C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
195/34.7
C12N 5/08 (2006.01) C12N 5/06 (2006.01)
Patent
CA 1309677
Form 24 ABSTRACT A novel B-cell line (designated WEE-1) has been artfully cloned from human peripheral blood. By innovative isolation procedures and specialty phenol-red free culture medium, the cloned cells grow into large clusters. Under light microscopy, these fusiform cells exhibit apical cytoplasmic projections, and irregular large nuclei. By electron microscopy (EM) the cell reveals abundant mitochondria, stratified rough endoplasmic reticulum (RER), ribosome lamella complex, and a heterochromatic thick-rimmed, irregular nucleus containing prominent nucleoli. The cells are positive for immunoglobulin (Ig) G and M heavy-chains, and Kappa (K) light-chain, and they express tartrate-resistant acid phosphatase (TRAP), Epstein-Barr virus (EBV) antigen, and abnormal karyotype of 45, X, -Y, +12q. When cell DNA was hybridized with recombinant probes specific for immunoglobulin heavy-chain joining region (JH), kappa and lambda light-chain constant regions (CK and C.lambda.), autoradiographs reveal rearranged JH, and CK genes and germ line configuration of C.lambda.. Upon immunophenotyping by flow cytometry, the WEE-1 cells express a sequence of human leucocyte antigen (HLA) class II antigens DR+DP+DQ+. These distinctive cells exhibit the CD19 and CD20 pan-B cell antigens CD23 and CD71 receptors for IgE and transferrin, but lack the common acute lymphocytic leukaemia antigen (CALLA or CD10), haematopoietic progenitor cell antigen (HPCA-1 or CD34), terminal deoxynucleotidyl transferase, and the common myelomonocytic and T-cell lineage antigens. The WEE-1 cell line is an invaluable tool for the study of B-cell development and differentiation, and acquisition of a non- toxic 21 kilodaltons (Kd) protein designated blood tumour necrosis factor (BTNF or TNF-.gamma.) for use in biological response modifications. Interaction of these unique cells with other haematopoietic cell lines will increase our comprehension of host defense and immunity, cell-cell communication, intracellular processing of proteins, and functional significance of histocompatibility.
577582
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