Neutralizing antibodies to granulocyte–macrophage colony‐stimulating factor, interleukin‐1α and interferon‐α but not other cytokines in human immunoglobulin preparations

Abstract

Summary Human immunoglobulin preparations are used therapeutically for various disorders. Such therapy is generally safe but adverse effects occasionally occur in recipients. It has been suggested that antibodies to cytokines present in clinical immunoglobulin products may contribute to undesirable effects in recipients. Therefore, we investigated intravenous and intramuscular immunoglobulin products for the presence of cytokine‐specific neutralizing antibodies. Using validated bioassays, we detected neutralizing activity against human granulocyte–macrophage colony‐stimulating factor (GM‐CSF), interferon‐α2a (IFN‐α2a) and interleukin‐1α (IL‐1α) in immunoglobulin products. We found no neutralization of granulocyte colony‐stimulating factor, macrophage colony‐stimulating factor, stem cell factor, IL‐1β, IL‐2, IL‐3, IL‐4, IL‐6, IL‐9, IL‐10, IL‐12, tumour necrosis factor‐α, oncostatin M (OSM) and IFN‐γ. Most batches which neutralized IFN‐α2a activity also neutralized other IFN‐α subtypes, IFN‐ω and IFN‐β. Most products (94%) neutralized the biological activity of GM‐CSF. No correlation between batches and their ability to neutralize bioactivities of GM‐CSF, IFN‐α2a and IL‐1α was found. This neutralizing activity could be traced to plasma pools used for manufacture of immunoglobulins. The neutralization was mediated by specific cytokine antibodies contained within immunoglobulin products as it was present in specific immunoglobulin G (IgG) fractions eluted from cytokine affinity chromatography columns. Specific binding of such IgG fractions to cytokines in immunoblots and in enzyme‐linked immunosorbent assays (ELISAs) was observed. This contrasts with the broad non‐specific recognition of cytokine proteins observed using unfractionated immunoglobulins in ELISAs. This is the first comprehensive study showing the presence of neutralizing antibodies against GM‐CSF, IL‐1α, or IFN‐α2a in immunoglobulin products.

Antibody-Mediated Protection in MurineCryptococcus neoformansInfection Is Associated with Pleotrophic Effects on Cytokine and Leukocyte Responses

“…Administration of polyclonal IgG has wide-ranging immunomodulatory effects, including modulation of Fc␥RIIB expression (50), stimulation of NO production (52), and down-regulation of multiple cytokines by monocytes and T cells in response to stimulation (3,4,46). In retrospect, the choice of polyclonal Ab was almost certainly not an appropriate control for a MAb, since polyclonal immunoglobulin preparations may also contain antibodies to chemokine receptors and cytokine-neutralizing antibodies (8,59). The issue of what is an appropriate control for specific Ab is extremely complex and depends largely on what question one is trying to answer.…”

Intravenous immunoglobulin prevents release of proinflammatory cytokines in human monocytic cells stimulated with procalcitonin

“…One mechanism of action of IVIG in infectious diseases is proposed to involve a variety of specific antibodies to bacterial cell components and toxins [16][17][18]. In addition, it has been reported that IVIG contains neutralizing antibodies against some cytokines such as granulocyte-macrophage colonystimulating factor, IL-1a and interferon-a2a [19]. In this study, we found that IVIG inhibited release of proinflammatory cytokines in human monocytic cells stimulated with PCT and might also contain a PCT-specific antibody.…”

Are neutralizing anti–GM-CSF autoantibodies present in all healthy persons?

“…In Uchida et al's study, 1 all healthy control sera tested were positive for anti-GM-CSF AA using an enzyme-linked immunosorbent assay. Although it is difficult to reconcile this observation with results from previous studies, [3][4][5]8 variations in assay design, format and serum factors may contribute to nonspecific reactivity and generate a false positive signal in the enzyme-linked immunosorbent assays. We believe that in demonstrating specificity, it is essential to show binding of GM-CSF to the F(abЈ) 2 fraction of anti-GM-CSF IgG and absence of binding of antibodies to other proteins (ie, non-GM-CSF).…”

“…6,7 Nevertheless, low-titer neutralizing anti-GM-CSF AAs have been found in human intravenous immunoglobulin (IVIg) products. 4,8 These originated from the plasma donated by relatively few donors having high levels of AA; a batch of IVIg that did not contain them was shown to originate from plasma pools devoid of neutralizing activity. 8 Paradoxically, Uchida et al's data 1 indicates that neutralizing anti-GM-CSF AAs are present in every individual, are bound to circulating GM-CSF and can regulate GM-CSF activities in vivo.…”

“…4,8 These originated from the plasma donated by relatively few donors having high levels of AA; a batch of IVIg that did not contain them was shown to originate from plasma pools devoid of neutralizing activity. 8 Paradoxically, Uchida et al's data 1 indicates that neutralizing anti-GM-CSF AAs are present in every individual, are bound to circulating GM-CSF and can regulate GM-CSF activities in vivo. Because the procedures used in this study to measure neutralizing anti-GM-CSF AAs are not accompanied by an explanation of the full complement of validation experiments and negative controls used, we have reservations about these conclusions.…”