interleukin-8 and Leukemia--Myeloid

The currently used National Cancer Institute (NCI) adverse events criteria for mucosal barrier injury (MBI) are insufficient for use in children. We searched for objective, easily measurable indicators for MBI in children with cancer.. In children with acute myeloid leukemia, various MBI-related clinical and laboratory tests were investigated, reflecting clinical severity (NCI symptomatic adverse events criteria (gold standard), daily gut score (DGS)), inflammation (plasma and fecal interleukin-8 (IL-8), fecal calprotectin), enterocytic loss (plasma citrulline, ratio fecal human DNA/total DNA) and intestinal permeability (sugar absorption tests).. Intestinal MBI as detected by the NCI adverse events criteria was found in 55% of chemotherapy cycles, correlating well with the continuous DGS (n = 55, rho = 0.581; P < 0.001). Intestinal cell loss as measured by the ratio fecal human DNA/total DNA and plasma citrulline correlated well with both NCI criteria (n = 61, rho = 0.357, P = 0.005 resp. n = 58, rho = -0.482; P < 0.001) and DGS (n = 54, rho = 0.352, P = 0.009 resp. n = 55, rho = -0.625; P < 0.001). Plasma IL-8 correlated strongly to plasma citrulline (n = 46, rho = -0.627; P < 0.001).. MBI was reflected by parameters indicating inflammation (IL-8) and cell loss (plasma citrulline, ratio fecal human DNA/total DNA). We conclude that plasma citrulline might be a good parameter for MBI. Further studies are needed to show whether plasma citrulline can be used as a marker for MBI in future research.

Topics: Acute Disease; Adolescent; Amsacrine; Antineoplastic Combined Chemotherapy Protocols; Biomarkers; Carbohydrates; Cell Death; Child; Child, Preschool; Citrulline; Cytarabine; Daunorubicin; DNA; Enterocytes; Etoposide; Feces; Female; Humans; Infant; Interleukin-8; Intestinal Absorption; Leukemia, Myeloid; Leukocyte L1 Antigen Complex; Male; Mitoxantrone; Models, Biological; Mucositis; Stomatitis

The immunomodulatory activities of recombinant human interleukin-11 (rhIL-11) were investigated in a clinical trial among patients with hematological malignancy, randomized to either rhIL-11 or placebo throughout chemotherapy. Daily serum concentrations of sTNFRI, IL-6, IL-8, TNFalpha, and CRP were measured. Higher sTNFRI levels [mean pg/ml (95% CI)] were detected in patients receiving rhIL-11 compared to placebo [1749.7 (1626-1882.9) versus 1038.5 (953.3-1131.3)] respectively (P = 0.01) for all 898 observations and during febrile days [2327.6 (2142.6-2528.2) versus 1308.9 (1163-1473.2), P = 0.12] and during days without infection [1406.6 (1266.1-1563) versus 871.3 (774.9-979.6), P < 0.001]. A similar pattern in CRP concentrations was observed. Multivariate analysis indicated rhIL-11 was associated with elevated sTNFRI or CRP independent of infectious episodes and other factors. 7 patients (all receiving placebo) of 40 had elevated TNFalpha levels. IL-6 and IL-8 levels were not substantially affected by rhIL-11. Bacteremia, fungal infections, and fever of unknown origin (FUO) were reduced in rhIL-11-treated patients. Given the role of sTNFRI in dampening the deleterious effects of a hyperactive TNFalpha environment, rhIL-11-induced upregulation of sTNFRI shedding is a potentially important mechanism for modulating immune and inflammatory responses in humans.

Topics: Adolescent; Adult; Anti-Inflammatory Agents; Biomarkers; C-Reactive Protein; Female; Humans; Interleukin-11; Interleukin-6; Interleukin-8; Leukemia, Myeloid; Lymphoma, Non-Hodgkin; Male; Middle Aged; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Prospective Studies; Receptors, Tumor Necrosis Factor, Type I; Recombinant Proteins; Tumor Necrosis Factor-alpha

Drug-induced liver injury (DILI) is thought to be involved in the participation of drugs that either directly affect the cell viability or elicit an immune response. However, there is limited information about the immune responses induced by drugs, including those drugs that are metabolically activated. In this study, we constructed an in vitro assay system to assess the involvement of immune-related factors induced by metabolic activation of drugs. To investigate whether CYP3A4-mediated metabolism of 10 hepatotoxic drugs is associated with immune-related responses, human monocytic leukemia THP-1 cells were co-incubated with CYP3A4 Supersomes. Cluster of differentiation (CD) 86 and CD54 expression levels on THP-1 cells were upregulated by treatment with albendazole and amiodarone (AMD), respectively, in the presence of CYP3A4. Additionally, N-desethylamiodarone (DEA), a major metabolite of AMD, upregulated the CD54 expression of THP-1 cells with CYP3A4. The release of interleukin (IL)-8 and tumor necrosis factor (TNF) α from THP-1 cells was significantly increased by the treatment of AMD or DEA with CYP3A4. Similarly, IL-8 and TNFα were also upregulated by the treatment of AMD and DEA with human liver microsomes, but were inhibited by adding ketoconazole to the cell culture. In this study, we first report that albendazole, AMD and DEA activate immune reaction when metabolically activated.

Topics: Albendazole; Amiodarone; B7-2 Antigen; Biotransformation; Cell Line, Tumor; Chemical and Drug Induced Liver Injury; Cytochrome P-450 CYP3A; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Ketoconazole; Leukemia, Myeloid; Microsomes, Liver; Monocytes; RNA, Messenger; Tumor Necrosis Factor-alpha; Up-Regulation

T cell targeting immunotherapy is now considered a possible strategy in acute myelogenous leukaemia (AML), and IFNgamma release may then contribute to the antileukaemic effects. We investigated the effects of IFNgamma on native human AML cells. Normal T cells could be activated to release IFNgamma in the presence of AML cells. Furthermore, high levels of CD119 (IFNgamma receptor alpha chain) expression were observed for all 39 patients examined. Receptor expression was decreased after exposure to exogenous IFNgamma, and receptor ligation caused Stat1 phosphorylation but no phosphorylation of the alternative messengers Erk1/2. The effect of exogenous IFNgamma on AML blast proliferation was dependent on the local cytokine network and IFNgamma (1) inhibited proliferation in the presence of exogenous IL1beta, GM-CSF, G-CSF and SCF; (2) had divergent effects in the presence of IL3 and Flt3 (65 patients examined); (3) inhibited proliferation in the presence of endothelial cells but had divergent effects in the presence of fibroblasts, osteoblasts and normal stromal cells (65 patients examined). IFNgamma increased stress-induced (spontaneous) in vitro apoptosis as well as cytarabine-induced apoptosis only for a subset of patients. Furthermore, IFNgamma decreased the release of proangiogenic CXCL8 and increased the release of antiangiogenic CXCL9-11. We conclude that IFNgamma can be released in the presence of native human AML cells and affect AML cell proliferation, regulation of apoptosis and the balance between pro- and antiangiogenic chemokine release.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Cell Proliferation; Chemokine CXCL9; Chemokines, CXC; Cytarabine; Endothelium, Vascular; Female; Fibroblasts; Flow Cytometry; fms-Like Tyrosine Kinase 3; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon gamma Receptor; Interferon-gamma; Interleukin-1beta; Interleukin-3; Interleukin-8; Leukemia, Lymphoid; Leukemia, Myeloid; Lymphocyte Activation; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Osteoblasts; Phosphorylation; Receptors, Interferon; Signal Transduction; STAT1 Transcription Factor; Stromal Cells; T-Lymphocytes; Tumor Cells, Cultured

Indirubin, a purple vegetable dye, is a traditional Chinese medicine for myelocytic leukaemia. Indirubin inhibits cyclin-dependent protein kinases (CDKs) and is present in human urine and serum. When indirubin was present during the neutrophilic differentiation of human myelocytic leukaemia HL-60 cells, it augmented superoxide production triggered by opsonized zymosan (OZ) by the terminally differentiated HL-60 cells. It also augmented the calcium response to OZ stimulation, and HL-60 cell chemotaxis evoked by interleukin-8 (IL-8, CXCL8) and formylpeptide. In addition, indirubin induced marked IL-8 release by the cells during differentiation and the cells differentiated with indirubin had typical neutrophilic properties, deformed nuclei and granules. Use of stable cloned HL-60 cells that contained a reporter vector for monitoring the activity of the transcription factor PU.1, which acts specifically at the stage of promyelocyte differentiation into neutrophils and monocytes, revealed that indirubin has a potent promoting activity on intracellular PU.1. Indirubin enhanced the expression of typical neutrophil proteins, including granulocyte-colony stimulating factor receptor, the beta2-integrin subunit CD18, the NADPH-oxidase subunit p47phox, and the IL-8 receptor CXCR1, all are controlled by PU.1. Indirubin also inhibited CDK2-dependent phosphorylation of retinoblastoma protein during neutrophilic differentiation. These results suggest that indirubin augments the neutrophilic differentiation of human myelocytic leukaemia HL-60 cells through inhibition of CDK2 and activation of PU.1.

Topics: CD18 Antigens; Cell Differentiation; Chemotaxis; Cyclin-Dependent Kinases; Dimethyl Sulfoxide; Dose-Response Relationship, Drug; Granulocyte Colony-Stimulating Factor; HL-60 Cells; Humans; Image Processing, Computer-Assisted; Immunoblotting; Indoles; Interleukin-8; Leukemia, Myeloid; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Phosphorylation; Receptors, Granulocyte Colony-Stimulating Factor; Receptors, Interleukin-8A; Retinoblastoma Protein; Stimulation, Chemical; Superoxides

Adherence of hematopoietic progenitor cells (HPCs) to stroma is an important regulatory step in megakaryocytic differentiation. However, the mechanisms through which megakaryocytic progenitors are inhibited by stroma are poorly understood. We examined the role of sulfated glycoconjugates, such as proteoglycans (PGs), on human bone marrow stroma (hBMS). To this end, PG structure was altered by desulfation or enzymatic cleavage. PGs participated in adhesion of human HPC, as desulfation resulted in about 50% decline in adhesion to hBMS. Heparan sulfate proteoglycans (HSPGs) were found to be responsible by showing about 25% decline in adhesion after pre-incubation of HPC with heparin and about 15% decline in adhesion after enzymatic removal of HSPGs from hBMS. Furthermore, PGs were involved in binding cytokines. Both desulfation and enzymatic removal of stromal HSPGs increased release of megakaryocytopoiesis-inhibiting cytokines, that is, interleukin-8 (IL-8, 1.9-fold increase) and macrophage inflammatory protein-1alpha (MIP-1alpha, 1.4-fold increase). The megakaryocytic output of HPC grown in conditioned medium of desulfated stroma was decreased to 50% of the megakaryocytic output in CM of sulfated stroma. From these studies, it can be concluded that PGs in bone marrow, in particular HSPGs, are involved in binding HPC and megakaryocytopoiesis-inhibiting cytokines. Bone marrow stromal PGs thus reduce differentiation of HPC toward megakaryocytes.

Topics: Acute Disease; Antigens, CD34; Blood Proteins; Bone Marrow; Cell Adhesion; Cell Differentiation; Cells, Cultured; Chemokine CCL3; Chemokine CCL4; Culture Media, Conditioned; Eosinophil Major Basic Protein; Hematopoietic Stem Cells; Heparitin Sulfate; Humans; Interleukin-8; Leukemia, Myeloid; Lymphoma, Non-Hodgkin; Macrophage Inflammatory Proteins; Megakaryocytes; Proteoglycans; Stromal Cells

All-trans retinoic acid (ATRA) has been shown to induce differentiation of human acute promyelocytic leukaemia (APL) cells and eventual elimination of the malignant clone. Matrix metalloproteinase-9 (MMP-9) is produced by neutrophils and its expression appears to be linked with myeloid cell differentiation. We investigated effects of ATRA on MMP expression in two human myeloid leukaemia cell lines, PL-21 and NB4. Both cells could differentiate into neutrophils after exposure to ATRA. Both the activity and antigen levels of MMP-9 were much higher in NB4 cells than in PL-21 cells. Stimulation with ATRA significantly increased MMP-9 levels approximately three- to fivefold in both PL-21 and NB4-conditioned media. MMP-9 mRNA levels increased in ATRA-treated cells and was almost in parallel with the increase in MMP-9 activity, suggesting that ATRA induced MMP-9 by activating its gene expression. ATRA can induce interleukin 8 (IL-8) in APL cells. IL-8, chemokine for neutrophils and a potent inducer of MMP-9, was also induced by ATRA in PL-21 cells. However, recombinant IL-8 did not induce MMP-9 expression. In addition, a neutralizing antibody against IL-8 did not inhibit ATRA-induced MMP-9 expression in either cell type. These observations suggest that ATRA can induce both MMP-9 and IL-8, but IL-8 is not involved in ATRA-induced MMP-9 expression. As MMP-9 can truncate and activate IL-8, simultaneous induction of MMP-9 and IL-8 by ATRA could activate leucocytes excessively, causing the hyper-inflammatory events in retinoic acid syndrome.

Topics: Antineoplastic Agents; Humans; Interleukin-8; Leukemia, Myeloid; Matrix Metalloproteinase 9; RNA, Messenger; Tretinoin; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Recently, the role of inter-leukin-8 (IL-8) in angiogenesis was reported. We consequently addressed here the question whether IL-8 produced by acute myeloid leukemia (AML) blasts might have a comparable function.. In 21 pediatric patients with AML the role of AML derived IL-8 in angiogenesis related processes were investigated. Therefore, IL-8 protein and mRNA expression were measured and endothelial cell (EC) migration and proliferation assays were performed. In addition, bFGF and VEGF mRNA expression were measured by RT-PCR.. In the supernatant of the AML blasts, IL-8 protein was present in a varying amount (median 0.86 microg/L, range: 0.1-320 microg/L) and confirmed by RT-PCR. Normal bone marrow mononuclear cells secreted a significant lower amount of IL-8 protein (median: 0.053 microg/L, range: 0.023-0.055 microg/L, P = 0.007). Seven of the 17 tested AML supernatants induced a varying low amount of EC proliferation compared to control media, which was not inhibited by anti-IL-8 antibodies. In contrast, in the EC migration assay, 15 out of the 17 AML supernatants tested, showed an increased EC migration (median fold increase: 1.97, range: 0.66-6.36, P = 0.002) compared to control medium. The increase in EC migration could partially be blocked by anti-IL-8 in 59% of the cases (18% decrease, range 0-62%, P = 0.003). Other contributors for the increase in EC migration were also determined. Vascular endothelial growth factor (VEGF) transcripts by RT-PCR were demonstrated in six out of the nine tested AML cases, while no transcripts for basic fibroblast growth factor (VEGF) could be shown.. Neutralizing anti IL-8 antibodies inhibit EC migration when stimulated with AML supernatant. This suggests a facilitating role for AML-derived IL-8 in an important step in angiogenesis.

Topics: Adolescent; Bone Marrow Cells; Case-Control Studies; Cell Differentiation; Cell Movement; Child; Child, Preschool; DNA Primers; Endothelial Growth Factors; Endothelium, Vascular; Female; Fibroblast Growth Factor 2; Gene Expression Regulation, Neoplastic; Humans; Infant; Interleukin-8; Leukemia, Myeloid; Lymphokines; Male; Neovascularization, Pathologic; Receptors, Interleukin-8A; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

To compare the effects of water-soluble polysaccharides, FI0-b, and its formic acid-modified derivative, FI0-b-H, on production of human proinflammatory cytokines.. The polysaccharides were modified by formic acid. Cytokine production was quantitated by radioimmunoassay. mRNA for cytokines was measured by semi-quantitative RT-PCR.. FI0-b and FI0-b-H 4, 40, and 400 mg/L significantly downregulated interleukin-1 alpha (IL-1 alpha) production by THP-1 cells induced by lypopolysaccharide (LPS) 1 or 10 mg/L and phorbol myristate acetate (PMA) 200 nmol/L. At lower stimulation with LPS 10 mg/L and PMA 200 nmol/L, both polysaccharides significantly upregulated tumor necrosis factor alpha (TNF alpha) production by THP-1 cells. However, at higher stimulation with LPS 100 mg/L and PMA 200 nmol/L, they downregulated TNF alpha production. FI0-b-H downregulated interleukin-8 (IL-8) production by THP-1 cells at a lower-dose of LPS 1 mg/L and PMA 200 nmol/L, but upregulated IL-8 production at a higher-dose of LPS 10 mg/L and PMA 200 nmol/L. Production of cytokines (IL-1 alpha and TNF alpha) was transcriptionally or post-transcriptionally regulated by FI0-b and FI0-b-H.. The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different stimulatory conditions in a dose-dependent manner. Compared with FI0-b, FI0-b-H has more marked effects on human proinflammatory cytokine production.

Topics: Adult; Cell Separation; Drugs, Chinese Herbal; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Leukemia, Myeloid; Leukocytes, Mononuclear; Mycelium; Polysaccharides; Reishi; RNA, Messenger; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Topics: Humans; Interleukin-8; K562 Cells; Leukemia, Myeloid

Serum levels of interleukin 8 (IL-8) were examined in eight patients with acute myeloid leukaemia during 16 courses of chemotherapy. The patients experienced 14 episodes of fever which occurred in periods with granulocyte counts < 0.5 x 10(9)/l. Febrile episodes were classified as bacteriologically defined infection (n = 6), clinically defined infection (n = 2), and unexplained fever (n = 6). IL-8 was detected in 18/25 (72%), 2/3 (67%) and 3/7 (43%) of the serum samples in the respective groups. In contrast, IL-8 was detected in 22/90 (24%) of the samples taken when no fever was present (P < 0.00003 versus bacteriologically defined infection). The median concentration of IL-8 in samples taken during febrile episodes was 194 ng/ml (range 0-6358 ng/ml) and 0 (range 0-5392 ng/ml) on days without fever (not significant). In three patients with infections caused by, respectively, Streptococcus sanguis, Acinetobacter calcoanitratus and Candida albicans, IL-8 rose to a peak levels and declined during recovery. We conclude that IL-I is released systemically during infections with gram-positive and gram-negative bacteria and Candida albicans in patients with acute myeloid leukaemia and peripheral granulocytopenia due to chemotherapy. However, IL-8 can also be detected when no sign of infection is present.

Topics: Acute Disease; Adult; Agranulocytosis; Antineoplastic Combined Chemotherapy Protocols; Female; Fever; Humans; Interleukin-8; Leukemia, Myeloid; Leukocyte Count; Male; Middle Aged; Opportunistic Infections

By chemical cross-linking experiments we show that at physiologically relevant concentrations IL-8 and NAP-2 monomers are in an equilibrium with dimers and even oligomers (KD approximately 300-800 nM). Oligomerization seems to be more prevalent for IL-8 than for NAP-2. The form in which IL-8 and NAP-2 bind to their specific receptors was analyzed in binding experiments with COS-1 cells expressing IL-8 receptor A or B in recombinant forms. Both receptors were cloned from the human myeloid leukemic cell line AML-193. Type A receptor had high affinity for IL-8 (KD approximately 4 nM) and low affinity for NAP-2 (KD > or = 700 nM), whereas the type B receptor was of equally high affinity (KD approximately 2 nM) for both IL-8 and NAP-2. However, IL-8 receptor B could bind specifically three to four times more IL-8 than NAP-2, and NAP-2 was a weak competitor for IL-8 binding to the same receptor. In addition, IL-8, but not NAP-2, could be cross-linked to dimers when bound to IL-8 receptor B. We suggest from these findings that IL-8, but not NAP-2, binds as a dimer and oligomer to IL-8 receptor.

Topics: Animals; beta-Thromboglobulin; Cell Line; Chlorocebus aethiops; Cloning, Molecular; Cross-Linking Reagents; Electrophoresis, Polyacrylamide Gel; Humans; Interleukin-8; Iodine Radioisotopes; Kinetics; Leukemia, Myeloid; Macromolecular Substances; Peptides; Receptors, Interleukin; Receptors, Interleukin-8A; Recombinant Proteins; Transfection; Tumor Cells, Cultured

T lymphocytes, macrophages, and oxidized low-density lipoprotein (Ox-LDL) are collocalized in early atherosclerotic lesions. Using a low-endotoxin in vitro system, we observed that Ox-LDL but not native LDL induced the production, by both freshly adherent human peripheral blood monocytes and human monocytic THP-1 cells, of the alpha chemokine interleukin (IL)-8, a potent chemoattractant for T lymphocytes. Marked IL-8 induction by Ox-LDL did not require IL-1 beta generation in THP-1 cells. Ox-LDL-induced chemokine production was selective, as Ox-LDL did not stimulate the production by THP-1 cells of the T-lymphocyte chemotactic beta chemokine macrophage inflammatory protein (MIP)-1 alpha. IL-8 induction increased in proportion to the extent of oxidation of LDL as measured by the content of lipid oxidation end products. To identify potentially active components of Ox-LDL, we tested malondialdehyde, an arachidonate-derived lipid oxidation product, and 9-hydroxyoctadecadienoic acid, an oxidation product of linoleate, the major polyunsaturated fatty acid in LDL, and observed that they induced IL-8 generation in the absence of Ox-LDL. Furthermore, when most free lipid oxidation products were removed from Ox-LDL by dialysis, some IL-8-inducing activity was released into the dialysate. However, the major IL-8-inducing activity was not dialyzable. To address the nature of the LDL particle modification required to induce IL-8, acetylated or malondialdehyde-treated native LDL particles were monitored for activity. Neither procedure rendered LDL capable of inducing IL-8. However, phospholipase A2-treated LDL induced THP-1 cell expression of IL-8.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acetylation; Chemotaxis, Leukocyte; Humans; Interleukin-1; Interleukin-8; Leukemia, Myeloid; Linoleic Acids; Linoleic Acids, Conjugated; Lipoproteins, LDL; Malondialdehyde; Monocytes; Oxidation-Reduction; Phospholipases A; Phospholipases A2; T-Lymphocytes; Thiobarbituric Acid Reactive Substances; Tumor Cells, Cultured

We looked for chemotaxin/interleukin 8 (CT/IL-8) activity in the culture fluids of 97 human leukemia cell lines and found it in two of the T cell lines, six of the myeloid cell lines, and one of the normal B-cell lines. It was particularly strong in the culture fluids of two cell lines. These cell lines secreted a chemotactic protein into the culture fluids under certain conditions of stimulation with phorbol-12-myristate 13-acetate (PMA), lipopolysaccharide, or hemagglutinin-P. A myeloid leukemia cell line, ML-1, secreted an inducible chemotaxin when stimulated with PMA (1 ng/ml) for 24 h. We purified the chemotaxin from ML-1 cell culture fluid using an improved procedure: concentration with DEAE-Sepharose CL-6B and CM-Sepharose CL-6B, CM-Sepharose column chromatography, and reverse-phase 5TMS-300 column on HPLC with the retention time coinciding with that of LUCT/IL-8 [Suzuki et al., 1989, J. Exp. Med. 169, 1895]. The yield was 200 micrograms protein from 6 liters of the culture fluid. The N terminus of CT/IL-8 was AVLPR-SAKELRXQXIKTYSK- - -, the same as that of LUCT/IL-8, which is constitutively secreted from lung giant cell carcinoma LU65C cells. The optimal concentration in the chemotactic activity of CT/IL-8, equivalent to that of bacterial chemotactic peptide fMet-Leu-Phe (10 nM), was found to be 5 nM. The results show that this chemotaxin is identical to LUCT/IL-8.

Topics: Amino Acid Sequence; B-Lymphocytes; Chemotactic Factors; Chemotaxis, Leukocyte; Chromatography, High Pressure Liquid; Humans; Interleukin-8; Leukemia, Myeloid; Molecular Sequence Data; Neutrophils; Sequence Homology, Amino Acid; T-Lymphocytes; Tumor Cells, Cultured

Interleukin-8 (IL-8), neutrophil activating peptide-2 (NAP-2), and growth regulated gene (GRO, also known as melanoma growth stimulatory activity) are members of a family of peptides which are chemotactic agents for inflammatory cells such as neutrophils. Receptors have been identified for IL-8, GRO and NAP-2 on human neutrophils and granulocytic cell lines, and it has been observed that these cytokines can cross-compete for binding to a common receptor. Using the recently characterized rabbit IL-8 receptor as a probe, two classes of cDNAs, termed type 1 and type 2, were isolated from a human neutrophil library. The type 1 receptor binds only IL-8 while the type 2 receptor binds IL-8, GRO and NAP-2 at high affinity when respective cDNAs are expressed in COS-7 cells. The two cDNAs encode proteins that have an amino acid sequence identity of 77% while the type 1 and 2 receptors have an identity of 84 and 74% with the rabbit IL-8 receptor. These receptors also show significant homology with receptors for other chemotactic agents and with potential coding regions from the human cytomegalovirus genome.

Topics: Amino Acid Sequence; Animals; beta-Thromboglobulin; Cell Line; Cells, Cultured; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; DNA; Dose-Response Relationship, Drug; Granulocytes; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Leukemia, Myeloid; Mice; Molecular Sequence Data; Neoplasm Proteins; Neutrophils; Peptides; Receptors, Cell Surface; Receptors, Cytokine; Receptors, Immunologic; Receptors, Interleukin-8A; Sequence Alignment; Sequence Homology, Amino Acid; Tumor Cells, Cultured

Purified leukemic cells from 30 acute myeloid leukemia (AML) cases at diagnosis were investigated for the presence of interleukin 8 (IL-8) mRNA by Northern blot analysis. IL-8 specific transcripts were detected in uncultured blasts in 14/30 cases, 10/14 from patients with M4-M5 and 4/14 from cases with M0-M3 morphology. The transcript expression was associated with the detection of IL-8 molecule in blast cells by immunostaining performed on cytospin preparations. After 24-hour culture, a strong up-regulation or the appearance in cases negative before culture of IL-8 mRNA was observed in all cases tested, and culture supernatants contained high amounts of IL-8. Our data demonstrate that leukemic cells in AML are equipped with the functional apparatus for IL-8 production. Since IL-8 displays a wide range of biological activities, including the regulation of some membrane molecules relevant to adhesion and migration processes, its production by AML blasts might be of relevance for the pattern of leukemic growth.

Topics: Acute Disease; Cell Division; Gene Expression; Humans; Interleukin-8; Leukemia, Monocytic, Acute; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukemia, Myelomonocytic, Acute; Leukemia, Promyelocytic, Acute; RNA, Messenger; Tumor Cells, Cultured