Quantitation and identification of human monocytic colony-stimulating factor in human serum by enzyme-linked immunosorbent assay

Abstract

An enzyme-linked immunosorbent assay (ELISA) system for the quantitation of human monocytic colony-stimulating factor (hM-CSF) was established, which was based on the “dual antibody immunometric sandwich” principle using horse and rabbit polyvalent antibodies against human urinary colony-stimulating factor (CSF-HU). The minimal detectable level of hM-CSF was 10 U/mL, and the assays showed good reproducibility. As measured by this method, the average serum hM-CSF level of 20 normal adults was 540 +/- 110 U/mL (range, 300 to 800 U/mL). The peak of hM-CSF measured by ELISA was identical to that measured by bioassay when semipurified CSF-HU was fractionated by reversed-phase high performance liquid chromatography (HPLC). This method detected two types of hM-CSF, which had approximate molecular weights of 85 Kd (CSF-HU) and 45 Kd in human serum and urine; the ratio of 85:45 Kd was very high in serum and the amounts of the two types were nearly equal in urine. After anticancer chemotherapy, the serum hM- CSF level of one half of the patients with hematological malignancy was elevated according to the reduction in neutrophil number, while it was almost in the normal range in the other half of the patients, indicating the possibility that anticancer chemotherapy damaged the hM- CSF-producing cells. This ELISA method may be useful for monitoring the serum hM-CSF level after anticancer chemotherapy.

High serum levels of M‐CSF and G‐CSF in Kawasaki disease

“…The M-CSF concentration was determined by an ELISA system as previously described (Hanamura et al, 1988). The minimal detectable level of M-CSF was 10 U/ml.…”

Cyclic change of cytokines in a patient with cyclic thrombocytopenia

“…Each cytokine was assayed with ELISA commercially available except for M-CSF and TPO (Boehringer Mannheim, Germany; Amersham, U.K.; R&D Systems Inc., Minneapolis; Endogen Inc., Cambridge, Mass., U.S.A.). M-CSF ELISA was performed as reported previously (Hanamura et al, 1988). Each value was the mean of wells in duplicate.…”

Serum macrophage colony-stimulating factor (M-CSF) level is elevated in patients with old cerebral infarction related to vascular damage

“…The serum M-CSF level was measured by an enzymelinked immunosorbent assay (ELISA) using antihuman M-CSF horse antibody, antihuman M-CSF rabbit antibody and horseradish perioxidase-conjugated anti-rabbit immunoglobulin G goat antibody, as reported previously [11]. The plasma levels of von Willebrand factor (vWF), thrombomodulin (TM), D-dimer products of crosslinked fibrin degradation products (D-dimer), thrombinantithrombin III (TAT) complex, prothrombin fragment 1+2 (F1+2), and plasmin-antiplasmin (PAP) complex were measured by ELISAs using anti-vWF (Boehringer Mannheim, Mannheim, Germany) for vWF, TM Panacela (Fuji Rebio, Tokyo, Japan) for TM, Enzygnost enzyme immunoassay (EIA) kit (Behringwerke, Marburg, Germany) for TAT, Enzygnost F1+2 (Behringwerke) for F1+2, the Dimertest EIA kit (AGEN Biomedical, Acacia Ridge, Queensland, Australia) for D-dimer, and an ␣ 2 -PI complex EIA kit (Teijin, Tokyo) for PAP.…”

“…The serum levels of M-CSF have been reported to be elevated in patients with various diseases including in-fectious diseases [11], idiopathic thrombocytopenic purpura [12], aplastic anemia [13], and ovarian cancer [14], as well as in pregnant women [15]. These elevated M-CSF levels may indicate the activated functioning of monocytes-macrophages in these diseases.…”