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Efficient cytokine detection: The Way Forward – 3rd July 2009

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If you have a question about this talk, please contact Dr Astrid Englezou.

The meeting will be held at The BioPark, Hertfordshire, UK and has CPD approval. The Chair: will be Dr Stephen Thompson, (King’s College London, UK)

Are you measuring or want to measure cytokines ?, trying to optimise your technique or do you want just to make sure you are going about it the right way? This meeting aims to summarise some of the options open to you and to discuss which techniques best suit your requirements. A series of presentations from experts working in the academic, clinical and commercial sectors will compare recent developments in technologies ranging from relatively “low-tech” assays such as ELISA , Elispot and bioassays through to the “high-tech” platform technologies of various multiplex cytokine detection systems. Their use in basic science, R&D and translational research will be discussed.

If you book this event you will get 30% off the EARLY REGISTRATION FEE for registration to the partner ELISPOT FOLLOW UP EVENT which will take place on the 15th Oct 2009. To find out more about this meeting go to www.regonline.co.uk/elispot09. On registering for Efficient cytokine detection: The Way Forward you will receive an email giving you details of how to receive your discount to the October ELISPOT event. The Agenda includes:

Cytokine detection in collagen-induced arthritis. Dr Richard Williams, Kennedy Institute of Rheumatology Division, Imperial College, UK Collagen-induced arthritis is a model of rheumatoid arthritis that has been used extensively to validate novel therapeutic targets. The main pathological features of the disease include synovitis, pannus formation and joint erosion. There is a great deal of interest in the development and testing of drugs with the capacity to modulate inflammatory pathways in arthritis. Hence, there is a need to monitor the effect of novel treatments on cytokine expression in vivo. This presentation will focus on the techniques used to quantify changes in cytokine expression following therapeutic intervention

Using ELIspot to detect rare antigen specific T cells . Dr Tim Tree, Kings College, London

Isolation of live Regulatory, Effector and interleukin-17-producing T cells using cytokine secretion – Dr John Campell – Miltenyi Biotec Ltd, Surrey, UK IL-17 producing T cells are currently the focus of much interest in the fields of inflammation and immunity. The IL-17-producing phenotype is somewhat plastic, and generation of IL-17-producing T cells in vitro is complex. Here I will demonstrate the detection and isolation of live IL-17-producing T cells direct from blood and spleen using the IL-17 secretion assay system. The phenotypes and functions of natural IL-17-producing cells will be discussed, along with the possibility of splitting the IL-17 and IFN -gamma secreting populations based on two-colour cytokine secretion.

IL-10, regulatory T cells and respiratory health: the role of the vitamin D pathway. Dr Catherine Hawrylowicz, Kings College, London CD4+Foxp3+ Treg and IL-10 secreting Treg are proposed to play a role in the control of immune homeostasis in the lung and may have therapeutic potential in allergic and asthmatic disease. Our studies are investigating pharmacological protocols to promote these regulatory T cell populations in allergic and asthmatic patients.

Application of cytokines assays in the biotechnology sector – from proof-of-principle to clinical trials. Dr James N Francis, Senior Vaccine Development Scientist, Immune Targeting Systems Ltd, London, UK The development of novel T-cell vaccines relies heavily on cytokine measurements for proof-of-principle experiments, product characterization and immunomonitoring of clinical trials. Multiple approaches to cytokine measurements are utilized during these processes. The ELI Spot remains the workhorse for most immunomonitoring of phase I and II vaccine clinical trials due to the assay’s high sensitivity and optimisation of these assays are crucial for success. In-house characterization of novel T-cell vaccines may utilse multiplex cytokine arrays and intracellular cytokine staining to determine the immunological profile of candidate vaccines. Practical aspects of cytokine measurement will be discussed during this presentation

Talk title to be confirmed – Professor Paul Lehman and Wenji Zhang

Talk title to be confirmed. Dr. Deborah A. O’Neil, NovaBiotics Ltd, Aberdeen, UK

Immunohistochemical visualization of cytokines and other small molecules. Dr Chris van der Loos, Academic Medical Center, Univ. of Amsterdam , The Netherlands The use of post-fixed cryostat tissue sections has been widely accepted as ‘gold-standard’ for testing primary antibodies in immunohistochemistry (IHC). However, IHC staining of unbound small molecules like cytokines, may result into false-positive plasma cells, whereas specific staining is lacking. Leaking of the cytokine molecule during post-fixation is most likely the cause of this problem. In contrast, pre-fixation does retain the small molecules much better. Based on these findings suggestions are made for positive controls from cells and the use of optimally formalin-fixed paraffin tissue sections. Furthermore, testing of more than one primary antibody is highly recommended.

Differing multiplex cytokine analysis platforms . Dr Gendie Lash, Institute of Cellular Medicine, Newcastle University Multiplex cytokine analysis technologies have become readily available in the last seven years. Two main formats exist: multiplex sandwich ELISA and bead based assays. While these have each been compared to individual ELIS As, there has been little direct comparison between the two formats. I will discuss the comparison between two multiplex sandwich ELISA assays (FAST Quant and SearchLight) and a bead based assay (UpState Luminex).

We invite abstract submissions. A prize of a year’s subscription to Nature Reviews Immunology at the conclusion of the day will be awarded to the best oral/poster presentation. We encourage you to submit abstracts for these sessions, which are designed to promote a balanced program of talks, delivered by both well-established, pre-eminent researchers in the field as well as those starting out on their careers.

If you would like to book a place please visit : www.regonline.co.uk/cytokines09

This talk is part of the euroscicon series.

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