Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV

Haifa H. Jabara, Lynda C. Schneider, Stuart K. Shapira, Caroline Alfieri, Curtis T. Moody, Elliott Kieff, Raif S. Geha, Donata Vercelli

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Abstract

EBV and rIL-4 induce T cell-independent IgE production by normal human B cells. We demonstrate here that EBV and IL-4 induced the synthesis of IgE by surface IgE-negative B cell precursors isolated by cell sorting. This result suggests that the induction of IgE by EBV and IL-4 results not merely from the expansion of a precommitted surface IgE-positive B cell population but more likely from IL-4-directed switching to IgE. At the molecular level, IL-4 and EBV induced the appearance of 2.0- and of 1.8-kilobase (kb) RNA bands, both of which hybridized with an 0.88-kb HinfI fragment spanning part of the C∈1 exon and the entire C∈2 exon. The 1.8-kb band but not the 2.0-kb band also hybridized with a cloned genomic 0.7-kb SmaI fragment located ≈2 kb upstream of C∈. Thus, EBV and IL-4 induced germ-line (1.8-kb) as well as mature (2.0-kb) C∈ transcripts. IL-4 by itself induced germ-line C∈ transcripts but not mature C∈ transcripts in purified normal B cells. IL-4 failed to induce IgE synthesis in established EBV B cell lines and failed to induce 2.0-kb mature C∈ transcripts but induced 1.8-kb germ-line C∈ transcripts. These data show that IL-4 is sufficient for the induction of C∈ germ-line transcription. In contrast, the transcription of mature ∈ mRNA requires an additional activating signal, provided by infection with EBV. Established EBV transformation results in a dissociation between germ-line C∈ transcription and the ability to undergo IgE switching in response to IL-4.

Original languageEnglish (US)
Pages (from-to)3468-3473
Number of pages6
JournalJournal of Immunology
Volume145
Issue number10
StatePublished - Nov 15 1990
Externally publishedYes

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Human Herpesvirus 4
Germ Cells
Interleukin-4
Immunoglobulin E
B-Lymphocytes
Exons
Epstein-Barr Virus Infections
B-Lymphoid Precursor Cells
RNA
T-Lymphocytes
Cell Line
Messenger RNA
Population

ASJC Scopus subject areas

  • Immunology

Cite this

Jabara, H. H., Schneider, L. C., Shapira, S. K., Alfieri, C., Moody, C. T., Kieff, E., ... Vercelli, D. (1990). Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV. Journal of Immunology, 145(10), 3468-3473.

Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV. / Jabara, Haifa H.; Schneider, Lynda C.; Shapira, Stuart K.; Alfieri, Caroline; Moody, Curtis T.; Kieff, Elliott; Geha, Raif S.; Vercelli, Donata.

In: Journal of Immunology, Vol. 145, No. 10, 15.11.1990, p. 3468-3473.

Research output: Contribution to journalArticle

Jabara, HH, Schneider, LC, Shapira, SK, Alfieri, C, Moody, CT, Kieff, E, Geha, RS & Vercelli, D 1990, 'Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV', Journal of Immunology, vol. 145, no. 10, pp. 3468-3473.
Jabara HH, Schneider LC, Shapira SK, Alfieri C, Moody CT, Kieff E et al. Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV. Journal of Immunology. 1990 Nov 15;145(10):3468-3473.
Jabara, Haifa H. ; Schneider, Lynda C. ; Shapira, Stuart K. ; Alfieri, Caroline ; Moody, Curtis T. ; Kieff, Elliott ; Geha, Raif S. ; Vercelli, Donata. / Induction of germ-line and mature C∈ transcripts in human B cells stimulated with rIL-4 and EBV. In: Journal of Immunology. 1990 ; Vol. 145, No. 10. pp. 3468-3473.
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abstract = "EBV and rIL-4 induce T cell-independent IgE production by normal human B cells. We demonstrate here that EBV and IL-4 induced the synthesis of IgE by surface IgE-negative B cell precursors isolated by cell sorting. This result suggests that the induction of IgE by EBV and IL-4 results not merely from the expansion of a precommitted surface IgE-positive B cell population but more likely from IL-4-directed switching to IgE. At the molecular level, IL-4 and EBV induced the appearance of 2.0- and of 1.8-kilobase (kb) RNA bands, both of which hybridized with an 0.88-kb HinfI fragment spanning part of the C∈1 exon and the entire C∈2 exon. The 1.8-kb band but not the 2.0-kb band also hybridized with a cloned genomic 0.7-kb SmaI fragment located ≈2 kb upstream of C∈. Thus, EBV and IL-4 induced germ-line (1.8-kb) as well as mature (2.0-kb) C∈ transcripts. IL-4 by itself induced germ-line C∈ transcripts but not mature C∈ transcripts in purified normal B cells. IL-4 failed to induce IgE synthesis in established EBV B cell lines and failed to induce 2.0-kb mature C∈ transcripts but induced 1.8-kb germ-line C∈ transcripts. These data show that IL-4 is sufficient for the induction of C∈ germ-line transcription. In contrast, the transcription of mature ∈ mRNA requires an additional activating signal, provided by infection with EBV. Established EBV transformation results in a dissociation between germ-line C∈ transcription and the ability to undergo IgE switching in response to IL-4.",
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AU - Alfieri, Caroline

AU - Moody, Curtis T.

AU - Kieff, Elliott

AU - Geha, Raif S.

AU - Vercelli, Donata

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N2 - EBV and rIL-4 induce T cell-independent IgE production by normal human B cells. We demonstrate here that EBV and IL-4 induced the synthesis of IgE by surface IgE-negative B cell precursors isolated by cell sorting. This result suggests that the induction of IgE by EBV and IL-4 results not merely from the expansion of a precommitted surface IgE-positive B cell population but more likely from IL-4-directed switching to IgE. At the molecular level, IL-4 and EBV induced the appearance of 2.0- and of 1.8-kilobase (kb) RNA bands, both of which hybridized with an 0.88-kb HinfI fragment spanning part of the C∈1 exon and the entire C∈2 exon. The 1.8-kb band but not the 2.0-kb band also hybridized with a cloned genomic 0.7-kb SmaI fragment located ≈2 kb upstream of C∈. Thus, EBV and IL-4 induced germ-line (1.8-kb) as well as mature (2.0-kb) C∈ transcripts. IL-4 by itself induced germ-line C∈ transcripts but not mature C∈ transcripts in purified normal B cells. IL-4 failed to induce IgE synthesis in established EBV B cell lines and failed to induce 2.0-kb mature C∈ transcripts but induced 1.8-kb germ-line C∈ transcripts. These data show that IL-4 is sufficient for the induction of C∈ germ-line transcription. In contrast, the transcription of mature ∈ mRNA requires an additional activating signal, provided by infection with EBV. Established EBV transformation results in a dissociation between germ-line C∈ transcription and the ability to undergo IgE switching in response to IL-4.

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