Development and Characterization of Monoclonal Antibodies Specific for Mouse and Human Fcγ Receptors

Tutt, Alison L.; James, Sonya; Laversin, Stéphanie A; Tipton, Thomas; Ashton‐Key, Margaret; French, Ruth R.; Hussain, Khiyam; Vaughan, Andrew T M; Dou, Liping; Earley, Alexander; Dahal, Lekh N.; Chen, Lu; Dunscombe, Melanie S.; Chan, H T Claude; Penfold, Christine A.; Kim, Jinny H.; Potter, Elizabeth A.; Mockridge, C. Ian; Roghanian, Ali; Oldham, Robert J.; Cox, Kerry L.; Lim, Sean H.; Teige, Ingrid; Frendéus, Björn; Glennie, Martin J.; Beers, Stephen A.; Cragg, Mark S.

doi:10.4049/jimmunol.1402988

Cited by 35 publications

(42 citation statements)

References 56 publications

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“…Interestingly, hFcgRIIB staining was higher on Ly-6C low ''patrolling'' monocytes than on Ly-6C hi ''classical'' monocytes from VG1543 mice (see Fig E3, D), as it is on the analogous populations in human blood, CD14 low CD16 hi ''patrolling'' monocytes, and CD14 hi '' classical'' monocytes (see Fig E3, E). Furthermore, spleen monocytes in human subjects 36 and VG1543 mice express significant levels of hFcgRIIB, reconciling hFcgRIIB expression on monocytes in this compartment between human donors and VG1543 mice. Macrophages from the peritoneum, but not from BAL fluid, of VG1543 mice were found to be positive for hFcgRIIB (Fig 2, B).…”

Section: Vg1543 Mice Exhibit Hfcgr Expression Patterns On Immune Cellmentioning

confidence: 87%

“…Only a fraction of donors we analyzed demonstrated hFcgRIIB expression on blood monocytes (see Fig E3, A, in this article's Online Repository at www.jacionline.org), which is consistent with its previously reported variable expression on CD14 lo monocytes and absence of expression on CD14 hi monocytes. 36 Thus VG1543 mice exhibit overexpression of hFcgRIIB on blood monocytes compared with that seen on human blood monocytes. Interestingly, hFcgRIIB staining was higher on Ly-6C low ''patrolling'' monocytes than on Ly-6C hi ''classical'' monocytes from VG1543 mice (see Fig E3, D), as it is on the analogous populations in human blood, CD14 low CD16 hi ''patrolling'' monocytes, and CD14 hi '' classical'' monocytes (see Fig E3, E).…”

Section: Vg1543 Mice Exhibit Hfcgr Expression Patterns On Immune Cellmentioning

confidence: 98%

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Mechanisms of anaphylaxis in human low-affinity IgG receptor locus knock-in mice

Gillis

Jönsson

Mancardi

et al. 2017

Journal of Allergy and Clinical Immunology

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Section: Vg1543 Mice Exhibit Hfcgr Expression Patterns On Immune Cellmentioning

confidence: 87%

Section: Vg1543 Mice Exhibit Hfcgr Expression Patterns On Immune Cellmentioning

confidence: 98%

Mechanisms of anaphylaxis in human low-affinity IgG receptor locus knock-in mice

Gillis

Jönsson

Mancardi

et al. 2017

Journal of Allergy and Clinical Immunology

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“…FACS analyses were performed using the FacsAria (BD Biosciences).For quantification of the Fcg receptor IV (FCGRIV) surface staining, pelleted cells were incubated for 30 min at 48C with rat anti-mouse CD16-2 (1:200; Clone AT137; AbDSerotec;Tutt et al, 2015). FACS analyses were performed using the FacsAria (BD Biosciences).For quantification of the Fcg receptor IV (FCGRIV) surface staining, pelleted cells were incubated for 30 min at 48C with rat anti-mouse CD16-2 (1:200; Clone AT137; AbDSerotec;Tutt et al, 2015).…”

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confidence: 99%

The microglial reaction signature revealed by RNAseq from individual mice

Hirbec

Marmai

Duroux-Richard

et al. 2018

Glia

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Microglial cells have a double life as the immune cells of the brain in times of stress but have also specific physiological functions in homeostatic conditions. In pathological contexts, microglia undergo a phenotypic switch called "reaction" that promotes the initiation and the propagation of neuro-inflammation. Reaction is complex, molecularly heterogeneous and still poorly characterized, leading to the concept that microglial reactivity might be too diverse to be molecularly defined. However, it remains unknown whether reactive microglia from different pathological contexts share a common molecular signature. Using improved flow cytometry and RNAseq approaches we studied, with higher statistical power, the remodeling of microglia transcriptome in a mouse model of sepsis. Through bioinformatic comparison of our results with published datasets, we defined the microglial reactome as a set of genes discriminating reactive from homeostatic microglia. Ultimately, we identified a subset of 86 genes deregulated in both acute and neurodegenerative conditions. Our data provide a new comprehensive resource that includes functional analysis and specific molecular markers of microglial reaction which represent new tools for its unambiguous characterization.

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“…The SPAR platform biosensor cells are designed to stably express both the modified mFcγRI-CD3ζ universal cell surface receptor and an aequorin reporter system. Fc receptors are expressed on the surface of hematopoietic cells and function as essential components of the antibody-mediated immune response and thus have been well characterized (28, 29). Each type of Fc receptor reacts specifically with a corresponding class of immunoglobulin: FcγRI receptors have been demonstrated to bind to their cognate antibodies with high affinity (28, 30, 31) though the affinity of the mFcγRI receptor for mouse antibodies varies with antibody subclass (32, 33).…”

Section: Discussionmentioning

confidence: 99%

Development of a Surface Programmable Activation Receptor system (SPAR): A living cell biosensor for rapid pathogen detection

Kittle¹,

Lwande²,

Williams³

et al. 2019

Preprint

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8Efficient pathogen detection is essential for the successful treatment and prevention of infectious 9 disease; however, current methods are often too time intensive to be clinically relevant in cases 10 requiring immediate intervention. We have developed a Surface Programmable Activation 11 Receptor (SPAR) diagnostic platform comprised of universal biosensor cells engineered for use 12 in combination with custom or commercial antibodies to achieve rapid and sensitive pathogen 13 detection. SPAR cells are stably transfected Jurkat T cells designed to constitutively express a 14 modified T cell mouse FcγRI receptor on the cell surface and a high level of the luminescent 15 reporter protein aequorin in the cytoplasm. The modified mFcγRI-CD3ζ receptor protein binds 16 with high affinity to the Fc region of any full-length mouse IgG2a and some IgG2 antibodies: 17 this allows customized target detection via the selection of specific antibodies. T-cell receptor 18 aggregation in response to target antigen binding results in signal transduction which, when 19 amplified via the endogenous T cell signal cascade, triggers the rapid intracellular release of 20 calcium. Increased Ca 2+ concentrations activate the expressed reporter protein aequorin resulting 21 in the immediate emission of detectable light. Testing demonstrates the accurate and specific 22 2detection of numerous targets including P. aeruginosa, E. coli O111, and E. coli O157. We 23 report that the SPAR biosensor cell platform is a reliable pathogen detection method that enables 24 the rapid identification of bacterial causative agents using standard laboratory instrumentation. 25The technology lends itself to the development of efficient point-of-care testing and may aid in 26 the implementation of effective and pathogen-specific clinical therapies. 27 Introduction 28The rapid and accurate identification of causative agents is critical to the prompt application of 29 directed, pathogen-specific antibiotic therapies. Effective and timely clinical intervention is 30 essential for the control of infectious disease as well as in the successful treatment of bacterial 31 infections. The observed increases in the frequency and severity of nosocomial infections (1) 32 and the increasing prevalence of antibiotic resistance induced by non-specific antibiotic use (2) 33 further highlight the need for informed antibiotic selection based upon precise and efficient 34 pathogen detection. 35 Current bacterial identification methods include both classic procedures and novel molecular 36 techniques. Traditional culture-based methods, while sensitive and reliable, are also labor-37 intensive and time-consuming and therefore often cannot provide definitive diagnostics within a 38 clinically relevant timeframe (3). Molecular diagnostic methods include immunological assays 39 such as ELISA (4), microarray immunoblot (5), or serological assays (6); nucleic acid-based 40 techniques including PCR (7), DNA sequencing (8), hybridization techniques (9), or DNA/RNA 41 microarrays (10);...

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Development and Characterization of Monoclonal Antibodies Specific for Mouse and Human Fcγ Receptors

Cited by 35 publications

References 56 publications

Mechanisms of anaphylaxis in human low-affinity IgG receptor locus knock-in mice

Mechanisms of anaphylaxis in human low-affinity IgG receptor locus knock-in mice

The microglial reaction signature revealed by RNAseq from individual mice

Development of a Surface Programmable Activation Receptor system (SPAR): A living cell biosensor for rapid pathogen detection

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