bluuebunny/crossref_metadata_2025 huggingface dataset:
Localization, Molecular Weight and Immunoglobulin Subclass Response to Aspergillus fumigatus Allergens in Acute Bronchopulmonary Aspergillosis
Authors: Leung Patrick S.C. , Gershwin Eric , Coppel Ross , Halpern Georges , Novey Harold , Castles James J.
Published: August 2009

A specimen of Aspergillus fumigatus was isolated from a patient with acute bronchopulmonary aspergillosis (ABPA). Cultures were allowed to propagate and were separated into spores and mycelium to > 95% homogeneity. Extracts of both the spores and mycelium were prepared and used for study by both dot blot analysis and immunoblotting to study the major allergens. By dot blot analysis, 22 of 22 patients with ABPA and none of 10 healthy controls reacted with mycelium when probed for both IgG and IgE anti-Aspergillus reactivity. Similar results were obtained when these extracts were separated on polyacrylamide gel electrophoresis and then probed; this included both IgG and IgE reactivity. Further, the reactivity to preparations of whole extract was absorbed with mycelium but not spores. Several distinct protein bands were detected with IgG, ranging from 30 to 110 kilodaltons (kD). In contrast, the majority of patients with ABPA, when studied for IgE reactivity, reacted only with a 70-kD protein, although 1 out of 22 patients reacted with a 30-kD protein. Subclass analysis of IgG reactivity demonstrated that 90% of reactive sera contained IgG2 and 73% IgG4 reactivity. IgG1 and IgG3 reactivity were present but less frequently detected. The use of immunoblotting will enable the identification of relevant allergens. Further, the demonstration that the allergenicity is located primarily in mycelium will allow more definitive studies of allergen isolation for studies of reactivity and, ultimately, cloning of the 70-kD protein and, finally, epitope mapping.
Vicia villosa Lectin Is a Mitogen for Mouse T Lymphocytes
Authors: Abrignani Sergio , Cammisuli Salvatore
Published: August 2009

Mouse T lymphocytes cultured in the presence of Vicia villosa lectin, usually considered to be non-mitogenic, proliferate and produce interleukin-2. V. villosa-induced proliferation does not correlate with the cell ability to adhere to V. villosa-coated dishes, this is evident from the fact that either V. villosa-adherent or -nonadherent cells are equally sensitive to the mitogenic effect of the lectin. Furthermore, the sugar N-acetyl-d-galactosamine, which is reported to be highly specific for V. villosa, does not reduce the lectin-induced proliferation. These data indicate that there is a difference in the carbohydrate moieties required for the adherence to V. villosa and its mitogenic effect. These results are discussed on the light of the current use of V. villosa lectin as marker for some T cell subpopulations.
Successful Contact Sensitization to Chromate in Mice
Authors: Mor S. , Ben-Efraim S. , Leibovici J. , Ben-David A.
Published: August 2009

Mice of the randomly bred strain ICR and of the inbred strain BALB/c could be successfully sensitized to chromate by repeated topical applications of a solution of potassium bichromate in dimethyl sulfoxide. The specificity of chromium sensitization was ascertained by ear tests, specific desensitization by intravenous injection of potassium bichromate and by histological ear sections showing marked infiltrates of leukocytes following ear application of chromium-dimethyl sulfoxide in already sensitized animals. It is concluded that mice may provide a useful experimental model for study of contact sensitivity to chromium.
Propagation and Characterization of Human Blood Basophils
Authors: Sorensen Lyle S. , Mullarkey Michael F. , Bean Michael A. , Mochizuki Diane Y. , Chi Emil Y. , Henderson William R.
Published: August 2009

Basophils were isolated and propagated in large numbers from the blood of patients with chronic myelogenous leukemia. Propagation over 4–6 weeks of culture was dependent upon a growth factor(s) other than interleukin-2 obtained from a lectin-stimulated clone of the Jurkat cell line. Evidence that these basophils were dividing during culture included an increase in both the number of basophils and the histamine content of the cultures over time, as well as ultrastructural studies that demonstrated basophil cell division. The cells also had the capacity to be stimulated in an IgE-dependent manner characteristic of basophils. Cultured basophils passively sensitized with IgE underwent noncytotoxic degranulation after stimulation with specific antigen. Antigen-stimulated basophils released histamine, leukotrienes B4 and C4 and other 5-lipoxygenase products of arachidonic acid metabolism. Culture models such as this may permit the propagation and purification of sufficient numbers of basophils to allow biochemical and immunological analyses of basophil physiology.
Comparative Study of IgG1 and IgE Antibody Mediated Homologous PCA in the Mouse Ear
Authors: Inagaki Naoki , Nagai Hiroichi , Koda Akihide
Published: August 2009

To characterize IgG1 antibody and IgE antibody mediated homologous passive cutaneous anaphylaxis (PCA) in the mouse ear, cross-desensitization was studied using a double-sensitizing technique. Mice were sensitized by injecting into their ears a mixture of two kinds of antibodies with distinct antigen specificities, and PCAs were elicited twice with specific antigens at an appropriate interval. The first PCA was elicited with one antigen free from Evans blue dye and the second with the other antigen in the presence of Evans blue to visualize the reaction. The amount of extravasated dye caused by the latter was determined colorimetrically and compared to that of control which did not receive the first challenge. In PCA mediated by IgG1 antibodies, elicitation of the first reaction significantly inhibited the second reaction evoked after 6, 12, or 24 h. Using IgE antibodies, elicitation of the first PCA also significantly inhibited the following reaction. Inhibition of the second reaction was observed 6 h to 12 days after the first reaction. Conversely, IgG1 antibody mediated PCA did not affect the following reaction caused by IgE antibody. These results strongly suggest that alteration of skin mast cells is smaller in IgG1 antibody than in IgE antibody mediated PCA, when reactions with a similar intensity are evoked. Therefore, it is also suggested that the slight reaction of mast cells initiated by IgG1 antibody might be potentiated by another mechanism in IgGl antibody mediated PCA.
Characteristics of the Mitogenic Activity Found in the Supernatant of Pokeweed Mitogen Stimulated Human Mononuclear Cells
Authors: Uiterdijk H.G. , Kornman-van den Bosch H.J. , de Vreede T.M. , de Bruijn A.M. , Klein F. , de Vries E.
Published: August 2009

Human peripheral blood mononuclear cells were stimulated with pokeweed mitogen (PWM), washed, and cultured in fresh mitogen-free culture medium; a second set of peripheral blood mononuclear cells was used to assess the mitogenic activity of supernatants of these cultures. The effects of this stimulation resembled those of a normal PWM stimulation, including the formation of blast cells, proliferation of lymphoid cells, and immunoglobulin synthesis. Our findings led us to perform experiments to investigate the nature of this mitogenic factor. The molecular weight of the mitogenic activity was found in the region of 700 kD. Additional experiments indicated that human α2-macroglobulin was not responsible for the high molecular weight of the mitogenic factor. Protein digestion or precipitation with trichloroacetic acid only partially abolished the mitogenicity of the mitogenic supernatant. Comparison of the molecular weight of the mitogenic factor with that of the PWM in culture medium showed that the mitogenic activity in the supernatants could be caused by PWM associated with a high molecular weight component.
Analysis of Rye Pollen (Secale cereale) Allergens Using Patients’ IgE, Immunoprint, Western Blot and Monoclonal Antibodies
Authors: Westphal W. , Becker W.-M. , Schlaak M.
Published: August 2009

Rye pollen (Secale cereale) was investigated by Immunoprint and Western Blot analysis. Using Immunoprint, more than 30 proteins could be shown to be allergens in terms of their IgE binding, and 9 were determined as major allergens. In Western Blot 17 allergens were discovered, 3 of them major allergens. 4 monoclonal antibodies raised against a pollen extract of 6 grasses showed cross-reactivity to rye pollen, thus indicating common epitopes. 1 of 3 monoclonal antibodies produced against an extract of rye pollen was only reacting to this pollen. Looking at the overall binding pattern of the 7 monoclonal antibodies, they showed partial identity to patients’ IgE in Immunoprint and Western Blot. The possibility of defining epitopes using preparative isolelectric focusing and monoclonal antibodies is discussed.
Effects of Tumour Necrosis Factor Alpha and Interleukin-1 Alpha and Beta on Human Neutrophil Migration, Respiratory Burst and Degranulation
Authors: Ferrante Antonio , Nandoskar Madhuri , Walz Alfred , Goh David H.B. , Kowanko Ingeborg C.
Published: August 2009

Recombinant human tumour necrosis factor alpha (rHuTNFα) was shown to inhibit human neutrophil migration in the presence or absence of a chemotactic gradient generated with the tripeptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), at doses of 20–100 U/106 cells. In contrast, neither recombinant human interleukin-1 alpha (rHuIL-1α), rHuIL-1β, human leucocyte-derived IL-lα (lHuIL-lα) nor lHuIL-1β contained neutrophil migration inhibition properties. However, both the interleukins (lHuIL-1α, lHuIL-1β and rHuIL-1α) and rHuTNFα stimulated a neutrophil respiratory burst and significantly elevated the neutrophil respiratory response to fMLP (measured as chemiluminescence and H2O2 production). The stimulatory effects were observed at doses of between 5 and 100 U/5 × 105 cells. A characteristic feature of the effects of the cytokines was the range of variation observed in neutrophil responses from different individuals. However, a concentration-related effect was observed with each experiment, delineating suboptimal, optimal and supra-optimal cytokine concentrations. Neutrophils treated with rHuTNFα and rHuIL-1α and washed free of exogenous cytokine retained the capacity to show an enhanced response to fMLP. Pretreatment of cells with cytochalasin B enhanced their response to fMLP, and this response was further increased if the cells had also been pretreated with the cytokines. The response to phorbol myristate acetate was also enhanced by rHuTNFα and rHuIL-1α. The effects of these cytokines on neutrophils could be abolished by boiling the preparation but not by treating it with polymixin B, suggesting that bacterial lipopolysaccharide was not responsible for the activity of these preparations. The rHuIL-1α increased the release of lysozyme, β-glucuronidase and myeloperoxidase initiated by cytochalasin B/fMLP, while rHuTNFα only increased lysozyme release.
Experimental Studies on Red Cell-Based Assays for Total IgE and Allergen-Specific IgE
Authors: Matejtschuk P. , Easter Gwen , Thorpe R. , Coombs R.R.A.
Published: August 2009

The potential of red cell-based assays for IgE and allergen-specific IgE has been examined using mouse monoclonal anti-human IgE antibodies and chimaeric human IgE anti-4-hydroxy-3-iodo-5-nitrophen-acetyl. Experiments concerned with developing a red cell IgE antibody-capture assay for allergen-specific IgE have pointed to the advantages of presenting the allergen on a second agglutinable red cell.
Role of IgG4 in ï list a mine Release from Human Basophil Leucocytes
Authors: Poulsen L.K. , Skov Stahl , Mosbech H. , Weeke B.
Published: August 2009

Several conflicting reports on the ability of IgG4 to mediate type I allergic reactions have appeared lately. We have developed a model system for testing this possibility, using passive sensitization of basophil leuocytes from normal individuals. At first, the system was optimized with regard to donor cells and anti-IgG4 antibody: among 3 normal individuals with cells showing high, medium and low responses to anti-IgE, only the high responder showed a reproducible, but low response to anti-IgG4. This response was consistent when testing anti-IgG4 from different sources, and could be potentiated by a phorbol ester TPA, although not up to the level of anti-IgE. The cells from the high responding donor and a monoclonal anti-IgG4 were selected for further studies. Serum pools from patients allergic to house dust mite (Dermatophagoides pteronyssinus) were used for passive sensitization. The pools contained allergen-specific IgE with or without concomitant IgG4 of the same specificity. The sensitized basophils reacted to anti-IgE and allergen, but to anti-IgG4 only to a small extent. However, when these serum pools were fractionated by affinity chromatography, only the IgE fractions and not the IgG4 fractions made the cells reactive towards specific allergen. It was still possible to elicit a reaction by triggering IgE with anti-IgE. whereas only a small reaction was seen, when IgG4-sensitized basophils were provoked with antí-IgG4. We conclude that IgG4 does not sensitize normal basophils.
The Suppressive Effect of Histamine on Anti-DNP Antibody Production in Mice
Authors: Okitsu-Negishi Shako , Fumsawa Shuichi , Yoshino Kazuya , Abe Toshiaki
Published: August 2009

To clarify the effect of histamine on antibody production, histamine was injected intravenously into DNP-KLH immunized C3H/HeN female mice. The number of anti-DNP plaque-forming splenic cells decreased in immunized and histamine-injected mice. Decreased PFC responses similarly occurred in recipient mice into which Lyt-2.1 T cells obtained from histamine-injected mice were transferred before immunization. The effect of histamine is not due to inhibition on plaque formation. The results indicated that histamine induced the augmentation of suppressor T cell (Ts) function and caused the decrease of antibody production in histamine-injected mice. The maximum activity of Ts in recipient mice was obtained from mice injected with histamine 3 days before the cell transfer. The suppressive activity of histamine was not antigen specific because Ts was induced in histamine-injected, nonimmunized mice. The treatment of histamine type 2 (H2) agonist showed decreased PFC responses in the mouse as well as histamine did. When mice were treated with an H2 antagonist followed by histamine, antibody production was not reduced. The above results indicated that histamine seemed to regulate antibody production through H2 receptors on lymphocyte membranes.
1,25-Dihydroxyvitamin D-Induced Suppressor Cells in Uremic versus Normal Lymphocytes
Authors: Modai D. , Weissgarten J. , Stelian J. , Golik A. , Averbukh Z. , Peller S. , Shaked U. , Kaufman S. , Tieder M.
Published: August 2009

1,25-Dihydroxyvitamin D3 (DHD) has been shown to suppress mitogen-induced blast transformation. This inhibition is abolished by prior elimination of adherent cells. Chronic renal failure is an immunodeficiency state on the one hand and is associated with abnormalities in vitamin D metabolism on the other. The effect of DHD on the induction of suppressor cells in uremic vs. normal peripheral blood mononuclear cells was investigated. Study groups included 16 chronically uremic patients and 16 age- and sex-matched controls. DHD induced suppressor cell activity in normal lymphocytes. However, no suppressor cell activity was observed in lymphocytes from the uremic patients preincubated with DHD. The origin of the responder cells (normal or uremic) did not affect the outcome. The results would suggest that monocyte-adherent suppressor cells from uremic subjects are either incapable of binding DHD or fail to mount a normal post-receptor intracellular chain of events culminating in suppressor activity.
IgA Immune Response Patterns to Gliadin in Serum
Authors: Mascart-Lemone F. , Cadranel S. , Van den Broeck J. , Dive C. , Vaerman J.P. , Duchateau J.
Published: August 2009

The incidence, titer and molecular size of anti-gliadin (GL) IgA were analyzed in sera from children with celiac disease (CD, n = 66), hospitalized control children (n = 64) and children with other malabsorption disorders (n = 103). Anti-GL IgA was assessed by solid-phase radioimmunoassay, and its molecular size was analyzed by sucrose density gradient ultracentrifugation. The median anti-GL IgA titer was significantly higher in untreated CD than in the other groups, with some overlap. Whereas 57% of anti-GL IgA were polymeric IgA (pIgA) in acute CD, only 17% were pIgA in children with other malabsorption disorders. These anti-GL pIgA consisted mainly of dimers with in some children, a low proportion of anti-GL secretory IgA (SIgA). A large proportion of anti-GL pIgA was associated with high total serum IgA. After the onset of a gluten-free diet (GFD), anti-GL pIgA disappeared more quickly from serum than mlgA. During the gluten challenge test, anti-GL IgA comprised 20% pIgA when they reappeared in serum, but 59% pIgA at the end of the test. The presence of anti-GL pIgA in serum seems related to chronic exposure to gluten of CD patients with a flat intestinal mucosa. These serum pIgA antibodies could result from a spillover from the intestinal mucosal synthesis into the circulation.
Proliferative Response of Human T Lymphocytes to a Vaccinal Preparation of Ribosomes from Streptococcus pyogenes
Authors: Millet Isabelle , Lafont Sergyl , Jeannin Michèle , Revillard Jean-Pierre , Normier Gérard , d’Hinterland Lucien Dussourd
Published: August 2009

The in vitro lymphocyte-activating properties of a ribosomal preparation of Streptococcus pyogenes were investigated. The preparation was mitogenic for human lymphocytes with a peak of 3H-thymidine incorporation occurring after 3–5 days of culture. The response was abolished by removal of CD3-positive cells and by alteration of accessory cells by exposure to L-leucine methyl ester. Most of the cells synthesizing DNA at the end of the culture expressed CD4 or CD8 but not CD20 antigens. No immunoglobulin synthesis was demonstrable. Although the same preparation was shown to be a T-independent polyclonal B-cell activator of murine cells, it preferentially triggers T cells in humans.
Hypocomplementemia and Human Immunodeficiency Virus Infection
Authors: Lin Robert Y. , Wildfeuer Olga , Franklin Monica M. , Candido Kenneth
Published: August 2009

The levels of C4 and C3 were measured and related to other immunological and clinical parameters in 44 patients with the human immunodeficiency virus (HIV) infection. Circulating immune complexes (C1q-CΙC) as measured by an enzyme-linked immunosorbent assay employing monoclonal antibody with specificity for bound C1q, and serum immunoglobulin G (IgG) concentrations were assessed simultaneously. Clinical parameters assessed included: (1) the presence of specific anti-infective medications; (2) the presence of hypotension and fluid administration, and (3) bacterial and specific opportunistic infections. Hypocomplementemia was observed in 25 of 46 sera for C3, and in 8 of 46 sera for C4. C1q-CIC increases were seen in 26 of 46 sera and hyper-IgG was present in 25 of 46 sera. Lower C3 concentrations were significantly associated with elevated C1q-CIC levels (p < 0.001). There were significant correlations between C1q-CIC levels and C3 concentrations (p = 0.0065, negative) and IgG levels (p = 0.0075, positive). C1q-CIC were significantly higher with serum p-24 antigen levels of 50 pg/ml or greater. These data demonstrate that elevated C1q-CIC and hypocomplementemia are both common in HIV-infected patients and may have significant relationships.
Natural Killer Cells in Chronic Leukemia
Authors: Sørskaar D. , Førre Ø. , Stavem P.
Published: August 2009

Twenty-two patients with chronic lymphocytic leukemia (CLL) and 14 patients with chronic myelogenous leukemia (CML) were studied with respect to natural killer (NK) cell activity and related cell markers (Leu 7 and Leu 11b). Significantly reduced NK cell activity was detected in peripheral blood from the patients with CLL. Similarly, a reduced number of cells with the markers Leu 7 and Leu 11b (CD 16) were detected in the same patients. Removal of the leukemic cells by centrifugation of cells forming rosettes with mouse erythrocytes led to an augmented, but not fully normalized, NK activity. This indicates that the low NK activity in CLL partly may be due to overgrowth of leukemic cells. However, in spite of the lymphocytic infiltration, the NK cell activity in the bone marrow of CLL patients did not differ significantly from that of normal controls. The patients with CML in the chronic phase as well as patients in the accelerated or blast phase also had a reduced NK activity. The finding that patients in the chronic phase had a reduced NK activity and normal numbers of Leu 11b (CD 16) positive cells, together with no detectable blasts in the peripheral blood, indicates that patients with CML may have an inherent NK cell defect. The highly reduced activity found in patients with the accelerated/blast form may in addition partly be due to overgrowth of leukemic cells. This low NK activity may be of importance in the development of chronic leukemia.
Influence of Long-Term Antigenic Stimulation Started in Young C57BL Mice on the Development of Age-Related Monoclonal Gammapathies
Authors: van den Akker T.W. , Brondijk R. , Radl J.
Published: August 2009

Long-term antigenic stimulation by multiple antigens (DNP conjugated to human serum albumin (DNP-HSA), ovalbumin and pneumococcal polysaccharide) without adjuvant in young C57BL mice resulted in the development of homogeneous immunoglobulins (H-Ig) during aging in frequencies higher than those in the control group. Our data showed that antigen-specific B-cell clones were at least in part responsible for this increased incidence of age-related monoclonal gammapathies (MG). Using in situ adsorption performed on Wieme’s agar plates and in immunoelectrophoresis, antibody activity to one of the immunizing agents (DNP-HSA) could be demonstrated within 10% of the in old mice appearing H-Ig components. This frequency was significantly different from the 0.3% of the H-Ig components of the aging control mice. The antigen-specific MG belonged most likely to the category of benign MG. These findings indicate that long-lasting antigenic stimulation contributes to the development of age-related B-cell proliferative disorders, namely of the benign MG.
Isolation and Immunological Characterisation of Allergens from Salivary Glands of the Australian Paralysis Tick Ixodes holocyclus
Authors: Gauci M. , Stone B.F. , Thong Y.H.
Published: August 2009

Radioimmunoassay and Western blot analysis showed that salivary gland extract (SGE) prepared from Ixodes holocyclus was a more concentrated source of allergens than whole body extract. Isolation and partial immunological characterisation of allergens was performed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of SGE, electroelution and Western blotting. Immunoblotting using allergic sera showed that partially purified SGE from I. holocyclus contained two common allergens of approximately 28 and 35 kilodaltons and several minor allergens. The two most common allergens gave radioimmunoassay ratios significantly greater than the less common allergens. The immunological purity of electroeluted allergens was confirmed by further SDS-PAGE, silver staining and Western blotting.
Measurement of IgG Subclasses in the Sera of Laboratory Workers Exposed to Rats
Authors: Lee D. , Edwards R.G. , Dewdney Janet M.
Published: August 2009

Human IgG antibody subclasses have been measured in the sera of workers exposed to rats, using a crude extract of rat urinary protein antigens, in an ELISA system. The antibody titres in individuals either with or without specific IgE were similar, with the exception of IgG4 where the mean level of this subclass was lower in those individuals with measurable titres of IgE (p < 0.01). Symptomatic individuals, with specific IgE, also had lower titres of IgG4 than the corresponding asymptomatic, IgE-positive subjects (p < 0.05). The frequency of positives in each subclass assay was similar in both groups. These findings suggest that higher levels of IgG4 may have a protective role.
Identification of a German Cockroach-Specific Allergen by Human IgE and Rabbit IgG
Authors: Helm R.M. , Bandele E.O. , Swanson M.C. , Campbell A.R. , Wynn S.R.
Published: August 2009

To further study the immunology of insect hypersensitivity, we identified and partially characterized the principal allergens in whole body German cockroach (WBGCR) (Blattella germanica) and compared this extract to whole body antigens prepared from other insects. WBGCR extract was fractionated over a calibrated Sephadex G-200 column; peak allergenic activity was contained in fraction 3 (GCR3), containing components with apparent molecular weights ranging from 12,500 to 75,000 daltons. The antigenicity, allergenicity, and specificity of GCR3 components were tested by using rabbit antisera raised to GCR3 or true armyworm (Pseudaletia unipuncta). Radioimmunoassay, cross-inhibition and immunoblot studies revealed, particularly in the IgE system, that GCR3 contained an allergen with a pI of 6.7 and MW of 36,000 daltons that was unique to WBGCR extracts and not present in other insect species, including true armyworm, caddis fly, lakefly, yellow jacket, or honeybee. This GCR3 component may represent a specific marker for the diagnosis of cockroach hypersensitivity in an insect-sensitive population of individuals.
ELISA Analysis of IgA Subclass Antibodies to Dietary Antigens
Authors: Kemp Michael , Husby Steffen , Larsen Mogens Lytken , Svehag Sven-Erik
Published: August 2009

Enzyme immunoassays for the quantitation of IgA1 and IgA2 antibodies to dietary antigens were developed. Serum IgA1 antibodies to bovine serum albumin (BSA) were detectable in 2/30 healthy adults, in 3/26 healthy children, and at high levels in 8/11 children with coeliac disease, without relation to gluten exposure. IgA1 antibodies to ovalbumin (OA) and beta-lactoglobulin (BLG) at high titers were seen in one coeliac child but were otherwise low or absent. IgA2 antibodies to BSA were detectable in 28/48 healthy subjects and in 8/11 coeliac children. IgA2 antibodies to OA and BLG were measurable in a few samples from each group. IgA1 antibodies to the gluten component glycgli were found at low levels in 15/56 normal sera, and anti-glycgli antibodies of the IgA2 subclass in 14/48 sera from healthy persons, also at low levels. IgA1 anti-glycgli antibodies were measurable in 5/11 sera from CD patients on a gluten-free diet. Elevated levels of IgA1 anti-glycgli antibodies were detected in all sera from CD patients challenged with gluten, except in 1 patient with a markedly reduced serum IgA level. In contrast, the IgA2 anti-glycgli antibody levels were unaffected. Thus, increased levels of IgA antibodies to dietary protein antigens in childhood coeliac disease were observed only within the IgA1 isotype.
Isolation and Characterization of Poa p I Allergens of Kentucky Bluegrass Pollen with a Murine Monoclonal Anti-Lol p I Antibody
Authors: Lin Zhengwei , Ekramoddoullah Abul K.M. , Kisil Fred T. , Hebert Jacques , Mourad Walid
Published: August 2009

The Poa p I allergens were isolated from the retentate fraction of a dialyzed preparation of an aqueous extract of Kentucky bluegrass pollen by means of a reverse immunosorbent prepared with a murine anti-Lol p I monoclonal antibody, Mab 290-A-167. By sodium dodecyl sulphate-polyacrylamide gel electrophoresis and preparative isoelectrofocusing, Poa p I was found to consist of a 35.8-kD component with an iso-electric point of 6.4 and a 33-kD component with one of 9.1 and designated as Poa p la (acidic) and Poa p lb (basic), respectively. The relative protein content of these components was estimated from the intensity of the stained bands following sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thus, Poa p la appeared to be the major protein constituent, and on Western immunoblot it also bound the monoclonal antibody to a greater extent than Poa p lb. On the other hand, Poa p lb was shown by Western immunoblot and autoradiographic analysis, to bind to a greater extent the IgE antibodies present in a pool of sera from grass-allergic individuals. Therefore, Poa p lb was considered as the major allergenic component of Poa p I. By competitive inhibition of the radioallergosorbent test, it was demonstrated that the Mab inhibited the binding of Poa p I allergens to human IgE antibodies to the extent of 70%. Hence, it is suggested that Mab and human IgE antibodies recognize identical or closely related determinants of Poa p I allergens.
Autoantibody to Human c-myc Oncogene Product in Autoimmune Patient’s Sera
Authors: Deguchi Yasuhiro , Negoro Shigeru , Kishimoto Susumu
Published: August 2009

Sera from autoimmune diseases including systemic lupus erythematosus, dermatomyositis, progressive systemic sclerosis and Sjögren’s disease, were examined for reactivity against human c-myc oncogene product. We first found the presence of the autoantibody to human c-myc oncogene product in sera from autoimmune diseases. No significant correlation between the presence of the anti-c-myc product antibody and disease activity, clinical disease types and other autoantibodies such as antinuclear factor, anti-DNA antibody, anti-RNP antibody and anti-Sm antibody were shown in this study.
Elution of Kidney-Bound Antibody by pH-Shift Does Not Discriminate between Antibodies Differing in Functional Affinity for Their Ligand
Authors: Shih Winston Y. , Clayman Michael D. , Kelly Carolyn J. , Hines William , Neilson Eric G.
Published: August 2009

Antibodies eluted from kidneys by traditional methods of pH shift have been used as reagents in a wide variety of experimental analyses without knowledge of whether their ligand affinity influenced their removal from parenchymal tissue. In the current study we employed two monoclonal antibodies, differing only in their functional affinity (high; K = 2.1 × 108/M and low; K = 6.2 × 106/M) to a common ligand found on the renal basement membrane, to evaluate whether a standard elution technique might selectively facilitate the removal of one antibody over the other. Our findings indicate, however, that the routine methods of elution by pH shift remove both antibodies equally well (41–48%), and without loss of paratypic function. These results provide new evidence that elution by pH shift can produce eluate antibodies which are not biased by preferred affinities.
Progesterone Assay by Inhibition of Reverse Passive Haemagglutination
Authors: Coombs R.R.A. , Kirk Denise , Saunders J. , Heap R.B. , Taussig M.J.
Published: August 2009

An assay for the steroid hormone progesterone is described based on the inhibition of the agglutination by progesterone-bovine serum albumin conjugate of red cells coupled with monoclonal antiprogesterone antibody (reverse passive haemagglutination). A low-affinity agglutination system produced the optimal sensitivity, capable of detecting less than 1 ng/ml steroid. This assay has the simplicity and sensitivity for a potential clinical test for placental or corpus luteum function. It also serves as a model for detection of other small ligands (drugs, hormones) by inhibition of reverse passive haemagglutination.
Detection of Antigens from Gram-Negative Bacilli in Urine of Children with Urinary Tract Infections
Authors: Kim Dong Soo , Gorzynski Eugene A. , Milgrom Felix
Published: August 2009

An enzyme immunoassay for detection of bacterial antigens in urine specimens from urinary tract infections (UTI) was developed. The antigens detected in infections by Enterobacteriaceae: Escherichia coli, Enterobacter aerogenes, and Proteus sp. were different from the enterobacterial common antigen (ECA). They were digested by trypsin and were characterized by remarkable thermostability; their molecular weights were approximately 34,000 and 31,000. They were detected by means of antisera to ECA or to suspension of E. coli. The frequency of positive results with urine specimens from enterobacterial UTI was 87–99%. In contrast, the urine specimens of UTI by Pseudomonas aeruginosa were positive in fewer than 20% with anti-E.coli sera, but were positive in more than 85% in tests with antiserum to P. aeruginosa. The possible diagnostic application of the described tests was discussed.
Development of Monoclonal Mouse Antibodies Specific for Allergenic Components in Ryegrass (Lolium perenne) Pollen
Authors: Smart I.J. , Heddle R.J. , Zola H. , Bradley J.
Published: August 2009

The development of monoclonal mouse antibodies against ryegrass (Lolium perenne) pollen allergens is described. Hybridoma colonies secreting antibodies specific for allergenic components were detected using an enzyme-linked immunosorbent assay. Positive colonies were cloned and expanded. The pollen components with which the monoclonal antibodies interact were identified and characterised following sodium dodecyl sulphate polyacrylamide gel electrophoresis and electrophoretic transfer to nitrocellulose. In this paper six monoclonal mouse antibodies are described. Three antibodies interact with a single molecule of between 30,000 and 35,000 daltons. One antibody interacts with a component of 16,000 daltons whereas the remaining two antibodies react with more than one component, one reacting with two components at 28,000 and 30,000, and the other with five components having molecular weights between 18,000 and 71,000 daltons.
Regulation of the Development of Plaque-Forming Cells to Bromelain-Treated Syngeneic Mouse Erythrocytes in Bone Marrow Cell Cultures
Authors: Garzelli C. , Forlani A. , Lombardi S. , Colizzi V. , Campa M. , Falcone G.
Published: August 2009

The development of plaque-forming cells (PFC) to bromelain-treated syngeneic mouse red blood cells (Br-MRBC) was studied in bone marrow cell (BMC) cultures. It was found that the number of marrow PFC to Br-MRBC does not show the typical spontaneous increase observed in spleen cell (SPC), or peritoneal cell (PC) cultures. The number of anti-Br-MRBC PFC was markedly increased by lipopolysaccharide (LPS), even in conditions in which cell proliferation was blocked by mitomycin C, suggesting the presence of high numbers of Br-MRBC-specific precursor cells, potentially capable of differentiating into autoantibody-producing cells, in the marrow. Moreover, the low levels of anti-Br-MRBC PFC were further reduced in the presence of concanavalin A (Con A). The addition of Con A-activated BMC to BMC, SPC, or PC cultures actively suppressed the development of anti-Br-MRBC PFC. Con A-activated BM suppressor cells were found to be Thy 1.2-negative, Ig-negative, nonadherent cells. A possible role for the BM suppressor cell in tolerance to self antigens is discussed.
Comparison of Antigens in the Rat MHC That Act as CML Determinants
Authors: Liebert Monica , Kunz Heinz W. , Gill, III Thomas J. , Cramer Donald V.
Published: August 2009

We have examined a variety of RT1 haplotypes using congenic strains of rats to investigate the comparative relationships among MHC antigens that have been reported to serve as cell-mediated lympholysis (CML) targets. These studies demonstrate that: (1) RT1.A antigens can act as CML determinants, and in some haplotypes they display identical serological and CML reaction patterns; (2) in some donor-recipient strain combinations RT1.A antigens may act as immunodominant determinants, preventing the generation of effectors capable of recognizing other antigens, and (3) a portion of the antigenic determinants that define the anti-CT CML response can be mapped to the same region of the rat MHC as the RT1.E locus.
Studies on the Antigenic Composition of Aqueous Cotton Dust Extracts
Authors: O’Neil Carol E. , Reed Margaret A. , Aukrust Lars , Butcher Brian T.
Published: August 2009

An immune response to cotton dust components has been proposed as a possible mechanism of byssinosis. To evaluate this possibility, specific precipitating antibodies against aqueous cotton dust extracts (ACDE) were raised in rabbits, and a reference pattern for crossed immunoelectrophoresis (CIE) was established using this extract. A significant portion of the immune response was directed against Alternaria tenuis, Aspergillus niger, and Fusarium solani, all documented contaminants of cotton dust. Preliminary crossed radio-immunoelectrophoretic studies demonstrated that some of these antigens are allergenic in man, suggesting that an IgE-mediated hypersensitivity against cotton dust, or a fungal component of the dust, may be important in byssinosis pathogenesis.
Hapten-Specific Help Mediated by an Anti-ABA T Cell Line
Authors: Kojima Asato , Thomas Wayne R. , Mottram Patricia L. , Miller Jacques F.A.P.
Published: August 2009

Long-term cultures of functional T cells were grown from mice injected with azobenzene arsonate (ABA) coupled spleen cells or mice painted with the diazonium salt of p-phenylarsonate. All cell lines produced delayed hypersensitivity to ABA. When tested for helper function in an antibody-forming cell assay only one line, AA3, helped plaque-forming cell formation. It was able to help anti-FLU response of FLU-HGG primed B cells to ABA-KLH-FLU. The help required linkage of the ABA and FLU determinants because cultures stimulated with a mixture of ABA-KLH and TNP-KLH-FLU did not show increased PFC. These experiments demonstrate hapten-specific help in a system which eliminates involvement of passively absorbed antibody in hapten recognition.
Decreased Sensitivity and Response of Isolated Tracheal Muscle to Methacholine and Histamine without Changing the Activity of Pulmonary Muscarinic Receptors in the Egg Albumin-Sensitized Guinea Pig
Authors: Cheng John B. , Townley Robert G.
Published: August 2009

Guinea pigs were sensitized by daily intraperitoneal injections of 100 mg of egg albumin for 3 consecutive days and sacrificed at 30–35 days after the last injection. The in vitro sensitivity and response of the sensitized tracheal muscle to methacholine and histamine were significantly less than those of littermate controls. However, there was no difference between the dissociation constants and concentrations of pulmonary muscarinic receptor binding sites of the control and sensitized guinea pigs. We conclude that subsensitivity and hyporesponsiveness to methacholine and histamine occur in the airway muscle of this guinea pig model of experimental asthma and suggest that the decreased reactivity to methacholine might be due to defects beyond the receptor.
Anti-Allergic Effects of Milrinone and Sulmazole in Isolated Rat Lungs in Comparison with Theophylline
Authors: Post Mark J. , te Biesebeek Jan Dirk , van Rooij Hans H. , Wemer Johan , Porsius Arijan J.
Published: August 2009

Milrinone and sulmazole, two recently developed drugs, inhibit specific fractions of the phosphodiesterase (PDE) isozyme system. Since theophylline aspecifically inhibits the PDE complex, we compared the effects of milrinone and sulmazole with those of theophylline on antigen-induced bronchoconstriction, vasoconstriction, mediator release and leukotriene production. In the isolated perfused and ventilated lung of actively sensitized rats, we elicited antigen-induced bronchoconstriction, vasoconstriction and release of mediators like histamine, 5-hydroxytryptamine (5-HT) and slow-reacting substance of anaphylaxis (SRS-A). Milrinone, sulmazole and theophylline inhibited antigen-induced bronchoconstriction and vasoconstriction in a dose-dependent manner with minor differences in potency. Antigen-induced release of preformed mediators like histamine and 5-HT was inhibited only at high concentrations of milrinone, whereas sulmazole failed to inhibit mediator release. Theophylline also failed to inhibit 5-HT release. However, SRS-A synthesis was markedly reduced by these drugs in relatively low concentrations. It is concluded that milrinone and sulmazole have anti-allergic effects similar to those of theophylline and that all three PDE inhibitors reduce SRS-A synthesis.
Histamine Release from Peripheral Blood Leukocytes with Purified Bee Venom Allergens: Effect of Hyperimmune Beekeeper Plasma
Authors: Clinton P.M. , Kemeny D.M. , Youlten L.J.F. , Lessof M.H.
Published: August 2009

The response of 15 strongly bee-venom-allergic patients to highly purified venom allergens was compared using skin prick test titration, peripheral blood leukocyte (PBL) histamine release and radioallergosorbent test with three highly purified bee venom allergens: phospholipase (PLA2), hyaluronidase (HYAL) and acid phosphatase (ACID P). Sensitivity to the three allergens ranked in the same order for all three tests and in each case PLA2 was found to the most potent allergen. In the presence of hyperimmune beekeeper plasma, maximum histamine release was reduced significantly for all three allergens (p < 0.001). Furthermore, hyperimmune beekeeper plasma increased the amount of allergen required for a comparable release of histamine (mean shift in dilution curve PLA2 917-fold; HYAL, 492-fold; ACID P, 61-fold). The release of histamine from whole blood was also compared with PBL + 10% normal human serum (NHS). For all three allergens maximum release was much lower from whole blood compared with washed cells + 10% NHS (p < 0.001). These data confirm PLA2 as the major bee venom allergen by all three tests. Hyperimmune beekeeper plasma reduces maximum histamine release and increases its threshold. Histamine release in response to ACID P appears harder to block with hyperimmune beekeeper plasma than that provoked by PLA2 or HYAL (p < 0.01). Whole blood releases less histamine and requires more allergen than washed cells, indicating that sensitivity of PBL in vivo is unlikely to be as great as washed PBL in vitro.
Relationship of Serum Venom-Specific IgE Titers to Clinical Aspects of Stinging Insect Allergy
Authors: Reisman Robert E. , DeMasi James M.
Published: August 2009

Fifty patients were randomly selected who had markedly elevated titers of serum venom-specific IgE (range 100–340 IU, mean 144 IU), and compared with 50 patients with low titers (range 0–50 IU, mean 25 IU). At the time serum was obtained, none of the patients had received venom immunotherapy (VIT). Analysis of the demographic data showed more children in the high-RAST group, and comparable matching of the groups for sex, presence of atopy, and culprit insect identification. The nature of the reaction preceding evaluation was similar in both groups. In the high-RAST group, there were 12 local, 15 mild systemic, and 23 severe systemic reactions. In the low-RAST group, there were 8 local, 15 mild systemic, and 27 severe systemic reactions. Other prior local reactions had occurred in 4 high-RAST and 4 low-RAST patients, and other prior systemic reactions had occurred in 3 high-RAST and 8 low-RAST patients. The incidence of systemic and local reactions following VTT was low in both groups. All but 1 patient in each group reached maintenance venom doses. Following initiation of VIT, 10 of 28 patients in the high-RAST group and 5 of 29 patients in the low-RAST group had a further rise in antibody titers. Patients in both groups showed a decrease in titers over a period of time, with or without VTT. Serum venom-specific IgG was greater in the high-RAST group (mean 9.4 U) than in the low-RAST group (mean 3.8 U). The absolute titers of serum venom-specific IgE appear unrelated to any specific feature of stinging insect sensitivity.
Prevention and Reversal of Aerosol LTD4-Induced Changes in Guinea Pig Pulmonary Mechanics by Wy-48252, an Orally Active LTD4/E4 Receptor Antagonist
Authors: Hand James M. , Auen Melissa A. , Chang Joseph , Englebach Isabella M.
Published: August 2009

Anesthetized male albino guinea pigs were prepared for recording changes in the pulmonary mechanics parameters, dynamic compliance (Cdyn) and airway conductance (Gaw). Two aerosol leukotriene D4 (LTD4) challenges (0.125 μg/ml, 3–7 breaths, 1 h apart) were administered via an ultrasonic nebulizer to each animal and, produced reductions in Cdyn that were approximately 60% of those of Gaw. Intraduodenal administration of the LTD4/E4 receptor antagonists Wy-48252 (30 min), Wy-45911 and Ly-171883 (15 min) produced dose-related inhibition of the second LTD4 challenge. From the data, ID50 values were calculated and, Wy-48252 was 8- to 17-fold more potent (mg/kg) than Wy-45911 or Ly-171883. Responses to histamine were not altered by the antagonists. In separate experiments, Wy-48252 (0.3 and 1 mg/kg i.v.) rapidly reversed aerosol LTD4-induced decreases of Cdyn and Gaw, but did not reverse pulmonary mechanics decreases produced by prostaglandin F2α. The results indicate that systemically administered LTD4 receptor antagonists can both prevent and reverse bronchoconstriction produced by aerosol LTD4 in the guinea pig.
Maternal Determinants of Neonatal Immune Response to Ovalbumin: Effect of Breast Feeding on Development of Anti-Ovalbumin Antibody in the Neonate
Authors: Okamoto Yoshitaka , Freihorst Joachim , Ogra Pearay L.
Published: August 2009

Inbred Brown-Norway female rats were immunized intraperitoneally with ovalbumin (OVA) or sham-immunized 14 days before and 10 days after mating. In subsequent studies with OVA, babies fed by immunized mothers, regardless of whether they were born from immunized or sham-immunized mothers, showed suppression of IgG, IgM and IgE anti-OVA responses. In additional studies, these babies developed OX-8-positive but W3/25-negative phenotypic suppressor T-cells specific for anti-OVA antibody production. However, these regulatory cells did not react with OVA itself when tested for in vitro proliferative response to OVA. Subsequent immunization of the neonates with OVA appeared to abrogate suppression of IgG and IgM antibody responses. However, maternally induced suppression of IgE persisted and was not influenced by subsequent immunization.
Modulation of Total IgE Levels in Serum of Normal and Athymic Nude BALB/c Mice by T Cells and Exogenous Antigenic Stimulation
Authors: Savelkoul H.F.J. , van den Akker T.W. , Soeting P.W.C. , van Oudenaren A. , Benner R.
Published: August 2009

Several different grades of T-system impairment were studied for their effects on the total serum IgE concentration in BALB/c mice. Homozygous athymic nu/nu mice and their heterozygous nu/+ littermates were compared for serum IgE levels while kept under either barrier-maintained or conventional conditions. The results show a paradox between the T-cell dependency of the IgE immune response and the increased levels of serum IgE in the absence of T cells. Both barrier-maintained and conventionalized nu/nu mice have at least two fold increased serum IgE levels as compared to nu/+ mice. With age, IgE levels increased faster and reached higher plateau values in nu/nu than in nu/+ mice. Moreover, after adult thymectomy of BALB/c mice the serum IgE levels increased up to 15-fold at 4 months of age, while infusion of immunocompetent T cells in nude mice resulted in a 2- to 5-fold decrease of the IgE level.