Simultaneous expression of germline gamma 1 and epsilon immunoglobulin heavy chain transcripts in single murine splenic B-cells

Goodman, DJ; Gerondakis, S

Author(s): Goodman, DJ; Gerondakis, S;
Details: Publication Year 1997-08,Volume 34,Issue #12-13,Page 919-927
Journal Title: MOLECULAR IMMUNOLOGY
Publication Type: Journal Article
Abstract: While successive isotype switching can occur from IgM to IgE via IgG1, little is known about the pattern of germline transcript expression in normal B cells committed to switching to multiple isotypes. In this study we define the relationship between germline transcript expression and immunoglobulin isotype expression and secretion in murine splenic B cells. Following 7 days stimulation with LPS and IL-4, 10% of single cells secrete IgE and of these only 1 in 10 secrete IgE alone. In contrast, when cells were stimulated with LPS and IL-4 for 48 hr prior to sorting into clonal cultures, 71% secreted IgE alone. In an attempt to isolate switch intermediates, IgG1(+)IgM(-) cells were sorted and upon re-culture secreted predominantly IgG1 alone or IgG1 and IgE. The frequency of cells expressing germline epsilon transcripts was 34% for surface IgG1(+)IgM(-) expressing cells and 14% for surface IgG1(-)IgM(+) negative cells. Furthermore, analysis of single surface IgG1(+)IgM(-) cells demonstrated that these cells can co-express germline gamma 1 and epsilon transcripts. Sorting of IgG1(-)IgM(+) cells according to surface expression of the integral membrane proteoglycan, Syndecan, demonstrated that IgM(+)Syndecan(+) cells had a lower frequency of germline transcript expression and a lower frequency of isotype switching (32%) compared to IgM(+)Syndecan(-) cells (83%). Collectively, these finding show that murine B cells can switch successively from IgM to IgE via IgG1 and that IgG1 expressing intermediates express germline transcripts. Furthermore, Syndecan expression appears to be linked to germline C-H transcription and isotype switch commitment. (C) 1998 Elsevier Science Ltd. All rights reserved.
Publisher: PERGAMON-ELSEVIER SCIENCE LTD
Keywords: CLASS SWITCH RECOMBINATION; MOLECULAR CHARACTERIZATION; MONOCLONAL-ANTIBODY; IMMUNE-RESPONSE; DEFICIENT MICE; CIRCULAR DNA; INTERLEUKIN-4; REGION; ALPHA; REARRANGEMENT
Publisher's Version: https://doi.org/10.1016/S0161-5890(97)00106-5
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 1997-08-01 12:00:00