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Upregulation of interleukin 8 by oxygen-deprived cells in glioblastoma suggests a role in leukocyte activation, chemotaxis, and angiogenesis.

Desbaillets I, Diserens AC, Tribolet N, Hamou MF, Van Meir EG - J. Exp. Med. (1997)

Bottom Line: In glioblastoma, it further localizes to oxygen-deprived cells surrounding necrosis.Hypoxic/anoxic insults on glioblastoma cells in vitro using anaerobic chamber systems or within spheroids developing central necrosis induced an increase in IL-8 messenger RNA (mRNA) and protein expression. mRNA for IL-8-binding chemokine receptors CXCR1, CXCR2, and the Duffy antigen receptor for chemokines (DARC) were found in all astrocytoma grades by reverse transcription/PCR analysis.These results support a model where IL-8 expression is initiated early in astrocytoma development through induction by inflammatory stimuli and later in tumor progression increases due to reduced microenvironmental oxygen pressure.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Tumor Biology and Genetics, Neurosurgery Department, University Hospital (CHUV), Lausanne, Switzerland.

ABSTRACT
Leukocyte infiltration and necrosis are two biological phenomena associated with the development of neovascularization during the malignant progression of human astrocytoma. Here, we demonstrate expression of interleukin (IL)-8, a cytokine with chemotactic and angiogenic properties, and of IL-8-binding receptors in astrocytoma. IL-8 expression is first observed in low grade astrocytoma in perivascular tumor areas expressing inflammatory cytokines. In glioblastoma, it further localizes to oxygen-deprived cells surrounding necrosis. Hypoxic/anoxic insults on glioblastoma cells in vitro using anaerobic chamber systems or within spheroids developing central necrosis induced an increase in IL-8 messenger RNA (mRNA) and protein expression. mRNA for IL-8-binding chemokine receptors CXCR1, CXCR2, and the Duffy antigen receptor for chemokines (DARC) were found in all astrocytoma grades by reverse transcription/PCR analysis. In situ hybridization and immunohistochemistry localized DARC expression on normal brain and tumor microvascular cells and CXCR1 and CXCR2 expression to infiltrating leukocytes. These results support a model where IL-8 expression is initiated early in astrocytoma development through induction by inflammatory stimuli and later in tumor progression increases due to reduced microenvironmental oxygen pressure. Augmented IL-8 would directly and/or indirectly promote angiogenesis by binding to DARC and by inducing leukocyte infiltration and activation by binding to CXCR1 and CXCR2.

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Detection of IL-8–binding  receptors in glioblastoma by immunohistochemistry. DARC (A) and factor VIII  (B) expression on microvascular cells of  glioblastoma No. 1069. DARC (C) and  preimmune serum (D) staining on nontumoral brain No. T265CN. CXCR1 (E)  and CD3 (F) staining on glioblastoma No.  906. CXCR2 staining on sections with  vessels (G) or in a perinecrotic region (H)  of glioblastoma No. 842. Examples of positive cells are shown by arrows.
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Figure 4: Detection of IL-8–binding receptors in glioblastoma by immunohistochemistry. DARC (A) and factor VIII (B) expression on microvascular cells of glioblastoma No. 1069. DARC (C) and preimmune serum (D) staining on nontumoral brain No. T265CN. CXCR1 (E) and CD3 (F) staining on glioblastoma No. 906. CXCR2 staining on sections with vessels (G) or in a perinecrotic region (H) of glioblastoma No. 842. Examples of positive cells are shown by arrows.

Mentions: The biological relevance of IL-8 expression in glioblastoma depends on the presence of cells with receptors capable of binding IL-8. Therefore, we evaluated whether IL-8 might have autocrine and/or paracrine functions in vivo. First, we examined CXCR1, CXCR2, and DARC mRNA expression by RT/PCR. CXCR1 mRNA was found in 5 of 5 grade II, 1 of 7 grade III, and 8 of 12 grade IV astrocytoma. CXCR2 mRNA was found in 5 of 5 grade II, 6 of 7 grade III, and 11 of 12 grade IV. DARC mRNA was found in 3 of 5 grade II, 5 of 7 grade III, and 10 of 12 grade IV (results not shown). This suggested that IL-8 may serve a biological function during the progression of human astrocytoma in vivo, either on tumor cells or on accessory cells. Next, we examined which cell types expressed IL-8–binding receptors in vivo by immunohistochemistry (representative stainings are shown in Fig. 4). Very interestingly, DARC was specifically expressed by microvascular cells in 5 of 6 grade II, 5 of 6 grade III, and 14 of 19 grade IV astrocytomas (Fig. 4 A) with a staining pattern similar to the one obtained for factor VIII, a microvascular marker (Fig. 4 B). DARC expression was also present on 5 of 5 nontumoral brains (Fig. 4 C). Control stainings with preimmune serum was negative on all samples (Fig. 4 D). In contrast, for both CXCR1 and CXCR2, isolated positive cells were found surrounding blood vessels. For CXCR1, 2 of 9 grade II, 1 of 3 grade III, and 5 of 18 grade IV were positive (Fig. 4 E), and for CXCR2, 1 of 9 grade II, 0 of 3 grade III, and 5 of 18 grade IV (Fig. 4 G). Staining of adjacent sections for CD3, a specific marker of T lymphocytes (Fig. 4 F), and CD15, a macrophage marker (not shown), showed infiltrates in these areas, suggesting expression by a subset of T lymphocytes and/or macrophages. Unfrequently, numerous CXCR2 positive cells were found close to necrosis (Fig. 4 H).


Upregulation of interleukin 8 by oxygen-deprived cells in glioblastoma suggests a role in leukocyte activation, chemotaxis, and angiogenesis.

Desbaillets I, Diserens AC, Tribolet N, Hamou MF, Van Meir EG - J. Exp. Med. (1997)

Detection of IL-8–binding  receptors in glioblastoma by immunohistochemistry. DARC (A) and factor VIII  (B) expression on microvascular cells of  glioblastoma No. 1069. DARC (C) and  preimmune serum (D) staining on nontumoral brain No. T265CN. CXCR1 (E)  and CD3 (F) staining on glioblastoma No.  906. CXCR2 staining on sections with  vessels (G) or in a perinecrotic region (H)  of glioblastoma No. 842. Examples of positive cells are shown by arrows.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199083&req=5

Figure 4: Detection of IL-8–binding receptors in glioblastoma by immunohistochemistry. DARC (A) and factor VIII (B) expression on microvascular cells of glioblastoma No. 1069. DARC (C) and preimmune serum (D) staining on nontumoral brain No. T265CN. CXCR1 (E) and CD3 (F) staining on glioblastoma No. 906. CXCR2 staining on sections with vessels (G) or in a perinecrotic region (H) of glioblastoma No. 842. Examples of positive cells are shown by arrows.
Mentions: The biological relevance of IL-8 expression in glioblastoma depends on the presence of cells with receptors capable of binding IL-8. Therefore, we evaluated whether IL-8 might have autocrine and/or paracrine functions in vivo. First, we examined CXCR1, CXCR2, and DARC mRNA expression by RT/PCR. CXCR1 mRNA was found in 5 of 5 grade II, 1 of 7 grade III, and 8 of 12 grade IV astrocytoma. CXCR2 mRNA was found in 5 of 5 grade II, 6 of 7 grade III, and 11 of 12 grade IV. DARC mRNA was found in 3 of 5 grade II, 5 of 7 grade III, and 10 of 12 grade IV (results not shown). This suggested that IL-8 may serve a biological function during the progression of human astrocytoma in vivo, either on tumor cells or on accessory cells. Next, we examined which cell types expressed IL-8–binding receptors in vivo by immunohistochemistry (representative stainings are shown in Fig. 4). Very interestingly, DARC was specifically expressed by microvascular cells in 5 of 6 grade II, 5 of 6 grade III, and 14 of 19 grade IV astrocytomas (Fig. 4 A) with a staining pattern similar to the one obtained for factor VIII, a microvascular marker (Fig. 4 B). DARC expression was also present on 5 of 5 nontumoral brains (Fig. 4 C). Control stainings with preimmune serum was negative on all samples (Fig. 4 D). In contrast, for both CXCR1 and CXCR2, isolated positive cells were found surrounding blood vessels. For CXCR1, 2 of 9 grade II, 1 of 3 grade III, and 5 of 18 grade IV were positive (Fig. 4 E), and for CXCR2, 1 of 9 grade II, 0 of 3 grade III, and 5 of 18 grade IV (Fig. 4 G). Staining of adjacent sections for CD3, a specific marker of T lymphocytes (Fig. 4 F), and CD15, a macrophage marker (not shown), showed infiltrates in these areas, suggesting expression by a subset of T lymphocytes and/or macrophages. Unfrequently, numerous CXCR2 positive cells were found close to necrosis (Fig. 4 H).

Bottom Line: In glioblastoma, it further localizes to oxygen-deprived cells surrounding necrosis.Hypoxic/anoxic insults on glioblastoma cells in vitro using anaerobic chamber systems or within spheroids developing central necrosis induced an increase in IL-8 messenger RNA (mRNA) and protein expression. mRNA for IL-8-binding chemokine receptors CXCR1, CXCR2, and the Duffy antigen receptor for chemokines (DARC) were found in all astrocytoma grades by reverse transcription/PCR analysis.These results support a model where IL-8 expression is initiated early in astrocytoma development through induction by inflammatory stimuli and later in tumor progression increases due to reduced microenvironmental oxygen pressure.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Tumor Biology and Genetics, Neurosurgery Department, University Hospital (CHUV), Lausanne, Switzerland.

ABSTRACT
Leukocyte infiltration and necrosis are two biological phenomena associated with the development of neovascularization during the malignant progression of human astrocytoma. Here, we demonstrate expression of interleukin (IL)-8, a cytokine with chemotactic and angiogenic properties, and of IL-8-binding receptors in astrocytoma. IL-8 expression is first observed in low grade astrocytoma in perivascular tumor areas expressing inflammatory cytokines. In glioblastoma, it further localizes to oxygen-deprived cells surrounding necrosis. Hypoxic/anoxic insults on glioblastoma cells in vitro using anaerobic chamber systems or within spheroids developing central necrosis induced an increase in IL-8 messenger RNA (mRNA) and protein expression. mRNA for IL-8-binding chemokine receptors CXCR1, CXCR2, and the Duffy antigen receptor for chemokines (DARC) were found in all astrocytoma grades by reverse transcription/PCR analysis. In situ hybridization and immunohistochemistry localized DARC expression on normal brain and tumor microvascular cells and CXCR1 and CXCR2 expression to infiltrating leukocytes. These results support a model where IL-8 expression is initiated early in astrocytoma development through induction by inflammatory stimuli and later in tumor progression increases due to reduced microenvironmental oxygen pressure. Augmented IL-8 would directly and/or indirectly promote angiogenesis by binding to DARC and by inducing leukocyte infiltration and activation by binding to CXCR1 and CXCR2.

Show MeSH
Related in: MedlinePlus