Clonal and molecular characteristics of the human IgE-committed B cell subset.
We have followed the pathway of the IgE-committed B lymphocyte from fresh, unstimulated peripheral blood, through EBV activation, transformation, and eventual cloning. Using cell sorting in conjunction with limiting dilution culture systems, we found that: (a) cells that are selected in the cell sorter and secrete IgE in culture are sIgM+/sIgD+. They secrete all three isotypes after EBV activation and continue to do so stably in culture; (b) individual IgE+ cells in culture coproduce IgM, IgD, and IgE and cytoplasmic Ig of each isotype can be detected in single cells; (c) no rearrangement was observed of VDJ to epsilon in any of six lines tested. DNA between the rearranged VDJ-mu and -epsilon appears to be overall intact, including a region 10.5 kb upstream and 18 kb downstream of the 2-kb epsilon coding region and; (d) mRNA of mu and epsilon species is of normal and comparable size. In contrast to IgG- and IgA-producing clones, multiple isotype expression appears to be both frequent and stable in cells committed to IgE production. We propose that IgE-committed cells represent a unique B cell sublineage whose differentiation pathway may be more strictly regulated than that of other isotypes with regard to the signals required for classical, deletional switch recombination that has been observed in rare IgE-producing myeloma cell lines.
MacKenzie
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Division of Immunology and Rheumatology, Hospital for Sick Children, Toronto, Ontario, Canada.
Dosch
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HM
D001402:B-Lymphocytes / Q000145:classification / Q000166:cytology*; D015152:Blotting, Northern; D002454:Cell Differentiation; D002469:Cell Separation; D002472:Cell Transformation, Viral; D002999:Clone Cells; D005455:Fluorescent Antibody Technique; D015322:Gene Rearrangement, B-Lymphocyte; D004854:Herpesvirus 4, Human; D006801:Humans; D007072:Immunoglobulin D / Q000096:biosynthesis; D007073:Immunoglobulin E / Q000096:biosynthesis*; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D015183:Restriction Mapping
D007072:Immunoglobulin D; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D012333:RNA, Messenger; D011947:Receptors, Antigen, B-Cell; D007073:Immunoglobulin E