Expression of transcripts for two interleukin 8 receptors in human phagocytes, lymphocytes and melanoma cells

B. Moser, L. Barella, S. Mattei, C. Schumacher, F. Boulay, M. P. Colombo, M. Baggiolini

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Abstract

Two cDNAs coding for distinct interleukin 8 (IL-8) receptors, IL-8R1 and IL-8R2 have been reported, and biochemical studies on human neutrophils have revealed two proteins (p70 and p44) that bind IL-8 with high affinity. We have cloned the cDNA coding for IL-8R1 from a library of differentiated HL-60 cells. Transfection of this cDNA into Jurkat cells resulted in the expression of high-affinity binding for IL-8 and two related cytokines, GROα and neutrophil-activating peptide 2 (K(d) 0.5-1.0 nM). Northern-blot analysis with the IL-8R1 cDNA as probe revealed abundant expression of transcripts of different size in human neutrophils and low-level expression of a single RNA species in HL-60 cells differentiated with dimethyl sulphoxide and retinoic acid. Because of the extensive nucleotide sequence similarity of the cDNAs for IL-8R1 and IL-8R2, the reverse-transcription PCR method was used for analysis of RNA expression in myeloid and lymphoid cells, 19 cell lines established from human primary melanomas or metastases, two melanocyte and one fibroblast cell lines. IL-8R1 mRNA transcripts were expressed at high levels in neutrophils, and to a lesser extent in blood monocytes and the myeloid cell lines, HL-60 and AML 193, but were not found in THP-1 cells, lymphocytes and Jurkat cells. IL-8R2 mRNA transcripts, by contrast, were found in all blood cells and related cell lines, as well as in all melanoma, melanocyte and fibroblast cell lines tested. As for IL-8R1, IL-8R2 mRNA expression was highest in neutrophils. These results suggest that IL-8R1 and IL-8R2 may both be involved in neutrophil activation by IL-8 and related cytokines, and presumably correspond to p70 and p44, the receptors that were identified biochemically. Possible IL-8 functions on lymphocytes and melanoma cells, e.g. chemotaxis and proliferation, must be independent of IL-8R1 and may be mediated by IL-8R2.

Original languageEnglish
Pages (from-to)285-292
Number of pages8
JournalBiochemical Journal
Volume294
Issue number1
Publication statusPublished - 1993

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Interleukin-8 Receptors
Lymphocytes
Phagocytes
Melanoma
Interleukin-8
Complementary DNA
Cells
Neutrophils
Cell Line
Jurkat Cells
HL-60 Cells
Melanocytes
Myeloid Cells
Messenger RNA
Fibroblasts
RNA
Cytokines
Blood
Neutrophil Activation
Chemotaxis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Moser, B., Barella, L., Mattei, S., Schumacher, C., Boulay, F., Colombo, M. P., & Baggiolini, M. (1993). Expression of transcripts for two interleukin 8 receptors in human phagocytes, lymphocytes and melanoma cells. Biochemical Journal, 294(1), 285-292.

Expression of transcripts for two interleukin 8 receptors in human phagocytes, lymphocytes and melanoma cells. / Moser, B.; Barella, L.; Mattei, S.; Schumacher, C.; Boulay, F.; Colombo, M. P.; Baggiolini, M.

In: Biochemical Journal, Vol. 294, No. 1, 1993, p. 285-292.

Research output: Contribution to journalArticle

Moser, B, Barella, L, Mattei, S, Schumacher, C, Boulay, F, Colombo, MP & Baggiolini, M 1993, 'Expression of transcripts for two interleukin 8 receptors in human phagocytes, lymphocytes and melanoma cells', Biochemical Journal, vol. 294, no. 1, pp. 285-292.
Moser, B. ; Barella, L. ; Mattei, S. ; Schumacher, C. ; Boulay, F. ; Colombo, M. P. ; Baggiolini, M. / Expression of transcripts for two interleukin 8 receptors in human phagocytes, lymphocytes and melanoma cells. In: Biochemical Journal. 1993 ; Vol. 294, No. 1. pp. 285-292.
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abstract = "Two cDNAs coding for distinct interleukin 8 (IL-8) receptors, IL-8R1 and IL-8R2 have been reported, and biochemical studies on human neutrophils have revealed two proteins (p70 and p44) that bind IL-8 with high affinity. We have cloned the cDNA coding for IL-8R1 from a library of differentiated HL-60 cells. Transfection of this cDNA into Jurkat cells resulted in the expression of high-affinity binding for IL-8 and two related cytokines, GROα and neutrophil-activating peptide 2 (K(d) 0.5-1.0 nM). Northern-blot analysis with the IL-8R1 cDNA as probe revealed abundant expression of transcripts of different size in human neutrophils and low-level expression of a single RNA species in HL-60 cells differentiated with dimethyl sulphoxide and retinoic acid. Because of the extensive nucleotide sequence similarity of the cDNAs for IL-8R1 and IL-8R2, the reverse-transcription PCR method was used for analysis of RNA expression in myeloid and lymphoid cells, 19 cell lines established from human primary melanomas or metastases, two melanocyte and one fibroblast cell lines. IL-8R1 mRNA transcripts were expressed at high levels in neutrophils, and to a lesser extent in blood monocytes and the myeloid cell lines, HL-60 and AML 193, but were not found in THP-1 cells, lymphocytes and Jurkat cells. IL-8R2 mRNA transcripts, by contrast, were found in all blood cells and related cell lines, as well as in all melanoma, melanocyte and fibroblast cell lines tested. As for IL-8R1, IL-8R2 mRNA expression was highest in neutrophils. These results suggest that IL-8R1 and IL-8R2 may both be involved in neutrophil activation by IL-8 and related cytokines, and presumably correspond to p70 and p44, the receptors that were identified biochemically. Possible IL-8 functions on lymphocytes and melanoma cells, e.g. chemotaxis and proliferation, must be independent of IL-8R1 and may be mediated by IL-8R2.",
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AU - Moser, B.

AU - Barella, L.

AU - Mattei, S.

AU - Schumacher, C.

AU - Boulay, F.

AU - Colombo, M. P.

AU - Baggiolini, M.

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AB - Two cDNAs coding for distinct interleukin 8 (IL-8) receptors, IL-8R1 and IL-8R2 have been reported, and biochemical studies on human neutrophils have revealed two proteins (p70 and p44) that bind IL-8 with high affinity. We have cloned the cDNA coding for IL-8R1 from a library of differentiated HL-60 cells. Transfection of this cDNA into Jurkat cells resulted in the expression of high-affinity binding for IL-8 and two related cytokines, GROα and neutrophil-activating peptide 2 (K(d) 0.5-1.0 nM). Northern-blot analysis with the IL-8R1 cDNA as probe revealed abundant expression of transcripts of different size in human neutrophils and low-level expression of a single RNA species in HL-60 cells differentiated with dimethyl sulphoxide and retinoic acid. Because of the extensive nucleotide sequence similarity of the cDNAs for IL-8R1 and IL-8R2, the reverse-transcription PCR method was used for analysis of RNA expression in myeloid and lymphoid cells, 19 cell lines established from human primary melanomas or metastases, two melanocyte and one fibroblast cell lines. IL-8R1 mRNA transcripts were expressed at high levels in neutrophils, and to a lesser extent in blood monocytes and the myeloid cell lines, HL-60 and AML 193, but were not found in THP-1 cells, lymphocytes and Jurkat cells. IL-8R2 mRNA transcripts, by contrast, were found in all blood cells and related cell lines, as well as in all melanoma, melanocyte and fibroblast cell lines tested. As for IL-8R1, IL-8R2 mRNA expression was highest in neutrophils. These results suggest that IL-8R1 and IL-8R2 may both be involved in neutrophil activation by IL-8 and related cytokines, and presumably correspond to p70 and p44, the receptors that were identified biochemically. Possible IL-8 functions on lymphocytes and melanoma cells, e.g. chemotaxis and proliferation, must be independent of IL-8R1 and may be mediated by IL-8R2.

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