"Detection of cytokines is often at the heart of medical research, but the multitude of techniques involved, and their complexity can make finding the right technique difficult. This meeting will give an insight into the latest advances in both established techniques, such as ELISPOT and FACS and the more recently developed mulitplex technology for both genes and proteins, such as the Luminex system. Talks will be given by both the companies developing these techniques and the academics using them". Chair: Dr Catherine Derry
09:00 – 09:25 Registration
09:25 - 09:30 Introduction by the chair – Dr Catherine Derry
09:30 – 09:50 Introduction of the first ELISPOT based diagnostic assay – Diagnosing TB in the 21st Century Dr Ian Durrant, Research Director, Oxford Immunotec Limited, UK
Tuberculosis has been poorly served for diagnostic purposes for many years. The current skin test is over 100 years old and is acknowledged as having a generally poor specificity and also in some cases a low sensitivity. The ELISPOT technique offers a route for development of an up-to-date blood based diagnostic assay that rapidly confirms active TB disease and also correlates with exposure to accurately diagnose latent TB infection. The ability to detect latent TB, and therefore identify at-risk individuals, is of increasing importance as more people are exposed to immunosuppressive therapies.
09:50 – 10:10 The Bio-Plex Suspension Array System: Multiplexed cytokine detection made easy Dr Alex Liversage, Biorad, UK
The Bio-Plex Suspension Array System is a multiplexed bead-based assay technology that allows for the multiplexing of up to 100 assays from a single microtitre plate well to dramatically increase sample throughput and data generation. Bio-Rad has developed an extensive but still growing panel of cytokine and phosphoprotein assays which in conjunction with the Bio-Plex System allow for simple and intuitive multiplexed cytokine or phosphoprotein analysis.
10:10 - 10:40 Discovery multiplex platform: Sensitive measurement of multiple cytokines in 25 ml sample volume Dr Tuc Ahman, MSD, UK
10:40 - 11:00 Morning tea/coffee
11:00 – 11:30 Developing reagents for detection of cytokines in large animals Dr Jayne Hope, Institute for Animal Health, Compton, Near Newbury, UK
There is a relative paucity of reagents to study immunological mechanisms in cattle. We have developed antibodies and recombinant proteins to assist our studies of cytokine expression in cattle.
11:30 – 12:00 Comparison of three multiplex cytokine analysis systems: Luminex, SearchLight and Fast Quant Dr Gendie Lash, University of Newcastle-upon-Tyne, UK
In my research group we are interested in uterine factors involved in regulating trophoblast invasion and spiral artery remodelling. In particular factors secreted by the major pregnant decidua leucocyte, uterine natural killer (uNK) cells, however while isolation of these cells is possible, low numbers are obtained. Wanted a technique to be able to measure several factors in the same sample. Came to Euroscicon meeting in 2003 to learn more about technologies having received grant money for a Luminex system. From that meeting trialed SearchLight system and later the FAST Quant system. For our own interest had run the same samples on all three systems, and were therefore in a position to compare them to each other – as opposed to single ELISA as has been done previously. This comparison study has just been published as a technical note in Journal of Immunological Methods, and to our knowledge is the first of its kind.
12:00 – 12:20 tba
12:20 - 13:15 Lunch in the exhibition area
13:15 – 13:45 Detection of HIV-1-Specific T Lymphocytes in HIV-1 Infected Individuals and Vaccine Recipient Dr Nesrina Imami, Department of Immunology, Imperial College London, Chelsea and Westminster Hospital , UK
In clinical nonprogressors (CNPs), CD4 T-cell orchestration of anti-HIV-1 immunity is mediated via a balanced proliferative ability and cytokine expression/production. Whilst CNPs make vigorous CD4 and CD8 T-cell proliferative IL-2-producing responses to all HIV-1 antigens, the functional defects in HIV-1-specific CD4 and CD8 T cells in chronic HIV-1 infection include an inability to proliferate and produce IL-2 in responses to HIV-1 antigens/peptides, although secretion of antiviral cytokines such as IFN? and TNF? are unimpaired. These important finding demonstrates that HIV-1-specific CD4 and CD8 T cells are not deleted/destroyed during chronic infection, but are actually present although unable to respond properly to HIV-1.
13:45 - 14:15 Development of subunit vaccines and specific reagents to detect tuberculosis in cattle: Antigen mining by comparative genomics and transcriptomics Dr Martin Vordermeier, TB Research Group, VLA-Weybridge
Bovine tuberculosis in cattle is an increasing economic problem in GB. The definition of antigens for the diagnosis of bovine tuberculosis, as well as for the development of effective vaccines for cattle are research priorities. To continue diagnosis alongside Mycobacterium bovis BCG-based vaccination regimens, it is necessary to define reagents that allow the discrimination of infected and vaccinated animals. Potential M. bovis-specific antigens were listed by comparative analysis of the genomes of M. bovis, BCG, M. avium, M. paratuberculosis, and Streptomyces coelicolor and by transcriptome analysis after bovine alveolar macrophage were infected with M. bovis or BCG. Potential antigens were then tested by applying a high-throughput peptide-based screening system measuring IFN-?. Promising antigens were further refined and formulated into diagnostic peptide cocktails for further evaluation. Refinement of the in silico strategy to select antigens will be discussed.
14:15 – 14:35 Cytokine Secretion Assay Technology to Isolate and Characterise Effector and Suppressor T Cells Dr John Campbell, Miltenyi Biotec
The Cytokine Secretion Assay system uniquely allows the isolation of T cells based on the cytokines they produce as a result of their function. Here I will concentrate on two very different applications – 1. Isolation of virus-specific effector cells, and their use as a cellular therapeutic. 2. Novel cellular stimuli to induce IL-10 production, the subsequent isolation of these cells, and examination of suppressor function.
14:35 – 15:00 Afternoon Tea
15:00 – 15:30 Multiplex cytokine detection in autoimmune disease Dr Graham Wallace, University of Birmingham, UK
15:30 – 16:00 Intracellular cytokine staining to analyse the function of tissue dendritic cells (DC) Dr Andrew Stagg, Imperial College, London, UK
A number of approaches are available to measure cytokine production by immune cells in vitro. These include analysis of culture supernatants (eg by ELISA), measurement of mRNA (eg by PCR) or intracellular staining and analysis by flow cytomtery. Each of these approaches has strengths and weaknesses. DC are key immunoregulatory cells but infrequent in tissues and difficult to obtain at high purity. Intracellular staining is powerful as it permits unambiguous identification of the cellular source of cytokine. The talk will describe how the challenges of intracellular staining of DC have been tackled in terms of experimental design and analysis of data.
16:30 Close
Registration fees
Standard fee - £440
Academic fee - £220
Student fee- £140
IBMS members fee - £199
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