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Article ; Online: MHCII restriction demonstrates B cells have very limited capacity to activate tumour-specific CD4

Guy, Thomas V / Terry, Alexandra M / McGuire, Helen M / Shklovskaya, Elena / Fazekas de St Groth, Barbara

2023 Volume 13, Issue 1, Page(s) 2290799

Abstract: There has been growing interest in the role of B cells in antitumour immunity and potential use ... of B cells to specifically activate tumour-specific CD4+ T cells ...

Abstract	There has been growing interest in the role of B cells in antitumour immunity and potential use in adoptive cellular therapies. To date, the success of such therapies is limited. The intrinsic capacity of B cells to specifically activate tumour-specific CD4+ T cells
MeSH term(s)	Mice ; Animals ; Antigen-Presenting Cells/physiology ; CD4-Positive T-Lymphocytes ; Lymphocyte Activation ; B-Lymphocytes ; Neoplasms
Language	English
Publishing date	2023-12-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	2645309-5
ISSN	2162-402X ; 2162-402X
ISSN (online)	2162-402X
ISSN	2162-402X
DOI	10.1080/2162402X.2023.2290799
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Article ; Online: Effects of storage time and temperature on highly multiparametric flow analysis of peripheral blood samples; implications for clinical trial samples.

Jerram, Amelia / Guy, Thomas V / Beutler, Lucinda / Gunasegaran, Bavani / Sluyter, Ronald / Fazekas de St Groth, Barbara / McGuire, Helen M

Bioscience reports

2021 Volume 41, Issue 2

Abstract: ... limited to, monocytes, NK cells, B cells, Treg cells, and naïve, central memory and effector memory CD4+ ...

Abstract	We sought to determine the effect of time and temperature of blood sample storage before preparation of human peripheral blood mononuclear cells (PBMCs) by Ficoll-hypaque density gradient centrifugation. Blood samples from healthy donors were stored at room temperature (RT) or refrigerated at 4°C before preparation of PBMCs. Cell yield and viability, and proportions of major cell populations within PBMCs, as determined by fluorescence flow cytometry, were assessed for both fresh and cryopreserved samples. Highly multiparametric mass cytometry was performed on cryopreserved PBMCs. We found that refrigeration had marked negative effects on subsequent PBMC yield. Storage at RT led to co-purification of low density neutrophils with PBMCs, but had no detectable effects on the proportions of multiple cell subsets including, but not limited to, monocytes, NK cells, B cells, Treg cells, and naïve, central memory and effector memory CD4+ and CD8+ T cells and CD45RA-positive terminal effector CD8+ T cells. Expression of a number of cell surface receptors, including CXCR5, CCR6, CXCR3 and TIGIT, but not CD247 was reduced after RT storage before PBMC preparation, and this effect correlated with the degree of low density neutrophil contamination. As such, when PBMC preparation cannot be undertaken immediately after blood draw, storage at RT is far superior to refrigeration. RT storage leads to neutrophil activation, but does not compromise measurement of PBMC subset distribution. However caution must be applied to interpretation of cytometric measurements of surface molecules such as chemokine receptors.
MeSH term(s)	Adult ; Female ; Flow Cytometry/methods ; Humans ; Lymphocytes/immunology ; Lymphocytes/metabolism ; Male ; Pilot Projects ; Receptors, Cell Surface/metabolism ; Specimen Handling ; Temperature ; Young Adult
Chemical Substances	Receptors, Cell Surface
Language	English
Publishing date	2021-02-18
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	764946-0
ISSN	1573-4935 ; 0144-8463
ISSN (online)	1573-4935
ISSN	0144-8463
DOI	10.1042/BSR20203827
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Article ; Online: Stable and Highly Efficient Antibody-Nanoparticles Conjugation.

Maddahfar, Mahnaz / Wen, Shihui / Hosseinpour Mashkani, Seyed Mostafa / Zhang, Lin / Shimoni, Olga / Stenzel, Martina / Zhou, Jiajia / Fazekas de St Groth, Barbara / Jin, Dayong

Bioconjugate chemistry

2021 Volume 32, Issue 6, Page(s) 1146–1155

Abstract: ... on the B cells. This work suggests a library of stabile, specific, and bioactive lanthanide-doped nanoconjugates ...

Abstract	Functional ligands and polymers have frequently been used to yield target-specific bio-nanoconjugates. Herein, we provide a systematic insight into the effect of the chain length of poly(oligo (ethylene glycol) methyl ether acrylate) (POEGMEA) containing polyethylene glycol on the colloidal stability and antibody-conjugation efficiency of nanoparticles. We employed Reversible Addition-Fragmentation Chain Transfer (RAFT) to design diblock copolymers composed of 7 monoacryloxyethyl phosphate (MAEP) units and 6, 13, 35, or 55 OEGMEA units. We find that when the POEGMEA chain is short, the polymer cannot effectively stabilize the nanoparticles, and when the POEGMEA chain is long, the nanoparticles cannot be efficiently conjugated to antibody. In other words, the majority of the carboxylic groups in larger POEGMEA chains are inaccessible to further chemical modification. We demonstrate that the polymer containing 13 OEGMEA units can effectively bind up to 64% of the antibody molecules, while the binding efficiency drops to 50% and 0% for the polymer containing 35 and 55 OEGMEA units. Moreover, flow cytometry assay statistically shows that about 9% of the coupled antibody retained its activity to recognize B220 biomarkers on the B cells. This work suggests a library of stabile, specific, and bioactive lanthanide-doped nanoconjugates for flow cytometry and mass cytometry application.
MeSH term(s)	Antibodies/chemistry ; Nanoparticles/chemistry ; Polymerization ; Polymers/chemistry
Chemical Substances	Antibodies ; Polymers
Language	English
Publishing date	2021-05-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1024041-x
ISSN	1520-4812 ; 1043-1802
ISSN (online)	1520-4812
ISSN	1043-1802
DOI	10.1021/acs.bioconjchem.1c00192
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Zs.A 3400: Show issues

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Article ; Online: The Role of Antigen-Competitive Dynamics in Regulating the Immune Response.

Pooladvand, Pantea / Kim, Peter S / Fazekas de St Groth, Barbara

Bulletin of mathematical biology

2021 Volume 83, Issue 5, Page(s) 40

Abstract: The clonal expansion of T cells during an infection is tightly regulated to ensure an appropriate immune response against invading pathogens. Although experiments have mapped the trajectory from expansion to contraction, the interplay between mechanisms ... ...

Abstract	The clonal expansion of T cells during an infection is tightly regulated to ensure an appropriate immune response against invading pathogens. Although experiments have mapped the trajectory from expansion to contraction, the interplay between mechanisms that control this response is not fully understood. Based on experimental data, we propose a model in which the dynamics of CD4+ T cell expansion is controlled through the interactions between T cells and antigen-presenting cells, where T cell stimulation is proportional to antigen availability, and antigen availability is regulated through downregulation of antigen by T cells. This antigen-dependent-feedback mechanism operates alongside an intrinsic reduction in cell proliferation rate that may also be responsible for slowing expansion. Our model can successfully predict T cell recruitment rates into division, expansion, and clonal burst size per cell when initial precursors are varied or when T cells are introduced late into an ongoing immune response. Importantly, the findings demonstrate that a feedback mechanism between T cells and antigen-presenting cells, along with a reduction in cell proliferation rate, can explain the ability of the immune system to adapt its response to variations in initial conditions or changes that occur later in the response, ensuring a robust yet controlled line of defence against pathogens.
MeSH term(s)	Antigens/immunology ; CD4-Positive T-Lymphocytes/cytology ; CD4-Positive T-Lymphocytes/immunology ; Cell Division ; Cell Proliferation ; Host-Pathogen Interactions/immunology ; Humans ; Immunity ; Models, Biological
Chemical Substances	Antigens
Language	English
Publishing date	2021-03-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	184905-0
ISSN	1522-9602 ; 0007-4985 ; 0092-8240
ISSN (online)	1522-9602
ISSN	0007-4985 ; 0092-8240
DOI	10.1007/s11538-021-00867-7
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Zs.B 754: Show issues

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Article ; Online: Faecal microbial transfer and complex carbohydrates mediate protection against COPD.

Budden, Kurtis F / Shukla, Shakti D / Bowerman, Kate L / Vaughan, Annalicia / Gellatly, Shaan L / Wood, David L A / Lachner, Nancy / Idrees, Sobia / Rehman, Saima Firdous / Faiz, Alen / Patel, Vyoma K / Donovan, Chantal / Alemao, Charlotte A / Shen, Sj / Amorim, Nadia / Majumder, Rajib / Vanka, Kanth S / Mason, Jazz / Haw, Tatt Jhong /

Tillet, Bree / Fricker, Michael / Keely, Simon / Hansbro, Nicole / Belz, Gabrielle T / Horvat, Jay / Ashhurst, Thomas / van Vreden, Caryn / McGuire, Helen / Fazekas de St Groth, Barbara / King, Nicholas J C / Crossett, Ben / Cordwell, Stuart J / Bonaguro, Lorenzo / Schultze, Joachim L / Hamilton-Williams, Emma E / Mann, Elizabeth / Forster, Samuel C / Cooper, Matthew A / Segal, Leopoldo N / Chotirmall, Sanjay H / Collins, Peter / Bowman, Rayleen / Fong, Kwun M / Yang, Ian A / Wark, Peter A B / Dennis, Paul G / Hugenholtz, Philip / Hansbro, Philip M

Gut

2024 Volume 73, Issue 5, Page(s) 751–769

Abstract: Objective: Chronic obstructive pulmonary disease (COPD) is a major cause of global illness and death, most commonly caused by cigarette smoke. The mechanisms of pathogenesis remain poorly understood, limiting the development of effective therapies. The ... ...

Abstract	Objective: Chronic obstructive pulmonary disease (COPD) is a major cause of global illness and death, most commonly caused by cigarette smoke. The mechanisms of pathogenesis remain poorly understood, limiting the development of effective therapies. The gastrointestinal microbiome has been implicated in chronic lung diseases via the gut-lung axis, but its role is unclear. Design: Using an Results: FMT alleviated hallmark features of COPD (inflammation, alveolar destruction, impaired lung function), gastrointestinal pathology and systemic immune changes. Protective effects were additive to smoking cessation, and transfer of CS-associated microbiota after antibiotic-induced microbiome depletion was sufficient to increase lung inflammation while suppressing colonic immunity in the absence of CS exposure. Disease features correlated with the relative abundance of Conclusion: The gut microbiome contributes to COPD pathogenesis and can be targeted therapeutically.
MeSH term(s)	Humans ; Mice ; Animals ; Pulmonary Disease, Chronic Obstructive/etiology ; Lung/metabolism ; Lung/pathology ; Pneumonia/etiology ; Inflammation/metabolism ; Carbohydrates/pharmacology
Chemical Substances	Carbohydrates
Language	English
Publishing date	2024-04-05
Publishing country	England
Document type	Randomized Controlled Trial ; Journal Article
ZDB-ID	80128-8
ISSN	1468-3288 ; 0017-5749
ISSN (online)	1468-3288
ISSN	0017-5749
DOI	10.1136/gutjnl-2023-330521
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Zs.A 160: Show issues

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Article ; Online: A High-Throughput Colorimetric Microplate Assay for Determination of Plasma Arginase Activity.

Smith, Natalie J / Maddahfar, Mahnaz / Gunasegaran, Bavani / McGuire, Helen M / Fazekas de St Groth, Barbara

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2620, Page(s) 273–286

Abstract: Arginase, an enzyme involved in the urea cycle, is gaining attention as a critical player in numerous chronic pathologies. Additionally, increased activity of this enzyme has been shown to correlate with poor prognosis in a range of cancers. Colorimetric ...

Abstract	Arginase, an enzyme involved in the urea cycle, is gaining attention as a critical player in numerous chronic pathologies. Additionally, increased activity of this enzyme has been shown to correlate with poor prognosis in a range of cancers. Colorimetric assays that measure the conversion of arginine to ornithine have long been used to determine the activity of arginase. However, this analysis is hindered by a lack of standardization across protocols. Here, we describe in detail a novel revision of the Chinard's colorimetric assay used to determine arginase activity. Dilution series of patient plasma are plotted to form a logistic function, from which activity can be interpolated by comparison to an ornithine standard curve. Inclusion of patient dilution series rather than a single point increases the robustness of the assay. This high-throughput microplate assay analyzes 10 samples per plate to produce highly reproducible results.
MeSH term(s)	Humans ; Arginase ; Colorimetry/methods ; Arginine ; Ornithine ; Plasma/chemistry
Chemical Substances	Arginase (EC 3.5.3.1) ; Arginine (94ZLA3W45F) ; Ornithine (E524N2IXA3)
Language	English
Publishing date	2023-04-24
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2942-0_29
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Article ; Online: Pembrolizumab for anaplastic thyroid cancer: a case study.

Aghajani, Marra Jai / Cooper, Adam / McGuire, Helen / Jeffries, Thomas / Saab, Jawad / Ismail, Kasim / de Souza, Paul / Bray, Victoria / Fazekas de St Groth, Barbara / Niles, Navin / Roberts, Tara Laurine

Cancer immunology, immunotherapy : CII

2019 Volume 68, Issue 12, Page(s) 1921–1934

Abstract: Blockade of the PD-1/PD-L1 pathway with targeted monoclonal antibodies has demonstrated encouraging anti-tumour activity in multiple cancer types. We present the case of a patient with BRAF-negative stage IVC anaplastic thyroid cancer (ATC) treated with ... ...

Abstract	Blockade of the PD-1/PD-L1 pathway with targeted monoclonal antibodies has demonstrated encouraging anti-tumour activity in multiple cancer types. We present the case of a patient with BRAF-negative stage IVC anaplastic thyroid cancer (ATC) treated with the anti-PD-1 monoclonal antibody, pembrolizumab, following radiographic progression on chemoradiation. Blood samples were collected prior to and at four time points during treatment with pembrolizumab. Mass cytometry was used to determine expression of relevant biomarkers by peripheral blood mononuclear cells. Faecal samples were collected at baseline and 4 weeks following treatment initiation; taxonomic profiling using 16S ribosomal RNA (rRNA) gene sequencing was performed. Following treatment, a marked expansion in CD20
MeSH term(s)	Antibodies, Monoclonal, Humanized/therapeutic use ; Antineoplastic Agents/therapeutic use ; B-Lymphocytes/immunology ; Bacteroides ; CD4-Positive T-Lymphocytes/immunology ; Feces/microbiology ; Humans ; Killer Cells, Natural/immunology ; Male ; Microbiota ; Middle Aged ; Programmed Cell Death 1 Receptor/antagonists & inhibitors ; RNA, Ribosomal, 16S/analysis ; Thyroid Carcinoma, Anaplastic/drug therapy ; Thyroid Neoplasms/drug therapy
Chemical Substances	Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; PDCD1 protein, human ; Programmed Cell Death 1 Receptor ; RNA, Ribosomal, 16S ; pembrolizumab (DPT0O3T46P)
Language	English
Publishing date	2019-10-22
Publishing country	Germany
Document type	Journal Article
ZDB-ID	195342-4
ISSN	1432-0851 ; 0340-7004
ISSN (online)	1432-0851
ISSN	0340-7004
DOI	10.1007/s00262-019-02416-7
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Article ; Online: Pediatric Burn Survivors Have Long-Term Immune Dysfunction With Diminished Vaccine Response.

Johnson, Blair Z / McAlister, Sonia / McGuire, Helen M / Palanivelu, Vetrichevvel / Stevenson, Andrew / Richmond, Peter / Palmer, Debra J / Metcalfe, Jessica / Prescott, Susan L / Wood, Fiona M / Fazekas de St Groth, Barbara / Linden, Matthew D / Fear, Mark W / Fear, Vanessa S

Frontiers in immunology

2020 Volume 11, Page(s) 1481

Abstract: ... B-cells and myeloid cells. Manual gating confirmed increased memory T-regulatory and central memory CD4+ ... T-cells, with altered expression of T-cell, B-cell, and dendritic cell markers. Conclusions: This study ...

Abstract	Epidemiological studies have demonstrated that survivors of acute burn trauma are at long-term increased risk of developing a range of morbidities. The mechanisms underlying this increased risk remain unknown. This study aimed to determine whether burn injury leads to sustained immune dysfunction that may underpin long-term morbidity. Plasma and peripheral blood mononuclear cells were collected from 36 pediatric burn survivors >3 years after a non-severe burn injury (<10% total body surface area) and from age/sex-matched non-injured controls. Circulating cytokine and vaccine antibody levels were assessed using multiplex immunoassays and cell profiles compared using a panel of 40 metal-conjugated antibodies and mass cytometry. TNF-α (1.31-fold change from controls), IL-2 (1.18-fold), IL-7 (1.63-fold), and IFN-γ (1.18-fold) were all significantly elevated in the burn cohort. Additionally, burn survivors demonstrated diminished antibody responses to the diphtheria, tetanus, and pertussis vaccine antigens. Comparisons between groups using unsupervised clustering identified differences in proportions of clusters within T-cells, B-cells and myeloid cells. Manual gating confirmed increased memory T-regulatory and central memory CD4+ T-cells, with altered expression of T-cell, B-cell, and dendritic cell markers. Conclusions: This study demonstrates a lasting change to the immune profile of pediatric burn survivors, and highlights the need for further research into post-burn immune suppression and regulation.
MeSH term(s)	Antibodies, Bacterial/blood ; Burns/immunology ; Child ; Child, Preschool ; Cytokines/metabolism ; Diphtheria-Tetanus-Pertussis Vaccine/immunology ; Female ; Humans ; Immunomodulation ; Immunophenotyping ; Infant ; Leukocytes, Mononuclear/immunology ; Male ; T-Lymphocyte Subsets/immunology ; T-Lymphocytes/immunology
Chemical Substances	Antibodies, Bacterial ; Cytokines ; Diphtheria-Tetanus-Pertussis Vaccine
Language	English
Publishing date	2020-07-21
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.2020.01481
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Article ; Online: Pro- and anti-tumour effects of B cells and antibodies in cancer: a comparison of clinical studies and preclinical models.

Guy, Thomas V / Terry, Alexandra M / Bolton, Holly A / Hancock, David G / Shklovskaya, Elena / Fazekas de St. Groth, Barbara

Cancer immunology, immunotherapy : CII

2016 Volume 65, Issue 8, Page(s) 885–896

Abstract: The primary immune role of B cells is to produce antibodies, but they can also influence T cell ... to define the role of B cells in antitumor immune responses, but despite this considerable body of work ... cancers, the presence of tumour-infiltrating B cells and tumour-reactive antibodies correlates ...

Abstract	The primary immune role of B cells is to produce antibodies, but they can also influence T cell function via antigen presentation and, in some contexts, immune regulation. Whether their roles in tumour immunity are similar to those in other chronic immune responses such as autoimmunity and chronic infection, where both pro- and anti-inflammatory roles have been described, remains controversial. Many studies have aimed to define the role of B cells in antitumor immune responses, but despite this considerable body of work, it is not yet possible to predict how they will affect immunity to any given tumour. In many human cancers, the presence of tumour-infiltrating B cells and tumour-reactive antibodies correlates with extended patient survival, and this clinical observation is supported by data from some animal models. On the other hand, T cell responses can be adversely affected by B cell production of immunoregulatory cytokines, a phenomenon that has been demonstrated in humans and in animal models. The isotype and concentration of tumour-reactive antibodies may also influence tumour progression. Recruitment of B cells into tumours may directly reflect the subtype and strength of the anti-tumour T cell response. As the response becomes chronic, B cells may attenuate T cell responses in an attempt to decrease host damage, similar to their described role in chronic infection and autoimmunity. Understanding how B cell responses in cancer are related to the effectiveness of the overall anti-tumour response is likely to aid in the development of new therapeutic interventions against cancer.
MeSH term(s)	Animals ; Antibodies/immunology ; B-Lymphocytes/immunology ; Disease Models, Animal ; Humans ; Mice ; Neoplasms/immunology
Chemical Substances	Antibodies
Language	English
Publishing date	2016-05-24
Publishing country	Germany
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	195342-4
ISSN	1432-0851 ; 0340-7004
ISSN (online)	1432-0851
ISSN	0340-7004
DOI	10.1007/s00262-016-1848-z
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Article ; Online: Experimental models to investigate the function of dendritic cell subsets: challenges and implications.

Hancock, D G / Guy, T V / Shklovskaya, E / Fazekas de St Groth, B

Clinical and experimental immunology

2013 Volume 171, Issue 2, Page(s) 147–154

Abstract: The dendritic cell (DC) lineage is remarkably heterogeneous. It has been postulated that specialized DC subsets have evolved in order to select and support the multitude of possible T cell differentiation pathways. However, defining the function of ... ...

Abstract	The dendritic cell (DC) lineage is remarkably heterogeneous. It has been postulated that specialized DC subsets have evolved in order to select and support the multitude of possible T cell differentiation pathways. However, defining the function of individual DC subsets has proven remarkably difficult, and DC subset control of key T cell fates such as tolerance, T helper cell commitment and regulatory T cell induction is still not well understood. While the difficulty in assigning unique functions to particular DC subsets may be due to sharing of functions, it may also reflect a lack of appropriate physiological in-vivo models for studying DC function. In this paper we review the limitations associated with many of the current DC models and highlight some of the underlying difficulties involved in studying the function of murine DC subsets.
MeSH term(s)	Animals ; Antigen Presentation ; Cell Communication ; Cell Differentiation ; Cell Lineage ; Dendritic Cells/immunology ; Humans ; Immune Tolerance ; Lymphocyte Activation ; Mice ; Models, Animal ; T-Lymphocyte Subsets/immunology ; T-Lymphocytes/immunology
Language	English
Publishing date	2013-01-03
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	218531-3
ISSN	1365-2249 ; 0009-9104 ; 0964-2536
ISSN (online)	1365-2249
ISSN	0009-9104 ; 0964-2536
DOI	10.1111/cei.12027
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