A DISTRIBUTION STOCK OF THE ESTABLISHED CELL LINES WILL BE PREPARED AND WILL BE MADE AVALAIBLE TO EUROPEAN RESEARCH CENTERS FOR R & D AND DIAGNOSTIC WORK.
THESE LINES WILL BE IMPORTANT FOR INDUSTRY TO DEFINE MOLECULES INVOLVED IN THE PATHOGENESIS OF THE DISEASE AND FOR IN VITRO TESTING OF DRUGS FOR FUTURE THERAPY.
A CATALOGUE OF CELL LINES TOGETHER WITH A COMPLETE HLA TYPING AND DETAILS OF DIAGNOSIS WILL BE PRODUCED.
Human blood lymphocytes are specialized for certain immune functions and express a number of surface markers. Among these, the human leucocyte antigens (HLA) are of utmost importance for the cellular individuality and immunocompetence of a given person. Since lymphocytes have a restricted life span, immortalization by Epstein-Barr virus (EBV) to become continuously in vitro growing cell lines and collection according to HLA phenotypes are essential for further immunogenetic studies.
In order to create a bank of immunogenetically defined cell lines, expertise in cell culture technology and in HLA typing using serological, cellular, biochemical and molecular genetics techniques are neccessary. Lymphocytes from probands proven to be homozygous for a particular HLA haplotype by family studies or to be a member of an HLA recombinant family, from patients suffering from leukaemia or HLA linked and associated diseases and from their relatives were immortalized by EBV, cryobiologically frozen and stored under liquid nitrogen. Since the HLA system is the most polymorphic gene system in man, a collection of cell lines must be relatively extensive to cover all recognized HLA specificities and thus serve as a reference for research on the fine analysis of HLA and other genetic systems.
2200 lymphoblastoid cell lines (LCL) from 296 families were established. This figure includes LCL from 241 HLA homozygotes of which 88 were analysed in international workshops on histocompatibility testing and 20 families with an informative intra-HLA recombination. Technical improvements for immunogenetic characterization of the LCL were made by molecular genetics techniques (eg polymerase chain reaction) and typing with HLA class II allele specific oligonucleotides, biochemical subtyping of HLA class I gene products by 1-dimensional isoelectric focussing and immunoblot, and nonradioactive (eg digoxigenin) labelling of HLA-DPA- and HLA-DPB-specific complementary deoxyribonucleic acid (cDNA ) probes. In affiliation with the European collection of animal cell cultures (ECACC), Salisbury, United Kingdom with 145 HLA homozygous reference LCL, these repositories form the European collection for biochemical research (ECBR).
COLLECTION OF EBV TRANSFORMED B CELL LINES FROM INDIVIDUALS KNOWN TO BE HOMOZYGOUS WITH RESPECT TO HLA SPECIFICITIES (A, B, C, DR, DQ, DP) AND OTHER MARKERS (C2, C4A, C4B, BF, GLO, ETC.); FROM PATIENTS AFFECTED WITH DISEASES PROVED OR SUPPOSED TO BE OF GENETIC ORIGIN, AND FROM THEIR FAMILIES. SELECTION AND EXTENSIVE HLA TYPING WILL BE PERFORMED OF FAMILIES WITH AT LEAST TWO PATIENTS AFFECTED WITH: COELIAC DISEASE, JUVENILE DIABETES, ANKYLOSING SPONDYLITIS, MULTIPLE SCLEROSIS, 21-HYDROXYLASE DEFICIENCY, COREA MAJOR, ALZHEIMER'S DISEASE OR OTHER GENETIC DISEASES, AND WITH AT LEAST ONE HEALTHY SIBLING AS A CONTROL FOR THE GENETIC ORIGIN.
Funding SchemeCSC - Cost-sharing contracts