The Technological Objectives of TRANS-NET can be summarised as :-

•  The development of microarray and bioinformatics platforms and microarrays for use in HSCT.

•  Cross-standardisation of complex technologies, i.e. chimerism analysis, skin explant model, immunohistochemistry.

•  Isolation and characterisation of novel peptides and subsequent functional analysis.

An example of a particular novel technological objective is the combined use of a skin explant assay and MHC expression profiling in a rat animal model which may lead to breakthroughs in the identification of new MHC genes of importance to determining outcome in HSCT.

•  New gene polymorphisms associated with good or poor prognosis or relapse of disease and or transplant success.

•  New markers for GvHD as opposed to viral or drug-induced adverse reactions .

•  Novel peptides/ therapies for use in pre-clinical trials.

Research Method - Background and Methodology of the Project

The multidisciplinary methodological approach to be employed to investigate the broad objectives of the project 1) identification of predictive and diagnostic markers, including novel peptides, and understanding GvL and GvHD responses and 2) training of researchers in application of results have been divided into 7 interacting WORKPACKAGES (WP's) and TASKS.

Each WP starts with an initial Training Task WP 1 ( Models of GvHD ) exploits the use of an in vitro human skin explant model, developed in Newcastle, for studying and predicting the occurrence of GvHD . The assay consists of incubating 7 day mixed lymphocyte culture (MLC) alloreactive donor cells with patient skin (for 3 days of co-culture) in vitro prior to transplant and assessing the graft versus host type alloreactivity (GvHR) of the donor cells by histopathological assessment. Histopathological assessment of the assay, due to FPIV collaboration, is reproducible between partners. All biopsies including those used for profiling experiments will be evaluated centrally by collaboration and exchange of slides, with sub contractor, Prof Sviland (Bergen). The assay is a novel and unique tool for studying the immunobiology, human alloreactivity and GvHD .