Γ-globulin Preparations Research Articles

Altered expression of cell membrane HLA-G molecules in mothers of children with inborn heart defects upon exposure to plasma gamma-globulin from multiparous women

High incidence of newborns with inborn heart defects (HD) and sufficient contribution of this disorder into perinatal, infant and pediatric mortality, as well as disability levels, even after radical surgical treatment determine significance of search for novel methods of prediction and prevention of appropriate HD risks at the stage of pregnancy planning. HLA-G is among key molecules participating in immune interactions between maternal microenvironment and embryos. Maximal antibody titers for HLA antigens is detected in multiparous women. However, the question remains open, which HLA molecules participate in blockage of immune inflammation in the mother-embryo system: either by maternal antibodies, or by donor immune globulins. Hence, the aim of our study was to obtain enriched γ-globulin preparation from blood of multiparous women and evaluation of its activity towards female HLA-G molecules. The γ-globulin fraction (GGF) was obtained from blood plasma of muliparous women by means of affine chromatography using DEAE Affi-Gel Blue system (BioRad, USA). We have formed 2 groups: a control group (14 healthy males), and experimental group of 14 women who gave birth to the children with inborn heart defects.The changes of HLA-G expression on lymphocytes exposed to GGF were evaluated according to a flow cytometry protocol with calculation of percentage values using appropriate quotients. Evaluation of functional activity exerted by the GGF concentrate in control group showed inhibition of HLA-G expression on CD3+ and CD--lymphocytes against effects of autologous serum. GGF effects in experimental group upon HLA-G expression were differently directed, e.g., GGF sufficiently inhibited membrane HLA-G expression on the CD3-negative lymphocytes, compared to control group. Meanwhile, GGF showed stimulatory effect upon CD3+-lymphocytes, or it did not show any inhibitory action. The preparation obtained may serve as prototype for intravenous infusion. Moreover, in future one may use such immunoglobulin preparations, both for immune prophylaxis of non-syndromal sporadic HDs at the pregragravidary stage, as well as at early terms of pregnancy. The therapeutic effect will achieved due to blockage of embryoblast HLA molecules available to recognition.

Regulation ofin vitro human lymphocyte blast transformation associated with γ-globulin fraction containing a carbohydrate component

Carbohydrate-enriched components were isolated from human serum γ-globulin by chelation of associated transition metals. Similar to the initial γ-globulin preparation, these components stimulatein vitro blast transformation of human lymphocytes from healthy donors and inhibit subsequent phytomitogen-induced lymphocyte blast transformation.

A Seral Epidemiological Study of HIV Transmitted through Human Seral ΓGlobulin Preparations

In order to study the potential risk of transferring HIV through human seral gamma-globulin preparations (immunoglobulin), indirect immunofluorescent antibody test (IFA) and Western Blot (WB) assay were applied to 343 random samples (sera) with previous injection of imported human seral gamma-globulins (Ig) positive for Human Immunodeficiency Virus (HIV) antibodies between 1981-1987 for the detection of HIV antibodies. All results were negative and tests on all 23 controls who had previously received Ig made in China also gave negative results. However all 12 batches of imported Ig collected from the above-mentioned users, were positive for HIV antibodies when tested by WB and IFA. This study shows that under normal conditions, human seral gamma-globulin does not transmit HIV.

Intravenous γ-Globulin Use in Children With Kawasaki Disease

A recent controlled American trial1 of treatment for Kawasaki disease compared the use of acetylsalicylic acid (aspirin) alone v aspirin at the same dose plus a single preparation of IV γ-globulin, which is not currently licensed in the United States. The dosage of IV γ-globulin was 400 mg/kg/d for four days, and the dosage of aspirin was 80 to 100 mg/kg/d for as many as 8 weeks. The results indicated that, if administered within ten days of onset of fever, the combined treatment significantly lowered the prevalence of coronary artery dilation and aneurysms detected at 2 and 7 weeks postenrollment. The IV γ-globulin-treated group showed significant and rapid reduction in fever, WBC count, and generalized inflammatory signs. Prior studies in Japan also showed benefit from IV γ-globulin.2 Because the etiology of Kawasaki disease is not known and the active component of the immune globulin has not been identified, it is not known whether all IV γ-globulin preparations currently licensed in the United States will be effective. Further studies are underway to compare different IV γ-globulin preparations and to determine the optimal dose and schedule of administration. Pediatricians should consider using IV γ-globulin and aspirin for the treatment of patients who meet the strict criteria for this syndrome.3 If IV γ-globulin is to be used, therapy should be initiated with any one of the products licensed in the United States and administered within ten days of the onset of illness at the dosage schedule described above. The efficacy of such therapy if initiated more than ten days after onset of illness or after aneurysms have been found has not yet been demonstrated.

Elimination of undesirable immunoglobulin contaminants including aggregated IgG from γ-globulin preparations by jackfruit lectin affinity chromatography

Jackfruit lectin, jacalin, has been characterized to combine with IgA of only IgAl subclass but not with IgA2 subclass and other immunoglobulins. However, we found in the present study that a lectin from another batch of jackfruit (Jacalin-H) showed a different nature in binding reaction. Jacalin-H combined with immunoglobulins of every class or subclass except monomer IgG. More interestingly, the Jacalin-H also combined with aggregated IgG. By means of the Jacalin-H affinity chromatography, we could efficiently eliminate aggregated IgG as well as immunoglobulins other than monomer IgG from commercial γ-globulin preparations. Jacalin-H affinity chromatography is easy, inexpensive and efficient in the elimination of undesirable immunoglobulin contaminants in the commercial γ-globulin preparations.

Treatment of pneumococcal postsplenectomy sepsis in the rat with human γ-globulin

A rat model was used to evaluate the possible effect on experimental postsplenectomy sepsis of a human γ-globulin preparation for intravenous use (Sandoglobulin). Sixty splenectomized male Sprague-Dawley rats were given 3 × 10 3 Streptococcus pneumoniae type 1 intravenously. Twelve of the animals received no treatment and all died, in contrast to 12 sham-operated controls which all survived the challenge. The remaining splenectomized rats were divided into four groups, each consisting of 12 animals. One group was given 120 mg human γ-globulin twice intraperitoneally (0.3 g/kg body wt), at 18 and 42 hr, after challenge; 10 of the 12 survived, in contrast to none of the 12 in the second group receiving 120 mg human albumin instead of γ-globulin ( P = 0.00003). When the injections were delayed to 24 and 48 hr, 9 12 γ-globulin-treated animals still survived, in contrast to 0 12 in the albumin group. These findings point to new possibilities for treatment and perhaps prevention of overwhelming postsplenectomy sepsis by administration of high doses of γ-globulins.

Effects of serum and serum components on the binding of DNA to thioglycollate-stimulated mouse peritoneal macrophages

Binding of ssDNA and dsDNA to thioglycollate-stimulated peritoneal macrophages derived from normal CBA mice was increased not only by ssDNA-specific antibody but also by albumin and γ-globulin preparations. Fresh mouse serum inhibited ssDNA binding, even if the ssDNA and serum were preincubated at 37°C. Heat decomplemented mouse serum also depressed DNA binding. These results are significant to interpretations of DNA clearance rate data from serum or blood, and subsequent inferences concerning blood DNA levels, autoimmune response, and systemic lupus erythematosus.

Properties of intact and univalent (Fab) antibodies raised against isolated, solubilized, mouse zonae pellucidae.

Intact and univalent antibodies were prepared against mechanically isolated mouse zonae pellucidae solubilized in a variety of ways (heat, low pH, SDS, urea and trypsin). The antisera bound avidly and specifically to solubilized iodinated zona antigens and the intact zona structure. When the concentrations of immunoreactive Fab material in the intact and univalent antibody preparations were equalized and compared for their ability to block the sperm-binding stage of fertilization, only the intact gamma-globulin preparations possessed antifertility activity. These results indicate that antibodies raised against intact solubilized zonae pellucidae block fertilization by cross-linking antigens on the outer zona surface, thereby indirectly masking the sperm receptor sites. The integrity of these surface components did not appear to be affected by solubilization procedures that disrupt non-covalent bonds (heating, low pH, SDS and urea) although they did appear to be adversely affected by trypsin treatment. None of the antisera tested contained antibodies directed against the sperm receptor site indicating that these critical components lack immunogenicity.

Effect of human plasma proteins on stabilisation of platelet anti-aggregatory activity of prostacyclin.

Following the demonstration that biological activity of prostacyclin is more stable in plasma than in buffer solutions at physiological pH, we investigated the possibility that a plasma protein may be responsible for this effect. The duration of platelet anti-aggregatory activity of prostacyclin in fatty acid-free human albumin solution was significantly longer than in buffer solutions. The duration of this 'protection' was proportional to the concentration of albumin; alpha, beta, and gamma-globulin preparations had no 'protective' effect. The degree of 'protection' by albumin solutions was lower than that by plasma despite identical albumin concentrations. This discrepancy may in part be explained by the tendency of plasma to become alkaline on standing, a change that would tend to stabilise prostacyclin. The clinical relevance of our findings is discussed.

IgG Subclasses in Human γ-Globulin Preparations for Intravenous Use and Their Reactivity with Staphylococcus Protein A

In two of four non-enzymatically treated γ-globulin preparations C Immunoglobulin Schura, Immunoglobulin SRK), the distribution of IgG subclasses was found to be close to that of normal human serum. In two other preparations (sulphonated and β-propiolactone- treated) lgG3 was not detectable by means of appropriate antiserum. The IgG residual portion of plasmin-treated γ-globulin was enriched in IgG2, while IgG3 was absent. In affinity chromatography on protein A Sepharose, IgG3 in the unbound and IgG1, IgG2 and IgG4 in the bound fractions were found in Immunoglobulins Schura and SRK. In the sulphonated preparation no IgG was found in the unbound fraction, while IgG1, IgG2 and IgG4 were eluted from the bound fraction. In β-propiolactone-treated γ-globulin IgG1, IgG2 and IgG4 were present in both fractions. The testing of reactivity of IgG subclasses with Staphylococcus protein A can supply important information about the state of the Fc part in immunoglobulin preparations.

Standard and Special Human Immune Serum Globulins as Therapeutic Agents

Human γ-globulin is a valuable and frequently prescribed biological used to provide passive immunity against microbial agents, toxins, or cells. γ-Globulin is most widely used for the short-term prevention of certain bacterial or viral diseases when active immunization is unavailable (eg, hepatitis A) or when active immunization has not been given before exposure (eg, tetanus, rabies). γ-Globulin is also indicated on a continuous basis in replacement therapy for persons with antibody immunodeficiency. It is also (mis)used by many physicians for colds, allergies, and recurrent infection despite lack of proved efficacy. Two types of γ-globulin preparations are available: standard human immune serum globulin (HISG) for general use, and special human immune serum globulins with a known antibody content for specific illnesses (Table I). In addition, certain animal sera and antitoxins are available for passive immunity (Table I). Although a discussion of these agents is beyond the scope of this review, it is important to emphasize that their use is associated with considerable risk of adverse side reactions (anaphylaxis and serum sickness). Accordingly, the physician should know the exact indications for their use, always institute sensitivity testing, and be ready to treat side reactions immediately.1 SOME PROPERTIES OF IgG GLOBULIN The immunoglobulins or "γ-globulins" are the proteins of the plasma and tissue made in lymphoreticular tissues which have antibody activity.2 Although there are six classes of immunoglobulin—IgG, IgM, IgA, IgD, IgE, and secretory IgA—only IgG is present in significant quantities in HISG. IgG is a glycoprotein with a molecular weight of 150,000 daltons distributed equally between the serum and the tissues.

Distribution of IgG Subclasses in Commercial and Some Experimental γ-Globulin Preparations

The IgG subclass distribution was determined in six commercial and in four experimental human γ-globulin preparations. The concentrations of IgG subclasses were measured in a modified radioimmunoassay using subclass-specific antisera. In commercial γ-globulins, the distribution of the subclasses corresponded roughly to the distribution in normal human serum. A considerable enrichment of the IgG 4 was found in experimental lots prepared either from the ethanol fraction III or from the rivanol-precipitable IgG.

EFFICACY OF PROPHYLACTIC GAMMA-GLOBULIN IN PREVENTING NON-A, NON-B POST-TRANSFUSION HEPATITIS

Of 279 cardiac-surgery patients receiving a mean of twelve transfusions, 47 had significantly increased transaminase concentrations 14 to 180 days postoperatively and 10 were icteric. Preoperatively, each patient randomly received high-titre HbsAb γ-globulin, normal γ-globulin, or placebo and was followed at intervals for 9 months. Only 3 patients had serological evidence of hepatitis-B infection. 3 additional patients had serological evidence of cytomegalovirus infection, while none had evidence of hepatitis-A or Epstein-Barr infection. Less icteric hepatitis occurred in patients receiving the γ-globulin preparations (p=0·003), and the overall frequency of hepatitis was significantly reduced when compared with recipients of placebo. The protective effects of the two γ-globulin preparations were not significantly different. Most post-transfusion hepatitis today is neither viral hepatitis type B nor type A, and its severity and transmission are reduced by pre-transfusion γ-globulin.

Some immunochemical properties of natural antiglobulin factors (homoreactants)

A naturally occurring antiglobulin factor against the Fab-fragment of homologous IgG, contained in the rabbit and human blood serum and γ-globulin preparations, was shown by gelfiltration on Sephadex G-200 to have a molecular weight of about 250,000 daltons. In man this factor does not pass through the placenta in normal pregnancy. Despite differences in the physicochemical and effector properties of 7S IgG and protein with homoreactant activity, the latter has the specific antigenic determinants of IgG. These observations suggest that a complex of IgG with another protein or nonprotein compound possesses homoreactant properties.

The Binding of Thyrotropin to Isolated Bovine Thyroid Plasma Membranes

Abstract Plasma membranes have been prepared from bovine thyroid cells and have been characterized as being reasonably free of other subcellular structures by electron microscopy and enzyme marker analysis. Bovine thyrotropin specifically binds to these membranes, and this binding can be correlated with adenylate cyclase activation. The binding of thyrotropin to the membranes is immediate, is inversely dependent on temperature, is inhibited by increasing salt concentrations, and is not affected by a range of mono-, di-, and trisaccharides. The thyroid-stimulating activity of the thyrotropin preparations is unaffected by incubation with membranes under conditions of the binding assay. Both the α and the β subunits of bovine thyrotropin bind to thyroid membranes, β better than α on a molar basis; only the β subunit, however, inhibits thyrotropin binding at the concentrations tested. Mouse tumor thyrotropin is a potent inhibitor of bovine thyrotropin binding; human thyrotropin is not as effective. Since γ-globulin preparations with or without long acting thyroid-stimulating activity inhibit thyrotropin binding only slightly at high γ-globulin concentrations, the long acting thyroid stimulator itself appears to have no effect on thyrotropin binding to thyroid membranes.

Recent Advances in our Understanding of Hepatitis B Infections

The development of highly sensitive hemagglutination (HA) assays for detection of hepatitis B antibody has made possible observation of primary anamnestic antibody responses in most individuals exposed to hepatitis B virus and the development and evaluation of a hyperimmune γ-globulin preparation. The hemagglutination inhibition assay, while more sensitive than previous techniques for the detection of hepatitis B antigen (HB-ag) carriers, does not presently appear suitable for use in routine screening of blood donors. However, use of this assay to subtype the antigenic specificities of HB-ag makes it a valuable identity test for ultrasensitive assays. Prospective serologic follow-up of children newly admitted to an institution for the mentally retarded and studies on HB-ag carriers and their families (by hemagglutination inhibition and HA techniques) have revealed that the ratio of infection to clinical disease in hepatitis B infections is greater than 28:1.

A radioimmunoassay of human TSH, employing a solid phase second antibody, and a purified globulin preparation for standardization of nonspecific protein interactions

A modified immunoassay system for measurement of h-TSH in serum has been developed. Differences between standards and samples in nonspecific protein-protein interactions were minimized by adding a purified bovine γ-globulin preparation to the standards. For separation of bound and unbound tracer hormone a solid phase second antibody, anti-rabbit γ-globulin, coupled to cellulose, was used. In 37 healthy adults mean serum TSH was 2.0 μU/ml (SD 1.53 μU/ml), whereas in 27 hyperthyroid and 45 primary hypothyroid patients 0.9 μU/ml (SD 1.78 μU/ml) and 59.5 μU/ml (SD 33.9 μU/ml) were found respectively. The assay system showed a sensitivity of 0.5 μU/ml. The coefficient of variation within assays was 9.5% at the level of 20 μU/ml, and between assays was 2.4% at the level of 6 μU/ml. Exogenous h-TSH was completely recovered.

Immunological properties of isolated protein fractions from Artemia ribosomes

Acid-soluble ribosomal proteins from cysts of Artemia salina were separated by high-resolution polyacrylamide gel electrophoresis at pH 4.3. Three distinct protein bands, occurring in different parts of the electrophoretic pattern, were used for immunization in rabbits, and the γ-globulin fractions of the antisera were prepared. These preparations produced precipitation lines in agarose gel with protein extracted from whole 80S ribosomes and from 60S and 40S ribosomal subunits. With γ-globulin preparations from non-immune or anti-ovalbumin sera no reactions were obtained. In gel diffusion experiments the three γ-globulin preparations were reacted with the corresponding antigens in homologous and heterologous combinations, as well as with other disc-electrophoretic fractions of Artemia ribosomal proteins. Precipitation lines were formed in all combinations. Immunological precipitates were also obtained with rat ribosomal protein. 125I-labelled γ-globulins against the ribosomal proteins reacted to a very limited extent with intact 80S ribosomes, and only slightly more with EDTA-treated ribosomes. The amino acid incorporating activity of the ribosomes was not impaired by preincubation with the γ-globulins.

Reduction and alkylation of immunoglobulins from normal, irradiated and splenectomized rabbits.

SummaryThe L chain patterns of reduced and alkylated γ-globulin preparations from splenectomized, irradiated and normal rabbits were compared by starch gel electrophoresis. Animals irradiated with their spleen shielded produced γ-globulin with L chains similar to those of splenectomized and normal rabbits. Thus, the L chains formed in the spleen apparently do not differ electrophoretically from the L chains of γ-globuln formed in other organs.I wish to thank Dr. F. Haurowitz, under whose supervision this study took place, for helpful discussion and suggestions, L. Hopper for technical assistance, and Dr. Charles D. Barnes for help and advice in irradiation of the animals.

Intravenous Administration of Modified Gamma Globulin

Persons injected intravenously with human γ-globulin may develop symptoms which include anxiety, flushing of the face, a constricting feeling in the chest, muscle pain in the trunk or thighs, nausea and vomiting, fever, and rarely collapse. 1-3 In one study seven of 54 subjects receiving 110 ml of 1.5% γ-globulin solution intravenously experienced a reaction. 1 However, readministration, if within 72 hours, was well tolerated even by those who previously had reacted unfavorably. Individual susceptibility as well as rate of infusion, total amount given and perhaps the specific γ-globulin preparation may be determinants in the frequency and severity of symptoms. Peculiarly, the incidence of systemic reactions was extremely high in those with antibody deficiency syndromes such as primary agammaglobulinemia. 1,4 By contrast, pepsin treated preparations of γ-globulin elicited few or no symptoms when given to people with a variety of diseases including agammaglobulinemia. 5-7 The present report describes symptoms and laboratory findings observed during