Human Gamma Globulin Preparations Research Articles

The history and evolution of immunoglobulin products and their clinical indications

The history of providing antibodies to treat diseases began in the 19th century with the discovery of tetanus and diphtheria toxins and the demonstration that immunity to tetanus and diphtheria infections could be transferred by immune sera. Characterization of the mediators of this immunity resulted in the discovery that antibodies are proteins that can be isolated and used to protect against infectious diseases. Development of a method to isolate antibodies from human plasma that could be safely injected into people initiated the development of human gamma globulin preparations to provide antibodies to patients with inherited antibody deficiencies. To overcome the limitations imposed by intramuscular injection of gamma globulin, intravenous gamma globulin preparations were developed that began to be used in a wide variety of clinical conditions. Thus the original clinical indication for infection prevention was expanded to several other indications that employ large doses to suppress inflammatory and autoimmune disorders. The most recent development in immunoglobulin therapy is the production of concentrated immune globulins for subcutaneous injection. Home infusions of subcutaneous immunoglobulin are increasingly used to treat immunodeficient patients and are being studied for other clinical applications.

Hidden high-avidity anti-DNA antibodies occur in normal human gammaglobulin preparations

An ability to detect hidden high-avidity DNA-binding protein in human gammaglobulin samples was investigated. Using ion exchange chromatography on QAE-Sephadex A-50 a highly positive Farr assay DNA-binding fraction was reproducibly isolated from several commercial normal human gammaglobulin preparations. The estimated dissociation constant had a value of 1.04 x 10(-11) M thus confirming high avidity protein-DNA complex formation. Anti-DNA antibodies (Ab) ELISA data revealed that immunoglobulin (Ig) G, IgM and IgA participated in the protein-DNA interaction. Inhibitory experiments involving a number of polynucleotides, synthetic and natural polyanions demonstrated that both double-stranded DNA (dsDNA) and heat-denatured DNA but not RNA inhibited the protein-3H-DNA binding. Generally, the inhibiting effect was more pronounced when purine base-containing polynucleotides and polydeoxy- rather than polyribonucleotides were used. Synthetic polyanions and normal human sera (NHS) also markedly reduced the binding. The presence of hidden high-avidity DNA-binding antibodies in normal gammaglobulin preparations was suggested.

クレブシェラ肺炎による敗血症性ショックの1例及び基礎的検討

A 51-year-old male, who had a history of excessive drinking and chronic hepatitis, was admitted to our hospital because of high fever and shock. Physical examinations, chest X-ray films and hemodynamic data revealed that he had progressed to septic shock due to pneumonia. Combination chemotherapy of latamoxef plus piperacillin was immediately started. After Klebsiella pneumoniae was isolated from bronchoalveolar lavage fluid, the antibiotics were changed to ceftizoxime plus amikacin. Furthermore human gamma globulin preparations and frozen fractional plasma were administered because of granulocytopenia and a decrease in complement. Gabexate mesylate, methylprednisolone (MP) and branched chain amino acids were given to prevent disseminated intravascular coagulation and/or multiple organ failure. With this intensive care, he recovered from shock and pneumonia. The effects of MP on the whole blood chemiluminescence (CL) were examined. Incubation of whole blood with 25, 50 or 100 micrograms/ml of MP for 10 to 60 minutes had no effects on the CL response. This indicates that MP does not affect the production of reactive oxygen species from phagocytic cells at concentrations comparable to those used in drug therapy.

Cell-mediated cytotoxicity-supporting activity of various human gammaglobulin preparations.

Antibody activity, especially that involved in the reaction of antibody-dependent cell-mediated cytotoxicity (ADCC), of five commercially available human gammaglobulin preparations (standard, pepsin-treated, plasmin-treated, polyethylene glycol-fractionated and S-sulfonated gammaglobulin) was measured. All these gammaglobulin preparations had high titers of hemagglutination inhibition and neutralizing antibody against measles virus. In ADCC reaction, the pepsin-treated gammaglobulin preparation showed no antibody activity. The standard gammaglobulin preparation showed weak activity only when highly diluted. The remaining three preparations showed high activity. Though the S-sulfonated gammaglobulin preparation showed no activity in ADCC reaction, it showed high activity after reconversion by means of oxidation and reduction in vitro. The plasmin-treated gammaglobulin preparation showed greater activity than the polyethylene glycol-fractionated preparation of the optimal concentration. In ADCC tests using the plasmin-treated gammaglobulin preparation, K cell activity was strongly inhibited by Hg (thimerosal), while, in those using the standard gammaglobulin preparation, the activity was hardly influenced by Hg, suggesting that the low ADCC activity of the standard gammaglobulin preparation of high concentrations was due to the inhibitory effect of aggregated immunoglobulin G molecules.

Protective Effects of Human Gamma-Globulin Preparations against Experimental Aerosol Infections of Mice with Bordetella pertussis

Protective Effects of Human Gamma-Globulin Preparations against Experimental Aerosol Infections of Mice with Bordetella pertussis

Protective effects of human gamma-globulin preparations against experimental aerosol infections of mice with Bordetella pertussis.

The protective effects of three types of pooled human gamma-globulin preparations (intact: GG, S-sulfonated: GGS, and pepsin-treated: GGP) for intravenous use against experimental aerosol infection of mice with Bordetella pertussis have been evaluated. The gamma-globulin preparations GG and GGS showed significant protective activity whereas GGP did not, as evaluated by survival numbers, body weight gains and suppression of leucocytosis. GG and GGS but not GGP also possessed neutralizing activity against leucocytosis-promoting and histamine-sensitizing activities of pertussis toxin (PT). Evaluation of protective activities of GG, GGS and GGP prepared from rabbit gamma-globulin, highly immunized with PT and not containing any anti-F-HA (filamentous or agglutinin antibodies, demonstrated that anti-PT-GG and anti-PT-GGS but not anti-PT-GGP protected against experimental infection by B. pertussis. These studies showed that the protective activity was correlated with anti-PT titre but that the Fc portion of the gamma-globulin molecule is necessary for actual protection against whooping cough by B. pertussis.

Protective effects of human gamma globulin preparations against experimental aerosol infections of mice with Bordetella pertussis

We evaluated the protective effects of the three types of pooled human gammaglobulin preparations (intact: GG, S-sulfonated: GGS, and pepsin-treated: GGP) for intravenous use against experimental aerosol infection of mice with B. pertussis. Gammaglobulin preparations, especially, GG and GGS showed significant protective activity but GGP did not at least as evaluated by survival numbers, body weight gains and repression of leukocytosis. Nutralizing activity for tetanus and diphtheria toxins did not vary among the three types of gammaglobulin preparations, whereas GG and GGS but not GGP possessed neutralizing activity against leukocytosis promoting activity of pertussis toxin. Evaluation of protective activities of GG, GGS, and GGP, prepared from rabbit gammaglobulin highly immunized with purified PT and not containing any and-F-HA and agglutinin antibodies, demonstrated that and-PT-GG and anti-PT-GGS but not anti-PT-GGP protected against experimental infection by B. pertussis. These results suggest that the protective activity was correlated with anti-PT titer but the Fc portion of the gammaglobulin molecule might be necessary for actual protection against whooping cough by B. pertussis.

Mechanism of Antibody‐Mediated Protection against Herpes Simplex Virus Infection in Athymic Nude Mice: Requirement of Fc Portion of Antibody

Experiments were performed to investigate the resistance of the host due to antibody-mediated mechanisms to herpes simplex virus (HSV) infection. Transfer of hyperimmune anti-HSV mouse serum inhibited the development of skin lesions and prolonged the survival of lethally HSV-infected nude mice. Relatively high concentrations of antibody were required to achieve this protection. Antisera prepared in heterologous animals were also effective, while administration of anti-cowpox virus serum or interferon provided no protection. This type of protection is therefore due to specific antibody and cannot be attributed to interferon. In order to delineate the requirement for antibody in antibody-mediated protection, human gamma globulin preparations were transferred to lethally HSV-infected nude mice. Transfer of intact human gamma globulin (GG) was effective in controlling infection. S-sulfonation of GG did not diminish the protective ability. However, purified F(ab')2 did not have any protective action even when it was administered frequently to maintain serum neutralizing antibody titer. GG was effective in C5-deficient mice lethally infected with HSV. These results indicate that in vivo antibody-mediated protection to HSV infection requires the Fc region of the intact IgG molecule and suggest that antibody-dependent cell-mediated cytotoxicity may be operative in vivo.

Protective effect of human gamma globulin preparations on experimental bacterial infections in mice

Untreated human gamma globulin (Ig), pepsin-treated Ig and purified F (ab') 2 were examined for their protective effects on experimental bacterial infections in mice. All these preparations were similarly effective in protecting mice from death after intraperitoneal administration of Streptococcus pneumoniae NCTC 7465, Escherichia coli 81 or Serratia marscescens OH 942. The protective effect of pepsin-treated Ig decreased significantly when the Ig was absorbed with the bacteria used for infection of mice, suggesting that the protective effect of Ig is due to a specific antibody. Administration of Ig preparations inhibited growth of bacteria in the blood stream of infected animals. In vitro experiments with a macrophage cell line showed that treatment of bacteria with Ig preparations including F (ab') 2 stimulated phagocytic activity of the macrophages.

新鮮血(癌家系以外)・ヒトγグロブリン製剤及び生薬(特にPS-K, W. T. T. C.)併用による癌免疫療法の試み

新鮮血(癌家系以外)・ヒトγグロブリン製剤及び生薬(特にPS-K, W. T. T. C.)併用による癌免疫療法の試み

IgG subclasses in human gamma-globulin preparations for intravenous use and their reactivity with staphylococcus protein A.

In two of four non-enzymatically treated gamma-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 beta-propiolactone-treated) IgG3 was not detectable by means of appropriate antiserum. The IgG residual portion of plasmin-treated gamma-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 beta-propiolactone-treated gamma-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.

Induction and mode of action of suppressor cells generated against human gamma globulin. I. An immunologic unresponsive state devoid of demonstrable suppressor cells.

A model of unresponsiveness to human gamma-globulin (HGG) which is maintained in the absence of demonstrable suppressor cells has been described. A/J mice were tolerized with deaggregated HGG purified from a variety of sources. The spleen cells from these tolerized mice were assessed for their ability to suppress the response of normal spleen cells to HGG when transferred into lethally irradiated mice. All of the HGG preparations obtained from commercial sources as Cohn fraction II of pooled, outdated plasma induced suppressor cells to HGG, although not of equal magnitude. However, suppressor cells could not be demonstrated in the spleens of mice tolerized with deaggregated HGG purified from the plasma of a healthy individual. This inability to detect suppression was independent of the method of purification of the HGG and of the time of assessment of the putative suppressor cells after tolerization. Similarly, deaggregated HGG isolated from an IgG1 lambda-myeloma protein induced unresponsiveness to HGG but did not stimulate demonstrable suppressor cells. These data suggest that suppressor T cells are not involved in the maintenance of tolerance to this antigen, although they may play a regulatory role in the immune response to HGG. Support for this concept was obtained by assessing the duration of unresponsiveness in the T and B lymphocytes of mice tolerized with the various HGG preparations. Mice tolerized with the HGG preparations that stimulated little or no suppression were among the last to recover responsiveness. Indeed, there was no consistent correlation between the level of suppressor cell activity and the degree of unresponsiveness in either the splenic T or B lymphocytes. Thus, although certain HGG preparations may provide a tool for the generation of antigen-specific suppressor T cells, the utilization of these suppressive preparations may be inappropriate for the investigation of the mechanisms of the induction and maintenance of the unresponsive state.

Distribution of IgG subclasses in commercial and some experimental gamma-globulin preparations.

The IgG subclass distribution was determined in six commercial and in four experimental human gamma-globulin preparations. The concentrations of IgG subclasses were measured in a modified radiommunoassay using subclass-specific antisera. In commercial gamma-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.

Improved compatibility of ALG therapy by application of aggregate free globulin.

Horse anti-human lymphocyte globulin (ALG) solutions contain, like human gamma-globulin preparations, a varying amount of IgG aggregates, which increase during longer periods of storage. Aggregate formation does not impair the immunosuppressive potency of ALG, as has been shown by prolongation of skin allograft survival time in primates. Deaggregated ALG, which can be obtained by ultracentrifugation at 100,000 g over 2 h, is less immunogenic than aggregate containing ALG. The clinical compatibility of ALG was significantly improved by ultracentrifugal deaggregation.

Kinin formation in human blood serum induced by cooling and by heat-aggregated human gamma globulin preparations, HAHGG

The original kinin research at the Middlesex Hospital Medical School concerned, for many years, the occurrence of plasma kinin when the human plasma touched foreign surface such as glass (Armstrong et al, 1952, 1957; Keele & Armstrong, 1964; Armstrong, 1968a). More recently the work has centered upon the spontaneous occurrence of kinins when the human plasma (heparinized or citrated) or serum is cooled, e.g. 370 – 00 C, without contact with glass (Armstrong & Mills, 1965; Armstrong et al, 1967; Armstrong, 1968a,c). Although not all kinins are pa in-producing (see Armstrong, 1968a,c) the kinin produced by cooling of the human serum is, and in this it resembles the kinin induced by glass exposure of the serum. Both the kinin formation induced by glass and that induced by the cold exposure of serum are Hageman Factor dependent reactions (Margolis, 1957, 1958a,b, 1960; Armstrong, 1968c; see also Armstrong et al, 1967; Armstrong, 1968a,b and 1969a). They thus resemble one another and also the kinin formation induced by antigen-antibody precipitates (Movat, 1967; Movat et al, 1968; Boreham & Goodwin, 1969) which appears to be a Hageman Factor dependent reaction also.