Natural and Therapy-Induced Anti-GM-CSF and Anti-G-CSF Antibodies in Human Serum

Abstract

Serum samples were obtained from patients with lymphoid and plasma cell malignancies who received after chemotherapy human recombinant GM-CSF or G-CSF. Sera from some patients revealed the presence of anti-cytokine antibodies, particularly after repetitive cytokine injections. Antibody Fab binding in a saturable manner by ELISA and Western immuno-blotting confirmed antibody specificity. Anti-cytokine antibodies were detected before the exogenous cytokine injections in some patients, but increasing antibody levels were found after one or subsequent treatments. Low levels of anti-GM-CSF and anti-G-CSF antibodies were also detected in a relatively large proportion (about 10-30%) of normal sera from different adult healthy volunteers who had never been treated before with exologous cytokines as well as from cord blood. EBV-immortalized cord blood derived B-cell cultures were also found to produce anti GM-CSF and/or anti-G-CSF antibodies with high frequency.

N‐ and O‐linked carbohydrates and glycosylation site occupancy in recombinant human granulocyte‐macrophage colony‐stimulating factor secreted by a Chinese hamster ovary cell line

“…A significant higher proportion of the unglycosylated glycopeptide I (GP I, Leu25-Arg30) was detected compared to nonoccupied glicopeptide II (GP II, Asp31-Arg58), suggesting a more complete N-glycosylation at site II in the total GM-CSF preparation. In the case of the N-terminal tryptic O-glycopeptide [SPSPSTQPWEHV NAIQEAR (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)], the form with two [Hex-HexNAc] moieties clearly dominates with 15% of mono and of triply O-glycosylated peptides being present. In order to confirm the results about the site occupancy with N-glycans, the N-linked oligosaccharides of the tryptic digest were liberated by PNGase F digestion and the mass spectrometric analysis was repeated.…”

Immune‐mediated neutropenia in the neonate

“…Taniguchi et al reported decreased levels of myeloid precursor cells in two adults with autoimmune neutropenia associated with long-term administration of rhG-CSF (70). In patients with leukemias and lymphomas on prolonged cytokine therapy, rising levels of antibodies against G-CSF and GM-CSF have been demonstrated (71). There are also concerns about the effect of rhG-CSF on bone mineral content, with reports of bone pain, unusual fractures and a higher incidence of osteopenia (72).…”

Recombinant human granulocyte-macrophage colony-stimulating factor: effect of glycosylation on pharmacokinetic parameters

“…As a heavily glycosylated member of the haematopoietic cytokine family, reports support the key role of the carbohydrate chains of hGM-CSF in terms of toxicity (Denzlinger el al. 1993;Dorr, 1993) and immunogenicity (Gribben et al 1990;Revoltella et al 1997). …”