BÜHLMANN CAST® Assays Technical Presentation

Slides:

Advertisements

Similar presentations

A. a). d). c). f). b).e). g). Figure S1. NAT2 expression in lymphocytes and monocytes from a healthy volunteer. PBMC from a healthy volunteer were simple.

Advertisements

Principles of Immunology Hypersensitivity and Allergy 4/11/06

Immunology مقدمة في علم المناعة.

Principles of Immunology Cells of the Immune System 1/26/06 “Those who have health, have hope. Those who have hope have everything.” Arabian proverb.

Immune Cells , Receptors, and Markers. Lymphoid Tissues and Organs.

Cells of inflammation and Immunity G. Wharfe 2005.

IgE mediated hypersensitivity

Introduction to Lab Ex. 24: Hypersensitivity. Response to antigens (allergens) leading to damage Require sensitizing dose(s) Introduction to Lab Ex. 24:

The Immune System memory and specificity Chapter 43

Humoral Immunity presented by Dr. MAHROO MIRAHMADIAN Professor of Immunology Tehran University of Medical Sciences.

L ECTURES 2014 KEFAH F. HASSOON L ECTURE N O. 1 Immune System Disorders Auto-immune Diseases Hypersensitivity reactions.

The immune response White Blood cell types. Myeloid stem cells Lymphoid cells Pluripotent stem cells (in bone marrow) Monocyte Mast cells Basophils Neutrophils.

Food Allergy & Food Intolerance. Allergy an abnormal reaction of the immune system to foreign (not infectious) material, leading to injury to the body.

Disorders of Immune System - Hypersensitivity Reactions: Immune response to exogenous antigens - Autoimmune diseases: Immune reactions against self antigens.

12 Immune Response to Biomaterials CHAPTER

NAJRAN UNIVERSITY College of Medicine NAJRAN UNIVERSITY College of Medicine Microbiology &Immunology Course Lecture No. 15 Microbiology &Immunology Course.

Ch 43 The Body’s Defense. Three lines of Defense: 1. External defenses 2. Internal (phagocytes, inflammation) 3. Specific defense: (lymphocytes) 1 & 2.

CATEGORY: IMMUNE DYSFUNCTION Anaphylaxis Tariq El-Shanawany, University Hospital of Wales, UK Anaphylaxis is a severe, life-threatening, generalised or.

Hypersensitivity MBBS- Batch 16 Remya.

Anaphylaxis Tariq El-Shanawany, University Hospital of Wales, UK

Basophils Karen Buckland, Imperial College London, UK

Mast Cells Y Y Y Y Y Y Y Y Y Y Y Y Y

Chapter 18 Immunological Disorders

Hypersensitivity reactions

Histamine, lipid mediators, cytokines

PANEL 1 All events Lymphocytes (FSC-A v SSC-A)

Selecting Allergen for BAT

Early suppression of basophil activation during allergen-specific immunotherapy by histamine receptor 2 Natalija Novak, MD, Nihal Mete, MD, Caroline.

Ch 43 The Body’s Defense.

Immune regulation Topics Humoral Immune Response Part II

Eosinophil Recruitement

CD8+ T lymphocytes and leukotriene B4: Novel interactions in the persistence and progression of asthma Erwin W. Gelfand, MD, Azzeddine Dakhama, PhD

Delayed clinical and ex vivo response to mammalian meat in patients with IgE to galactose-alpha-1,3-galactose Scott P. Commins, MD, PhD, Hayley R. James,

Odelya E. Pagovich, MD, Bo Wang, MD, Maria J

Making the Most of In Vitro Tests to Diagnose Food Allergy

IgE mediated hypersensitivity

Expression of IL-4 receptor α on smooth muscle cells is not necessary for development of experimental allergic asthma Frank Kirstein, PhD, William G.C.

The who, where, and when of IgE in allergic airway disease

Peanut oleosins associated with severe peanut allergy—importance of lipophilic allergens for comprehensive allergy diagnostics Christian Schwager, Dipl.

Naive T cells sense the cysteine protease allergen papain through protease-activated receptor 2 and propel TH2 immunity Genqing Liang, PhD, Tolga Barker,

Eosinophils contribute to the resolution of lung-allergic responses following repeated allergen challenge Katsuyuki Takeda, MD, PhD, Yoshiki Shiraishi,

Mass cytometry profiling the response of basophils and the complete peripheral blood compartment to peanut Leticia Tordesillas, PhD, Adeeb H. Rahman,

Direct monitoring of basophil degranulation by using avidin-based probes Régis Joulia, PhD, Claire Mailhol, MD, Salvatore Valitutti, MD, Alain Didier,

Ellen Mueller Fox, PhD, Marina N

Thymic stromal lymphopoietin activation of basophils in patients with allergic asthma is IL-3 dependent Brittany M. Salter, BSc, John Paul Oliveria,

Mucosal Immunology of Food Allergy

CELLS INVOLVES IN IMMUNITY UTTARAN MODHUKALYA 1. CONTENT 1. INTRODUCTION 2. HEMATOPOIETIC STEM CELL 3. LYMPHOCYTE CELL 4. MYELOID CELL 5. REFERENCE 6.

Patients Allergic to Fish Tolerate Ray Based on the Low Allergenicity of Its Parvalbumin Tanja Kalic, MSc, Francoise Morel-Codreanu, MD, Christian Radauer,

Role of IgE in autoimmunity

Yilin Qi, MSc, Darwin J. Operario, PhD, Christopher M

Individual IL-3 priming is crucial for consistent in vitro activation of donor basophils in patients with chronic urticaria Thomas Gentinetta, MSc, Tatjana.

Heterogeneity of mast cells and T cells in the nasal mucosa

1 What is the function of the blood cell at the tip of the arrow:

Department of Pathology

Oktay Kirak, MD, Gert Riethmüller, MD

Mohamed H. Shamji, PhD, FAAAI, Stephen R. Durham, MD, FRCP

Sarbjit S. Saini, MDa, Jennifer J

Sara Paveglio, PhD, MS, Erin Bennett, MS, Kelly L. Hawley, PhD, Adam P

Ellen Mueller Fox, PhD, Marina N

Molecular and cellular mechanisms of allergic disease

Utility of Total Serum IgE in Allergy

Microarray immunoassay: Association of clinical history, in vitro IgE function, and heterogeneity of allergenic peanut epitopes Wayne G. Shreffler, MD,

Clinical safety of Food Allergy Herbal Formula-2 (FAHF-2) and inhibitory effect on basophils from patients with food allergy: Extended phase I study

Thymic stromal lymphopoietin does not activate human basophils

Erythrocyte What type of cell is this?.

The future of biologics: Applications for food allergy

Volume 13, Issue 2, Pages (October 2015)

Human BAT in allergic and normal control subjects.

Inflammatory and immune cells involved in asthma.

Presentation transcript:

BÜHLMANN CAST® Assays Technical Presentation 03/06/2016

Type 1 Allergy Mechanism APC Sentinel Cells Allergens ‘Insults’ sIgE antibodies T Lymphocytes B Lymphocytes/Plsma Cells Specific Immune Response Activated & Degranulated Mast Cells Basophils Effector Cells Mediator Release: Histamin, Leukotriens Lab Training 16 - MR

BÜHLMANN CAST®: Functional IVD Cell Based Assays in Allergy Diagnosis The CAST® assay is the diagnostic support of the allergologist to predict the patient allergic reaction to a substance in a safe blood test. With CAST® assay we have standardized an in vitro functional assay to evaluate and quantify the potential of an allergen to activate the patient specific basophils, the circulating effector immune cells in allergy. Activated & Degranulated Basophils CAST® Allergens Basophil Activation Test (BAT) Flow CAST® Mediator Release: Leukotrienes Immunoassays CAST® ELISA Lab Training 16 - MR

Flow CAST® Principle: CCR3 / CD63 a) BÜHLMANN Allergen a) Selection Marker anti-CCR3-PE: Detects all basophils whithin the blood samples Activation Marker anti-CD63-FITC: Detects only basophils which have been activated by the allergen b) Lab Training 16 - MR

Flow CAST® Principle: CCR3 / CD63 Stimulation Control anti-FcεRI mAb: Immunological activation of the basophils: Mimics the cross-linking of two bound IgE with the allergen directly via FcεRI receptor Stimulation Control fMLP (tripeptide): Non-immunological activation of the basophils: The mechanism is not known. Detects the viability of the basophils. e) Lab Training 16 - MR

Flow CAST® Samples per Patient PB Patient Background (PB) Stimulation Buffer Blood Staining Reagent PC1 PC2 Patient Controls (PCs) Stimulation control (PC1: anti-FcεRI mAb; PC2: fMLP) Blood Staining Reagent Ax Allergen Allergen x Blood Staining Reagent Lab Training 16 - MR

Flow CAST® Analysis Basophils Eosinophils Neutrophils Lymphocytes FSC (rel. volume) Lymphocytes Monocytes SSC (rel. granularity) Lab Training 16 - MR

Flow CAST® Analysis Basophils Eosinophils Neutrophils Lymphocytes FL2+; SSClow FL2+; SSChigh Basophils Eosinophils CCR3-PE (FL2) Neutrophils Lymphocytes Monocytes SSC (rel. granularity) Lab Training 16 - MR

Flow CAST® Analysis Activated & degranulated basophils FL1+FL2- CD63 CD203c Activated & degranulated basophils CD63-FITC (FL1) FL1- FL2- FL1-FL2+ Not activated basophils CCR3-PE (FL2) Lab Training 16 - MR

Flow CAST® Analysis Patient Background (PB) ~ 2% Patient Control (PC) FSC (rel. volume) CCR3-PE (FL2) CCR3-PE (FL2) CD63-FITC (FL1) Patient Control (PC) (a-FceRI) SSC (rel. granularity) SSC (rel. granularity) CCR3-PE (FL2) Lab Training 16 - MR

Flow CAST® Analysis Patient Background (PB) ~ 2% Patient Control (PC) FSC (rel. volume) CCR3-PE (FL2) CCR3-PE (FL2) CD63-FITC (FL1) Patient Control (PC) (a-FceRI) >10% sample is evaluable SSC (rel. granularity) SSC (rel. granularity) CCR3-PE (FL2) Lab Training 16 - MR