Biotherapy 4: 147-153, 1992. © 1992 Kluwer Academic Publishers. Printed in the Netherlands.
Levels of human serum granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor under pathological conditions Fusayuki Omori ~, Seiichi Okamura 2, Kazuya Shimoda 2, Takeshi Otsuka l, Mine Harada ~ & Yoshiyuki Niho ~
1The First Department of Internal Medicine; 2Cancer Center, Faculty of Medicine, Kyushu University, Maidashi 3-1-I, Higashi-ku, Fukuoka 812, Japan Received 10 June 1991; accepted 21 June 1991
Key words: enzyme-linked immunosorbent assay, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor Abstract
Levels of serum granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colonystimulating factor (GM-CSF) in patients with various leukocyte disorders were estimated by enzymelinked immunosorbent assay (ELISA). Some cases of acute myelogenous leukemia and aplastic anemia showed elevated serum levels of G-CSF and/or GM-CSF, whereas almost all of 23 healthy controls showed G-CSF and GM-CSF levels lower than 100 pg/ml. High levels of both types of CSF were noted in patients with granulocytosis due to infection. These levels became lower after resolution of the infection. Daily changes in serum CSF levels were also examined in a patient with autoimmune neutropenia, and it was found that the peripheral neutrophilic granulocyte count changed almost in parallel with the serum G-CSF level but not with GM-CSF, following the pattern with a delay of about 4-5 h, suggesting the possibility that G-CSF mainly regulates peripheral neutrophil circulation.
Introduction
Materials and methods
Granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are known to be powerful regulators of hematopoietic progenitors [1-5], and have already been applied in clinical trials [6-9]. However, reports about serum CSF levels have been scarce [10, 11] and there seems no study investigating simultaneously both G-CSF and GM-CSF levels in biological samples. Recently, we reported enzyme-linked immunosorbent assay (ELISA) methods for measuring the levels of G-CSF and GM-CSF [12, 13]. In this article, we report the kinetics of G-CSF and GM-CSF observed simultaneously in various disease states, and suggest some possible roles of these CSFs.
Serum samples Blood specimens were collected from each patient in the morning before breakfast, except where otherwise described, and the sera after centrifugation were stored at -20°C until the time of assay. All the blood collections were performed with informed consent.
Hematological diseases There were nine cases of acute myelogenous leukemia (AML). The types of AML, as diagnosed by the FAB criteria, included M1, 2 cases; M2, 2 cases; M3, 2 cases; M4, 2 cases; M5, 1 case. There were also four cases of chronic myelogenous leukemia (CML), four cases of acute lymphoblastic leukemia (ALL) and one
148 case of chronic lymphocytic leukemia (CLL). In addition, six patients had aplastic anemia, one had myelofibrosis and four had polycythemia vera. None of the patients had received any therapy or shown an apparent evidence of infection at the time of blood sampling. Twenty-three healthy adults, 21 to 45 years old, 10 males and 13 females, who had neither apparent disease manifestations nor abnormal white blood cell counts, were also selected for the normal controls.
Infections Eight patients with infective diseases (two with pneumonia, three with urinary tract infections, one with cholecystitis and two with bronchitis), who had absolute neutrophilic granulocyte counts (ANC) of more than 5000//xl, were examined. The sera were collected during the active phase of the disease and also after the resolution accompanied with normal ANC following suitable antibiotic therapies.
Lymphoma with chemotherapy Two patients with non-Hodgkin's lymphoma were examined. Patient 1 was a 32-year-old man with the diffuse medium cell type, and patient 2 was a 37-year-old man with the diffuse large cell type. Neither patient had any apparent evidence of infection during the examination periods. Sera were collected every few days from the beginning of CHOP therapy (cyclophosphamide, doxorubicin hydrochloride, vincristine and prednisolone), and the ANC were examined on the same days.
Autoimmune neutropenia Sera were collected every few hours from an 83-year-old woman with autoimmune neutropenia, and ANC estimation was performed at the same time. The patient had no apparent evidence of infection. Serum collection was started at 08.00 h before breakfast and done continuously every few hours until the following morning. The patient took periodic food three times and rested quietly in the hospital.
previously [12, 13]. The methods involved sandwich immunoassay with anti-G-CSF or anti-GMCSF murine monoclonal antibodies on the solid phase, and anti-G-CSF or anti-GM-CSF rabbit polyclonal antibodies for the second layer. The sensitivities to detect the lowest activities, were both equivalent to 50 pg/ml recombinant G-CSF [14] and recombinant GM-CSF [15]. Results
Healthy controls The G-CSF levels were below the sensitivity level of the assay (
Levels of human serum granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor under pathological conditions Fusayuki Omori ~, Seiichi Okamura 2, Kazuya Shimoda 2, Takeshi Otsuka l, Mine Harada ~ & Yoshiyuki Niho ~
1The First Department of Internal Medicine; 2Cancer Center, Faculty of Medicine, Kyushu University, Maidashi 3-1-I, Higashi-ku, Fukuoka 812, Japan Received 10 June 1991; accepted 21 June 1991
Key words: enzyme-linked immunosorbent assay, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor Abstract
Levels of serum granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colonystimulating factor (GM-CSF) in patients with various leukocyte disorders were estimated by enzymelinked immunosorbent assay (ELISA). Some cases of acute myelogenous leukemia and aplastic anemia showed elevated serum levels of G-CSF and/or GM-CSF, whereas almost all of 23 healthy controls showed G-CSF and GM-CSF levels lower than 100 pg/ml. High levels of both types of CSF were noted in patients with granulocytosis due to infection. These levels became lower after resolution of the infection. Daily changes in serum CSF levels were also examined in a patient with autoimmune neutropenia, and it was found that the peripheral neutrophilic granulocyte count changed almost in parallel with the serum G-CSF level but not with GM-CSF, following the pattern with a delay of about 4-5 h, suggesting the possibility that G-CSF mainly regulates peripheral neutrophil circulation.
Introduction
Materials and methods
Granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are known to be powerful regulators of hematopoietic progenitors [1-5], and have already been applied in clinical trials [6-9]. However, reports about serum CSF levels have been scarce [10, 11] and there seems no study investigating simultaneously both G-CSF and GM-CSF levels in biological samples. Recently, we reported enzyme-linked immunosorbent assay (ELISA) methods for measuring the levels of G-CSF and GM-CSF [12, 13]. In this article, we report the kinetics of G-CSF and GM-CSF observed simultaneously in various disease states, and suggest some possible roles of these CSFs.
Serum samples Blood specimens were collected from each patient in the morning before breakfast, except where otherwise described, and the sera after centrifugation were stored at -20°C until the time of assay. All the blood collections were performed with informed consent.
Hematological diseases There were nine cases of acute myelogenous leukemia (AML). The types of AML, as diagnosed by the FAB criteria, included M1, 2 cases; M2, 2 cases; M3, 2 cases; M4, 2 cases; M5, 1 case. There were also four cases of chronic myelogenous leukemia (CML), four cases of acute lymphoblastic leukemia (ALL) and one
148 case of chronic lymphocytic leukemia (CLL). In addition, six patients had aplastic anemia, one had myelofibrosis and four had polycythemia vera. None of the patients had received any therapy or shown an apparent evidence of infection at the time of blood sampling. Twenty-three healthy adults, 21 to 45 years old, 10 males and 13 females, who had neither apparent disease manifestations nor abnormal white blood cell counts, were also selected for the normal controls.
Infections Eight patients with infective diseases (two with pneumonia, three with urinary tract infections, one with cholecystitis and two with bronchitis), who had absolute neutrophilic granulocyte counts (ANC) of more than 5000//xl, were examined. The sera were collected during the active phase of the disease and also after the resolution accompanied with normal ANC following suitable antibiotic therapies.
Lymphoma with chemotherapy Two patients with non-Hodgkin's lymphoma were examined. Patient 1 was a 32-year-old man with the diffuse medium cell type, and patient 2 was a 37-year-old man with the diffuse large cell type. Neither patient had any apparent evidence of infection during the examination periods. Sera were collected every few days from the beginning of CHOP therapy (cyclophosphamide, doxorubicin hydrochloride, vincristine and prednisolone), and the ANC were examined on the same days.
Autoimmune neutropenia Sera were collected every few hours from an 83-year-old woman with autoimmune neutropenia, and ANC estimation was performed at the same time. The patient had no apparent evidence of infection. Serum collection was started at 08.00 h before breakfast and done continuously every few hours until the following morning. The patient took periodic food three times and rested quietly in the hospital.
previously [12, 13]. The methods involved sandwich immunoassay with anti-G-CSF or anti-GMCSF murine monoclonal antibodies on the solid phase, and anti-G-CSF or anti-GM-CSF rabbit polyclonal antibodies for the second layer. The sensitivities to detect the lowest activities, were both equivalent to 50 pg/ml recombinant G-CSF [14] and recombinant GM-CSF [15]. Results
Healthy controls The G-CSF levels were below the sensitivity level of the assay (
