells are implicated in the pathogenesis of autoimmune inflammatory diseases such as rheumatoid arthritis. A subset of CXCR5<sup>-</sup> T cells, termed T peripheral helper (Tph) cells, which drive B cell differentiation, have been identified in ectopic lymphoid structures in established rheumatoid arthritis synovial tissue. Here, we aimed to characterise these in treatment-naïve, early rheumatoid arthritis to determine whether these cells accumulate prior to fully established disease. Methods: Fresh dissociated tissue and peripheral blood mononuclear cell (PBMC) suspensions were stained with Zombie UV, followed by anti-CD45RO, PD-1, CD3, ICOS, CD8, CD4, CD20, CXCR5, TIGIT and CD38 antibodies prior to analysis. For histology, rheumatoid arthritis synovial sections were prepared for Opal multispectral immunofluorescence with anti-CD45RO, CD20, PD-1 and CXCR5 antibodies. Images were acquired on the Perkin Elmer Vectra V.3.0 imaging system and analysed using InForm Advanced Image Analysis software. Results: Flow cytometry revealed T cell infiltration in the rheumatoid arthritis synovium with differential expression of PD-1, CD45RO, ICOS, TIGIT and CD38. We observed a higher frequency of PD1<sup>hi</sup>CXCR5<sup>-</sup> Tph in rheumatoid arthritis synovial tissue and PBMCs versus controls, and no significant difference in T follicular helper cell frequency. Microscopy identified a 10-fold increase of Tph cells in early rheumatoid arthritis synovial follicular and diffuse regions, and identified Tph adjacent to germinal centre B cells. Conclusions: These data demonstrate that PD-1<sup>hi</sup> Tph cells are present in early rheumatoid arthritis, but not osteoarthritis synovium, and therefore may provide a target for treatment of patients with early rheumatoid arthritis.
MeSH term(s)	Humans ; Programmed Cell Death 1 Receptor/metabolism ; Leukocytes, Mononuclear/metabolism ; Leukocytes, Mononuclear/pathology ; T-Lymphocytes, Helper-Inducer/metabolism ; T-Lymphocytes, Helper-Inducer/pathology ; Synovial Membrane/metabolism ; Receptors, CXCR5/metabolism ; Arthritis, Rheumatoid ; Osteoarthritis/pathology
Chemical Substances	Programmed Cell Death 1 Receptor ; Receptors, CXCR5
Language	English
Publishing date	2022-10-18
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2812592-7
ISSN	2056-5933 ; 2056-5933
ISSN (online)	2056-5933
ISSN	2056-5933
DOI	10.1136/rmdopen-2022-002563
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Article ; Online: Facilitating THP-1 macrophage studies by differentiating and investigating cell functions in polystyrene test tubes.

Small, Annabelle / Lansdown, Nikki / Al-Baghdadi, Marwah / Quach, Alex / Ferrante, Antonio

Journal of immunological methods

2018 Volume 461, Page(s) 73–77

Abstract	Macrophage cell lines are a useful model to explore the properties of primary macrophages. However, a major limitation in the use of these cells is that when they are differentiated, they become adherent and hence present with the same limitation as natural macrophages. The cells need to be detached and are often subjected to detachment techniques such as detachment buffers containing proteolytic enzymes or scraping with a rubber 'policeman'. These steps are time-consuming, reduce cell yields as well as cell viability and function. We have therefore investigated the possibility of differentiating the human macrophage THP-1 cell line in polystyrene FACS tubes to enable cells to be directly used for investigations by flow cytometry. Here we demonstrate that when the human macrophage cell line THP-1 are cultured in FACS tubes with phorbol myristate acetate added, they undergo differentiation into macrophages, assessed morphologically and by autofluorescence expression, in a similar manner to those cultured in tissue culture dishes. The cells can be readily washed and adjusted in concentration by centrifugation in the same tubes and can be directly tested for expression of cell surface markers and function by flow cytometry. This avoids the use of either detachment reagents or physical cell scraping. Consequently, we showed that the tube culture method results in increased cell yield and viability compared to those subjected to detachment procedures. The tube method generated functional macrophages which expressed the complement receptors, CR3 and CR4, and effectively phagocytosed complement opsonised Staphylococcus aureus via these receptors.
MeSH term(s)	Cell Differentiation/drug effects ; Cell Differentiation/immunology ; Cell Survival/drug effects ; Cell Survival/immunology ; Humans ; Integrin alphaXbeta2/immunology ; Macrophage-1 Antigen/immunology ; Macrophages/immunology ; Phagocytosis ; Polystyrenes/chemistry ; Staphylococcus aureus/immunology ; THP-1 Cells ; Tetradecanoylphorbol Acetate/pharmacology
Chemical Substances	Integrin alphaXbeta2 ; Macrophage-1 Antigen ; Polystyrenes ; Tetradecanoylphorbol Acetate (NI40JAQ945)
Language	English
Publishing date	2018-07-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2018.06.019
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Article ; Online: Human Dendritic Cells Express the Complement Receptor Immunoglobulin Which Regulates T Cell Responses.

Munawara, Usma / Perveen, Khalida / Small, Annabelle G / Putty, Trishni / Quach, Alex / Gorgani, Nick N / Hii, Charles S / Abbott, Catherine A / Ferrante, Antonio

Frontiers in immunology

2019 Volume 10, Page(s) 2892

Abstract: The B7 family-related protein V-set and Ig containing 4 (VSIG4), also known as Z39Ig and Complement Immunoglobulin Receptor (CRIg), is the most recent of the complement receptors to be identified, with substantially distinct properties from the classical ...

Abstract	The B7 family-related protein V-set and Ig containing 4 (VSIG4), also known as Z39Ig and Complement Immunoglobulin Receptor (CRIg), is the most recent of the complement receptors to be identified, with substantially distinct properties from the classical complement receptors. The receptor displays both phagocytosis-promoting and anti-inflammatory properties. The receptor has been reported to be exclusively expressed in macrophages. We now present evidence, that CRIg is also expressed in human monocyte-derived dendritic cells (MDDC), including on the cell surface, implicating its role in adaptive immunity. Three CRIg transcripts were detected and by Western blotting analysis both the known Long (L) and Short (S) forms were prominent but we also identified another form running between these two. Cytokines regulated the expression of CRIg on dendritic cells, leading to its up- or down regulation. Furthermore, the steroid dexamethasone markedly upregulated CRIg expression, and in co-culture experiments, the dexamethasone conditioned dendritic cells caused significant inhibition of the phytohemagglutinin-induced and alloantigen-induced T cell proliferation responses. In the alloantigen-induced response the production of IFNγ, TNF-α, IL-13, IL-4, and TGF-β1, were also significantly reduced in cultures with dexamethasone-treated DCs. Under these conditions dexamethasone conditioned DCs did not increase the percentage of regulatory T cells (Treg). Interestingly, this suppression could be overcome by the addition of an anti-CRIg monoclonal antibody to the cultures. Thus, CRIg expression may be a control point in dendritic cell function through which drugs and inflammatory mediators may exert their tolerogenic- or immunogenic-promoting effects on dendritic cells.
MeSH term(s)	Biomarkers ; Coculture Techniques ; Cytokines/metabolism ; Dendritic Cells/immunology ; Dendritic Cells/metabolism ; Gene Expression Regulation ; Humans ; Immunity, Cellular/genetics ; Immunomodulation ; Immunophenotyping ; Receptors, Complement/genetics ; Receptors, Complement/metabolism ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism ; T-Lymphocytes/immunology ; T-Lymphocytes/metabolism
Chemical Substances	Biomarkers ; Cytokines ; Receptors, Complement
Language	English
Publishing date	2019-12-10
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2019.02892
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