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Development of in-vitro tests for drug allegernicity and b cell switching to ige synthesis

Objective



The aim of this proposal is to develop in-vitro tests that are predictive of the capacity of drugs to induce allergic reactions in man. The processes that lead to antibody production by B cells in response to primary drug allergen exposure, culminating in IgE synthesis, will be modelled in-vitro using lymphoid cells of human origin. The methods to be used will draw extensively on novel in-vitro technology. Ultimately the aim is to simplify the system, for example by the use of cell lines, such that it may be easily developed as a test for screening of drugs for allergenicity.
The project is divided into 5 interactive Work Packages tWP) that are designed to meet the above scientific objectives. Throughout the project beta-lactam antibiotics will be used as model compounds since they are known to conjugate spontaneously to proteins to generate allergenic epitopes without requirement for metabolic activation.
In WP 1 the plasma protein and cellular peptide targets for drug conjugation will be characterised using immunochemical and molecular biological techniques. In particular the possibility that beta-lactam drugs may conjugate directly to MHC class II molecules will be investigated. These conjugates will be made available to WP 2-5.
In WP 2 the responses of antigen-presenting cells to drugs and drug-derived peptides will be studied in terms of cytokine expression and surface adhesion molecule expression.
The responses of drug-specific human T cells in terms of patterns of cytokine expression, surface adhesion molecule expression and activation of transcription factors will be studied in WP 3.
In WP 4 T cell clones will be derived from drug-allergic patients. Patterns of cytokine and adhesion molecule expression will be studied, and compared with primary cultures.
Optimal conditions for generation and activation of drug- specific T cells will be established in WP 3 and WP 4 and subsequently these cells and techniques will be transferred to WP 5.
In WP 5 primary B cell antibody production to drugs will be induced using B cells stimulated via CD40 in the presence of interleukins-4 and -13 and composite peptides incorporating B cell and T cell drug-derived epitopes. The signals required for IgE switching will be identified. This system will provide the basis for development.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

THE UNIVERSITY OF LIVERPOOL
Address
Ashton Street
L69 3GE Liverpool
United Kingdom

Participants (5)

Bibra International
United Kingdom
Address
Woodmansterne Road
SM5 4DS Carshalton
LUND UNIVERSITY
Sweden
Address
33,Sölvegatan 33
220 07 Lund
Servizio Andaluz de Salud
Spain
Address
S/n,camino Antequera
29010 Malaga
Universita degli Studi di Firenze
Italy
Address
Viale Morgagni 85
50134 Firenze
University College Dublin
Ireland
Address
Fosters Avenue Blackrock
4 Dublin