Involvement of interferon-γ and macrophage colony-stimulating factor in pathogenesis of haemophagocytic lymphohistiocytosis in adults

Summary We investigated the role of monocyte/macrophage-activating cytokines in pathogenesis of haemophagocytic lymphohistiocytosis (HLH) in 21 adult patients. Sera from patients with active HLH contained extremely high levels of macrophage colony-stimulating factor (M-CSF) and of interferon-γ (IFN-γ). These levels returned to almost normal during remission. Neither interleukin-4 nor granulocyte/macrophage colony-stimulating factor could be detected. Active HLH sera also contained high concentrations of inflammatory monokines, such as interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α). Serum concentrations of soluble CD8 and soluble interleukin-2 receptor were extremely high during active HLH, and returned to virtually normal levels during remission. Circulating CD2⁺ T-cells obtained from patients with active HLH spontaneously secreted M-CSF and IFN-γ in vitro , whereas circulating monocytes did not produce detectable levels of both M-CSF and IFN-γ, but produced high levels of IL-6 and TNF-α. These findings suggest that IFN-γ and M-CSF at least partly from T-cells, such as CD8⁺ T-cells, might contribute to activation of monocytes or histiocytes, resulting in the up-regulated monokine production and haemophago-cytosis in HLH.     

Virus-associated haemophagocytic syndrome in previously healthy adults

Abstract: The clinical characteristics of 15 adult patients with virus-associated haemophagocytic syndrome (VAHS) were studied. The patients were 3 males and 12 females with a mean age of 39.5 years (range 20 to 67 years). Seven patients (mean age 48.6 years) were immunosuppressed by drugs or as a result of having malignant or autoimmune disease. Eight patients (mean age 31.6 years) had no underlying diseases. The prognosis of the patients with immunosuppression was poor, as previously reported, and 3 of them died. In younger adult patients with this syndrome who had no underlying immunosuppressive diseases, the prognosis was good even without therapy. The sera from the patients in both groups contained extremely high levels of macrophage colony-stimulating factor (M-CSF) and slightly elevated levels of tumour necrosis factor-α (TNF-α). These findings suggested that VAHS, can occur in presumably healthy adults and that VAHS can be a more benign condition than previously believed. M-CSF and TNF-α may play an important role in the development of the syndrome in both groups.     

Parvovirus B19-associated haemophagocytic syndrome in healthy adults

Between August 1992 and August 1994 we found seven adult patients with haemophagocytic syndrome associated with human parvovirus B19 (HPV) infection. Five of the patients were previously healthy but the other two had underlying immunosuppressive disease; the former five patients recovered spontaneously without any treatment. Eight cases of HPV-associated haemophagocytic syndrome have been reported; six of them were children and the remainder were adults with immunosuppressive disease or haematological disorder. Our study suggests that HPV is not a rare cause of virus-associated haemophagocytic syndrome (VAHS). Moreover, HPV-associated haemophagocytic syndrome could occur more frequently than previously thought, not only in children and adults with underlying disease but also in adults, even in good health.     

Successful use of granulocyte colony-stimulating factor to correct neutropenia in chronic lymphocytic leukaemia

A case of wound infection secondary to profound neutropenia associated with chronic lymphocytic leukaemia is described. Both the neutropenia and infection resolved with the use of granulocyte colony-stimulating factor. This agent may be useful in other similar patients.     

Recombinant human granulocyte colony-stimulating factor (rhG-CSF; filgrastim) treatment of clozapine-induced agranulocytosis

Abstract. After 10 weeks of treatment with clozapine, severe agranulocytosis was diagnosed in a 33-year-old female. The patient was treated with filgrastim (granulocyte colony-stimulating factor [G-CSF]) 5 μg kg^?1 day^?1. The neutrophil count was 0.234 × 10⁹ l^?1 on admission, with a further decrease the next day to < 0.050 × 10⁹ l^?1, and this complete agranulocytosis continued for 10 days. As no response was obtained after 1 week the dosage of filgrastim was increased to 10 μg kg^?1 day^?1 with immediate improvement. A rapid and pronounced leucocytosis developed with maximal value of neutrophil granulocytes (including immature forms) of 33.108 × 10⁹ l^?1 on day 12 after admission. The patient only had minor infectious complications during the neutropenic period. In conclusion, early treatment with filgrastim seems warranted in severe cases of clozapine-induced agranulocytosis. A dosage of 10 μg kg^?1 day^?1 can be recommended.     

Role of granulocyte colony-stimulating factor as adjunct therapy for septicemia in children with acute leukemia

The granulocyte colony-stimulating factor (G-CSF) has been shown to accelerate recovery from severe neutropenia and to decrease the incidence of documented infections after intensive chemotherapy in cancer patients. However, the routine prophylactic use of G-CSF is expensive. This study was conducted to determine the role of G-CSF as adjunct therapy for septicemia following neutropenia caused by chemotherapy in children with acute leukemia. Fifty consecutive episodes of septicemia were studied involving 34 episodes of Gram-negative, 7 episodes of Gram-positive, 5 episodes of polymicrobial bacterial septicemia, one episode of fungemia, and 3 episodes of disseminated fungal infection. In the first 25 episodes, G-CSF was not used (group A). For the next 16 episodes, G-CSF 200 μg per square meter per day subcutaneously was given immediately after the septicemia was documented until the absolute neutrophil count was maintained at more than 1,500 per cubic millimeter (group B). Thereafter, G-CSF at the same dose as that of group B was prophylactically used in all the children who received high-dose cytosine arablnc-side-containing regimens. Nine episodes of septicemia occurred (group C). The incidences of mortality per episode of septicemia in groups A, B, and C were 12.0% (3/25), 12.5% (2/16) and 0% (0/9), respectively. Statistically, there was no difference between the three groups overall and in pair-wise comparisons (all P > 0.5). The durations of G-CSF administration in group B ranged from 6 to 26 days with a median of 12 days and the durations of G-CSF administration in group C ranged from 10 to 23 days with a median of 19 days. With or without G-CSF, there may be no significant difference in the mortality of septicemia following neutropenia caused by chemotherapy in children with acute leukemia.     

粒细胞集落刺激因子对大鼠颈总动脉损伤后新生内膜增生的影响

目的探讨粒细胞集落刺激因子（G-CSF）对动脉粥样硬化（AS）大鼠颈总动脉球囊损伤后再狭窄的影响。方法Wistar大鼠48只,随机分为3组:普通饮食组（普食组,n=6）,高脂饮食组（高脂组,n=42）。高脂组又分为:G-CSF组（n=21）和生理盐水组（NS组,n=21）。高脂饮食后1个月用HE染色及透射电镜证明AS形成情况。G-CSF组和NS组分别于高脂饮食后1个月给予G-CSF或生理盐水干预6d,然后行左颈总动脉球囊损伤术,术后14、21、90d分别用扫描电镜、HE染色及图像分析观察血管内皮形成及内膜的增生情况。结果高脂组大鼠高脂饮食后1个月可见AS的早期改变,行球囊损伤后可见大鼠颈总动脉内膜逐渐增厚,但是G-CSF组明显小于NS组,G-CSF组损伤血管内皮较快恢复连续性,G-CSF组内膜与中膜面积比值（Ai/Am）远小于NS组（P<0.01）。结论G-CSF可以加速AS大鼠损伤血管的再内皮化、抑制血管损伤后再狭窄的发生。     

Clinicopathological spectrum of haemophagocytic syndrome in Epstein-Barr virus-associated peripheral T-cell lymphoma

Haemophagocytic syndrome (HS) is frequently observed in Epstein-Barr virus-associated peripheral T-cell lymphoma (EBV-PTCL) and represents a major cause of death. In this communication we have further analysed the spectrum of HS in 12 patients with EBV-PTCL. The patients could be divided into three groups according to the time of onset of HS during the clinical course of PTCL. Group I patients (four cases) had HS as the initial clinical manifestation. All four patients were initially suspected to have malignant histiocytosis (MH) but a MH-like PTCL was later diagnosed. Group II patients (six cases) developed HS at the time of lymphoma relapse. Four of them belonged to the angioinvasive type PTCL. Group III patients (two cases) developed HS at clinical remission; both were angioinvasive type PTCL. Nine patients had serological evidence suggesting active EBV infection. The clinical course after the onset of HS was generally fulminant in each group with a median survival of only 44 d despite combination chemotherapy and/or empirical therapy with high-dose immunoglobulin and corticosteroids in six patients. In conclusion, HS represents a severe complication of EBV-PTCL. Although most patients develop HS at a time of active lymphoma, the syndrome may occur when the lymphoma is in remission. Because of the poor outcome, early diagnosis and a new modality of treatment for HS associated with EBV-PTCL should be pursued in future.     

Assessment of the value of treatment with granulocyte colony-stimulating factor in children with acute lymphoblastic leukemia: a randomized clinical trial

Abstract: The present trial was designed to test the effects of G-CSF on the duration of the second phase of induction chemotherapy in children with newly diagnosed acute lymphoblastic leukemia (ALL). A total of 32 patients were assigned randomly to a group that received (14 patients; group A) or a group that did not receive (18 patients; group B) G-CSF (10 g/kg/day subcutaneously and daily) throughout of the second phase of induction therapy. One of 14 (7.1%) patients in group A and 2 of 18 (11.1%) patients in group B completed the course of chemotherapy within the planned time. The median length of this phase was 37 days (range, 29 to 65; mean, 40; SD, 8.6) for patients in group A and 36 days (range, from 29 to 55; mean, 38; SD, 7.4) for those in group B, and the difference was not statistically significant. The number of days during which patients had granulocyte counts of less than 2 × 10⁹/l, the number of febrile episodes of unknown origin, the number of bacterial and fungal infections and the number of days of hospitalization did not differ in a statistically significant manner between the two groups. Our data suggest that G-CSF supportive therapy may be unnecessary in children with neutropenia of short duration, for whom the risk of infection is low.     

Effects of granulocyte colony-stimulating factor upon coxsackievirus B3 myocarditis in mice

Granulocyte colony-stimulating factor is a potent activatorof mature granulocytes, and subsequently enhances superoxiderelease. The purpose of this study was to investigate the effectsof granulocyte colony-stimulating factor upon murine coxsackievirusB3 myocarditis in relation to free radical-mediated cardiacdamage. Two-week-old, male, C₃H/He mice were inoculated intraperitoneallywith coxsackievirus B3. Gramulocyte colony-stimulating factor20 µg.kg^–1. day^–1, polyethylene glycol-conjugatedsuperoxide dismutase (an enzyme catalyzing the conversion ofO₂^- to H₂O₂) 1 x 10³ U.kg^–1 day^–1 and granulocytecolony-stimulating factor 20 µg. kg^–1 day^–1,plus polyethylene glycol-conjugated superoxide dismutase 1 x10³ U.kg^–1. day^–1, were injected subcutaneouslydaily on days 0 to 14. Treated groups were compared to the infected,untreated group. The survival rate in the polyethylene glycol-conjugatedsuperoxide dismutase group was higher than that of the untreatedgroup on day 14, but on day 7, cardiac pathology was not significantlydifferent among the four groups. On day 14, the scores of cellularinfiltration, myocardial necrosis and calc were lower in thepolyethylene glycol-conjugated superoxide disnuitase group andin the granulocyte colony-stimulating factor plus polyethyleneglycol-conjugated superoxide dismutase group than in the untreatedgroup. The myocardial virus titres on days 7 and 14 did notd sign y among the four groups. The number of total white bloodcell and neutrophil counts were signifIcantly greater in thegranulo cyte colony-stimulating factor group than in the untreatedgroup on day 7. Taken altogether with the previous reports andpresent evidence that the administration of granulocyte colony-stimulatingfactor did not exacerbate coxsackievirus B3 myocarditis, itmay be that oxygen-free radicals appeared to be derived notfrom leukocytes but from other components in this experimentalmodel of myocarditis, whereas the myocardium was inflamed withleukocytes.     

Effects of granulocyte colony-stimulating factor upon coxsackievirus B3 myocarditis in mice

Granulocyte colony-stimulating factor is a potent activatorof mature granulocytes, and subsequently enhances superoxiderelease. The purpose of this study was to investigate the effectsof granulocyte colony-stimulating factor upon murine coxsackievirusB3 myocarditis in relation to free radical-mediated cardiacdamage. Two-week-old, male, C₃H/He mice were inoculated intraperitoneallywith coxsackievirus B3. Gramulocyte colony-stimulating factor20 µg.kg^–1. day^–1, polyethylene glycol-conjugatedsuperoxide dismutase (an enzyme catalyzing the conversion ofO₂^- to H₂O₂) 1 x 10³ U.kg^–1 day^–1 and granulocytecolony-stimulating factor 20 µg. kg^–1 day^–1,plus polyethylene glycol-conjugated superoxide dismutase 1 x10³ U.kg^–1. day^–1, were injected subcutaneouslydaily on days 0 to 14. Treated groups were compared to the infected,untreated group. The survival rate in the polyethylene glycol-conjugatedsuperoxide dismutase group was higher than that of the untreatedgroup on day 14, but on day 7, cardiac pathology was not significantlydifferent among the four groups. On day 14, the scores of cellularinfiltration, myocardial necrosis and calc were lower in thepolyethylene glycol-conjugated superoxide disnuitase group andin the granulocyte colony-stimulating factor plus polyethyleneglycol-conjugated superoxide dismutase group than in the untreatedgroup. The myocardial virus titres on days 7 and 14 did notd sign y among the four groups. The number of total white bloodcell and neutrophil counts were signifIcantly greater in thegranulo cyte colony-stimulating factor group than in the untreatedgroup on day 7. Taken altogether with the previous reports andpresent evidence that the administration of granulocyte colony-stimulatingfactor did not exacerbate coxsackievirus B3 myocarditis, itmay be that oxygen-free radicals appeared to be derived notfrom leukocytes but from other components in this experimentalmodel of myocarditis, whereas the myocardium was inflamed withleukocytes.     

Serum granulocyte colony-stimulating factor levels in patients with chronic neutropenia of childhood: modulation of G-CSF levels by myeloid precursor cell mass

The serum G-CSF levels of eight patients with severe congenital neutropenia (SCN) were found to be significantly higher than those of 22 patients with chronic benign neutropenia (CBN). The relative number of cells expressing the G-CSF receptor in light density bone marrow cells (LDBMC) was lower in patients with SCN than in patients with CBN or in normal subjects. When recombinant human G-CSF was incubated with LDBMC, G-CSF levels were decreased by LDBMC from normal subjects and CBN patients, but not by those from SCN patients. Serum G-CSF concentrations, which are affected by mature neutrophils, may also be modulated by myeloid precursor cells in the bone marrow.     

Functional interaction between mutations in the granulocyte colony-stimulating factor receptor in severe congenital neutropenia

Most severe congenital neutropenia (SCN) cases possess constitutive neutrophil elastase mutations; a smaller cohort has acquired mutations truncating the granulocyte colony-stimulating factor receptor (G-CSF-R). We have described a case with constitutive extracellular G-CSF-R mutation hyporesponsive to ligand. Here we report two independent acquired G-CSF-R truncation mutations and a novel constitutive neutrophil elastase mutation in this patient. Co-expression of a truncated receptor chain restored STAT5 signalling responses of the extracellular G-CSF-R mutant, while constitutively-active STAT5 enhanced its proliferative capacity. These data add to our knowledge of SCN and further highlight the importance of STAT5 in mediating proliferative responses to G-CSF.     

Successful treatment of chronic severe neutropenia with weekly recombinant granulcyte-colony stimulating factor

Daily treatment for symptomatic chronic neutropenia with recombinant granulocyte-colony stimulating factor (rhG-CSF) filgrastim is costly and sometimes causes neutrophillia. We report the use of weekly filgrastim in a 40-year-old man with life-long symptomatic neutropenia. Baseline neutrophil counts were <1×10⁹/l 60% of the time, and fell below 0.5×10⁹/l for 7 d periods every 22 d. Following 1 year of weekly filgrastim treatment, the absolute neutrophil count was maintained >1×10⁹/l (averaging 2×10⁹/l) and the frequency and severity of symptoms were reduced by 85%. Therefore the benefits of filgrastim for the treatment of at least one form of chronic severe neutropenia can be derived from weekly rather than daily doses.     

Phase II study of glycosylated recombinant human granulocyte colony-stimulating factor after HLA-identical sibling bone marrow transplantation

Background: The lengthy period of neutropenia which follows allogeneic bone marrow transplantation (BMT) results in significant morbidity and some mortality. Recombinant human granulocyte colony-stimulating factor (rhuG-CSF) effectively reduces neutropenia and morbidity when given after autologous BMT, but has not been adequately investigated in allografts. Aims: To assess the tolerability, safety and efficacy of rhuG-CSF after allogeneic BMT. Methods: rhuG-CSF was administered to 13 adult patients with haematological malignancies after HLA-identical sibling BMT. Five μg/kg of rhuG-CSF was given daily by subcutaneous bolus injection, commencing four hours after marrow infusion and continuing until the neutrophil count was ≥ 1.0 × 10⁹/L on three consecutive days. Graft-versus-host disease (GVHD) prophylaxis was cyclosporin and short-course methotrexate (days 1, 3, 6 and 11). Prophylactic intravenous (IV) antibiotics were administered from the onset of neutropenia. The control group consisted of patients with comparable diagnoses, transplanted before and after the current study using identical supportive care and GVHD prophylaxis policies. Results: Although time to recovery of the neutrophil count to >0.1 × 10⁹/L was similar, the rhuG-CSF-treated patients experienced accelerated recovery to > 0.5 × 10⁹/L, which occurred at a median of 15 days (range 11–21) after marrow infusion in study patients compared to 18.5 days (range 14–41) in the controls (p = 0.04). No significant differences were detected in any of the indices of transplant-related morbidity examined, including the number of days of fever, the incidence of culture-positive infections, the usage of antibiotics, the requirement for parenteral nutrition and IV morphine, the maximum severity of mucositis and GVHD, and the day of discharge. Conclusion: Within the context of this study, rhuG-CSF had limited impact on the clinical outcome of HLA-identical sibling BMT. (Aust NZ J Med 1994; 24: 541–546.)     

Characterization of granulocyte colony-stimulating factor receptor expressed on human lymphocytes

We have studied the characterization of granulocyte colony-stimulating factor receptor (G-CSFR) in human lymphocytes. About one-third to one-quarter of the B lymphocytes from peripheral B-cell sources displayed G-CSF binding on the two-colour immunofluorescence study. The rate of G-CSFR-expressing (G-CSFR+) B cells was higher in bone marrow and cord blood than in peripheral blood, spleen and tonsil. G-CSFR expression was greater in the surface immunoglobulin D (IgD)-positive (sIgD+) B-cell population, but scarce in the sIgD- B-cell population. In tonsil, G-CSFR+ B cells were present among the cells with naive B and germinal-centre B phenotypes, but those with memory B phenotype were rarely found on triple-colour immunofluorescence analysis. Mitogen-activated, but not resting, T lymphocytes also showed G-CSF binding. Several continuous T- and B-cell lines expressed functional G-CSFR, because the addition of G-CSF enhanced the proliferative response of these cell lines. A sequence analysis of G-CSFR mRNA isoforms obtained from the T and B cells revealed that G-CSFR was derived from class I and class IV mRNA. Our results indicated that G-CSFR was constitutively expressed on the B-cell surface and was inducible in T cells.     

COP-BLAM regimen combined with granulocyte colony-stimulating factor and high-grade non-Hodgkin's lymphoma

Abstract: The clinical efficacy of COP-BLAM chemotherapy combined with human recombinant granulocyte colony-stimulating factor (G-CSF) was evaluated in 104 previously untreated patients with non-Hodgkin's lymphoma (NHL). According to the method of Laurence et al., a modified COP-BLAM regimen was administered every 21 days. G-CSF was added when the granulocyte count fell below 1000 × 10⁹/l. Ninety-eight of 104 (94.2%) patients achieved a complete remission; the 4-year survival rate was 82.4% with a median duration of observation of 26 months. Survival was significantly longer in patients with low serum LDH levels, B-cell type or low CRP or in earlier clinical stages, than in patients with high serum LDH levels, T-cell type or higher CRP levels or in advanced clinical stages. The mean duration of administration of G-CSF was 5.4 days. Twelve patients developed infections during treatment. The adverse effects of G-CSF included interstitial pneumonia, bone pain and fever. Patients administered COP-BLAM combined with G-CSF achieved a high rate of remission and had a low incidence of infection. Nearly all the patients could be treated in 21-day cycles. The results suggest that G-CSF combined with COP-BLAM was effective in treating NHL, because the patients can tolerate a higher dose of the anticancer agents.     

Absence of mutations in the granulocyte colony-stimulating factor (G-CSF) receptor gene in patients with myelodysplastic syndrome/acute myeloblastic leukaemia occurring after treatment of aplastic anaemia with G-CSF

The development of myelodysplastic syndrome/acute myeloblastic leukaemia (MDS/AML) has been reported in patients with aplastic anaemia (AA) after administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF). Similarly, patients with severe congenital neutropenia (SCN) have an increased risk of developing MDS/AML after treatment with rhG-CSF. Point mutations in the G-CSF receptor gene are found in about 20% of SCN patients who are predisposed to MDS/AML. We investigated the occurrence of mutations in the G-CSF receptor in eight patients with AA who developed MDS/AML. No mutations were detected around the cytoplasmic domain of the gene in our patients, indicating that the mechanisms of clonal evolution to MDS/AML in patients with AA might be different from those with SCN.