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PubMed1975

Studies on the stability of antilymphocyte globulins.

An account is given of the effect of storage under various conditions on the physical and immunosuppressive properties of antilymphocyte globulins. No loss of in vivo immunosuppressive potency was shown by horse antimouse lymphocyte globulin when stored for up to 2 years in a liquid, frozen, or lyophilised state. Similar studies on horse antihuman lymphocyte globulin showed no loss of in vitro activity, as measured by an immunofluorescence test. Ultracentrifugal analyses showed no increase in aggregate when antilymphocyte globulin was stored liquid or freeze-dried at 4 degrees C. Storage experiments at abnormally high temperatures demonstrated that under these conditions freeze-dried horse antihuman lymphocyte globulin was stabilised by the presence of glucose (4.6%w/v). Antilymphocyte globulin lyophilised in the presence of glucose showed improved solubility properties as compared with material dried from saline.

Animals↗
PubMed2002

CD3 monitoring of antithymocyte globulin therapy in thoracic organ transplantation.

BACKGROUND: Antithymocyte globulin is frequently used as a component of induction therapy in thoracic organ transplantation. This study evaluates the utility of monitoring peripheral CD3 lymphocytes to rationally adjust antithymocyte globulin therapy in this patient population. METHODS: A total of 17 heart and 19 lung transplant recipients received antithymocyte globulin (ATGAM or thymoglobulin) as induction therapy or to treat steroid-resistant acute or chronic rejection. Absolute CD3 counts were maintained between 50 and 100 cells/microl. RESULTS: With CD3 monitoring, the doses of antithymocyte globulin were reduced from 10-15 mg/kg to 1-5 mg/kg during the course of therapy. The total amount of antithymocyte globulin given to each CD3 monitored patient was reduced by 48%. Dose reduction did not alter the number of acute rejection or infectious episodes, and hematological side effects were infrequent. CONCLUSION: CD3 monitoring of antithymocyte globulin therapy in thoracic organ recipients reduced the amount of drug received by each patient, while maintaining CD3 counts less than 100 cells/microl.

Acute Disease↗
PubMed1984

Intracellular transport and posttranslational cleavage of oat globulin precursors.

The synthesis, transport, and posttranslational processing of reserve globulin in Avena sativa L. seeds were studied by pulse-chase labeling. Developing oat seeds were labeled with radioactive sulfate and tissue homogenates were used for globulin extraction.Two globulin precursors (58-62 kilodaltons) were labeled after 1 hour pulse. The alpha and beta globulin subunits appeared between 2 and 10 hours later, while simultaneously the 58 to 62 kilodaltons polypeptides gradually disappeared. This confirmed a precursor-product relationship. In a second pulse-chase experiment, the tissue extracts were fractionated on a sucrose gradient. The major portion of radioactivity was initially (1 hour pulse) associated with the endoplasmic reticulum. However, a significant amount of radioactivity shifted from the endoplasmic reticulum to protein bodies after 20 hours chase, suggesting the transport of the newly synthesized proteins. Protein bodies isolated from pulse-chased seeds were analyzed for the arrival of the newly synthesized globulin. Labeled precursors were detected after 2 hours chase and gradually disappeared. The alpha and beta subunits appeared during the same chase period and assembled into a 12S oligomer.The data indicated that oat globulin was synthesized as two large precursors which were transported from endoplasmic reticulum into protein bodies where they were processed to the alpha and beta subunits forming a 12S oligomer.

Journal Article↗
PubMed1985

Biosynthesis and Intracellular Transport of 11S Globulin in Developing Pumpkin Cotyledons.

In vitro studies to explore the biosynthesis of 11S globulin developing cotyledons of pumpkin (Cucurbita sp.) demonstrated that 11S globulin is synthesized on membrane-bound polysomes. M(r) of the translation products (preproglobulin) synthesized by the poly(A)(+)-RNA isolated from developing cotyledons were determined to be 64,000 and 59,000, which are larger than those of the mature globulin subunit (62,000 and 57,000). Preproglobulin is then cotranslationally processed by cleavage of the signal peptide to produce proglobulin. In vivo pulse-chase experiments showed the sequential transformation of the single-chain proglobulin to mature globulin subunit (disulfide-linked doublet polypeptides) indicating posttranslational modification of the proglobulin.Subcellular fractionation of the pulse-chased intact cotyledons showed that the [(35)S]methionine label is detectable in proglobulin in rough endoplasmic reticulum shortly after the pulse label. With time, the labeled proteins move into other cellular fractions: proglobulin in the density = 1.24 grams per cubic centimeter fractions after 30 minutes and mature globulin subunit associated with protein bodies after 1 to 2 hours. The distribution of proglobulin in sucrose density gradients did not correspond with those of catalase (microbody marker) or fumarase (mitochondria marker). An accumulation of proglobulin occurred in the density = 1.24 grams per cubic centimeter fractions, whereas the mature globulin was scarcely detectable in this fraction. In contrast, proglobulin was not detected by immunochemical blotting analysis in the protein bodies prepared under the mild conditions from cotyledon protoplasts. The results suggest that the d = 1.24 grams per cubic centimeter fractions are engaged in the translocation of proglobulin into the protein bodies.

Journal Article↗
PubMed1988

Molecular characterization of oat seed globulins.

We have isolated full-length cDNA clones that encode oat (Avena sativa) seed storage globulin mRNAs from a cDNA library in the expression vector lambda gtll. The longest of these clones, pOG2, has an 1840-base pair insert that encodes a complete precursor subunit with a signal peptide of 24 amino acids followed by an acidic polypeptide of 293 amino acids and a basic polypeptide of 201 amino acids. Near the C terminus of the acidic polypeptide are four repeats of a highly conserved, glutamine-rich octapeptide. Other oat globulin cDNA clones contain five of these repeats. Nucleotide sequence comparisons between these clones indicate that the genes encoding these proteins are highly conserved. We estimate there to be 7 to 10 genes for the oat globulin per haploid genome. Comparisons of amino acid sequences show that the oat globulin is 30 to 40% homologous with storage globulins of legumes and about 70% homologous with the rice seed storage globulin (glutelin).

Journal Article↗
PubMed1971

Glomerular basement membrane--reactive antibody in anti-lymphocyte globulin.

Equine immunoglobulin was detected along the glomerular basement membrane of three human homograft recipients who had been treated with equine anti-lymphocyte globulin. Anti-lymphocyte globulins, given these patients, were obtained by immunization of horses with lymphocytes from human spleens and/or lymph nodes and contained glomerular basement membrane-reactive antibodies. Quantitative paired-label isotope experiments (in rats) demonstrated that 30-170 mug/ml of kidney-fixing antibodies were present in these preparations. The anti-lymphocyte globulins formed a line of identity with a sheep anti-human glomerular basement serum when reacted against collagenase-solubilized human glomerular basement membrane in double diffusion in agar. The renal fixation of these antibodies was blocked by absorption with human glomerular basement membrane, but not by buffy-coat leukocytes, indicating that they were directed specifically toward antigens in the basement membrane and were not cross-reacting anti-lymphocyte antibodies. Anti-lymphocyte globulin preparations for human use were studied for glomerular basement membrane-reactive antibodies by a direct immunofluorescent assay in rats. Anti-lymphocyte globulin from 13 of 20 horses, and 7 of 10 serum pools from horses immunized with lymphocytes derived from solid lymphoid organs (spleen, thymus, lymph node, tonsil), contained glomerular basement membrane-reactive antibodies. Sera from 18 horses injected with thoracic duct cells or cultured lymphoblasts had no glomerular basement membrane-reactive antibodies. An equine anti-human thymus serum containing glomerular basement membrane-reactive antibodies, which produced fatal glomerulonephritis in monkeys, was shown to cause both immediate and delayed glomerular injury in monkeys after intravenous injection. The reaction of this antibody with glomerular basement membrane in vivo was associated with little complement deposition in spite of the fact that the antibody could fix complement. This lack of glomerular complement fixation resulted from almost complete in vivo decomplementation of the monkeys receiving this anti-lymphocyte globulin.

Adult↗
PubMed1999

Supplemental immune globulins in sepsis.

Intravenous immune globulins are widely used as supplemental treatment of sepsis, septic shock and systemic inflammation in the critically ill, although this indication has at best been validated in part. Likely beneficial mechanisms of action may include the improvement of serum bactericidal activity due to neutralizing and opsonizing immunoglobulin (Ig)G- and IgM-antibodies, as well as stimulation of phagocytosis and neutralization of bacterial endo- and exotoxins; another attractive mode of action may represent immune globulin-mediated modification and specific suppression of proinflammatory cytokine release from endotoxin- and superantigen-activated blood cells. For the "entire group of patients with sepsis and septic shock" a reduction in mortality by intravenous immune globulin could not be documented; however, in the score-based immunoglobulin in sepsis (SBITS)-study with 653 patients included, a moderate improvement in sepsis morbidity and multiple organ dysfunction syndrome was demonstrated. In defined sepsis subgroups, a reduction in mortality by intravenous immune globulin has been seen in individual small, not yet confirmed trials. Finally, the incidence of some severe infections in well characterized "patients at risk" and "operations at risk" is reduced by intravenous immune globulin prophylaxis. Thus, intravenous immune globulin is not a "magic bullet"of sepsis treatment, but it may reduce morbidity and thereby represent a useful piece of stone in the therapeutic mosaic of sepsis treatment.

Controlled Clinical Trials as Topic↗
PubMed1991

Increased urinary free cortisol and decreased serum corticosteroid-binding globulin in polycystic ovary syndrome.

The adrenal participation in the pathogenesis of polycystic ovary syndrome is still under debate. In order to reappraise androgen and glucocorticoid secretion in this disease, we measured serum androstenedione, dehydroepiandrosterone-sulphate, total and free testosterone, sex hormone-binding globulin, LH, FSH, PRL, cortisol, corticosteroid-binding globulin, and urinary free cortisol in 45 women with polycystic ovary syndrome and 27 controls, subdivided in obese and normal-weight subjects. Androstenedione, total and free testosterone were significantly increased, whereas sex hormone-binding globulin tended to be reduced in patients with polycystic ovary syndrome compared with controls, reaching a significant difference between obese patients and matched controls. Free testosterone and sex hormone-binding globulin were significantly increased and reduced, respectively, in obese compared with normal-weight patients. Urinary free cortisol and serum corticosteroid-binding globulin were significantly increased (p less than 0.001) and decreased (p less than 0.005), respectively. Urinary free cortisol exceeded the upper limit of the normal range in 50% of our patients. No appreciable differences were found in PRL and cortisol levels. Besides confirming a hyperandrogenic state, our findings point to an overactivity of the hypothalamic-pituitary-adrenal axis with subsequent diminution of corticosteroid-binding globulin in polycystic ovary syndrome. They also indicate that urinary free cortisol is not a reliable index in differentiating polycystic ovary syndrome from Cushing's disease.

Adult↗
PubMed1998

The induction by estrogen of rat alpha 2u-globulin gene expression in mouse L-cells.

Expression of the rat alpha 2u-globulin gene family is regulated in the adult male liver by a number of hormones, including growth hormone, thyroid hormone and several steroids. Upon injection into ovariectomized females, estrogens first induce alpha 2u-globulin expression and then suppress this gene after several days of hormone administration. To study this phenomenon, we developed a mouse L-cell line that expressed the human estrogen receptor. High levels of rat alpha 2u-globulin transcript were induced in stable transfectants of this line carrying a cloned alpha 2u-globulin gene, following exposure to 17 beta-estradiol. Since this induction was inhibited by cycloheximide, the response to estrogen, as to other steroids, appears to be secondary. Using genes with variously deleted 5'-upstream regions, sequences responsible for this induction were located between -730 bp and -223 bp relative to the start of transcription. Examination of the DNA in this region revealed that an estrogen receptor element was located at -590 bp in an area that is highly conserved in most known alpha 2u-globulin genes. Administration of both dexamethasone and estrogen produced a synergistic effect in this system. The induction of alpha 2u-globulin RNA by estrogen in L-cells may re-capitulate the initial response to estrogen in vivo, and therefore represents a good model system to seek the identity of the other factors required to effect full induction.

Animals↗
PubMed1982

Thyroid-stimulating antibodies in Grave's disease and the effect of thyrotrophin-binding globulins on their determination.

Globulin preparations from the sera of 104 untreated patients with Grave's disease have been tested for their thyroid-stimulating antibody (TSAb) activities. Eighty-one of the samples (78%) were positive in the assay for the thyrotrophin-binding inhibitory immunoglobulins, 48 samples (46%) contained human thyroid adenyl cyclase stimulators (HTACS) and 71 (68%) contained human thyroid stimulators (HTS) measured as stimulation of colloid droplet formation in human thyroid slices. All 104 samples were positive in one or other of the assays, 29 (28%) were positive in all three assays and 38 (37%) in two. All samples were tested for their specific TSH-binding characteristics, 40 (38%) possessed 'B-type' binding sites (previously characterized as TSH-binding sites with low affinity but high capacity for the ligand) but the remaining 64 samples (62%) were no different from normal control samples and had 'A-type' binding sites (high affinity but low capacity binding sites for TSH). Samples without detectable thyrotrophin-binding inhibitory immunoglobulin did not contain B-type TSH-binding globulins. Globulins exhibiting B-type binding were more active in the HTACS and HTS assays. The B-type TSH-binding globulins have a characteristic, dose-dependent reducing effect on the human thyroid adenyl cyclase stimulation by TSH whereas A-type globulins do not. Globulins exhibiting B-type TSH-binding may therefore have a significant effect on assays for TSAb activities. The method used to measure TSAb have been reviewed from this point of view.

Adenylyl Cyclases↗
PubMed1979

Infertility in the turkey. I. Effects of anti-sperm immune globulins on fertilizing ability of turkey spermatozoa.

Two experiments were conducted to study the effects of globulins, isolated from sera of turkey hens immunized against semen, on the fertilizing ability of spermatozoa and the hatchability of eggs. Sperm were incubated in sera from hens immunized against semen and in sera from non-immunized hens as well as in globulin solutions isolated from these various sera. In neither experiment did sera from control hens or gamma globulins isolated from their sera affect the fertilizing ability of sperm. In contrast, all sera from immunized hens drastically reduced the fertilizing ability of sperm. In Experiment 1, .75% gamma globulin solutions from two hens did not depress fertility but a 1.5% solution from one of the immunized hens significantly (P less than .05) reduced it. In this experiment none of the treatments had a significant effect on hatchability. Antisemen globulin solutions of 2.25 and 4.50% from 12 hens significantly reduced the fertilizing ability of sperm in dose related fashion. The higher of the two levels suppressed it to the level of whole sera. Hatchability of fertile eggs was significantly reduced by the anti-semen sera and globulins isolated from it in the second experiment.

Animals↗
PubMed1979

Passive-active immunity from hepatitis B immune globulin. Reanalysis of a Veterans Administration cooperative study of needle-stick hepatitis. The Veterans Administration Cooperative Study Group.

The mechanism of action of hepatitis B immune globulin (HBIG) and immune serum globulin was sought in a reanalysis of a Veterans Administration cooperative study on needle-stick exposure to hepatitis B surface antigen (HBsAg)-positive blood. Sera from 296 exposed persons were tested for HBsAg, antibody to HBsAg (anti-HBs), and antibody to hepatitis B core antigen (anti-HBc) by radioimmunoassay. Type B hepatitis developed in three HBIG (2%) and in 12 ISG (8%) recipients. In contrast, subclinical infection (development of HBsAg or anti-HBs and anti-HBc without symptoms or jaundice) developed in 16 HBIG (10%) but only six immune serum globulin (4%) recipients. Thus, infection occurred equally in both groups but was more likely to be subclinical in HBIG recipients, indicating that HBIG permitted development of passive-active immunity to type B hepatitis. An additional 53 immune serum globulin recipients (36%) but only one HBIG recipient developed anti-HBs alone, without hepatitis, HBsAg, or anti-HBc. This response was more compatible with immunization by HBsAg than with infection. Ultracentrifugation analysis revealed occult HBsAg in the immune serum globulin but not the HBIG, indicating that some immune serum globulin preparations contain HBsAg and can induce active immunity to type B hepatitis.

Hepatitis B↗
PubMed1990

[The comparative efficacy of splenectomy and antilymphocyte globulin therapy in patients with aplastic anemia].

Analysis of the clinical efficacy and long-term survival of patients with grave and not grave forms of aplastic anemia, subjected to splenectomy or treated with antilymphocytic globulin has shown that in grave aplastic anemia, the clinical efficacy of antilymphocytic globulin is higher than the efficacy of the operative treatment methods. The 30-month survival can be attained in 42% of the patients given globulin and in 28% of the patients after splenectomy. In patients with grave aplastic anemia, it is recommended that the treatment be instituted from the use of antilymphocytic globulin in a dose of 15 mg/kg bw daily for 5 days, provided there are no contraindications. In patients with not grave aplastic anemia, the results of splenectomy and therapy with antilymphocytic globulin are approximately the same: the 5-year survival amounts to 94 and 93%, respectively. Therefore, the use of globulin may be recommended as a method of choice for the above patients' group, since surgical approaches to the treatment are still coupled with a high risk for the patient's life.

Actuarial Analysis↗
PubMed1986

Modulation of alpha 2-acute phase globulin serum concentration in turpentine- and immune complex-challenged rats by dexamethasone and adrenaline: evidence for a synergistic mode of action.

The acute phase response (APR) induced by immune complexes or turpentine and modulated by dexamethasone as well as adrenaline was investigated in intact, sham-adrenalectomized and adrenalectomized rats. The serum level of alpha 2-acute phase globulin was examined by means of electroimmunodiffusion, and the complex serum protein profile was studied using the crossed-IE. Dexamethasone administration increased the alpha 2-acute phase globulin concentration weakly over a wide dose range whereas rats dosed with adrenaline responded strongly over a small dose range. Turpentine but not immune complex challenge provoked an almost complete acute phase protein profile as well as a moderate alpha 2-acute phase globulin elevation in adrenalectomized rats. After combined administration of the prophlogistic agents and dexamethasone or adrenaline a remarkably high alpha 2-acute phase globulin level was found (up to 14 mg/ml with about 0.05 mg/ml as base level). From these results a synergistic mode of action can be suggested. With crossed-IE some of the proteins could by divided in proteins responding in a glucocorticoid-dependent (e.g. alpha 2-acute phase globulin, haptoglobin) and -independent fashion (alpha 1-acute phase globulin), respectively. It can be concluded that glucocorticoids, catecholamines as well as cytokines are the main candidates for triggering the acute phase response; however, additional factors may also be of importance.

Adrenalectomy↗
PubMed1985

Intravenous hyperimmune globulin prophylaxis against cytomegalovirus interstitial pneumonitis after allogenic bone marrow transplantation.

In an attempt to reduce the incidence of lethal cytomegalovirus (CMV) interstitial pneumonitis after allogenic bone marrow transplantation 49 patients were randomized in a multicenter controlled study to receive either CMV-hyperimmune globulin or a control immune globulin with low anticytomegalovirus titer. Immune globulin was administered intravenously 6 times with 20 days interval, starting on day 7 before transplantation. Patients receiving CMV hyperimmune globulin or control immune globulin were comparable with regard to age, diagnosis, pretransplant anti-CMV titer, incidence of graft-versus-host disease and transfusions. In each group, the incidence of histologically proven CMV interstitial pneumonitis during the first 110 days post BMT was recorded. Six of 23 patients in the control group versus 1 of 26 in the CMV hyperimmune globulin group died of CMV interstitial pneumonitis (p less than 0.05). No significant effect on idiopathic pneumonitis or survival was observed.

Bone Marrow Transplantation↗
PubMed1986

Antiepileptic effects of globulin-N, an intact human immunoglobulin and its tissue-distribution in kindled cats.

The antiepileptic effects of globulin-N, an intact human immunoglobulin was examined, using the amygdaloid kindled animals prepared by the method of Goddard et al. [1969] and Wada et al. [1974 b]. In the non-treated kindled cats, kindled convulsion and after-discharge (AD) were simultaneously created at 24-hour intervals by electrically stimulating generalized seizure-triggering threshold (GST) in amygdala, hippocampus and neocortex. The kindled cats, treated intravenously with 200 mg/kg of globulin-N, the generalized convulsion and AD produced by GST-stimulation disappeared entirely from 30 min to 8 days in 8 out of 10 cases. Furthermore, kindled cats treated with phenobarbital and phenytoin showed inhibition of GC and AD by GST-stimulation for 24-48 hours. Globulin-N was determined to have a plasma half-life of about 10 days in the kindled cats, and was presented immunohistochemically in the visceral organ, brain and spinal cord. The administered globulin-N passed through the BBB easily in the kindled cats with GST stimulation induced epileptic seizures, and was apparently taken up by the nerve and glial cells in the cerebral cortex and other deep structures of the central nervous system. These results may not only yield clues for the elucidation of the antiepileptic mechanisms of globulin-N, but may also support the clinical use of globulin-N in patients with intractable epilepsy [Ariizumi et al. 1982], including epileptic psychosis.

Amygdala↗
PubMed1984

[Antiepileptic effects of globulin-N as an intact human immunoglobulin and its tissue-distribution in kindled cats].

The antiepileptic effects of Globulin-N as an intact human immunoglobulin were examined using the amygdaloid kindled animals, which was prepared by the method of Goddard et al (1969) and Wada et al (1974). In the non-treated kindled cats, generalized convulsion (GC) and after-discharge (AD) in amygdala, hippocampus and neocortex was caused synchronously by electrical stimulation with generalized seizure-triggering threshold (GST) at constant intervals of 24 hours. By contrast, in kindled cats treated intravenously with 200 mg/kg Globulin-N, the GC and AD due to the GST-stimulation disappeared entirely for the period 4 to 8 days after medication in 8 out of 10 cases. Further, kindled cats treated with anticonvulsant, phenobarbital or phenytoin inhibited GC and AD to GST-stimulation until 24-48 hours after medication. On the other hand, the plasma half-life of Globulin-N was determined as about 10 days in the kindled cats, and Globulin-N was distributed immunohistochemically not only in the visceral organ-tissues but also in the tissues of the brain and spinal cord. The administered Globulin-N or 14C-human immunoglobulin pass through the blood-brain barrier easily in kindled cats with GST induced-epileptic seizure, and was apparently taken up by the nerve cells and glial cells in the cerebral cortex and other extensive deep structure involved in the central nervous system. These results seem to support the clinical usefulness of Globulin-N in patients with epilepsy (Ariizumi et al, 1982) and also epileptic psychosis.

Amygdala↗
PubMed1994

Immune globulin therapy in allogeneic bone marrow transplant: a critical review.

Various immune globulin products have been utilized in allogeneic bone marrow transplant (BMT) in an effort to decrease the incidence of cytomegalovirus (CMV) infection, infection due to other pathogens and graft versus host disease (GVHD). Controlled trials regarding the use of prophylactic immune globulin have been reviewed. Differences among products and use of various dosing regimens complicates the comparison of the results of one trial to another. Risk factors, such as donor and recipient CMV serology, GVHD prophylaxis, the use of white blood cell transfusions and conditioning regimens, vary significantly, further complicating the analysis. Autologous BMT patients do not appear to benefit from prophylactic globulin therapy. The effectiveness of immune globulin in the prevention of CMV disease in allogeneic BMT is questionable; other measures, including prophylactic ganciclovir and screening of blood products appear to be of greater value. Immune globulin inconsistently decreases the incidence of infection due to pathogens other than CMV. The best established role for prophylactic immune globulin in allogeneic BMT is in the prevention of GVHD.

Bone Marrow Transplantation↗