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Biomedical subjects

G J Gleich

Publications and source records attributed to G J Gleich.

At least 361 records · Page 20Linked to original sources
PubMed1981

Preseasonal intranasal immunotherapy with nebulized short ragweed extract.

We determined the effect of preseasonal intranasal short ragweed (SRW) immunotherapy in a double-blind, nonpaired, 20-wk study involving 33 SRW-sensitive patients. Patients were selected on the basis of an elevated IGE serum antibody level, a positive intradermal skin test, and a positive intranasal challenge to SRW antigen. SRW-treated patients sprayed SRW solutions intranasally six times a day for 12 wk preseasonally. Placebo-treated patients used nebulized solutions containing buffer or histamine that were interchanged randomly throughout this period. The SRW-treated group reported more preseasonal symptoms than the placebo-treated group (p less than 0.003); however, during the SRW pollination season, the SRW-treated group reported significantly less sneezing, nasal congestion, rhinorrhea, red/itchy eyes, itchy nose/throat, and cough/wheeze. Supplemental antihistamine usage was similar in both groups. The treatment did not affect serum IgE antibody levels to crude SRW, AgE Ra3, or Ra5 in either group at any time during the study. No significant production of IgG antibody to SRW was seen in either group. One SRW-treated patient developed acute sinusitis after 2 wk of treatment; otherwise no side effects other than symptoms of hay fever were noted. Although intranasal SRW immunotherapy may offer an effective and less costly alternative to parenteral immunotherapy, reduction in hay fever symptoms during the pollination season was achieved at the expense of provoking these symptoms during the preceding weeks.

Administration, Intranasal↗
PubMed1981

Isolation and characterization of Russian thistle (Salsola pestifer) pollen allergens.

The radioallergosorbent test (RAST) was utilized to identify allergenically active molecules in an extract of Russian thistle (Salsola pestifer) pollen. Two glycoproteins (RT1 and RT2) were isolated by ion-exchange chromatography, preparative flat-bed electrofocusing, and gel filtration chromatography. These highly purified proteins were similar as judged by discontinuous polyacrylamide gel electrophoresis (PAGE), by PAGE in urea-acetate, and by immunodiffusion analysis. In Ouchterlony double-diffusion, rabbit antiserum to crude pollen extract showed a line of identity between RT1 and RT2. Each allergen possessed a single polypeptide chain, with molecular weight of 39,00 and 42,00 for RT1 and RT2 respectively. Both allergens contained 9.0% +/- 0.5% carbohydrate and 10.6% +/-0.2% nitrogen. Both RT1 and RT2 eluted from a calibrated Sephadex G-100 column near the ovalbumin marker, but the elution volume of RT1 was slightly greater than that of RT2. Amino acid analysis of RT1 and RT2 showed that there was no difference in composition between the two proteins. The proteins showed different isoelectric points (RT1 = 6.7; RT2 6.2). They had similar skin reactivity by Prausnitz-Kustner testing, showed similar potency and identical allergenic qualities as inhibitors in the RAST, and were more potent on a mass basis than crude extract in the inhibition of the reaction between solid-phase crude extract and IgE antibodies in the RAST. RT1 and RT2 represent 0.014% and 0.010% by mass, respectively, of the pollen. These results suggest that RT1 and RT2 are important allergens in Russian thistle pollen. These results suggest that RT1 and RT2 are important allergens in Russian thistle pollen and that these proteins are immunologically identical.

Allergens↗
PubMed1981

Aging and serum immunoglobulin E levels, immediate skin tests, RAST.

The changes of the serum IgE levels, specific immediate skin-test responses, and RAST measurements with age were evaluated. A total of 331 unrelated individuals were studied, consisting of 166 subjects with ragweed allergic rhinitis and/or asthma, 67 with idiopathic (intrinsic) asthma, and 98 who appeared in good health with no clinical evidence of atopic diseases. All subjects were evaluated by history and physical examination, intradermal skin testing to the common aeroallergens, measurements of IgE antibody to common aeroallergens with the RAST, and serum IgE levels. Results demonstrated a significant decrease in serum IgE levels with aging in atopic individuals. This decline was exponential in character. In addition, a tendency for RAST and immediate type skin-test responses for selected antigens and histamine to decrease with age was observed.

Adolescent↗
PubMed1981

Eosinophil granule major basic protein and Charcot-Leyden crystal protein in human tears.

We measured the levels of major basic protein and Charcot-Leyden crystal protein in tears from patients with ventral keratoconjunctivitis or miscellaneous inflammatory or noninflammatory ocular conditions, and from normal subjects. Patients with vernal keratoconjunctivitis had significantly increased levels of both proteins in their tears compared with the other subjects tested; levels of major basic protein seemed to correlate with the severity of the disease. The levels of Charcot-Leyden crystal protein paralleled those of major basic proteins.

Eosinophils↗
PubMed1981

Purification of human eosinophil-derived neurotoxin.

Human and animal eosinophils contain a powerful neurotoxin that causes selective neuronal and axonal damage to white matter of cerebellum and spinal cord of experimental animals when injected intrathecally. This reaction is termed the "Gordon phenomenon." We purified the eosinophil-derived neurotoxin from eosinophil-rich leukocyte suspensions or eosinophil granules from four patients with various hypereosinophilic syndromes. A single protein with an average molecular weight of 18,400 was isolated by sequential chromatography on Sephadex G-50 columns and analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of column fractions. The purified eosinophil-derived neurotoxin from the cells of these patients retained the potent neurotoxic activity of the crude eosinophil or eosinophil granule extracts in experimental animals. These animals developed the syndrome of stiffness and ataxia progressing to severe paralysis characteristic of the Gordon phenomenon. Histologic examination of the brains of animals injected with purified eosinophil-derived neurotoxin confirmed the characteristic widespread loss of Purkinje cells and severe spongiform vacuolation in the white matter of cerebellum, brain stem, and spinal cord. We have established the location of eosinophil-derived neurotoxin in the eosinophil granule and have shown that it is distinct from several other eosinophil proteins, the granule major basic protein, and the Charcot-Leyden crystal protein (lysophospholipase).

Animals↗
PubMed1981

Elevated serum levels of the eosinophil granule major basic protein in patients with eosinophilia.

A radioimmunoassay was established for the human eosinophil granule major basic protein (MBP). The mean level of MBP in sera from 105 normal control patients was 454 ng/ml, whereas in a sample of 188 patients with various forms of diseases, including the hypereosinophilic syndrome, levels as high as 14,000 ng/ml were measured. Serum levels of MBP did not correlate with eosinophil counts in normal subjects, but a positive correlation was seen in patients with eosinophilia; the patients with eosinophil counts greater than 350/mm3 generally showed increased levels of MBP. Many patients with skin disease and normal eosinophil counts had elevated levels of serum MBP. Monomer MBP has a molecular weight of 9,300, but in sera of patients with eosinophilia, the MBP activity was of high molecular weight, greater than 50,000. Analyses of serum by Sephadex G-200 and by electrofocusing suggest that MBP is not simply polymerized, but rather is bound to a larger carrier molecule. Monomeric MBP can be isolated from serum by reduction of serum with dithiothreitol, alkylation with iodoacetamide, and acidification to pH 2 followed by fractionation on Sephadex G-50 at pH 2. Under these conditions, up to 80% of the MBP emerges in monomeric form. The results indicate that eosinophil granule proteins circulate in blood covalently bound to serum proteins, and that elevated concentrations of serum MBP are present in some diseases associated with eosinophilia.

Alkylation↗
PubMed1981

Destruction of bloodstream forms of Trypanosoma cruzi by eosinophil granule major basic protein.

Human eosinophils are known to engage in an antibody-dependent cell-mediated cytotoxicity reaction causing destruction of virulent bloodstream forms of Trypanosoma cruzi. A similar cytotoxic effect was found in this work to be produced by the major basic proteins (MBP) purified from human and guinea pig eosinophil granules. Killing of T. cruzi by these proteins was concentration-dependent, with significant cytotoxicity observed at concentrations as low as 1 x 10(-5) M. Basicity appeared to be an important, but not the only, property required for the MBP molecule to destroy T. cruzi, since highly basic proteins such as arginine-rich histone and cytochrome C were inactive under the same conditions. However, other basic proteins, poly-L-arginine and protamine, lacked cytotoxicity at concentrations which were effective for MBP (1 x 10(-5) M), but killed the flagellates at higher concentrations (5 x 10(-5) M). Furthermore, heparin, an anionic molecule, effectively inhibited the cytotoxic effect of both human and guinea pig MBP on T. cruzi. Heating MBP at 56 degrees C for 4 hours, a treatment which causes the loss of reactivity of MBP with specific antibodies, effectively inhibited the lytic effect on the parasites. In contrast, heating had no effect on the cytotoxic effects of protamine or poly-L-arginine. Specific antiserum to MBP caused a marked reduction in the extent of trypanosome killing by MBP when added to reaction mixtures. The present results suggest that eosinophil-mediated killing of T. cruzi may be due to the discharge of basic granule components by the effector cells which are directly toxic for the parasite.

Animals↗
PubMed1981

Measurement of IgG blocking antibodies by interference in the radioallergosorbent test.

We previously found that sera of patients immunized with ragweed pollen extract contained a factor that interfered with the binding of IgE antibodies to solid-phase allergens in the radioallergosorbent test (RAST). We now describe an assay, RAST interference, to measure this factor, and we present evidence that the factor is IgG blocking antibody. Sera from immunized allergic patients were heated at 56 degrees C for 4 hr to destroy heat-labile Fc determinants on IgE and were tested for their ability to prevent binding of additional IgE antibody to solid-phase allergens in the RAST. Eight of 10 sera from allergic immunized patients gave RAST interference dose-response curves that did not differ from the arbitrary standard. The factor causing interference showed specificity for the immunizing antigen, was heat-stable, eluted from Sephadex G-200 in the 7S peak, was present only in sera of immunized patients, and rose after initiation of immunization. These results indicated that RAST interference can be used to measure IgG blocking antibodies with the same reagents employed for the measurement of IgE antibodies, provided the antiserum to IgE is specific for the heat-labile FC determinants on IgE.

Antibodies↗
PubMed1981

Elevated levels of the eosinophil granule major basic protein in the sputum of patients with bronchial asthma.

The eosinophil granule major basic protein (MBP) is toxic to parasites and mammalian cells. Because eosinophilia is characteristic of asthma, we tested the effect of MBP on bronchi and assayed sputa for this protein. We found that MBP damaged bronchial epithelium in vitro and produced changes that mimicked those in asthma. Radioimmunoassay of sputa from 100 consecutive patients with respiratory diseases revealed MBP levels above 0.1 mug/ml in 13 patients, and 11 of these had asthma. In 15 patient hospitalized for asthma, MBP levels of sputum were markedly elevated. Treatment with bronchodilators and glucocorticoids caused an increase peak expiratory flow rate, a reduction in blood eosinophils, and a decrease in the serum and sputum levels of MBP. The results indicate that eosinophil granule constituents are released into the bronchi in asthma and that measurement of sputum MBP may be useful in identifying asthma. The possibility that the eosinophil damages bronchial epithelium in asthma is discussed.

Asthma↗
PubMed1980

The heat stability of short ragweed pollen extract and the importance of individual allergens in skin reactivity.

A short ragweed pollen extract that was heated in a boiling-water bath remained skin reactive for some allergic individuals and lost reactivity for others. Comparison of heated and unheated extract by immunodiffusion showed that AgE, AgK, and Ra3 could not be detected in the boiled extract but Ra5 was present. By inhibition of the radioallergosorbent test the concentration of Ra5 remained essentially unchanged, but the concentrations of AgE and Ra3 were greatly diminished. The sera of these ragweed-allergic individuals were analyzed for IgE antibodies to AgE, Ra3, and Ra5. Skin reactivity in 17 ragweed-allergic individuals to the heated extract correlated significantly with the levels of IgE antibodies to Ra5. However, the large number of individuals reacting to the boiled extract indicates that Ra5 is not likely to be the only heat-stable allergen. Skin reactivity of the unheated extract did not correlate with IgE antibody levels to any allergen. The results underscore the importance of individual allergens in the materials used for skin testing.

Allergens↗
PubMed1980

Measurement of histamine: a quality control study.

This study evaluated the ability of various laboratories to accurately quantitate histamine in samples of plasma and buffered saline that contained known amounts of histamine. Histamine was dissolved in buffered saline and in plasma and these solutions were lyophilized glass ampules. Sealed ampules were sent to laboratories for analysis of their histamine content by one or more of the following methods: (1) the double-isotope dilution enzymatic method, (2) single-isotope enzymatic method, and (3) manual and automated fluorometry. We analyzed the same solutions in our laboratory by the double-isotope dilution enzymatic method over a 6-mo period; we found a coefficient of variation averaging 26% for these samples over that period of time and our values agreed with theoretical values within 12%. Twelve laboratories analyzed histamine by the double-isotope dilution enzymatic assay. The results revealed a marked variation among laboratories both for the determination of histamine in buffer and, more strikingly, for the determination of histamine in plasma. Three laboratories determined histamine by the single-isotope dilution enzymatic method and one reported results rather close to the standard, while the others reported results that were clearly different from the standards. Five laboratories measured histamine by automated fluorometry and three by manual fluorometry; again, there was marked variation among the results. Overall, these findings indicate that measurements of histamine by different laboratories vary greatly. Thus, absolute values for histamine in biologic specimens in the literature must be regarded with caution.

Chemical Phenomena↗
PubMed1980

Interference in ragweed pollen and honeybee venom radioallergosorbent tests.

We studied sera from patients sensitive to short ragweed (SRW) and honeybee venom (HBV) to investigate serum factors able to interfere with the measurement of IgE antibody levels by the radioallergosorbent test (RAST). We heated sera to destroy IgE antibodies and tested them for interference in the RAST. Heating sera for 4 hr at 56 degrees C destroyed up to 98% of the IgE antibody activity. After immunotherapy sera from patients sensitive to SRW and HBV produced striking interference in the RAST. The interference was most marked in the RAST employing 50-microgram quantities of microcrystalline cellulose-linked allergens, but it was also evident in the RAST employing 500-microgram quantities of such allergens and in the commercial RAST in which allergen is linked to paper disks. The interfering substance eluted from diethylaminoethyl (DEAE) cellulose in the IgG fraction. The interference could be eliminated by increasing the relative quantity of solid-phase allergen; RAST interference was not detected when SRW extract was linked to Sepharose 4B. The results indicate that serum factors, presumably IgG antibodies, produce interference in the RAST. Thus immunotherapy studies that measure IgE antibody levels by the RAST must consider the possibility that IgG antibodies can appear to depress IgE antibody levels. Furthermore, because commercially available RAST disks are susceptible to RAST interference, they must be used with caution for the diagnosis of allergy in patients whose sera may contain significant quantities of IgG antibodies.

Animals↗