Ig heavy chain expression and class switching in vitro from an allele lacking the 3' enhancers DNase I-hypersensitive hs3A and hs1,2

Shireen Saleque, Mallika Singh, Barbara K. Birshtein

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The murine Ig heavy chain (IgH) 3' regulatory region contains four enhancers: hs3A, hs1,2, hs3B, and hs4. Various studies have suggested a role for these enhancers in regulating IgH expression and class switching. Here we assess the role of hs3A and hs1,2 in these processes by exploiting a naturally occurring deletion of these enhancers from the expressed, C57BL/6 allele of the F1 pre-B cell line, 70Z/3. Equivalent μ expression in 70Z/3 and 18-81 (which has an intact 3' region) indicated that hs3A and hs1,2 were not essential for μ expression at the pre-B cell stage. To further examine the role of hs3A and hs1,2 in IgH function at the plasma cell stage, we fused 70Z/3 with the plasmacytoma NSO. Electromobility shift assay analysis of the 70Z/3-NSO hybrids revealed a transcription factor complement conducive to the activation of the 3' enhancers. Despite the lack of enhancers, hs3A and hs1,2, the level of μ RNA and protein in the 70Z/3-NSO fusion hybrids was substantially elevated relative to its pre-B parent and comparable with that observed in a number of μ-producing spleen cell hybridomas. Additionally, ELISAspot assays showed that the 70Z/3-NSO hybrid underwent spontaneous class switching in culture to IgG1 at a frequency comparable with that of most hybridomas. These results indicate that hs3A and hs1,2 are not essential for high levels of IgH expression or for spontaneous class switching in a plasma cell line.

Original languageEnglish (US)
Pages (from-to)2791-2803
Number of pages13
JournalJournal of Immunology
Volume162
Issue number5
StatePublished - Mar 1 1999

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Immunoglobulin Class Switching
Immunoglobulin Heavy Chains
Deoxyribonuclease I
Alleles
B-Lymphoid Precursor Cells
Hybridomas
Plasma Cells
Cell Line
Plasmacytoma
Nucleic Acid Regulatory Sequences
Transcription Factors
Spleen
Immunoglobulin G
RNA
In Vitro Techniques
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Ig heavy chain expression and class switching in vitro from an allele lacking the 3' enhancers DNase I-hypersensitive hs3A and hs1,2. / Saleque, Shireen; Singh, Mallika; Birshtein, Barbara K.

In: Journal of Immunology, Vol. 162, No. 5, 01.03.1999, p. 2791-2803.

Research output: Contribution to journalArticle

Saleque, Shireen ; Singh, Mallika ; Birshtein, Barbara K. / Ig heavy chain expression and class switching in vitro from an allele lacking the 3' enhancers DNase I-hypersensitive hs3A and hs1,2. In: Journal of Immunology. 1999 ; Vol. 162, No. 5. pp. 2791-2803.
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