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PubMed1994

Gc globulin (vitamin D-binding protein) increases binding of low concentrations of C5a des Arg to human polymorphonuclear leukocytes: an explanation for its cochemotaxin activity.

The chemotactic activity of native human C5a des Arg is enhanced significantly by the normal serum and plasma protein Gc globulin (vitamin D-binding protein). Gc globulin attaches to sialic acid residues within the oligosaccharide chain of C5a des Arg to form a complex with potent chemotactic activity for human PMN. We investigated the mechanism whereby this phenomenon may occur and found that Gc globulin enhanced the binding of low concentrations of [125I]C5a des Arg to PMN, but had no effect on C5a-induced displacement of bound [125]C5a des Arg. Gc globulin bound to PMN, and probably acted as a C5a des Arg chaperon. Thus, it appears that Gc globulin, by complexing to C5a des Arg, increases the number of C5a des Arg molecules per unit of PMN membrane without affecting its affinity of binding. This phenomenon provides a plausible explanation for the enhancing effect of Gc globulin on the chemotactic activity of low concentrations of native human C5a des Arg.

Chemotaxis, Leukocyte↗

PubMed1995

Treatment of systemic and renal-limited vasculitic disorders with pooled human intravenous immune globulin.

Idiopathic crescentic glomerulonephritis is characterized by an absence of immunohistological evidence of immune deposits, often with evidence of segmental glomerular necrosis. Such pauciimmune crescentic glomerulonephritis is the most common renal manifestation seen in patients with Wegener's granulomatosis, polyarteritis nodosa, and glomerulonephritis associated with other systemic vasculitic disorders (i.e., Churg-Strauss syndrome). Recently, the idiopathic crescentic glomerulonephritides, either in renal-limited form or in association with other systemic vasculitic disorders, were found to have in common a serologic marker, antineutrophil cytoplasmic autoantibodies. These cytoplasmic and perinuclear antineutrophil cytoplasmic autoantibodies are specific for constituents of neutrophil primary granules and monocyte lysosomes. As serologic markers for vasculitic disorders, they are also felt to be directly involved in the pathogenesis of necrotizing vascular injury. In vitro, both perinuclear and cytoplasmic antineutrophil cytoplasmic autoantibodies are capable of causing cytokine-primed neutrophils to undergo degranulation and respiratory burst, releasing toxic oxygen species and lytic enzymes. Anti-idiotype antibodies which inhibit antineutrophil cytoplasmic autoantibodies in vitro, in a V region-dependent manner, are found in pooled human gamma-globulin preparation. Intravenous immune globulin infusions in vivo have produced dramatic improvements in the necrotizing vascular injury produced by antineutrophil cytoplasmic autoantibodies, and a rapid reduction in these autoantibody levels is seen post-intravenous immune globulin infusion in most patients. The proposed mechanisms of action of intravenous immune globulin in vasculitic disorders include Fc-dependent mechanisms, and F(ab')2-dependent mechanisms are likely important. Intravenous immune globulin infusions appear to have an important place in the management of the necrotizing vascular injury. Blinded, randomized, placebo-controlled trials will be necessary to establish definitely intravenous immune globulin as a therapeutic option in vasculitic disorders.

Antibodies, Antineutrophil Cytoplasmic↗

PubMed1985

Cutaneous T cell lymphoma. Lymphocyte phenotype analysis after anti-thymocyte globulin therapy.

In an attempt to explore new or alternative therapy for refractory cutaneous T cell lymphoma, a patient with advanced disease refractory to conventional chemotherapy was treated with anti-thymocyte globulin. The therapy was associated with tolerable and manageable toxicities. Phenotypic analysis of neoplastic cells in blood and tissue was used to follow the disease response during the trial. The therapy resulted in resolution of skin lesions and disappearance of circulating cells with neoplastic phenotype. Marked improvement in skin infiltrate and restoration of normal skin architecture seen in skin biopsy specimens after anti-thymocyte globulin were prominent correlative findings. Lymphocytes with normal T cell phenotype appeared after anti-thymocyte globulin therapy, although their mitogenic response was subnormal. The infused anti-thymocyte globulin was found to localize in the skin and bind the neoplastic cells in circulation. The anti-thymocyte globulin-bound cells, with a half-life of 4.9 days, could still be detected up to 14 days after completion of anti-thymocyte globulin therapy.

Antibodies, Monoclonal↗

PubMed1976

Thyroxine-binding globulin biosynthesis in isolated monkey hepatocytes.

Thyroxine-binding globulin biosynthesis was demonstrated in hepatocytes isolated from normal adult Rhesus monkeys. Dispersed cells were obtained by in situ liver perfusion with collagenase, hyaluronidase and EDTA. Conditions for optimum cell survival and incorporation of radioactive leucine into newly synthesized proteins were defined. Protein synthesis, and specifically thyroxine-binding globulin synthesis, were shown to continue throughout the incubation period, while cell survival remained high (75% excluded trypan blue after 6h). Incubation medium, cytosol and a particulate fraction (extracted with digitonin) were analyzed for thyroxine-binding globulin. After extensive dialysis and purification by affinity chromatography, newly synthesized thyroxine-binding globulin was identified by specific double-antibody immunoprecipitation and by immunodiffusion and immunoelectrophoresis with autoradiography. Newly synthesized thyroxine-binding globulin was present after 4 h of incubation. After 6 h, the total synthesized had increased to 150% of the 4 h value, while the fraction present in the medium and increased to 300%, indicating probable thyroxine-binding globulin secretion

Animals↗

PubMed1986

Antitoxin in human pertussis immune globulins.

The level of antitoxin i.e. neutralizing antibodies to pertussis toxin, or lymphocytosis promoting factor, was determined in six pertussis immune globulin preparations from different manufactures. A comparison with antitoxin levels after natural pertussis disease in adults showed that pertussis immune globulins did not contain more antitoxin than convalescent phase sera, i.e. they had very low antitoxin content for specific immune globulins. Agglutinin and anti-FHA titres were relatively higher in immune globulins, probably reflecting a difference between the antibody response elicited by whole cell vaccines used for hyperimmunization in immune globulin production and by natural disease. The low antitoxin content of currently available pertussis immune globulin preparations could explain the inefficacy or conflicting results obtained with these products in prophylaxis and therapy of whooping cough.

Agglutination Tests↗

PubMed1983

Comparison of human thyroxine-binding globulin purification by affinity chromatography procedures.

Three procedures for the isolation of thyroxine-binding globulin from human serum, using affinity chromatography on triiodothyronine (T3) linked to Sepharose (A), thyroxine (T4) linked to Sepharose (B) or T3 linked to epoxy-Sepharose (C) as the first purification step, were compared. With the use of additional purification steps, the three procedures yielded pure thyroxine-binding globulin without desialylation. With procedure A, the initial binding of T4-binding globulin to T3-Sepharose was very low, yielding a poor final recovery (17%). Procedure B gave the highest yield (35%) after a three-step purification, with a low T4 content (0.15-0.30 mol/mol). Procedure C also gave a high yield (28%) after only two purification steps, with a T4 content greater than 0.7 mol/mol. The microheterogeneity of T4-binding globulin obtained with these three procedures was demonstrated by isoelectric focusing: five major bands were observed between pH 4.1 and 4.6, and intermediate faint bands (often doublets) in the same pH range. However, with procedures A and C, the most acidic bands (pH 4.10-4.20) were always absent. Thyroxine-binding globulin was preincubated with radioactively labelled T3 or T4 and the hormone-protein complex was analyzed by isoelectric focusing. The binding of T3--compared to that of T4--was reduced in the most acidic protein subspecies. These results suggest differences in the thyroid hormone binding properties of the various subspecies of human T4-binding globulin.

Chromatography, Affinity↗

PubMed1987

Purification and partial characterization of a corticosteroid-binding globulin from hamster serum.

Our objective was to characterize and purify the corticosteroid-binding proteins in hamster pregnancy serum. When [3H]cortisol-labeled pregnancy and proestrous serum were subjected to native polyacrylamide gel electrophoresis, a single peak of specific steroid-binding activity was detected in each, with identical electrophoretic mobility. The steroid-binding affinity (Ka = 1.07.10(8) M-1 for cortisol) is typical of corticosteroid-binding globulin from other species, but the steroid-binding specificity (cortisol greater than testosterone greater than progesterone) is not. An ultraviolet photoaffinity-labeling protocol was developed using 17 beta-hydroxy-4,6-[1,2-3H]androstadiene-3-one ([3H]androstadienolone), permitting analysis of ultraviolet photoaffinity-labeled proestrous and pregnancy serum by two-dimensional polyacrylamide gel electrophoresis and fluorography. Both sera contained the same labeled protein species. Corticosteroid-binding globulin was purified from pregnancy serum by DEAE-cellulose chromatography followed by steroid affinity chromatography on androstadienolone-17 beta-hemisuccinate-ethylenediamine-AffiGel 10. The purified protein (Mr = 62,250; pI = 3.95; n = 1; Stokes radius = 3.5; S = 4-5) was determined to be a glycoprotein. When analyzed by gel filtration and two-dimensional polyacrylamide gel electrophoresis, purified corticosteroid-binding globulin behaved the same as in unfractionated serum, and when ultraviolet photoaffinity-labeled with [3H]androstadienolone, purified corticosteroid-binding globulin produced the same fluorogram spot pattern seen in unfractionated serum. A specific corticosteroid-binding globulin antiserum was raised in rabbits, and this antiserum reacted with a single spot in Western blots of unfractionated serum. Thus, hamster pregnancy serum was determined to have one corticosteroid-binding protein. This protein is identical to the corticosteroid-binding globulin found in proestrous serum, with a higher titer in pregnancy serum. No other steroid-binding component is observed in hamster serum.

Animals↗

PubMed1990

Interaction between corticosteroid binding globulin and activated leukocytes in vitro.

The interaction between human corticosteroid binding globulin and activated leukocytes is restricted to the granulocyte population, and is characterized by specific proteolytic cleavage of corticosteroid binding globulin which markedly reduces its steroid binding activity. A direct interaction between corticosteroid binding globulin and the activated cells appears to enhance this event, and does not involve cellular internalization of corticosteroid binding globulin or its proteolytic degradation products, which resemble those obtained after incubation of corticosteroid binding globulin with neutrophil elastase. These data suggest that corticosteroid binding globulin interacts with elastase on the surface of activated neutrophils, and may promote glucocorticoid delivery to these cells during inflammation.

Blotting, Western↗

PubMed1984

Improved results of cadaver renal transplantation with azathioprine, prednisone and antilymphoblast globulin.

From 1980 to 1982, 100 consecutive cadaver renal transplants were performed. All but 2 recipients received preoperative transfusion and all received an initial 2-week course of antilymphoblast globulin. A prospective controlled evaluation of high versus low maintenance prednisone, and antilymphoblast globulin versus intravenous methylprednisolone for first rejection therapy was done. Over-all 1-year graft and patient survivals were 77 and 96 per cent, respectively. Graft survival was equal in the high and low steroid groups. Antilymphoblast globulin was as effective as intravenous methylprednisolone in reversing first rejections. Graft survival was improved with better donor-recipient matched grafts. We conclude that excellent results can be obtained in transfused cadaver renal allograft recipients managed with azathioprine, prednisone and antilymphoblast globulin. The regimen of prophylactic antilymphoblast globulin, low maintenance prednisone and antilymphoblast globulin alone for first rejections is immunologically effective and steroid sparing.

Antilymphocyte Serum↗

PubMed2001

Novel hydrophobic ligand-containing hydrogel membrane matrix: preparation and application to gamma-globulins adsorption.

In this study, phenylalanine as a hydrophobic ligand was covalently attached to the co-monomer methacrylochloride. Then, poly(2-hydroxyethylmethacrylate-co-methacrylamidophenyalanine) [poly(HEMA-MAPA)] membranes were prepared by UV-initiated photopolymerization of HEMA and methacrylamidophenyalanine. The gamma-globulins adsorption onto these affinity membranes from aqueous solutions containing different amounts of gamma-globulins at different pH was investigated in a batch system. The gamma-globulins adsorption capacity of the membranes was increased as the ligand density on the membrane surface increase. The non-specific adsorption of the gamma-globulins on the pHEMA membranes was negligible. The adsorption phenomena appeared to follow a typical Langmuir isotherm. The maximum adsorption capacity (q(m)) of the poly(HEMA-MAPA4) membrane for gamma-globulins was 2.37 mg g(-1) dry membrane. The equilibrium constant (k(d)) value was found to be 1.61x10(-1) mg ml(-1). More than 87% (up to 100%) of the adsorbed gamma-globulins were desorbed in 120 min in the desorption medium containing 50% ethylene glycol in 1.0 M NaCl.

Journal Article↗

PubMed2001

Behaviour of a protein isolate from rapeseed (Brassica napus) and its main protein components - globulin and albumin - at air/solution and solid interfaces, and in emulsions.

The behaviour of a rapeseed protein isolate (RI) and its main protein components - globulin (RG) and albumin (RA) - in adsorbed and spread monolayers, as well as in emulsions was investigated. Tensiometry, film-pressure area and Langmuir-Blodgett-techniques, and emulsion parameters were used to characterise the behaviour of the rapeseed proteins at various interfaces. The adsorption isotherms for albumin showed a plateau value for the surface pressure (Pi(e)) of 11.6 mN/m at the low critical association concentration (CAC) of 5.6x10(-8) g/ml. Both values were found to be distinctly higher for the globulin and the protein isolate. The isotherms of a mixture of RG and RA, which corresponds to the composition of RI, seems to be a superimposition of the isotherms of RA and RG. Contact angle measurements showed that all samples used were able to form LB-layers and to make hydrophilic glass surfaces more hydrophobic and vice versa. The changes in contact angle were more pronounced on hydrophobic glass surfaces. Monolayer and emulsion characteristics are dominated by the interfacial properties of albumin. The maximum film pressure reached by globulin was only about 8 mN/m. The globulin also possesses the lowest emulsifying activity. From the mean molecular area calculated for spread globulin, it is concluded that the globulin maintains its globular conformation at surfaces, which explains the low surface activity. Albumin and the protein isolate were highly surface active in monolayers and emulsion formation. The slightly different interfacial behaviour of the protein isolate compared with the corresponding mixture is probably due to additional effects of non-protein components and a partially denatured state of the protein.

Journal Article↗

PubMed1990

Monoclonal IgM rheumatoid factor-like anti-globulins enhance the inhibitory effects of Plasmodium falciparum-specific monoclonal antibodies in vitro.

Monoclonal IgM rheumatoid factor-like anti-globulins were produced by in vitro stimulation of naive BALB/c spleen cells with lipopolysaccharide, and by hyperimmunization of mice with merozoites of Plasmodium falciparum, followed by fusion of the spleen cells to mouse myelomas. In vitro, these anti-globulins augmented the inhibitory effects of P. falciparum-specific polyclonal mouse sera and monoclonal IgG1 and IgG2b antibodies by binding to Fc fragments of IgG molecules attached to blood-stage parasites. In some instances, the presence of anti-globulins correlated with an increase in the number of schizonts which failed to disperse merozoites. In other cases, parasitaemia remained low in the absence of the schizont inhibition phenomenon, suggesting that anti-globulins contribute to host cell protection not only by agglutinating merozoites, but also by increasing the density of the antibody coat surrounding the parasites, thus interfering with parasite receptor-erythrocyte ligand interactions. The anti-globulins were not inhibitory when added to parasite cultures containing IgG not specific for P. falciparum. These results may help explain the function of IgM anti-globulins found at elevated serum levels in some patients with malaria or other chronic infectious diseases.

Animals↗

PubMed1998

Effect of inclusion of cashew globulin (Anacardium occidentale) to a casein diet on lipid parameters in rats.

The effect of inclusion of cashew globulin to a casein diet on lipid metabolism was studied in rats fed diets with two levels of cashew globulin meal. Inclusion of cashew globulin to a casein diet produced lower levels of total cholesterol, triacylglycerol and phospholipids in the serum and tissues and lower levels of serum lipoprotein cholesterol. There was decreased cholesterogenesis in the liver as evidenced by decreased activity of HMG CoA reductase and decreased release of lipoproteins into circulation. Rats fed cashew globulin along with casein also showed higher activity of LPL in the heart and adipose tissue and higher activity of LCAT. Increased hepatic diversion of cholesterol to bile acid synthesis and increased excretion of bile acids and sterols were also observed in these groups. Activity of glucose-6-phosphate dehydrogenase and malic enzyme was decreased in rats fed cashew globulin along with casein. This study demonstrates that cashew globulins included in the diet of rats are able to alter lipid metabolism which results in lower levels of lipid parameters in the serum and tissues.

Adipose Tissue↗

PubMedDate not supplied

Resolution of red cell compatibility testing problems in patients receiving anti-lymphoblast or anti-thymocyte globulin.

Passively acquired hemagglutinating antibodies may be detected in the serum of patients receiving horse anti-lymphoblast globulin (ALG) or anti-thymocyte globulin (ATG). Thirty-seven patients receiving ALG following renal transplantation were studied. Eight patients monitored daily all developed positive direct antiglobulin tests (DAT) and positive red cell antibody screening tests. Fifteen of 32 recipients developed red cell antibodies reactive at room temperature in saline, 4 of 32 in albumin at 37 degrees C, and 33 of 37 in the antiglobulin test. Horse globulin was detected on the red cells of all six recipients tested with rabbit anti-horse globulin. Ether eluates prepared from the red cells of 20 patients showed no specificity for common red cell antigens. Anti-human globulin (AHG), absorbed with ALG-coated red cells to remove the component in the AHG which was crossreacting with horse globulin, was used successfully for antibody screening and identification, direct antiglobulin testing, and/or the antiglobulin crossmatching of 27 ALG and ATG recipients, including five with red cell alloantibodies.

Animals↗

PubMed1975

Hepatitis B "immune" globulin: effectiveness in prevention of dialysis-associated hepatitis.

A randomized, double-blind, multicenter study of hepatitis prevention by immune serum globulin with high anti-HBs titer ("hepatitis B immune globulin") was carried out among 318 new patients and 296 staff members of renal dialysis units. Three milliliters of high titer globulin, repeated at four months, was compared with equal doses of intermediate or normal titer globulin. Among staff members, the cumulative percentages developing hepatitis or HBs Ag, or both, within eight months were 6.9, 11.7, and 11.1 in the high, intermediate, and normal titer groups respectively. The lower incidence associated with high titer globulin was not significant (P greater than 0.05). However, among the patients the respective percentages were 7.9, 21.3, and 23.1 and the lower incidence in the high titer globulin group was significant.

Antibodies↗

PubMed1990

Randomized controlled trial of a monoclonal antibody against the interleukin-2 receptor (33B3.1) as compared with rabbit antithymocyte globulin for prophylaxis against rejection of renal allografts.

Interleukin-2 is a major growth factor for activated T lymphocytes, and antibodies reacting with the Tac-chain component of the interleukin-2 receptor can prevent allograft rejection in animals. Because Tac chains are expressed only on a small fraction of activated lymphocytes, monoclonal antibodies against the interleukin-2 receptor may offer a more specific means of immunosuppression than polyclonal antilymphocyte globulin in prophylaxis against graft rejection. Therefore, we compared the immunosuppressive effect of 33B3.1, a rat monoclonal antibody against the human Tac chain, with the effect of a rabbit polyclonal antithymocyte globulin in a randomized study of 100 recipients of first renal transplants. Injections of 33B3.1 (10 mg per day) were tolerated well, whereas major side effects in 15 of 47 patients (32 percent) receiving antithymocyte globulin required discontinuation of treatment before day 14. The incidence of rejection episodes was not statistically different in the two groups at days 14, 30, 60, and 90 after transplantation. Patient and graft survival was also equal in the two groups at one year (96 and 85 percent, respectively, in both groups), and graft function was similar. The total number of infectious episodes within the first three months was lower in the 33B3.1 group than in the antithymocyte group (47 vs. 72). The drop in peripheral-blood lymphocyte concentrations was significantly larger in the patients treated with antithymocyte globulin. The level of circulating Tac-chain-bearing lymphocytes remained below 1 percent during 33B3.1 treatment, as compared with 4 to 5 percent during antithymocyte-globulin treatment (P not significant). We conclude that 33B3.1 is as effective as antithymocyte globulin in the prevention of renal-transplant rejection, and its use results in fewer infections and side effects.

Adolescent↗

PubMed1992

A randomized trial comparing intravenous immune globulin and plasma exchange in Guillain-Barré syndrome. Dutch Guillain-Barré Study Group.

BACKGROUND: The subacute demyelinating polyneuropathy known as Guillain-Barré syndrome improves more rapidly with plasma exchange than with supportive care alone. We conducted a multicenter trial to determine whether intravenous immune globulin is as effective as the more complicated treatment with plasma exchange. METHODS: To enter the study, patients had to have had Guillain-Barré syndrome for less than two weeks and had to be unable to walk independently. They were randomly assigned to receive either five plasma exchanges (each of 200 to 250 ml per kilogram of body weight) or five doses of a preparation of intravenous immune globulin (0.4 g per kilogram per day). The predefined outcome measure was improvement at four weeks by at least one grade on a seven-point scale of motor function. RESULTS: After 150 patients had been treated, strength had improved by one grade or more in 34 percent of those treated with plasma exchange, as compared with 53 percent of those treated with immune globulin (difference, 19 percent; 95 percent confidence interval, 3 percent to 34 percent; P = 0.024). The median time to improvement by one grade was 41 days with plasma exchange and 27 days with immune globulin therapy (P = 0.05). The immune globulin group had significantly fewer complications and less need for artificial ventilation. CONCLUSIONS: In the acute Guillain-Barré syndrome, treatment with intravenous immune globulin is at least as effective as plasma exchange and may be superior.

Follow-Up Studies↗

PubMed2006

Rabbit antithymocyte globulin versus basiliximab in renal transplantation.

BACKGROUND: Induction therapy reduces the frequency of acute rejection and delayed graft function after transplantation. A rabbit antithymocyte polyclonal antibody or basiliximab, an interleukin-2 receptor monoclonal antibody, is most commonly used for induction. METHODS: In this prospective, randomized, international study, we compared short courses of antithymocyte globulin and basiliximab in patients at high risk for acute rejection or delayed graft function who received a renal transplant from a deceased donor. Patients taking cyclosporine, mycophenolate mofetil, and prednisone were randomly assigned to receive either rabbit antithymocyte globulin (1.5 mg per kilogram of body weight daily, 141 patients) during transplantation (day 0) and on days 1 through 4 or basiliximab (20 mg, 137 patients) on days 0 and 4. The primary end point was a composite of acute rejection, delayed graft function, graft loss, and death. RESULTS: At 12 months, the incidence of the composite end point was similar in the two groups (P=0.34). The antithymocyte globulin group, as compared with the basiliximab group, had lower incidences of acute rejection (15.6% vs. 25.5%, P=0.02) and of acute rejection that required treatment with antibody (1.4% vs. 8.0%, P=0.005). The antithymocyte globulin group and the basiliximab group had similar incidences of graft loss (9.2% and 10.2%, respectively), delayed graft function (40.4% and 44.5%), and death (4.3% and 4.4%). Though the incidences of all adverse events, serious adverse events, and cancers were also similar between the two groups, patients receiving antithymocyte globulin had a greater incidence of infection (85.8% vs. 75.2%, P=0.03) but a lower incidence of cytomegalovirus disease (7.8% vs. 17.5%, P=0.02). CONCLUSIONS: Among patients at high risk for acute rejection or delayed graft function who received a renal transplant from a deceased donor, induction therapy consisting of a 5-day course of antithymocyte globulin, as compared with basiliximab, reduced the incidence and severity of acute rejection but not the incidence of delayed graft function. Patient and graft survival were similar in the two groups. (ClinicalTrials.gov number, NCT00235300 [ClinicalTrials.gov].).

Acute Disease↗