Identification of upstream signals regulating interleukin-6 gene expression during in vitro treatment of human B cells with pokeweed mitogen

Dharminder Chauhan, Surender Kharbanda, Hiroshi Uchiyama, Mitsuyoshi Urashima, Ruben C Fragoso, Jyoti Sen, Donald W. Kufe, Kenneth C. Anderson

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Abstract

The present studies have characterized the regulation of interleukin-6 (IL-6) gene expression during pokeweed mitogen (PWM)-driven human B-cell differentiation. PWM induced an early and transient increase in the expression of immediatel-early response genes of the jun/fos leucine zipper family (c-jun, jun B, c-fos, and fos B). The induction of c-jun mRNA by PWM was concentration dependent. Nuclear run-on assays showed that PWM treatment is associated with an increased rate of c-jun gene transcription. The induction of c-jun mRNA precedes the induction of IL-6 gene expression and IL-6 secretion by the B cells. c-Jun antisense, but not sense, oligodeoxynucleotide (ODN) significantly decreases PWM-related B-cell (1) proliferation; (2) IL-6 mRNA induction; (3) IL-6 secretion; and (4) nuclear extract binding to AP-1 in electrophoretic mobility shift assay. In contrast, c-Fos antisense ODN did not effect either IL-6 mRNA induction or IL-6 secretion triggered in B cells by PWM. The results further show activation of c-Raf-1 kinase in PWM-treated B cells. Raf-1 acts upstream to mitogen- activated protein (MAP) kinase; therefore, studies were performed to assay for MAP kinase activation in these cells. The results show an increase in phosphorylation of myelin basic protein (MBP) and c-Jun 'Y' peptide in PWM- treated B cells. Taken together, these findings suggest that PWM is able to initiate an intracytoplasmic signaling cascade in normal human splenic B cells, which, at least in part, involves serine/threonine protein kinases. These results show transient induction of immediate-early response genes in B cells and support a potential role for the c-jun gene product in regulation of IL-6 transcription and secretion.

Original languageEnglish (US)
Pages (from-to)2243-2252
Number of pages10
JournalBlood
Volume84
Issue number7
StatePublished - Oct 1 1994
Externally publishedYes

Fingerprint

Pokeweed Mitogens
Gene expression
Interleukin-6
B-Lymphocytes
Cells
Gene Expression
jun Genes
Proto-Oncogene Proteins c-raf
Assays
Messenger RNA
Oligodeoxyribonucleotides
Transcription
Mitogen-Activated Protein Kinases
Chemical activation
fos Genes
In Vitro Techniques
Leucine Zippers
Electrophoretic mobility
Phosphorylation
Immediate-Early Genes

ASJC Scopus subject areas

  • Hematology

Cite this

Chauhan, D., Kharbanda, S., Uchiyama, H., Urashima, M., Fragoso, R. C., Sen, J., ... Anderson, K. C. (1994). Identification of upstream signals regulating interleukin-6 gene expression during in vitro treatment of human B cells with pokeweed mitogen. Blood, 84(7), 2243-2252.

Identification of upstream signals regulating interleukin-6 gene expression during in vitro treatment of human B cells with pokeweed mitogen. / Chauhan, Dharminder; Kharbanda, Surender; Uchiyama, Hiroshi; Urashima, Mitsuyoshi; Fragoso, Ruben C; Sen, Jyoti; Kufe, Donald W.; Anderson, Kenneth C.

In: Blood, Vol. 84, No. 7, 01.10.1994, p. 2243-2252.

Research output: Contribution to journalArticle

Chauhan, D, Kharbanda, S, Uchiyama, H, Urashima, M, Fragoso, RC, Sen, J, Kufe, DW & Anderson, KC 1994, 'Identification of upstream signals regulating interleukin-6 gene expression during in vitro treatment of human B cells with pokeweed mitogen', Blood, vol. 84, no. 7, pp. 2243-2252.
Chauhan, Dharminder ; Kharbanda, Surender ; Uchiyama, Hiroshi ; Urashima, Mitsuyoshi ; Fragoso, Ruben C ; Sen, Jyoti ; Kufe, Donald W. ; Anderson, Kenneth C. / Identification of upstream signals regulating interleukin-6 gene expression during in vitro treatment of human B cells with pokeweed mitogen. In: Blood. 1994 ; Vol. 84, No. 7. pp. 2243-2252.
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abstract = "The present studies have characterized the regulation of interleukin-6 (IL-6) gene expression during pokeweed mitogen (PWM)-driven human B-cell differentiation. PWM induced an early and transient increase in the expression of immediatel-early response genes of the jun/fos leucine zipper family (c-jun, jun B, c-fos, and fos B). The induction of c-jun mRNA by PWM was concentration dependent. Nuclear run-on assays showed that PWM treatment is associated with an increased rate of c-jun gene transcription. The induction of c-jun mRNA precedes the induction of IL-6 gene expression and IL-6 secretion by the B cells. c-Jun antisense, but not sense, oligodeoxynucleotide (ODN) significantly decreases PWM-related B-cell (1) proliferation; (2) IL-6 mRNA induction; (3) IL-6 secretion; and (4) nuclear extract binding to AP-1 in electrophoretic mobility shift assay. In contrast, c-Fos antisense ODN did not effect either IL-6 mRNA induction or IL-6 secretion triggered in B cells by PWM. The results further show activation of c-Raf-1 kinase in PWM-treated B cells. Raf-1 acts upstream to mitogen- activated protein (MAP) kinase; therefore, studies were performed to assay for MAP kinase activation in these cells. The results show an increase in phosphorylation of myelin basic protein (MBP) and c-Jun 'Y' peptide in PWM- treated B cells. Taken together, these findings suggest that PWM is able to initiate an intracytoplasmic signaling cascade in normal human splenic B cells, which, at least in part, involves serine/threonine protein kinases. These results show transient induction of immediate-early response genes in B cells and support a potential role for the c-jun gene product in regulation of IL-6 transcription and secretion.",
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