Human Intravenous Immunoglobulin Preparations Research Articles

Cross-Reactivity of Antibodies in Intravenous Immunoglobulin Preparation for Protection against SARS-CoV-2.

Severe cases of COVID-19 continue to put pressure on medical operations by prolonging hospitalization, occupying intensive care beds, and forcing medical personnel to undergo harsh labor. The eradication of SARS-CoV-2 through vaccine development has yet to be achieved, mainly due to the appearance of multiple mutant-incorporating strains. The present study explored the utility of human intravenous immunoglobulin (IVIG) preparations in suppressing the aggravation of any COVID-19 infection using a SARS-CoV-2 pseudovirus assay. Our study revealed the existence of IgG antibodies in human IVIG preparations, which recognized the spike protein of SARS-CoV-2. Remarkably, the pretreatment of ACE2/TMPRSS2-expressing host cells (HEK293T cells) with IVIG preparations (10 mg/mL) inhibited approximately 40% entry of SARS-CoV-2 pseudovirus even at extremely low concentrations of IgG (0.16-1.25 mg/mL). In contrast, the antibody-dependent enhancement of viral entry was confirmed when SARS-CoV-2 pseudovirus was treated with some products at an IgG concentration of 10 mg/mL. Our data suggest that IVIG may contribute to therapy for COVID-19, including for cases caused by SARS-CoV-2 variants, since IVIG binds not only to the spike proteins of the virus, but also to human ACE2/TMPRSS2. An even better preventive effect can be expected with blood collected after the start of the COVID-19 pandemic.

Naturally Acquired Human Immunity to Pneumococcus Is Dependent on Antibody to Protein Antigens.

Naturally acquired immunity against invasive pneumococcal disease (IPD) is thought to be dependent on anti-capsular antibody. However nasopharyngeal colonisation by Streptococcus pneumoniae also induces antibody to protein antigens that could be protective. We have used human intravenous immunoglobulin preparation (IVIG), representing natural IgG responses to S. pneumoniae, to identify the classes of antigens that are functionally relevant for immunity to IPD. IgG in IVIG recognised capsular antigen and multiple S. pneumoniae protein antigens, with highly conserved patterns between different geographical sources of pooled human IgG. Incubation of S. pneumoniae in IVIG resulted in IgG binding to the bacteria, formation of bacterial aggregates, and enhanced phagocytosis even for unencapsulated S. pneumoniae strains, demonstrating the capsule was unlikely to be the dominant protective antigen. IgG binding to S. pneumoniae incubated in IVIG was reduced after partial chemical or genetic removal of bacterial surface proteins, and increased against a Streptococcus mitis strain expressing the S. pneumoniae protein PspC. In contrast, depletion of type-specific capsular antibody from IVIG did not affect IgG binding, opsonophagocytosis, or protection by passive vaccination against IPD in murine models. These results demonstrate that naturally acquired protection against IPD largely depends on antibody to protein antigens rather than the capsule.

P012 Human IVIG preparations contain antibody to native and denatured HLA

Aim Human intravenous immunoglobulin (IVIG) is frequently used for treatment of antibody mediated rejection. Little is known regarding the risk of passively transferred HLA antibody (Ab) in these products. Our aim was to test commercial IVIG preparations for presence and level of HLA Ab and specificity. Methods Thirty IgG preparations derived from plasma collected in the US, Europe and Canada (CSL Behring: 6 Hizentra, 20% protein; 24 Privigen, 10% protein) were tested for HLA class I/II Ab in Luminex single antigen bead assays. Reagents were treated with Adsorb Out, then tested at various dilutions. Testing was performed with 2 vendor kits. Furthermore, preparations were tested on the C1q screening assay and after acid treatment. Results Initial testing showed HLA Ab reactivity (MFI >2000) at 1:10 but negative by 1:50. Regardless of protein concentration, IgG preparations had similar reactivity patterns. Following acid reduction, most class I beads remained positive with one vendor kit, suggesting the remaining reactivity was towards denatured and not native HLA molecules. As shown in the Table, class I Ab reactivity after acid treatment was limited to a few HLA specificities. Class I results with a second vendor kit were negative. Class II initial testing with one kit showed a similar reactivity pattern in 20/30 samples, mainly DR4+DR18, plus a few additional positive reactions; testing with the second kit gave MFI values Conclusions Ab reactivity in IgG preparations was low titer regardless of the percentage protein. All positive reactions were observed with One Lambda and not Immucor kits, and most positive class I reactions were towards denatured HLA molecules. Class II reactivity appeared to be non-specific. We conclude that these IgG preparations do not pose a risk for passively transferred HLA Ab that can potentially be harmful to the allograft. Research support and test materials provided by CSL Behring, LLC. Class I Luminex reagents provided by Immucor. M. Berger: Employee; Company/Organization; CSL Behring, LLC. M. Spycher: Employee; Company/Organization; CSL Behring AG. A. Zeevi: Grant/Research Support; Company/Organization; CSL Behring, LLC.

Preparation of commercial quantities of a hyperimmune human intravenous immunoglobulin preparation against an emerging infectious disease: the example of pandemic H1N1 influenza

The recent H1N1 pandemic provided an opportunity to conceptually assess the possibility of rapidly providing a "hyperimmune" human immunoglobulin (H-IVIG) to an emerging infectious disease, in useful quantities with respect to public health. Commercial-scale H-IVIG production from plasma collected from donors convalescent from or vaccinated against pandemic influenza A (H1N1) virus is described. A special protocol was implemented for the collection, processing, and shipment of plasma from previously qualified source plasma donors, self-identifying as convalescent from or vaccinated against H1N1 influenza. A licensed IVIG manufacturing process was utilized for the preparation of two commercial lots of approximately 50 kg 10% human IVIG preparation in total. The H1N1 hemagglutination inhibition and neutralization antibody titers of the resulting H-IVIG preparations were determined and compared with standard preparations. Twenty-six plasma collection centers participated in the protocol. Donor enrollment exceeded 300 donors per week and within 30 days of protocol deployment plasma was being collected at a rate of more than 2000 L/week. Manufacture of both H-IVIG lots was unremarkable and both lots met the requirements for commercial release and the bulk of the product was distributed in normal commercial channels. Examination of plasma pools and final IVIG product confirmed pandemic H1N1 antibody titers substantially higher than those collected before the emergence of the pandemic H1N1 virus. This work demonstrates the feasibility of producing a H-IVIG preparation at large scale relatively rapidly, with a significant enrichment in antibodies to the H1N1 influenza, achieved by donor self-identification.

Intravenous Immunoglobulin: Properties, Mode of Action and Practical Use in Dermatology

Since they were first administered to patients with antibody deficiency disorders over 50 years ago, human intravenous immunoglobulin preparations have been used successfully to treat a rapidly increasing number of autoimmune and inflammatory disorders, among which are a series of cutaneous autoimmune and inflammatory diseases. These include dermatomyositis, Kawasaki's disease, a number of autoimmune bullous diseases, severe adverse drug reactions, and other autoimmune and/or allergic conditions, such as atopic dermatitis. Although only a minority of these indications (dermatomyositis, Kawasaki's disease) are officially registered or based on double-blind, placebo-controlled clinical studies, the observed efficacy and safety profile of currently available intravenous immunoglobulin sometimes makes this a treatment of choice for initiation of therapy or for replacement of more toxic alternatives, such as systemic immunosuppressive medications. The increasing use of intravenous immunoglobulin has been associated with further understanding of its mechanism(s) of action, clinical manipulation and associated side-effects, as well as the introduction of improved or new types of intravenous immunoglobulin. This paper reviews the current knowledge of the mode of action of intravenous immunoglobulin, its reported therapeutic effects in cutaneous disease, its mode of administration and safety profile, and compares the currently available intravenous immunoglobulin preparations.

Inactivation of parvovirus B19 by liquid heating incorporated in the manufacturing process of human intravenous immunoglobulin preparations

Several reports have suggested the possible transmission of human parvovirus B19 (B19) through the administration of plasma derivatives that had undergone virus inactivation by various types of heat treatment. However, none of the reports evaluated and discussed the inactivation of B19 by the heat treatment that is implemented in the individual manufacturing processes of such products. The present study evaluated the ability to inactivate B19 of liquid-heat treatment at 60 degrees C for 10 h that was incorporated in the manufacturing process of intravenous human immunoglobulin preparations. The results showed that B19 was rapidly inactivated under the conditions used for the liquid-heat treatment.

Opsonic activity assessment of human intravenous immunoglobulin preparations against drug-resistant bacteria.

We have used the ability of opsonized bacteria to stimulate luminol-enhanced chemiluminescence (CL) of human polymorphonuclear leukocytes (PMN) to examine the opsonic capabilities of commercially available human intravenous immunoglobulin (i.v.Ig) preparations. The method was tested against 14 strains of drug-resistant gram-positive bacteria (including methicillin-resistant Staphylococcus aureus, hetero-vancomycin-resistant S. aureus, vancomycin-resistant enterococci, penicillin-resistant Streptococcus pneumoniae), and 23 strains of gram-negative bacteria (including extended-spectrum beta-lactamase-producing bacteria, metallo-beta-lactamase-producing bacteria, beta-lactamase-negative ampicillin-resistant Haemophilus influenzae). An Fc-intact i.v.Ig preparation treated with polyethylene glycol (PEG) was evaluated for opsonization effectiveness against these bacteria in vitro. The opsonization of these organisms was enhanced by an Fc-intact i.v.Ig, and the opsonic activity was dose dependent. A pepsin-treated i.v.Ig preparation exhibited poor opsonic activity for all bacteria tested. These results suggest that Fc-intact i.v.Ig, which augments opsonic activity against various drug-resistant bacteria, will be a useful addition to the treatment of severe bacterial infections in immunocompromised patients with impaired serum opsonic capacity.

In Vitro Cytotoxic Effects of Stabilizing Sugars within Human Intravenous Immunoglobulin Preparations against the Human Macrophage THP-1 Cell-line

Intravenous immunoglobulin (IVIg) is a safe and effective therapy for the treatment of primary and secondary humoral immune deficiencies and autoimmune disorders. Both minor and more serious side effects may occur following IVIg administration in approximately 1-15% of infusions and stabilizing sugars found in IVIg preparations may contribute some of these. In this report, we aimed to determine the cytotoxic effects of IVIg as compared with four stabilizing sugars (glucose, sucrose, maltose and D-sorbitol) found in IVIg preparations on human monocyte-macrophages. The human THP-1 macrophage cell-line was used as a model to determine the effects of stabilizing sugars and IVIg preparations on cell viability and growth. The sugars differentially affected the viability of THP-1 cells. In experiments using doses of the sugars commonly found in IVIg preparations, cell viability and proliferation was unaffected when compared with doses of IVIg typically administered to patients (5 mg/ml). However, in an LDH-release cell lysis assay that measures changes in cell permeability, glucose (50 mg/ml) induced significant release of LDH as compared with complete IVIg (5 mg/ml, p<0.0001). Intranucleosomal DNA fragmentation was not detected at therapeutically relevant doses of IVIg. This suggested that THP-1 cell death was not due to apoptosis. We conclude that osmotic stress mediated by the sugars at high doses promoted THP-1 cell death. We propose that IVIg per se is not cytotoxic to the autonomously growing human THP-1 cell-line but rather, the stabilizing sugars used in the preparations are the cytotoxic factors. This observation was evident when preparations of IVIg were used at high concentrations but not at levels one would associate with clinically relevant doses of IVIg.

Human Intravenous Immunoglobulin Preparation and Virus Inactivation by Pasteurization and Solvent Detergent Treatment

ABSTRACT Human intravenous immunoglobulin (IVIG) solutions were prepared by two different methods and compared to each other. The crude immunoglobulin fraction obtained from Cohn-Oncley fractionation of plasma was further purified and subjected to virus inactivation, either by polyethylene glycol precipitation and pasteurization at 60°C for 10 hours, or by ion exchange chromatography and solvent/detergent treatment.

Intravenous immunoglobulin preparation increases myoplasmic calcium concentration by activating the dihydropyridine–ryanodine receptor complex

A human intravenous immunoglobulin preparation (IVIg) released Ca 2+ from the sarcoplasmic reticulum of cultured human skeletal muscle cells in a dose-dependent manner. Blocking the dihydropyridine–ryanodine receptor complex abrogated the IVIg-mediated Ca 2+ response, whereas inhibition of the voltage-operated Na +-channels or acetylcholine receptors did not. This effect of IVIg was not mediated by its main component, the IgG molecules, and differed between preparations from different manufacturers. Heating destroyed the activity. Data shows that an unidentified serum protein present in IVIg can influence human muscle cells by an effect on the dihydropyridine receptor. This phenomenon may be important in interpreting the (side) effects of IVIg in neuromuscular diseases.

High-Dose Intravenous Immunoglobulin Modifies Complement-Mediated In Vivo Clearance

AbstractThe mechanism of effect of high-dose intravenous immunoglobulin (IVIG) therapy in immune cytopenias is incompletely known. One of the leading theories ascribes the short-term effects of IVIG to the competition of infused IVIG for Fc receptors, thereby inhibiting IgG-mediated clearance. Using a system independent of IgG-Fc receptor interactions, we examined another potential mechanism of IVIG action. Guinea pigs were infused with a human IVIG preparation at 600 mg/kg/day for two consecutive days. Parallel groups of animals were treated with the same volume and/or concentration of saline and albumin. Clearance of IgM-sensitized guinea pig erythrocytes, which is wholly complement dependent, was significantly retarded in animals treated with high-dose IVIG. The effect was specific for IVIG, since human albumin (as a second foreign protein) failed to change the clearance of IgM-sensitized guinea pig erythrocytes. Experiments in which IVIG- treated animals were subjected to pre- and posttreatmentclearance studies revealed heterogeneity among individual animals in respect to their response to IVIG infusions. Decrease of available plasma complement components did not account for the effect, since both C3 and CH50 values remained unchanged after IVIG treatment, despite rising levels of IVIG in sera of treated animals. The results of in vitro C3 uptake studies and the effect of IVIG on clearance of preopsonized cells suggest that IVIG produces a kinetic depression of C3 uptake and modifies the process of complement fragment deposition on erythrocytes. A generalized effect on mononuclear phagocytes is less likely but cannot be wholly ruled out. These studies establish another potential mechanism of IVIG action and suggest extension of its use to other complement-mediated diseases.

High-dose intravenous immunoglobulin modifies complement-mediated in vivo clearance

The mechanism of effect of high-dose intravenous immunoglobulin (IVIG) therapy in immune cytopenias is incompletely known. One of the leading theories ascribes the short-term effects of IVIG to the competition of infused IVIG for Fc receptors, thereby inhibiting IgG-mediated clearance. Using a system independent of IgG-Fc receptor interactions, we examined another potential mechanism of IVIG action. Guinea pigs were infused with a human IVIG preparation at 600 mg/kg/day for two consecutive days. Parallel groups of animals were treated with the same volume and/or concentration of saline and albumin. Clearance of IgM- sensitized guinea pig erythrocytes, which is wholly complement dependent, was significantly retarded in animals treated with high-dose IVIG. The effect was specific for IVIG, since human albumin (as a second foreign protein) failed to change the clearance of IgM- sensitized guinea pig erythrocytes. Experiments in which IVIG-treated animals were subjected to pre- and posttreatment clearance studies revealed heterogeneity among individual animals in respect to their response to IVIG infusions. Decrease of available plasma complement components did not account for the effect, since both C3 and CH50 values remained unchanged after IVIG treatment, despite rising levels of IVIG in sera of treated animals. The results of in vitro C3 uptake studies and the effect of IVIG on clearance of preopsonized cells suggest that IVIG produces a kinetic depression of C3 uptake and modifies the process of complement fragment deposition on erythrocytes. A generalized effect on mononuclear phagocytes is less likely but cannot be wholly ruled out. These studies establish another potential mechanism of IVIG action and suggest extension of its use to other complement-mediated diseases.

Topically administered immunoglobulin reduces pulmonary respiratory syncytial virus shedding in owl monkeys.

A human intravenous immunoglobulin preparation containing neutralizing antibodies to respiratory syncytial virus (RSV) was utilized for intratracheal and topical treatment of laboratory-induced RSV infections in owl monkeys (Aotus trivirgatus). Topically administered antibodies induced significant reductions in RSV shedding in the lower respiratory tract.

Functional antibacterial activity of a human intravenous immunoglobulin preparation: in vitro and in vivo studies.

The functional antibacterial activity of an intravenous immunoglobulin preparation (Swiss Red Cross Immunoglobulin, SRK-Ig) was tested both in vitro and in vivo. Using type III group B streptococci as a model system, the intravenous immunoglobulin preparation was shown to enhance phagocytosis and killing of the bacteria by human neutrophils. There was good activity to several type III strains, and for efficient opsonization both antibody and complement were required. In an animal model of neonatal group B streptococcal sepsis, passively administered immunoglobulin enhanced survival with 27/37 (73%) treated animals surviving compared to only 3/16 (19%) controls. These studies demonstrate that the intravenous immunoglobulin preparation SRK-Ig has retained functional activity and may have potential value for prophylaxis or therapy of certain bacteria infections.

Functional Antibacterial Activity of a Human Intravenous Immunoglobulin Preparation: in vitro and in vivo Studies

Functional Antibacterial Activity of a Human Intravenous Immunoglobulin Preparation: in vitro and in vivo Studies