Successful treatment of neutropenia in T-LGL leukemia (Tγ-lymphocytosis) with granulocyte colony-stimulating factor

Summary Severe neutropenia is a common feature in patients with T-large granular lymphocytic (LGL) leukemia. Neutropenia often causes severe infections and septicemia, thus representing a major cause of morbidity and mortality in this disease. We have treated two outpatients with T-LGL leukemia who had severe neutropenia (neutrophils <0.2×10⁹/l) successfully with G-CSF (5g/kg daily, s.c). After 10 days of treatment the neutrophil count was within the normal range and a severe oral infection healed rapidly. We conclude that G-CSF therapy is able to normalize the neutrophil count in T-LGL leukemia within a few days and that it can be used to treat severe infections in these patients even on an outpatient basis.     

Adsorptive granulocyte and monocyte apheresis for refractory Crohn’s disease: an open multicenter prospective study

Background Active Crohns disease (CD) is often associated with elevated levels of platelets, granulocytes, and monocytes that are activated and resistant to apoptosis. The level of neutrophils in the intestinal mucosa has been quantitatively related to the severity of intestinal inflammation in CD. We postulated that patients with CD that is refractory to conventional medications might respond to a reduction of granulocytes and monocytes by adsorptive apheresis.Methods Twenty-one patients with a CD activity index (CDAI) of 200–399 and unresponsive to standard medication, which included nutritional intervention, received granulocyte and monocyte adsorptive apheresis (GCAP) as an adjunct to their ongoing medication. GCAP was performed with an Adacolumn, which adsorbs granulocytes, monocytes, and a small fraction of lymphocytes (FcR and complement receptor-bearing leucocytes). Patients received one GCAP session/week for 5 consecutive weeks. CDAI, International Organization for the Study of Inflammatory Bowel Disease (IOIBD), and IBD questionnaire (IBDQ) scores were evaluated.Results During the initial conventional/nutritional therapy, no significant improvement was seen in any patient. However, at week 7 of GCAP therapy, significant improvements in CDAI, IOIBD, and IBDQ scores were observed. The CDAI, IOIBD, and IBDQ scores before GCAP were 275.6 ± 54.2, 3.4 ± 1.4, and 152 ± 22, respectively. The corresponding values after GCAP were 214.8 ± 89.2 (P = 0.0005), 2.54 ± 1.5 (P = 0.0224), and 165 ± 29 (P = 0.0327), respectively.Conclusions GCAP could be effective for inducing remission and improving quality of life in patients with active CD that is refractory to conventional therapy.     

Leukocyte numbers correlate with plasma levels of granulocyte–macrophage colony-stimulating factor in sickle cell disease

Despite a clear role for leukocytes in modulating the pathophysiology of sickle cell disease (SCD), the mechanism by which leukocyte numbers are increased in this disorder remains unclear. Hypothesizing that the chronic inflammatory state, elicited by adhesive interactions involving various cell types, might underlie leukocytosis, we measured plasma levels of proinflammatory or myeloid cytokines that play a role in leukocytosis and examined their correlations with leukocyte numbers in patients with SCD. Our studies found that, although plasma levels of granulocyte–macrophage colony-stimulating factor (GM-CSF), interleukin 3, and macrophage colony-stimulating factor are elevated in steady-state patients with SCD, only plasma GM-CSF levels are positively correlated with the numbers of total leukocytes, neutrophils, monocytes, and eosinophils, regardless of whether they received hydroxyurea. GM-CSF levels were significantly decreased in patients on hydroxyurea therapy. These data suggest a role of GM-CSF in leukocytosis of SCD. In contrast, plasma levels of granulocyte colony-stimulating factor, a major cytokine that induces leukocytosis due to bacterial infection, were lower than those of control subjects. These results indicate that elevated GM-CSF levels may contribute, at least in part, to high leukocyte numbers in SCD. As plasma GM-CSF levels were decreased in patients on hydroxyurea therapy, hydroxyurea may decrease leukocyte numbers by reducing circulating GM-CSF levels. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Synthesis of granulocyte–macrophage colony-stimulating factor as homogeneous glycoforms and early comparisons with yeast cell-derived material

Granulocyte–macrophage colony-stimulating factor (GM-CSF) is a medicinally important glycoprotein, used as an immunostimulant following bone-marrow transplant. On the basis of reports of its potential utility as an anticancer vaccine adjuvant, we undertook to develop a synthetic route toward single-glycoform GM-CSF. We describe herein a convergent total synthesis of GM-CSF aglycone and two homogeneous glycoforms. Analytical and biological studies confirm the structure and activity of these synthetic congeners.Granulocyte–macrophage colony-stimulating factor (GM-CSF) is a secreted glycoprotein that promotes cellular growth and proliferation. GM-CSF signaling initiates a cascade that culminates in the production of white blood cells through stem-cell stimulation in the bone marrow (Fig. 1) (1). Therapeutically, this glycoprotein is valued for its properties as an immunostimulant; GM-CSF impacts the production, differentiation, and function of dendritic cells through potentiation of the CD4⁺ T-cell response (2, 3) and is regularly administered to patients undergoing chemotherapy or autologous bone-marrow transplant. GM-CSF is approved in Europe as the aglycone (Molgramostim) (4) and in the United States as a glycosylated, mutant form (Sargramostim). At present, glycosylated GM-CSF is obtained exclusively via recombinant technologies using yeast (Sargramostim) or Chinese hamster ovary (CHO) cell (Regramostim) technologies, which yield complex mixtures of glycoforms. The glycan heterogeneity reflects a lack of specificity in CHO-cell posttranslational glycosylation. The degree of GM-CSF glycosylation has been reported to affect the in vivo properties of the glycoprotein (5, 6); the aglycone may cause increased adverse side effects, perhaps due to its enhanced susceptibility to truncation pathways (7).Open in a separate windowFig. 1.GM-CSF 3D structure and fully glycosylated GM-CSF sequence (1). The ribbon structure of GM-CSF is based on the X-ray of GM-CSF aglycone expressed by Escherichia coli.In light of our longstanding interest in synthetic anticancer and HIV vaccines (8), we took note of reports suggesting that GM-CSF might serve as a useful vaccine immunoadjuvant. Administration of GM-CSF results in robust potentiation of the immune response, and clinical studies suggest that the glycoprotein may hold promise as an adjuvant for anticancer vaccines (9, 10). However, studies to date have used recombinantly derived GM-CSF mixtures, and results have been inconclusive (9); perhaps such translational issues might be addressed in a more informative fashion with homogeneous GM-CSF agents. In a more speculative line of inquiry, we also wonder whether appendage of tumor-associated carbohydrate antigens to the protein backbone might yield powerful new anticancer vaccine candidates (11). Even beyond these fascinating medicinal questions, we recognized in GM-CSF a synthetically compelling glycoprotein target. With these considerations in mind, we undertook to design a modular route to homogeneous GM-CSF glycoforms. In this endeavor, we would rely upon key methodological and strategic advances from many groups, including ours, in the synthesis of complex glycoproteins. The past decade has indeed witnessed a dramatic maturation of the field of “biologics” through chemical synthesis. Our recent synthesis of homogeneous erythropoietin, bearing complex glycan domains at each native position, is illustrative of the growing power of chemical synthesis to facilitate the study of medicinally relevant biologic targets (12–14). We describe herein the synthesis of homogeneous GM-CSF glycoproteins through a convergent route that offers ready access to a menu of glycoforms for further study.Structurally, GM-CSF consists of 127 aa with multiple sites of glycosylation (Fig. 1). Two N-linked glycans are located at Asn²⁷ and Asn³⁷ (15, 16). Interestingly, neither the position nor the extent of O-linked glycosylation has been unambiguously established. Studies suggest a range from two (Ser⁷ and Ser⁹/Thr¹⁰) to four (Ser⁵, Ser⁷, Ser⁹, and Thr¹⁰) sites of glycosylation (17). The tertiary structure is strongly influenced by two cross-linked disulfide bonds.     

Bovine granulocyte chemotactic protein-2 is secreted by the endometrium in response to interferon-tau (IFN-τ)

Interferon-tau (IFN-τ) is secreted by the bovine conceptus and may regulate synthesis of uterine endometrial cytokines to provide an environment that is conducive to embryo development and implantation. Interferon-τ stimulates secretion of an 8-kDa uterine protein (P8) in the cow. P8 was purified, digested to yield internal peptides, and partially sequenced to determine identity. Two internal peptides had 100% (13-mer) and 92% (12-mer) amino acid sequence identity with bovine granulocyte chemotactic protein-2 (bGCP-2). Bovine GCP-2 is an α-chemokine that acts primarily as a potent chemoattractant for granulocyte cells of the immune system. A peptide was synthesized based on a region of bGCP-2 that overlapped with a P8 peptide amino acid sequence, coupled to keyhole limpet hemocyanin, and used to generate high titer polyclonal antiserum in sheep. Western blots revealed that bGCP-2 was not released by endometrium from day 14 nonpregnant cows, but was released in response to 25 nM IFN-τ (p<0.05). Uterine GCP-2 exhibited high affinity to heparin agarose, a characteristic shared by all α chemokines. This is the first report describing presence of GCP-2 in the uterine endometrium and regulation by IFN-τ. The regulation of bGCP-2 by IFN-τ may have important implications for cytokine networking in the uterus during pregnancy. Also, the regulation of inflammation and angiogenesis by bGCP-2 working together with other cytokines may be integral to establishing early pregnancy and implantation in the cow.     

Efficiency and safety analysis of Plerixafor combined with granulocyte colony-stimulating factor on autologous hematopoietic stem cell mobilization in lymphoma

<正>Objective To evaluate the advantages and safety of Plerixafor in combination with granulocyte colonystimulating factor(G-CSF) in autologous hematopoietic stem cell mobilization of lymphoma.Methods Lymphoma patients who received autologous hematopoietic stem cell mobilization with Plerixafor in combination with G-CSF or G-CSF alone were obtained.The clinical data,the success rate of stem cell collection,hematopoietic reconstitution,and treatment-related adverse reactions between the two ...     

Incidence of neutropenia and use of granulocyte colony-stimulating factors in multiple myeloma: is current clinical practice adequate?

Although immunomodulatory drugs, alkylating agents, corticosteroids, protease inhibitors, and therapeutic monoclonal antibodies improve multiple myeloma outcomes, treatment burden is still an issue. Neutropenia is a known complication of cytotoxic cancer therapy and is often associated with infections; it is an important consideration in myeloma given the fact that patients often have a weakened immune system. The risk of febrile neutropenia increases with severe and persisting neutropenia. Recombinant granulocyte colony-stimulating factors (G-CSFs) are commonly used to reduce the incidence, duration, and severity of febrile neutropenia. Here, we review the risk and management of neutropenia associated with new and commonly used anti-myeloma agents. Few papers report the use of G-CSF in patients with multiple myeloma receiving anti-cancer treatments, and fewer describe whether G-CSF was beneficial. None of the identified studies reported G-CSF primary prophylaxis. Further studies are warranted to evaluate the need for G-CSF prophylaxis in multiple myeloma. Prophylaxis may be particularly useful in patients at high risk of prolonged severe neutropenia.     

Passive anaphylaxis and IgE antibody production are compromised in tumor necrosis factor- and in granulocyte–macrophage colony stimulating factor-deficient mice

BackgroundA number of recent studies has demonstrated a critical role for mast cells and mast cell-derived cytokines, especially tumour necrosis factor (TNF), in the control of host defense mechanisms during inflammation. In the presesnt study, we investigated whether TNF-deficient (TNF^−/−) and granulocyte-macrophage colony stimulating factor (GM-CSF)-deficient (GM-CSF^−/−) mice expressed defects in normal mast cell function.MethodsBecause the first step in the passive cutaneous anaphylactic (PCA) reaction is fixation of the antibody to mast cells, we tried to obtain a PCA in TNF^−/− and GM-CSF^−/−mice.ResultsWhile an anti-dinitrophenyl IgE monoclonal antibody induced a strong PCA reaction in wild-type mice, it was not possible to obtain a PCA reaction in either TNF^−/− or GM-CSF^−/− mice. We next examined whether mast cells were present in these mice and if so, did they have functional FcεRI receptors on their surface. The number of mast cells in smears from the peritoneal fluid of the TNF^−/− and GM-CSF^−/− mice was similar to that seen in wild-type mice. However, the expression of FcεRI on mast cells from the peritoneal fluid of TNF^−/− and GM-CSF^−/− mice, measured by either rosetting assay or FACScan analysis, was compromised compared with wild-type mice. Previous studies have established that defects in FCεRI expression often have found that IgE production was compromised in both TNF^−/− and GM-CSF^−/− mice.ConclusionsThe observed defects may partially explain the immunodeficiency of these cytokine-deficient animals during infection.     

Interferon-α suppressed granulocyte colony stimulating factor production is reversed by CL097, a TLR7/8 agonist

Background and Aim: Neutropenia, a major side‐effect of interferon‐α (IFN‐α) therapy can be effectively treated by the recombinant form of granulocyte colony stimulating factor (G‐CSF), an important growth factor for neutrophils. We hypothesized that IFN‐α might suppress G‐CSF production by peripheral blood mononuclear cells (PBMCs), contributing to the development of neutropenia, and that a toll‐like receptor (TLR) agonist might overcome this suppression. Methods: Fifty‐five patients who were receiving IFN‐α/ribavirin combination therapy for chronic hepatitis C virus (HCV) infection were recruited. Absolute neutrophil counts (ANC), monocyte counts and treatment outcome data were recorded. G‐CSF levels in the supernatants of PBMCs isolated from the patients and healthy controls were assessed by enzyme‐linked immunosorbent assay following 18 h of culture in the absence or presence of IFN‐ α or the TLR7/8 agonist, CL097. Results: Therapeutic IFN‐α caused a significant reduction in neutrophil counts in all patients, with 15 patients requiring therapeutic G‐CSF. The reduction in ANC over the course of IFN‐α treatment was paralleled by a decrease in the ability of PBMCs to produce G‐CSF. In vitro G‐CSF production by PBMCs was suppressed in the presence of IFN‐α; however, co‐incubation with a TLR7/8 agonist significantly enhanced G‐CSF secretion by cells obtained both from HCV patients and healthy controls. Conclusions: Suppressed G‐CSF production in the presence of IFN‐α may contribute to IFN‐α‐induced neutropenia. However, a TLR7/8 agonist elicits G‐CSF secretion even in the presence of IFN‐α, suggesting a possible therapeutic role for TLR agonists in treatment of IFN‐α‐induced neutropenia.     

Adalimumab therapy following granulocyte and monocyte adsorptive apheresis in a patient with Crohn’s disease accompanied by chronic myeloid leukemia

A 52-year-old woman was diagnosed with Crohn??s disease (CD) of the large intestine in May 2001. Her disease was accompanied by the development of chronic myelogenous leukemia (CML) in December 2003. Remission of her CML has been maintained up to the present with tyrosine kinase inhibitors. Clinical and endoscopic remission of the patient??s CD was maintained with salazosulfapyridine (3000?mg/day) and occasional prednisolone (??20?mg/day) from 2001 to 2010. However, in December 2010 the patient complained of abdominal pain and diarrhea more than 10 times a day. Endoscopy showed serpiginous (snake-like) ulcers in the transverse colon and aphthous ulcers in the sigmoid colon. Intensive granulocyte and monocyte adsorptive apheresis (GMA) (two sessions per week, total of ten sessions) was performed, and the CD activity index (CDAI) decreased from 259 to 175. Six adalimumab injections were administered to improve the remaining inflammatory mucosa. Two months after induction therapy with adalimumab, the CDAI decreased from 175 to 107 without side effects. Endoscopy revealed mucosal healing of the colonic inflammatory lesions. We experienced a case of a patient with CD accompanied by CML. We successfully treated the patient by a combination of intensive GMA and adalimumab.     

Additive effects of PI3-kinase and MAPK activities on NB4 cell granulocyte differentiation: potential role of phosphatidylinositol 3-kinase γ

PURPOSE: In acute promyelocytic leukemia (APL) the chromosome translocation t(15;17) resulting in the PML-RARalpha fusion protein is responsible for a blockage of myeloid differentiation. In this study we investigated the expression of different Phosphatidylinositol 3-kinase (PI3K) isoforms during granulocyte differentiation of NB4 cells induced by all-trans-retinoic acid (ATRA), 9-cis-retinoic acid (9cisRA) or retinoic acid receptor (RAR) agonists. METHODS: NB4 cells were analysed for their ability to differentiate into granulocytic lineage by the use of ATRA, 9cisRA or RAR agonists. Expression of signalling proteins was investigated by western blot and real-time PCR. PI3K activity was determined by in vitro kinase assays. RESULTS: Co-treatment of NB4 cells with either LY294002 to inhibit PI3Ks or PD98059 in order to suppress MEK activity led to significant reduction of CD11b surface expression during ATRA, 9cisRA or the RARalpha agonist Ro40-6055 dependent NB4 cells granulocyte differentiation. We also show that only the G-protein coupled receptor activated PI3Kgamma isoform demonstrates up-regulated protein and mRNA expression during myeloid differentiation of NB4 cells via RARalpha and RARbeta-dependent mechanism. Furthermore, activation of MAPK cascade including phosphorylation of MEK increases during retinoid induced differentiation of NB4 cells. Interestingly, protein kinase assays of immunoprecipitated PI3Kgamma revealed a protein of about 50 kDa that is phosphorylated when NB4 cells were treated with the RARalpha agonist Ro40-6055. CONCLUSION: Collectively, our data suggest additive effects of PI3K and MAPK activity on ATRA-dependent NB4 cells granulocyte differentiation.     

Interleukin-2 and granulocyte–macrophage–colony-stimulating factor immunomodulation with high-dose chemotherapy and autologous hematopoietic stem cell transplantation for patients with metastatic breast cancer

Immunomodulation with cytokines was used to improve the result of high-dose chemotherapy (HDC)/autologous hematopoietic stem cell transplantation (AHST). We examined the use of IL-2 and growth factors for mobilization, ex vivo activation of peripheral blood stem cell (PBSC) and maintenance therapy after HDC/AHST in metastatic breast cancer. Eligible patients with metastatic breast cancer for HDC/AHST were assigned to 1 of 3 protocols for PBSC mobilization: G-CSF (group 1); IL-2 + G-CSF (group 2); or IL-2 + G-CSF + GM–CSF (group 3). HDC with cyclophosphamide, carmustine and thiotepa was given from day ?7 to ?5. PBSCs were treated ex vivo with IL-2 for 24 h and reinfused on day 0. Maintenance therapy included low-dose IL-2, followed by 2 courses of intermediate-dose IL-2. GM–CSF was given from day 1 until neutrophil recovery. Thirty-four patients (10 in group 1, 14 in group 2, and 10 in group 3) were included. Comparable numbers of CD34⁺ cells were collected from all 3 groups; incremental increases of CD3⁺ cells were collected from groups 1 to 2 and to 3 (p = 0.03). Major adverse effects from IL-2 were fever, hypotension and fatigue; no treatment-related mortality was seen. At a median follow-up of 790.5 days (range 150–2,722 days), median progression-free survival was 434 days and median overall survival was 1,432 days. Estimated 3-year progression-free and overall survival rates were 31 and 57%. Our study suggested that the use of IL-2 and growth factors immunomodulation with HDC/AHST was feasible with comparable survival rates.     

In enterovirus 71 encephalitis with cardio-respiratory compromise, elevated interleukin 1β, interleukin 1 receptor antagonist, and granulocyte colony-stimulating factor levels are markers of poor prognosis

Background.?Enterovirus 71 (EV71) causes large outbreaks of hand, foot, and mouth disease (HFMD), with severe neurological complications and cardio-respiratory compromise, but the pathogenesis is poorly understood. Methods.?We measured levels of 30 chemokines and cytokines in serum and cerebrospinal fluid (CSF) samples from Malaysian children hospitalized with EV71 infection (n?=?88), comprising uncomplicated HFMD (n?=?47), meningitis (n?=?8), acute flaccid paralysis (n?=?1), encephalitis (n?=?21), and encephalitis with cardio-respiratory compromise (n?=?11). Four of the latter patients died. Results.?Both pro-inflammatory and anti-inflammatory mediator levels were elevated, with different patterns of mediator abundance in the CSF and vascular compartments. Serum concentrations of interleukin 1β (IL-1β), interleukin 1 receptor antagonist (IL-1Ra), and granulocyte colony-stimulating factor (G-CSF) were raised significantly in patients who developed cardio-respiratory compromise (P?=?.013, P?=?.004, and P?100 at admission being the most accurate prognostic marker for death (P?相似文献   

Effect of interleukin-3, stem cell factor and granulocyte–macrophage colony-stimulating factor on committed stem cells, long-term culture initiating cells and bone marrow stroma in a one-step long-term bone marrow culture

Received: May 26, 1999 / Accepted: October 8, 1999