Chromosome mapping of biological pathways by fluorescence-activated cell sorting and cell fusion: human interferon gamma receptor as a model system

• Published in: Somatic cell and molecular genetics 1988-11, Vol.14 (6), p.583-592
• Main Authors: JUNG, V, JONES, C, RASHIDBAIGI, A, GEYEER, D. D, MORSE, H. G, WRIGHT, R. B, PESTKA, S
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Blotting, Northern; Cell Fusion; Cell Separation - methods; Chromosome Mapping - methods; chromosomes; Chromosomes, Human, Pair 21; Chromosomes, Human, Pair 6; Classical genetics, quantitative genetics, hybrids; Flow Cytometry - methods; Fundamental and applied biological sciences. Psychology; gamma -interferon; genes; Genetics of eukaryotes. Biological and molecular evolution; HLA Antigens - genetics; Human; Humans; Hybrid Cells; Receptors, Immunologic - genetics; Receptors, Interferon; RNA, Messenger - genetics
• ISSN: 0740-7750; 1572-9931
• DOI: 10.1007/BF01535312

Human chromosome 6 encodes both the interferon gamma receptor as well as the class I major histocompatibility complex antigens, HLA-A, -B, and -C. However, the presence of chromosome 6 in somatic cell hybrids is insufficient to confer sensitivity to human interferon gamma (Hu-IFN-gamma) as assayed by class I HLA induction; it is necessary for both human chromosomes 6 and 21 to reside in the hybrid to generate a response to Hu-IFN-gamma. Treatment of such a hamster-human hybrid, Q72-18, with Hu-IFN-gamma induces the class I HLA antigens. Q72-18 cells selected by fluorescence-activated cell sorting for the loss of class I HLA induction also lost human chromosome 21. Fusions of such cells to a hybrid that contains only human chromosome 21 reconstitutes HLA antigen induction by Hu-IFN-gamma. Furthermore, fusions of hybrids containing a translocated human chromosome 6q and the HLA-B7 gene to a line containing only human chromosome 21 or a translocated 21q also reconstitutes HLA-B7 mRNA and antigen induction by Hu-IFN-gamma. Thus the segregation of cells on the basis of a biological effect by fluorescence-activated cell sorting and reconstitution by hybrid fusion provides a strategy by which some biological pathways can be mapped at a chromosomal level.