Friday, July 03, 2009 9:00 am - 5:00 pm
BioPark
Broadwater Road
Welwyn Garden City, Hertfordshire AL7 3AX
United Kingdom
Talks include:
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.
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 ELISpot 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
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 ELISAs, 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
See www.regonline.co.uk/cytokines09 for further information
