Hematopoietic cytokines: from biology and pathophysiology to clinical application.
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Acute Disease↗
Biomedical subjects
R Mertelsmann
Publications and source records attributed to R Mertelsmann.
The significance of serum levels of soluble 60kDa receptors for tumor necrosis factor in patients with Hodgkin's disease.
Soluble forms of the two molecular species of the cell surface receptors (Rs) for tumor necrosis factor (TNF) have been detected in normal urine and serum including type I and type II TNF-Rs. Using enzyme-linked immunosorbent assay we have determined type I 60 kDa sTNF-R levels in the sera of 45 age- and sex-matched healthy subjects and 106 patients with Hodgkin's disease (HD). HD patients were either previously untreated (n = 76) or were in complete remission for at least 3 years after remission induction treatment (n = 30). The mean +/- SD concentrations of the 60 kDa type sTNF-R were significantly higher in HD patients than in healthy controls (1.32 +/- 0.19 ng/ml versus 0.6 +/- 0.13 ng/ml; p < 0.001). The extent of increase correlated with the disease stage. Soluble 60 KDa TNF-Rs were found to be significantly higher in stage III and IV (1.42 +/- 0.21 ng/ml) than in stages I and II (1.08 +/- 0.15 ng/ml). Patients with B-symptoms (n = 33) had higher levels (1.67 +/- 0.20 ng/ml) than patients without systemic symptoms (1.02 +/- 0.11 ng/ml; p < 0.001). In 52 patients evaluable for response, the complete remission (CR) rate of patients with 60 kDa sTNF-Rs < 1.2 ng/ml was higher (88%) than in those with 60 kDa sTNF-Rs > 1.2 ng/ml (64%; p < 0.01). A significant increase in serum levels of 60 kDa sTNF-R levels was also observed in HD patients in long-standing CR (1.04 +/- 0.10 ng/ml). Our data suggest that the pretreatment serum concentration of 60 kDa sTNF-Rs in HD may bear prognostic relevance. Increased 60 kDa sTNF-R levels seen in HD patients in remission may point to the defect in cellular immunity characteristic of HD patients.
Adolescent↗
Allogeneic family-unrelated bone marrow transplantation in acute and chronic myeloid leukemia.
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Acute Disease↗
Effect of long-term treatment with recombinant human interleukin-3 in patients with myelodysplastic syndromes.
In a phase II study, involving nine patients with refractory anemia or refractory anemia with ring sideroblasts, the effects of treatment with recombinant human interleukin-3 (IL-3) on hematopoietic function were assessed. Doses of IL-3 ranging from 60 micrograms/m2 during weeks 1-6 to 125 micrograms/m2 during weeks 7-12 were administered as subcutaneous bolus injections three times per week for 12 weeks. Platelet counts increased in six patients. Platelet increase correlated with stable or decreased serum tumour necrosis factor alpha (TNF-alpha) levels, while an increase of TNF-alpha levels during IL-3 therapy occurred in patients with no change or a decrease of platelet counts. Leukocyte counts increased in two patients and reticulocytes in three, without an effect on hemoglobin levels. Morphological analysis of the bone marrow revealed an expansion of the myeloid compartment in seven of eight evaluable patients, mainly due to stimulation of the precursor cells. No improvement of the in vitro growth of hematopoietic progenitor cells was observed. Sequential cytogenetic analyses indicate that IL-3 treatment does not act preferentially on either the cytogenetically abnormal or the normal clones. These results suggest that long-term treatment with low-dose IL-3 stimulates megakaryopoiesis with increase of platelet counts, but that additional later-acting cytokines probably will be required to augment neutrophil and erythrocyte counts.
Aged↗
High concentrations of the interleukin-1 receptor antagonist in serum of patients with Hodgkin's disease.
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Hodgkin Disease↗
A switch toward demethylation is associated with the expression of myeloperoxidase in acute myeloblastic and promyelocytic leukemias.
Expression of numerous genes encoding myeloid-specific proteins is tightly regulated during normal myeloid differentiation. This heterogeneous group of genes thus offers a tool for dissection of different maturational steps of myelopoiesis. We have previously shown that the human myeloperoxidase (MPO) gene is modified at numerous CpG residues in normal myeloid cells and in myeloid cell lines in a development-specific and expression-associated manner. In the present work, we have applied this type of methylation analysis to primary leukemia myeloid cell samples. We found that expression of MPO messenger RNA (mRNA) is grossly limited to leukemias of late myeloblastic and promyelocytic stages; expression is thus associated with progressive demethylation in the 5' region of the MPO gene. This modification was not global. Low-level induction of MPO mRNA expression in very immature leukemic cells using phorbol ester was not accompanied by progressive demethylation. This type of methylation analysis of a myeloid-specific gene may yield a molecular indicator for different types of myeloid leukemias.
Blotting, Northern↗
Expression of bcl-2 in Burkitt's lymphoma cell lines: induction by latent Epstein-Barr virus genes.
The bcl-2 oncogene blocks programmed cell death (apoptosis). Epstein-Barr virus (EBV) can immortalize B lymphocytes into continuously growing lymphoblastoid cell lines (LCL) by the coordinate expression of at least 9 latent genes (EBV nuclear antigen [EBNA] 1-6, latent membrane protein [LMP], and terminal proteins [TP] 1 and 2). We analyzed transcription and expression of bcl-2 and latent EBV genes in Burkitt's lymphoma (BL) cell lines with a germinal center phenotype (group I) as well as activated BL cell lines (group III) and LCLs. We found high expression of bcl-2 as well as the full spectrum of latent EBV genes in LCLs and activated group III BL cell lines. Group I BL cells expressed little or no bcl-2, EBNA-2, and LMP. Superinfection with nondefective EBV or an EBNA-2-defective virus as well as transfection with EBNA-2- or LMP-carrying vectors into the EBV-negative cell lines RAMOS, DG75, U698, or BJAB induced upregulation of bcl-2 expression. The strongest effect on bcl-2 was obtained by transfection with LMP, or infection with the nondefective virus. No change of bcl-2 expression was observed with EBNA-1. Our data indicate that the immortalization capacity of EBV and the growth advantage of EBV-positive compared with EBV-negative BL cells in vitro may predominantly be mediated via induction of bcl-2 and the main effectors are EBNA-2 and LMP.
Antigens, Viral↗
Synergy of interleukin 3 and tumor necrosis factor alpha in stimulating clonal growth of acute myelogenous leukemia blasts is the result of induction of secondary hematopoietic cytokines by tumor necrosis factor alpha.
Colony growth of leukemic colony-forming units (L-CFU) obtained from patients with primary acute myelogenous leukemia stimulated with recombinant human interleukin 3 (rh IL-3) is significantly potentiated when recombinant human tumor necrosis factor alpha (rh TNF-alpha) is present in cultures. The costimulatory activity of tumor necrosis factor (TNF) alpha is dose dependent and maximum at TNF-alpha concentrations of 10 ng/ml. At high density, L-CFU proliferatively respond to TNF-alpha stimulation in the absence of exogenous rh IL-3. Studies of the mechanism of action of rh TNF-alpha on acute myelogenous leukemia L-CFU growth suggest that TNF-alpha acts by inducing release of growth stimulatory hematopoietic cytokines by the leukemic cells themselves, including IL-1 alpha, IL-1 beta, Granulocyte-macrophage colony-stimulating factor (CSF), granulocyte CSF, and IL-6. Treatment of L-CFU cultures, with neutralizing antibodies to IL-1 alpha, IL-1 beta, granulocyte-macrophage CSF, granulocyte CSF, and IL-6 to eliminate the endogenous source of these factors is associated with significant inhibition of the synergistic interplay of TNF-alpha and IL-3.
Drug Synergism↗
Mobilization of peripheral blood progenitor cells by sequential administration of interleukin-3 and granulocyte-macrophage colony-stimulating factor following polychemotherapy with etoposide, ifosfamide, and cisplatin.
We report on the requirements that have to be met to combine a standard-dose chemotherapy regimen with broad antitumor activity with the mobilization of peripheral blood hematopoietic progenitor cells. Thirty-two cancer patients were given a 1-day course of chemotherapy consisting of etoposide (VP16), ifosfamide, and cisplatin (VIP; n = 46 cycles), followed by the combined sequential administration of recombinant human interleukin-3 (rhIL-3) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF). Control patients received GM-CSF alone or were treated without cytokines. Maximum numbers of peripheral blood progenitor cells (PBPC) were recruited on day 13 to 17 after chemotherapy, with a median of 418 CD34+ cells/microL blood (range, 106 to 1,841) in IL-3/GM-CSF-treated patients, 426 CD34+/microL (range, 191 to 1,380) in GM-CSF-treated patients, and 46 CD34+/microL (range, 15 to 148) in patients treated without cytokines. In parallel, there was an increase in myeloid (10,490 colony-forming unit-granulocyte-macrophage [CFU-GM]/mL blood; range, 1,000 to 23,400), as well as erythroid (10,660 burst-forming unit-erythroid [BFU-E]/mL blood; range, 3,870 to 24,300) and multipotential (840 CFU-granulocyte, erythrocyte, monocyte, megakaryocyte [GEMM]/mL blood; range, 160 to 2,070) progenitor cells in IL-3 plus GM-CSF-treated patients. In GM-CSF-treated patients, significantly less precursor cells of all lineages were mobilized, particularly multipotential progenitors (400 CFU-GEMM/mL blood; range, 200 to 2,150). Only small numbers of CD34+ cells and clonogenic progenitor cells could be recruited in intensively pretreated patients. Our data document that after standard-dose chemotherapy-induced bone marrow hypoplasia, IL-3 plus GM-CSF can be used to recruit PBPC, which might shorten the hematopoietic recovery after high-dose chemotherapy in chemosensitive lymphomas or solid tumors.
Adult↗
Interleukin-1 beta (IL-1 beta) expression in human blood mononuclear phagocytes is differentially regulated by granulocyte-macrophage colony-stimulating factor (GM-CSF), M-CSF, and IL-3.
In this report we show that recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and rh macrophage (M)-CSF induce accumulation of interleukin-1 beta (IL-1 beta) mRNA in blood-derived mononuclear phagocytes (MNP). GM-CSF and M-CSF treatment of MNP is also associated with IL-1 beta secretion. Regulation of GM- and M-CSF-induced IL-1 beta mRNA expression involves transcriptional and posttranscriptional mechanisms. However, the action of IL-3 on synthesis of IL-1 beta mRNA differs from that of other CSFs: While GM-CSF and M-CSF induce binding activity of the nuclear factor (NF) kappa B, IL-3 treatment of MNP has no profound effect on NF kappa B binding to DNA. Moreover, IL-3 decreases the transcription rate of the IL-1 beta gene and has only little effect on stability of IL-1 beta mRNA, which is increased by GM- and M-CSF. However, IL-3 enhances M-CSF-induced accumulation of IL-1 beta mRNA by unknown posttranscriptional means that may relate to an increased expression of M-CSF receptor (ie, c-fms) mRNA, detectable in mononuclear phagocytes on exposure to IL-3.
Base Sequence↗
Clonal analysis of bcr-abl rearrangement in T lymphocytes from patients with chronic myelogenous leukemia.
The cytogenetic hallmark of chronic myelogenous leukemia (CML) is the Philadelphia chromosome (Ph1), which reflects a chromosomal translocation t(9;22) and a rearrangement of the ABL and bcr genes. This marker is found in all cells arising from the same malignant precursor cell and can be detected in CML cells of the myeloid, monocytic, erythroid, and B-lymphocyte lineage. It is, however, controversial as to whether T lymphocytes of CML patients carry this gene rearrangement. An answer to this question would clarify whether the translocation in CML occurs in a pluripotent hematopoietic stem cell or in a precursor cell already committed to certain lineages, but not the T-cell lineage. To address this question, we established T-cell clones from peripheral venous blood cells of four patients with CML and screened these clones for bcr-abl fusion transcripts by means of polymerase chain reaction and Southern blot analysis. In four T-cell clones of three of these patients, the bcr-abl transcript could be detected. None of 12 T-cell clones of the fourth patient disclosed detectable bcr-abl amplification product. Both CD4+ as well as CD8+ clones displayed fused bcr-abl sequences. These data imply that in CML some but not all T lymphocytes may originate from the Ph1-positive stem cell.
Blotting, Southern↗
Secretion of granulocyte-macrophage colony-stimulating factor by human blood monocytes is stimulated by engagement of Fc gamma receptors type I by solid-phase immunoglobulins requiring high-affinity Fc-Fc gamma receptor type I interactions.
Despite reports on the secretion of granulocyte-macrophage-colony-stimulating factor (GM-CSF) by murine peritoneal macrophages in response to inflammatory stimuli, the ability of human monocytes to generate this growth factor has remained doubtful. Neither endotoxin, phorbol compounds, nor inflammatory cytokines have been shown to elicit GM-CSF by these cells. Our present studies indicate that exposure of monocytes to solid-phase murine IgG2a, but not to murine IgG1 and thus cross-linkage of the 72-kDa Fc gamma RI results in transcription of the GM-CSF gene, accumulation of stable GM-CSF mRNA and finally in release of biologically active GM-CSF protein. Cross-linking of Fc gamma RI by a murine anti-Fc gamma RI monoclonal antibody and goat anti-mouse antibody failed, however, to stimulate GM-CSF release. This suggests that high affinity Fc-Fc gamma RI interactions are required for induction of expression of GM-CSF by monocytes.
Animals↗
Leukotriene B4 transcriptionally activates interleukin-6 expression involving NK-chi B and NF-IL6.
Leukotriene B4 (LTB4) is a notable participant in inflammation and chemotaxis. It is, however, still unclear whether LTB4 acts in this regard directly or indirectly by stimulating the release of chemotactic and inflammatory cytokines. Here we report that LTB4 induces synthesis of interleukin (IL)-6 by human blood monocytes through transcriptional activation of the IL-6 gene. We furthermore demonstrate that this process involves activation of the transcription factor NF-chi B and, to a lesser extent, of NF-IL6, while the activity of the transcription factor AP-1, shown to otherwise confer IL-6 inducibility, appeared to be unaffected by LTB4. Involvement of NF-chi B and NF-IL6 in induction of IL-6 transcription by monocytes was demonstrated using deleted forms of the IL-6 promoter. Activation of the IL-6 promoter by LTB4 was not only associated with accumulation of the respective transcripts but resulted in synthesis of functional IL-6 protein as well. In addition, LTB4 mediated transactivation of a heterologous promoter construct containing the NF-chi B or the NF-IL6 enhancer, but not the AP-1 enhancer. The signaling events mediating this effect appeared to involve the release of H2O2, since LTB4 failed to induce NF-chi B or NF-IL6 in the presence of the scavenger of H2O2, N-acetyl-L-cysteine.
Binding Sites↗
Interleukin-3 and granulocyte-macrophage colony-stimulating factor in combination: clinical implication.
The multilineage hematopoietic effects of IL-3 appear to be most important for its clinical use comprising especially leucocyte and platelet responses. This was demonstrated to be dose dependent characterising doses of 250 to 500 micrograms/m2/day subcutaneously as hematopoietic effective and well tolerable. Since preclinical data suggest synergism between IL-3 and GM-CSF hematopoietic effects of their sequential administration were evaluated in 15 patients with preserved hematopoietic function. An enhanced stimulation of megakaryopoiesis combined with more pronounced stimulation of granulopoiesis than IL-3 alone could be demonstrated. The cytokine combination of IL-3 and GM-CSF was thus used during chemotherapy induced myelosuppression. Data indicates beyond the known amelioration of myelosuppression by GM-CSF additional chances to enhance platelet recovery which can be of important clinical impact.
Antineoplastic Combined Chemotherapy Protocols↗
Stimulation of growth of human malignant lymphoma and lymphoid leukemia cells in vitro.
We have analyzed cultures of malignant lymphoma cells and cells from patients with acute lymphoid leukemia in methylcellulose for their ability to from colonies. Clonogenic growth was examined in the presence or absence of fetal calf serum (FCS), platelet-poor plasma (PPP), medium conditioned by phytohemagglutinin-stimulated leukocytes (PHA-LCM), or irradiated allogeneic bone marrow stroma cells. Cells from 25 lymphoma patients--17 with non-Hodgkin's lymphoma (NHL), eight with Hodgkin's disease (HD)--and from 19 patients with acute lymphocytic leukemia (ALL) were investigated. We show that colony growth can be obtained in a minority of cases (in 3 NHL, 5 HD, and 2 ALL) and that the use of FCS and allogeneic irradiated stroma cells may be required for optimal colony formation.
Cell Division↗
Application of basic science to hematopoiesis and treatment of disease. 15th Bristol-Myers Squibb Symposium on Cancer Research held at the Fred Hutchinson Cancer Research Center, Seattle, November 4-5, 1991.
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Cell Differentiation↗
Detection of allele-specific expression of N-RAS oncogenes in human leukaemia cells.
We report analysis of allele-specific expression of N-RAS transcripts in myeloid leukaemic cells and cell lines. Expression was assessed by an assay of reverse-transcription/PCR combined with differential hybridization with mutation-specific oligonucleotides. In cells from all patients with acute myeloid leukaemia (AML) and in myeloid cell lines HL-60 and THP-1, expression of both the wild-type allele and the abnormal allele altered by a point mutation could be detected, albeit not always at comparable levels. This might be due for instance to allelic exclusion. The assay described provides a means of analysing the degree of expression of dominant oncogenes.
Acute Disease↗
Elevated circulating levels of tumor necrosis factor predict unresponsiveness to treatment with interferon alfa-2b in chronic myelogenous leukemia.
PURPOSE: The study was undertaken to analyze circulating tumor necrosis factor (TNF) levels in patients with chronic-phase chronic myelogenous leukemia (CML) undergoing interferon (IFN) alfa-2b therapy, and to correlate pretreatment serum levels of TNF with response to IFN alfa-2b therapy. PATIENTS AND METHODS: Fourteen patients with CML in chronic phase were treated with recombinant human IFN alfa-2b for 7 to 39 months. RESULTS: In eight patients IFN alfa-2b treatment failed due to lack of hematologic response. A complete or partial hematologic remission was achieved in the remaining six patients, of whom two patients experienced a complete cytogenetic response. Retrospective analysis of serum samples obtained from all patients before the onset of IFN alfa-2b administration revealed that levels (mean +/- SEM) of circulating TNF were higher (P less than .001) in the group of patients who did not respond to IFN alfa-2b treatment (157 +/- 15 U/mL) than in the responders (10.3 +/- 4 U/mL) or healthy control subjects (9.1 +/- 3 U/mL). However, there was no correlation between TNF serum levels and other patient characteristics at study enrollment including age, sex, duration of disease, performance status, splenomegaly, WBC count, platelet count, hemoglobin value, prior therapy, and prognostic category. CONCLUSION: These findings indicate that circulating levels of TNF are increased in a subset of patients with chronic-phase CML and that this elevation is associated with poor response to IFN alfa-2b therapy.
Adult↗