bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunogenicity of Hapten-Carrier Conjugates Associated with Liposomes
Authors: Neveu P.J.
Published: August 2009

Immunogenicity of liposome-entrapped hapten-carrier conjugates was studied in guinea pigs. Animals immunized intravenously with a subimmunogenic dose of a hapten-carrier complex entrapped in liposomes exhibited cutaneous anaphylactic reactions to the hapten, even when the liposomes were treated with trypsin, but not delayed hypersensitivity (DH) reactions to the carrier. Animals immunized with a mixture of empty liposomes and antigen showed anaphylactic reactions to the hapten that appeared to be elicited by antigen adsorbed on the outer membrane of liposomes. The induction of anaphylaxis to the hapten by liposome-associated antigen was not due to an adjuvant property of liposomes. DH reactions to the carrier after injection of a liposome-entrapped subimmunogenic dose of a hapten-carrier conjugate could be observed only after activation of macrophages by Bacillus Calmette-Guérin.
Expansion of a T Lymphocyte Subpopulation (CD3+,4––,8––) after Immunodepression Associated with Disseminated Histoplasmosis
Authors: Lehmann Paul F. , Donabedian Haig , Sawyer Tom
Published: August 2009

A major subpopulation of T lymphocytes bearing a high density of CD3 (T3/Leu 4) with no detectable CD4 (T4/Leu 3a) or CD8 (T8/Leu 2a) was found in a patient with cardiomyopathy. Previously the patient had developed disseminated histoplasmosis which had been associated with a profound anergic state. The proportion of CD3+ lymphocytes that bore the phenotype CD3+,4––,8–– has been increasing in the absence of reactivation of the histoplasmosis. The CD3+,4––,8–– lymphocytes bound anti-CD2 but did not bind WT31, a monoclonal antibody specific for the αβ heterodimer of the T cell antigen receptor.
Characterization of the Main IgE-Binding Components of Cat Dander
Authors: Duffort O. , Carreira J. , Lombardero M.
Published: August 2009

Cat dander extract (CDE) was fractionated by preparative isoelectric focusing (pIEF). All the fractions showed allergenic activity, but the most acidic ones (pH range 3.3–4.3) had the highest specific activity. These fractions also had the highest cat allergen 1 content and a partial antigenic identity as determined by fused rocket immunoelectrophoresis. Immunoblotting of the pIEF fractions and the crude CDE after analytical IEF showed the presence of native proteins with IgE-binding ability all along the pH range 3.3–6.2. However, when the IgE-immunoprecipitated components of the most acidic pIEF fractions were focused in the presence of urea, a neat band was found at pH3.9. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of CDE proved the presence of, at least, 9 polypeptides with affinity to IgE, those with molecular weights of 18,000 and 22,000 daltons being the most active ones. These polypeptides presumably belong to cat allergen 1 since they were found in the most acidic pIEF fractions as well as in the crossed immunoelectrophoresis arc corresponding to this allergen. Furthermore, we found a group of active proteins with molecular weights of 10,000, 22,000, 25,000, 50,000 and 65,000 daltons (cat albumin) and with isoelectric points between 3.3 and 6.2.
Hydrocortisone Both Decreases the Up-Regulation of Complement Receptors CR1 and CR3 and the Ingestion Process of Human Granulocytes
Authors: Forslid Jan , Hed Jan
Published: August 2009

We have studied the effect of hydrocortisone on the complement receptor expression (CR1 and CR3) of human granulocytes during the up-regulation phase and the following stable period when exposed to N-formyl-methionyl-leucyl-phenylalanine (fMLP) or the medium alone at 37 °C. The receptor expression was also correlated to the C3bi-mediated phagocytosis with special reference to attachment and ingestion. The results showed that after incubation with fMLP the increased expression of CR1 and CR3 is accompanied by an increased ingestion, but not attachment, of complement-opsonized yeast particles. This increase was significantly lower with regard both to attachment and ingestion, as well as receptor expression if hydrocortisone was present during the up-regulation phase. However, the addition of hydrocortisone after the up-regulation of fMLP-treated granulocytes decreased the ingestion of particles but not the CR1 and CR3 expression. The results indicate a membrane effect of hydrocortisone that affects both the receptor mobilization and ingestion mechanism of the phagocytes.
Functional Heterogeneity among Cytotoxic Clones Derived from Natural Killer Cells
Authors: Christmas S.E. , Moore M.
Published: August 2009

Clones were obtained from highly purified populations of human peripheral blood natural killer (NK) cells propagated in the presence of interleukin-2 and phytohaemagglutinin. Almost all clones were cytotoxic against standard NK targets and many were also able to kill the B lymphoblastoid cell line BSM. This latter property was not necessarily a result of the incorporation of this cell line into the feeder mixture used to derive the clones. In most cloning experiments there was a high degree of concordance between the killing of the NK targets K562 and Molt 4 by panels of clones. In some cases this extended to the killing of BSM targets but in other instances there was no relationship or even an inverse correlation between killing of BSM and other targets. In a single cloning experiment there was no relationship between killing of BSM and Raji targets. In some cases a panel of clones could be divided into two or more distinct groups based on their differential activity towards BSM and K562. Such differences were not solely due to inter-donor variation. These findings were extended by cold target inhibition experiments in which at least three types of clone were identified. In one group of clones, which was nonreactive towards BSM, cold BSM significantly enhanced the killing of K562 in a dose-dependent fashion. These experiments provide evidence for a limited degree of functional heterogeneity among clones derived from human peripheral blood NK cells.
Application of the First International Standard of Dermatophagoides pteronyssinus (House Dust Mite) in the Evaluation of Allergen Extracts Produced from Two Different Source Materials
Authors: Maasch H.J. , Wahl R. , Fuchs T.
Published: August 2009

The first international standard (IS) of the house dust mite Dermatophagoides pteronyssinus was used to compare side-by-side two allergen extracts prepared from two different house dust mite sources, namely whole mite cultures (WMC) and purified mite bodies (PMB), by employing several biochemical and immunochemical methods. In most methods employed, IS and WMC resembled more to each other than to PMB. These similarities comprehend protein contents, protein patterns, antigen and allergen patterns as well as total in vitro activity obtained in RAST inhibition (RI). The differences between WMC and PMB could be reproduced in different batches. Standardization in relation to IS of D. pteronyssinus yields activity ratios ranging from 2.4 to 70.1 between WMC and PMB depending on the variation of RI employed, indicating quantitative differences between the two preparations. When coupled to BrCN acivated paper disks and employed in direct RAST, both preparations yielded almost identical binding values of specific IgE. Also in skin prick test, WMC and PMB produced very similar results in 12 mite-allergic patients. It was not possible to differentiate between the two preparations when used in concentrations which are usually employed for diagnostic purposes. These findings demonstrate the diversity of allergens in mite extracts derived from different sources and reveal the problems which are involved with the use of the WHO D. pteronyssinus IS.
Inhibition of Histamine Release from Rat Mast Cells by Botulinum C2 Toxin
Authors: Böttinger Heiner , Reuner Karl-Heinrich , Aktories Klaus
Published: August 2009

The effects of botulinum C2 toxin, which ADP-ribosylates actin, were studied on the stimulated histamine release of rat peritoneal mast cells. Treatment of mast cells with botulinum C2 toxin for 4 h inhibited histamine release stimulated by compound 48/80 or MCD-peptide maximally by about 50%. A half-maximal and maximal inhibition occurred at about 30 and 450 ng/ml botulinum C2 toxin. At similar concentrations the toxin ADP-ribosylated actin in intact mast cells. Botulinum C2 toxin largely impaired the histamine release stimulated by 12-O-tetradecanoylphorbol-13-acetate but not by the ionophore A23187. The data indicate that botulinum C2 toxin partially inhibits the stimulated histamine release and suggest that actin is involved in the stimulus-secretion coupling in mast cells.
Role of Lymphocytes in the Modulation of the Immunogenicity of Bone Marrow: A Rat Model
Authors: Wottge H.-U. , Müller-Ruchholtz W.
Published: August 2009

In a rat model we describe the role T lymphocytes play in the immunogenicity of bone marrow. Mainly they modulate the MLC-like stimulatory cell potential and the MHC class II antigen expression of nonlymphoid bone marrow cells via the release of newly detected specific soluble factors and immunologically unspecifîc ones. These findings are discussed in relation to the graft rejection encountered in clinical bone marrow transplantation following certain T depletion protocols.
Mediators of C5a-Induced Bronchoconstriction in the Guinea Pig
Authors: Regal Jean F. , Bell Randy L.
Published: August 2009

The effect of intravenous injection of C5a on pulmonary resistance and dynamic lung compliance was determined in anesthetized, artificially respirated guinea pigs. A mixture of C5a plus C5ades arg was purified from yeast-activated guinea pig serum and is referred to as C5a. Intravenous injection of C5a caused a dose-related bronchoconstriction as evidenced by a decrease in compliance and increase in resistance. Conversion of the C5a in the mixture to C5ades arg by carboxypeptidase B digestion did not significantly alter the magnitude of the bronchoconstriction. Pharmacological antagonists were employed to determine if histamine, acetylcholine or products of the arachidonate metabolism were mediators of C5a-induced bronchoconstriction. The histamine H1 antagonist pyrilamine inhibited the C5a-induced bronchoconstriction, suggesting the involvement of histamine. The cholinergic receptor antagonist atropine in combination with pyrilamine caused an inhibition of the C5a-induced increase in resistance but not compliance, suggesting acetylcholine does not play a major role in C5a-induced bronchoconstriction beyond its known role in participating in histamine-induced bronchoconstriction. Involvement of arachidonate metabolites was suggested by the ability of the cyclooxygenase inhibitor, indomethacin, to prevent the C5a-induced bronchoconstriction. Because indomethacin also caused a delay in the leukotriene C4 (LTC4)-induced bronchoconstriction, the participation of peptidoleukotrienes in the C5a-induced bronchoconstriction could not be ruled out. The leukotriene antagonists FPL 55712 and L-649,923 were evaluated for their specificity in inhibiting LTC4-induced bronchoconstriction. FPL 55712 was nonselective since it inhibited prostaglandin D2 and histamine-induced bronchoconstriction as well as LTC4-induced bronchoconstriction. L-649,923 inhibited only the LTC4-induced bronchoconstriction and was without effect on the C5a-induced bronchoconstriction, suggesting that peptidoleukotrienes are not important mediators of C5a-induced bronchoconstriction. Using radioimmunoassay, the change in peptidoleukotriene levels detected in plasma during C5a-induced bronchoconstriction was not significantly different from 0. Thus, these studies have quantitated the C5a-induced decrease in dynamic lung compliance and increase in pulmonary resistance and suggest that histamine and cyclooxygenase products, but not peptidoleukotrienes, play a major role in C5a-induced bronchoconstriction.
Suppression of the Murine Anti-Trimellityl (TM) IgE Response: Induction of B Cell Tolerance by Conjugates of TM and Polyvinyl Alcohol
Authors: Wei B-Y. , Holford-Strevens V. , Sehon A.H.
Published: August 2009

It was previously shown that administration of conjugates of trimellitic anhydride with polyvinyl alcohol (TM-PVA) could suppress the IgE anti-TM immune response of mice sensitized with a TM-ovalbumin conjugate. In the present study, the existence of TM-specific B cell tolerance was shown by cell transfer experiments in which splenic B cells from mice treated with TM-PVA failed to interact with either the helper T (Th) cells of carrier primed recipients or with Th cells derived from carrier-primed donors. In contrast to previous findings from this laboratory indicating that tolerogenic conjugates of PVA and the 2,4-dinitrophenyl group led to both B cell tolerance and activation of suppressor T (Ts) cells, no evidence was obtained for the induction of Ts cells by TM-PVA. Thus, the induction of demonstrable Ts cells by hapten-PVA conjugates may depend on some property conferred by the haptenic group.
Involvement of PAF-Acether in Anaphylactic Bronchoconstriction Induced in Guinea Pigs by Aerosolized Antigen
Authors: Lagente Vincent , Cirino Maria , Desquand Stéphanie , Lefort Jean , Vargaftig Boris
Published: August 2009

Bronchoconstriction following the aerosolisation of PAF-acether or antigen to guinea pigs induces autodesensitization, but the responses to direct spasmogenic substances are not modified. Bronchoconstriction by PAF-acether is not reduced when it is aerosolized to passively sensitized animals previously desensitized by repeated inhalations of the allergen (ovalbumin). In contrast, when passively sensitized animals are initially desensitized to PAF-acether by repeated inhalations of this agonist, ovalbumin aerosolization induces a bronchoconstriction which is significantly reduced when compared with the response obtained in nondesensitized animals though, in this case, the response to aerosolized histamine is not modified. Thus, PAF-acether is released during intrapulmonary anaphylactic shock induced by aerosolized ovalbumin and can be a prime candidate for its development.
On the Nature of Natural Thymocytotoxic Antibodies
Authors: Sandberg G. , Stenvinkel C.
Published: August 2009

The thymocytotoxic activity of human sera against guinea pig cells was earlier shown to be mediated by IgM and a heat-labile serum factor, presumably complement. It is not known if such natural cytotoxic activity represents background activity of preexisting clones of immunoglobulin-producing cells, cross-reacting antibodies appearing after immunization, physiological immune regulatory molecules, or components of an immune network. We have therefore examined the presence of thymocytotoxic IgM molecules in normal adult and neonatal sera and in a number of diseases which affect the lymphoid and immune system. The thymocytotoxic effect of serum was measured in different dilutions, both directly and after heat inactivation of the sera and supplementation with a standard amount of IgM-depleted serum, which is inactive in itself but provides a fixed amount of the heat-labile cofactor. The cytotoxic IgM was present in various amounts in all sera tested, although in neonates very small amounts were found. No specific aberration in toxic activity was seen in rheumatoid arthritis or a number of hematological diseases. In general, the cytotoxic activity correlated well with the total amount of IgM. However, in cases of chronic lymphocytic leukemia, idiopathic thrombocytopenic purpura and immunocytoma aberrant cytotoxic activities were found, but to ascertain a connection between these diseases and the factor would require a more extensive follow-up study. The results indicate that the naturally occurring thymocytotoxic IgM is widespread and may reflect a clone of B cells which is activated by an endogenous stimulus, or by some ubiquitous exogenous immunogen.
Studies on Murine IgE with Monoclonal Antibodies
Authors: Hirano Takao , Miyajima Hiroaki , Kitagawa Hiroshi , Watanabe Naohiro , Azuma Miyuki , Taniguchi Osamu , Hashimoto Hiroshi , Hirose Shun-ichi , Yagita Hideo , Furusawa Shuichi , Ovary Zoltan , Okumura Ko
Published: August 2009

Rat monoclonal antibodies were constructed by fusion of immunized rat spleen cells with a non-secreting mouse myeloma cell. Two monoclonal antibodies (6HD5 and HMK-12) were selected for further study. Both reacted with various IgE molecules of different specificities and different allotypes, but did not react with immunoglobulins of other isotypes and with light chains. These antibodies were therefore anti-isotypic (IgE) and not anti-allotypic or anti-idiotypic. It was shown by competition studies that these antibodies recognize different epitopes on the FcRε fragment. A sensitive ELISA for the quantitation of murine IgE was developed with these monoclonal antibodies; the sensitivity was between 2 and 250 ng/ml for detection of serum IgE levels. Good correlation was obtained with protein amounts as determined by enzyme-linked immunosorbent assay (ELISA) and passive cutaneous anaphylaxis (PCA) activities. Both monoclonal antibodies were used to study anaphylactic reactions elicited by IgE antibodies. Both could inhibit PCA reactions and both could elicit reverse PCA reactions.
Variation in the Expression of Macrophage Fcε Receptors in Relation to Experimental Rat Schistosome Infection
Authors: Pestel Joël , Joseph Michel , Dessaint Jean-Paul , Capron André
Published: August 2009

The relationship between IgE and rat peritoneal macrophages during the course of Schistosoma mansoni infection was examined. Immune macrophages exhibited an IgE-dependent schistosomulicidal activity mainly between the 5th and the 7th weeks. At this period the percentages of anti-lgE rosettes whose variations appeared consecutive to the variations in the serum IgE level were higher. Following incubation in a serum-free medium immune macrophages could release membrane-associated IgE, to a certain extent, and consequently formed rosettes with IgE-coated erythrocytes, indicating the existence of Fcε receptors. Thus, in terms of rosettes, their number varied in the course of the disease. In fact IgE-rich serum (i.e. at day 42) as IgE molecules could induce this macrophage Fcε receptor. Moreover when transferred to syngeneic rats in associaton with parasite-specific IgE Fcε R2+ macrophages led to some protection against challenge infection. All the data reported here emphasize the determining role of macrophages and IgE in immunity to schistosomiasis.
Lymphocyte-Neutrophil Interactions: Opposite Effects of Interleukin-2 and Tumour Necrosis Factor-Beta (Lymphotoxin) on Human Neutrophil Adherence
Authors: Seow W.K. , Thong Y.H. , McCormack J.G. , Ferrante A.
Published: August 2009

Human neutrophil adherence was enhanced by recombinant human tumour necrosis factor-β (TNF/J) but suppressed by recombinant human interleukin-2 (IL-2). The opposite effects of these two lymphokines were observed over a range of concentrations consistent with their other biological activities, occurred within 15 min of incubation, and were still evident after 60 min. Pretreatment of neutrophils with both IL-2 and TNFβ resulted in adherence values intermediate between the values obtained with the individual lymphokines. IL-2 suppressed the stimulatory effects of both the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) and the phorbol ester phorbol myristate acetate (PMA). The combination of TNFβ with either FMLP or PMA produced enhancement of neutrophil adherence which exceeded that of either agent alone. These effects of the lymphokines were not due to endotoxin contamination since their effects were sensitive to heating and insensitive to polymyxin B treatment. These experiments provide further evidence for the critical role of these lymphokines in the regulation of acute and chronic inflammatory processes.
Antigens of Euparipha pisana (Snail)
Authors: Amoroso Saverio , Cocchiara Roberta , Locorotondo Giovanni , Parlato Alessandra , Lampiasi Nadia , Albeggiani Giuseppe , Falagiani Paolo , Geraci Domenico
Published: August 2009

The data obtained in this study suggest that eating Euparipha pisana (snail), a common food in Mediterranean countries, could give serious allergic reaction such as asthma. We describe here the identification and partial characterization of allergenic molecules form this new source. An aqueous extract of snail was obtained by homogenization in distilled water, centrifugation, dialysis and defatting with ethyl ether. Skin prick test (SPT) performed with the snail extract on 70 subjects allergic to the more common allergens of the Mediterranean area gave a SPT positivity in 61% of the subjects tested, with a mean value of histamine-equivalent prick (HEP) equal to 0.81 ± 0.25 (n = 43), while no SPT-snail-positive reactions were obtained by using the same extract on 30 not allergic subjects. To ascertain if such a sensitivity was IgE-mediated, sera from SPT-snail-positive subjects were analyzed by RAST, coupling the snail extract to polystyrene balls and to paper discs. 19% of the sera tested were RAST-positive, mean value of binding 4.8 ± 2.8% (n = 13), while when using sera from SPT-snail-negative subjects, the RAST mean value was 0.49 ± 0.18% (n = 27). Histamine release (HR) was also performed. Basophils prepared from SPT-snail-positive subjects were incubated with a snail extract. All of the SPT-snail-positive subjects gave a significant value of HR, mean value 21.8 ± 7% using 1 μg of snail extract (n = 16), while 1.41 ± 1.1% (n = 10) was the mean value obtained when SPT-snail-negative subjects were analyzed. The components of the snail extract, fractionated by sodium dodecylsulfate polyacryl-amide gel electrophoresis and then transferred to nitrocellulose membrane, were studied by electroblotting using 10 sera collected from SPT- and RAST-snail-positive subjects. After a reaction with 125I-antihuman IgE and autoradiography, 6 bands were capable of binding human IgE, ranging in molecular weight value from 12,000 to over 66,000 daltons. Finally, a strong cross-reactivity was detected by means of RAST inhibition, between the snail and the Parietaria judaica, Dermatophagoides pharinae and Poa pratensis allergens. The results obtained with four different methods provide evidence that the snail is a new source of allergenic molecules, and leads us to more extensive studies of this type of food allergy.
Interleukin-1-Like Activity Produced by Hybrids Constructed with Epstein-Barr-Virus-Transformed Human B Lymphocytes and Mouse Myeloma Cells
Authors: Garzelli Carlo , Puglisi Carla , Pacciardi Anna , Tarantino Enrico
Published: August 2009

Peripheral blood B lymphocytes from a donor with positive tuberculin skin test reaction were transformed into lymphoblastoid cell lines by Epstein-Barr virus and then fused by polyethylene glycol with mouse myeloma cells. Human-mouse hybrid cells producing human IgM monoclonal antibody to purified protein derivative of tuberculin were isolated, and the concentrated supernatant of one of these cell hybrids was tested for the capacity of interfering with DNA synthesis of human and mouse lymphocytes. The hybrid cell supernatant was found to contain soluble factors that increased DNA synthesis in unstimulated human and mouse lymphocytes and that, conversely, decreased DNA synthesis in concanavalin-A-stimulated cells. Gel filtration experiments showed that these antagonistic activities were due to at least two different factors, one of which resembled human interleukin-1 in biochemical and biological properties.
The Complement Membrane Attack Complex Stimulates the Prostanoid Production of Cultured Glomerular Epithelial Cells
Authors: Hänsch Gertrud M. , Betz Margarita , Günther Jeanette , Rother Klaus O. , Sterzel Bernd
Published: August 2009

Incubation of cultured rat glomerular epithelial cells (GEC) with sublytic amounts of the purified complement components C5b6, C7, C8 and C9 greatly stimulated the release of the prostanoids prostaglandin E (PGE) and thromboxane B2. Incubation of GEC with C5b-8 was also stimulatory, whereas omission of C7 abolished the enhanced prostanoid production. These effects were dose-dependent. The increased release of PGE was biphasic with peaks at 5 min and 24 h of incubation. The second peak could be prevented by treatment with cycloheximide, suggesting its dependence on protein synthesis. The observations on cultured GEC provide evidence that terminal complement components alter the metabolism of glomerular cells, resulting in increased production of prostanoids. The results are consistent with the concept that deposition of nonlytic amounts of complement in the glomerular capillary wall may affect the GEC in vivo and may indirectly contribute to abnormalities of the glomerular filter as it is seen in glomerular disease.
Immunological Suppression of Delayed Hypersensitivity Responses in Mouse Lungs as Reflected by Numbers of Mononuclear Cells, Mast Cells and Mucus-Producing Cells
Authors: Enander I. , Nygren H. , Ahlstedt S.
Published: August 2009

The suppression of delayed hypersensitivity (DH) in the lung as reflected by the appearance of mononuclear cells, mast cells and mucus-producing cells was studied in Balb/c mice. Immunosuppression was induced by intravenous and peroral administration of picrylsulfonic acid (PSA) in mice subsequently sensitized with picrylchloride (PiCl). The animals exhibited a decreased DH reactivity as assessed by ear thickness increase compared with mice sensitized but not exposed to PSA pretreatment. In mice exposed to PSA intravenously (suppressed) and sensitized with PiCl there was a decrease in the number of mononuclear cells in the lung after challenge, compared to mice sensitized and challenged only. Similarly, the mast cell and mucus-producing cell numbers were slightly lower in animals immunosuppressed intravenously with PSA compared with sensitized mice. Such a decrease in the numbers of mononuclear cells, mast cells and mucus-producing cells in the lung was not seen in animals treated with PSA perorally, although these animals exhibited suppressed DH reaction in the ears. The present results indicate that induction of mononuclear cells and to some extent mast cells and mucus-producing cells in the lung relates to the DH reaction and its regulation.
Ultrastructural Localization of IgE in Peripheral Blood Monocytes from Atopies
Authors: Turner K.J. , Marzo A. , Holt B.J. , Wharton A. , Papadimitriou J.M. , Cameron K.J. , Siemensma N.P. , Holt P.G.
Published: August 2009

Monocyte-enriched preparations derived from peripheral blood leukocytes of atopies were probed via a cocktail comprising peroxidase-conjugated (Fab1)2 fragments of two monoclonal antibodies against human IgE. Reaction product indicative of intracellular IgE was identified by electron microscopy in both large and small vacuoles, and at high magnification exhibited a characteristic granular deposition pattern consistent with highly concentrated (perhaps insolublized) material. IgE-containing vacuoles were observed with comparable frequency to those containing IgG, despite the > 10,000-fold relative excess of the latter in serum suggesting highly selective uptake of IgE by the monocytes. These results are similar to those reported recently for IgA in human milk macrophages.
Comparison of Aqueous Grain Dusts, Cotton Dusts, and Mold Extracts by Crossed Immunoelectrophoretic Techniques
Authors: O’Neil Carol E. , Butcher Brian T. , Chan Henry , Chan-Yeung Moira
Published: August 2009

To evaluate the possibility that an immune response is involved in the pathogenesis of grain dust or cotton dust induced respiratory disease, 1 grain dust extract (A) was compared with 4 similar grain dust extracts, a cotton dust extract, and a mixture of fungal preparations, using crossed immunoelectrophoretic techniques and rabbit anti-A sera. All extracts demonstrated a significant number of shared antigenic determinants. These data suggest that grain and cotton dust may induce disease via a common pathogenic mechanism and putative agent(s).
Nonresident Macrophages Are Absent from Glomeruli in IgA Nephropathy
Authors: Béné Marie C. , Bolle-Chanal Marie H. , Got Michel , Faure Gilbert C.
Published: August 2009

The presence of nonresident cells of monocyte-macrophage lineage was investigated in 50 renal biopsies, including 20 from patients suffering from IgA nephropathy. Histochemical and immunological techniques were used to identify, respectively, nonspecific esterase and peroxidase activities and HLA class II molecules. Although intraglomerular phagocytes were observed in all control biopsies, small numbers were seen in only two samples from patients with IgA nephropathy. This feature is consistent with an impaired clearance of immune complexes, possibly leading to their deposition in the mesangium.
Allergenic Cross-Reactivity of Several Strains of Trichinella in Mice and Rats by the Passive Cutaneous Anaphylaxis Technique
Authors: Santamarina M.T. , Leiro J. , Alvarez F. , Sanmartín Durán M.L. , Ubeira F.M.
Published: August 2009

The cross-reactivity of antiparasite IgE antibody responses induced by allergenic antigens obtained from Trichinella spiralis and Trichinella pseudospiralis was determined by the passive cutaneous anaphylaxis (PCA) technique in BCF1 mice infected with T. spiralis, Trichinella nelsoni, Trichinella nativa, and T. pseudospiralis and in rats infected with T. spiralis and T. pseudospiralis. Our results demonstrate that when the antigen used for the PCA challenge is derived from muscular L1 larvae of T. spiralis, high IgE antibody titres can be detected from sera of animals infected with T. spiralis, T. nelsoni, or T. nativa, but not with T. pseudospiralis, during the entire life cycle of the parasite. However, when homologous antigens are used in the PCA test, we obtained comparable values of IgE titres in rats and mice infected with both T. spiralis and T. pseudospiralis strains. These results suggest the existence of a high degree of immunologic identity between the allergenic antigens of T. spiralis, T. nelsoni, and T. nativa strains, but not T. pseudospiralis.
Lack of Induction of IgE and IgG Antibodies to Yeast in Humans Immunized with Recombinant Hepatitis B Vaccines
Authors: Wiedermann G. , Scheiner O. , Ambrosch F. , Kraft D. , Kollaritsch H. , Kremsner P. , Hauser P. , Simoen E. , André F.E. , Safary A.
Published: August 2009

Yeast-derived hepatitis B vaccines (partially purified or highly purified) at different dosages and a plasma-derived hepatitis B vaccine (control) were injected into 50 young volunteers 3 times at monthly intervals. Before and 4 weeks after this series of immunizations, blood samples were drawn and tested for presence of IgE and IgG antibodies against yeast antigens. No rise in IgE antibodies against Saccharomyces cerevisiae antigens nor in IgG antibodies against Candida albicans antigens was found. Together with former clinical results this underlines the experience that type I and type III reactions against putative yeast contaminants apparently play no major role after immunization with recombinant hepatitis B vaccines.
Does ‘Sugar’ Permeability Reflect Macromolecular Absorption? A Comparison of the Gastro-Intestinal Uptake of Lactulose and Beta-Lactoglobulin in the Neonatal Guinea Pig
Authors: Weaver L.T. , Coombs R.R.A.
Published: August 2009

Intestinal closure to cow’s milk beta-lactoglobulin occurs within 6 days of birth in the guinea pig. Passive intestinal permeability to lactulose persists through the suckling period. The uptake of small water-soluble markers does not reflect macromolecular absorption, and has no place in the measurement of immunologic protein handling by the gut.
Characteristics of Five Monoclonal Antibodies to Major Allergens of the Short Ragweed Pollen
Authors: Shen Horng-Der , Chang Lu-Yin , Su Song-Nan , Han Shou-Hwa
Published: August 2009

Monoclonal antibodies against major allergens of the short ragweed pollen were produced by fusion of NS-1 cells with splenic cells from BaLB/C mice that had been immunized separately with the major allergens, AgE-B+E-C and AgK, of the short ragweed pollen. These monoclonal antibodies were detected by enzyme-linked immunosorbent assay and further characterized by immunoblot analysis using the crude extract and highly purified allergens of the ragweed pollen. Three monoclonal antibodies obtained by immunization with AgE-B+E-C, designated as 36-6, 27-2 and 48-5, reacted mainly with the beta (36-6) and alpha (27-2) subunit of AgE and both AgE and AgK (48-5), respectively. Two monoclonal antibodies obtained by immunization with AgK, 4-7 and 8-5, had a similar reactivity with AgE-B+E-C, AgK, AgK-A and AgK-B. In addition, however, antibody 4-7 also reacted with AgE-B2 as well as the 36- and the 24- to 22-kilodalton antigens of the crude extract. All 5 monoclonal antibodies were characterized as IgG1 subclass. Besides, monoclonal antibody 48-5 also showed cross-reactivity to components of sage pollen. Beyond academic interests, the monoclonal antibodies described here may also be useful in clinical allergy.
Prophylactic and Therapeutic Effects of Ciclosporin A in Murine Schistosomiasis mansoni: Studies on Bisexual and Unisexual Infections and the Hepatic Inflammatory Response
Authors: Smith S.W.G. , Chappell L.H. , Thomson A.W. , MacGowan A.G. , Simpson J.G.
Published: August 2009

Ciclosporin A (CsA), administered subcutaneously as 5 daily injections of 50 mg kg––1, reduced the numbers of Schistosoma mansoni perfused from MF1 mice at 7 weeks post-infection. The timing of drug administration revealed that the antischistosomal effects were greater when CsA treatment coincided with or was within a few days of infection with the parasite. CsA exerted a clear prophylactic effect, which decreased with time and was virtually abolished by 4 months pre-infection. Adult worms treated in vivo were partially susceptible to CsA. In addition to its antiparasite action, CsA reduced hepatosplenomegaly due to schistosomiasis and diminished the granulomatous inflammatory response of mice to parasite eggs in the liver. The mode of action of CsA is not understood but evidence is presented that supports the proposition that the anti-parasite effects are perhaps host-mediated.
Immunogenicity and Allergenicity Studies on Two Beta-Lactam Structures, a Clavam, Clavulanic Acid, and a Carbapenem: Structure-Activity Relationships
Authors: Edwards R.G. , Dewdney Janet M. , Dobrzanski R.J. , Lee D.
Published: August 2009

Two newer β-lactam-containing structures, a clavam, clavulanic acid, and a carbapenem, MM22383, have been studied for their intrinsic immunogenicity and allergenicity. Clavulanic acid has a very low immunogenic and allergenic potential, in contrast to MM22383 which is a contact sensitiser in guinea pigs and an immunogen in rabbits. Evidence for the allergenic potential of MM22383 in man through occupational exposure is also presented. Consideration of the chemistry of these two compounds with respect to their reactivity with protein provides a rationale for the marked difference in their behaviour. The importance of stable hapten-protein conjugates and epitope density is discussed in relation to immunogenicity.
Increased Helper Cell Activity of NZB Mice against H-2-Identical Allogeneic Cells
Authors: Müller-Quernheim J. , Botzenhardt U. , Steldern D.v. , Lefkovits I.
Published: August 2009

The T cells of NZB mice become hyperreactive after stimulation with minor histocompatibility (MIH) antigens. This hyperreactivity has previously been demonstrated only for cytotoxic T cells of NZB, although there was some evidence for an increase of their T-helper cell activity facilitating the response. Here we report a quantitative analysis of T-cell help and help of T-cell subpopulations against autologous, MIH, and H-2 antigens in a limiting dilution assay. After stimulation of NZB T cells with autologous and H-2 antigens, the T-helper cell frequencies did not differ from that of normal mice. After stimulation with MIH antigens however, Lyt 1+2+ T-cells of NZB showed a higher response than those of BALB/c origin. The same difference was seen after prestimulation with ConA or the specific antigen. This demonstrates that NZB-helper cells are more easily activated by weak antigenic differences, and it is possible that this contributes to the prevalence of autoimmune disease in this strain.
Regulation of IgE-Dependent Antiparasite Functions of Rat Macrophages and Platelets by Nedocromil Sodium
Authors: Thorel T. , Joseph M. , Vorng H. , Capron A.
Published: August 2009

The IgE-dependent stimulation of mononuclear phagocytes and blood platelets can be measured by antiparasite cytotoxicity, oxygen-mediated chemiluminescence and, in macrophages, lysosomal enzyme activity. Using these parameters, the present study demonstrated an inhibition by nedocromil sodium of the IgE-mediated triggering of the nonmast cell inflammatory populations in the rat. These observations suggest that nedocromil sodium may be of value in the modulation of IgE-dependent cell activation inducing the release of inflammatory mediators.
Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate
Authors: Lee Ha-Baik , Rommel Fred , Adkinson, Jr. Franklin
Published: August 2009

An enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antihuman γ-chain antibody and a fluorogenic substrate has been developed for quantitation of IgG-blocking antibodies in human serum. Generation of fluorescent product was linear with time to 60 min. Using optimal conditions the ELISA was sensitive to < 1 ng/ml of specific IgG to short ragweed pollen. The assay demonstrated consistently parallel dilution curves with 51 sera (mean interdilutional coefficient of variation = 8.8%). Reproducibility was determined by constructing precision profiles for intra and interassay variation for the entire working range of the assay. Intraassay CVs ranged from a mean of 13% at threshold to < 5% at higher antibody concentration. Interassay reproducibility similarly ranged from 18 to 10%. In this assay the effect of serum dilution on nonspecific binding was minimal and specific binding of 4–10 ng IgG antibody to the antigen-adsorbed wells was largely complete (75.8 ± 4.8%) and highly specific ( > 98%). This application of ELISA for ragweed IgG antibody measurement has performance specifications equal or superior to previously developed radioimmunoassay and ELISA systems.
Beneficial Effect of Captopril on Systemic Lupus erythematosus-Like Disease in MRL 1pr/1pr Mice
Authors: Herlitz Hans , Tarkowski Andrej , Svalander Christian , Volkmann Reinhard , Westberg Gunnar
Published: August 2009

MRL 1pr/1pr (MRL/1) mice exhibit a disease similar to systemic lupus erythematosus (SLE) in humans. To investigate the influence of antihypertensive treatment on this disease, four groups of MRL/1 mice were treated with the angiotensin-converting enzyme inhibitor captopril (n = 25), with the sympathetic blocker bretylium (n = 15), and with cyclophosphamide (n = 10). Thirty-five mice did not receive any treatment and served as controls. Survival rate, blood pressure, incidence of proteinuria and hematuria, renal pathology, lymphoid hyperplasia and dermatitis were studied. The survival at the age of 36 weeks was significantly improved by captopril as compared to controls (60 vs. 25%, p = 0.035). The cyclophosphamide group showed no mortality at that time and the bretylium group did not differ from the control group. Captopril and bretylium reduced systolic blood pressure significantly while cyclophosphamide was without effect. Captopril and cyclophosphamide diminished significantly the glomerular damage with less proliferative changes and a decreased incidence of proteinuria. The bretylium-treated animals also exhibited an improved renal pathology index but they did not differ from the controls with respect to proteinuria and hematuria. Lymphoid hyperplasia and dermatitis were decreased only by captopril and cyclophosphamide. It is concluded that captopril improves survival in SLE disease of MRL/1 mice, counteracting lymphoid hyperplasia, renal disease, dermatitis and decreasing arterial blood pressure.
Detection of IgE Antibodies to a Wide Range of Insect Species in Subjects with Suspected Inhalant Allergies to Insects
Authors: Baldo B.A. , Panzani R.C.
Published: August 2009

Sera obtained from subjects diagnosed on the basis of case history and skin tests as having inhalant allergies to insects, were tested for the presence of IgE antibodies to antigens from the house fly Musca domestica, a blowfly Calliphora stygia, the common clothes moth Tineola bisselliella, warehouse moth Ephestia cautella, cockroach Blattella germanica, carpet beetle Anthrenus verbasci and silverfish Ctenolepisma longicaudata. Approximately one-third of the sera reacted with extracts from all seven species, over half the sera reacted with four of the extracts and only 3 sera proved negative to all of the extracts. Twenty-six of the sera also contained IgE antibodies that reacted with the house dust mite Dermatophagoides farinae. Tests with a further eleven species of flies from the order Diptera and with extracts of the grain borer Rhyzopertha dominica, locust Chortoicetes terminifera and Bogong moth Agrotis infusa revealed strong IgE antibody responses to all of the species. Electroblotting studies revealed up to 15 IgE-binding components in extracts from 11 different insect species including 6 species of flies. Some IgE-binding insect components with similar electrophoretic mobilities indicated the possible presence of common allergens in extracts from different species. In particular, blots of each of the fly extracts showed a dense IgE-binding component of MW approximately 37,000 daltons. ‘Pan allergy’ to insects may occur in subjects who have been sensitized to one or a few insects and allergenic similarities may extend to at least some other noninsect members of the phylum Arthropoda.
The Glycoprotein Allergen Ag-54 (C1a h II) from Cladosporium herbarum
Authors: Swärd-Nordmo Marit , Paulsen Berit Smestad , Wold Jens Kristian
Published: August 2009

The carbohydrate moiety of an important allergen, Ag-54 in Cladosporium herbarum was studied by alkaline-borohydride treatment, gel filtration, high-performance liquid chromatography, methylation analysis, gas liquid chromatography and mass spectrometry. The Ag-54 protein core possessed a very limited number of sugar chains. The carbohydrate moiety consisted mainly of one large highly branched polysaccharide chain which accounted for nearly 75% of the total molecular weight of the glycoprotein. The carbohydrate moiety is made up of D-mannose and D-glucose units in pyranose form having D-galactofuranose side chains attached. Mannose is both 1,2- and 1,6-linked, while glucose is 1,4- and 1,6-linked. Some of the 1,6-linked galactofuranose side chains are bound through C-2 of the 1,6-linked mannose units, and the rest to C-3 of 1,6-linked mannose and 1,2-linked mannose units. A few oligoglucosidic chains of approximately 4 glucose units are also attached to the protein.
The Glycoprotein Allergen Ag-54 (C1a h II) from Cladosporium herbarum
Authors: Swärd-Nordmo Marit , Paulsen Berit Smestad , Wold Jens Kristian
Published: August 2009

The carbohydrate-protein linkage of the allergen Ag-54 in the mould Cladosporium herbarum was studied after alkaline-borohydride treatment of the glycoprotein. By incorporating tritium into the peptide-linked monosaccharide through the alkaline scission of the linkage, it was demonstrated that the highly branched galactomannan moiety of Ag-54 was linked to the protein through mannose. Threonine was identified as the carbohydrate-linked amino acid. In addition, a few glucose units were peptide-linked, but the corresponding amino acids were not identified. When deglycosylated with trifl·uoromethanesulfonic acid, Ag-54 retained only 5% of its allergenic activity as compared to the native Ag-54 when tested by rocket immunoelectrophoresis and autoradiography. Immunological identity between native and deglycosylated Ag-54 was observed. Deglycosylation also resulted in reduced stability towards denaturation by heat or urea.
Antibody-Dependent Cell-Mediated Cytotoxicity against Tamm-Horsfall Protein in Acute Pyelonephritis
Authors: Tomizawa Shigeru , Kjellson Björn , Fasth Anders , Hagberg Marianne , Jodal Ulf , Kaijser Bertil , Söderström Tommy , Hanson Lars Å.
Published: August 2009

The cytotoxic activity of leukocytes from humans and rats with pyelonephritis were examined in an antibody-dependent cell-mediated cytotoxicity assay (ADCC) with CrCl3-treated erythrocytes coated with Tamm-Horsfall (TH) as target cells. The specificity of the ADCC was confirmed by absorption with TH urinary glycoprotein and inhibition of the ADCC activity seen with polyclonal rabbit anti-TH antisera by monoclonal mouse antibodies. The ADCC activity detected in children with acute pyelonephritis was low in the initial phase of the disease, but increased significantly 9 days after the start of antibacterial treatment. In rats with experimental pyelonephritis, ADCC activity decreased significantly with increased duration of infection. Depletion of cells adhering to carbonyl iron led to higher ADCC activity. During the course of the infection the difference in ADCC activity between effector cell preparations depleted using carbonyl iron and those not depleted decreased. The decreased ADCC activity demonstrated during acute pyelonephritis may point to mechanisms operating to diminish the risk of tissue damage.
The Major Allergen of the House Dust Mite, Dermatophagoides pteronyssinus, Is Synthesized and Secreted into Its Alimentary Canal
Authors: Thompson S.J. , Carswell F.
Published: August 2009

The natural production of Der p1, the major faecal allergen from the house dust mite (Dermatophagoides pteronyssinus), was investigated. Mite cultures grown on radiolabelled substrate were examined at intervals over a period of 21 days and incorporation of radiolabel into the mite proteins measured over the time course. Mite proteins were extracted, purified by chromatofocussing and characterized by immunodiffusion, SDS-PAGE, electroblotting. and autoradiography. The faecal allergen, Der p1, did not incorporate a significant level of label until the 14th day (p < 0.05). The results suggest that Der p1 is a protein synthesized and excreted by the house dust mite.
Antibodies Reactive with Streptococcal Peptidoglycan-Polysaccharide Complexes in Rheumatic Fever, Subacute Bacterial Endocarditis and Tuberculosis
Authors: Evans Helen B. , van der Merwe P.L. , Strachan A.F. , Johnson P.M.
Published: August 2009

Serum antibodies reactive with streptococcal peptidoglycan-polysaccharide complexes (PG-PS) have been estimated by enzyme-linked immunosorbent assay in patients with acute rheumatic fever (ARF), tuberculosis (TB) and subacute bacterial endocarditis (SBE) compared with normal age- and sex-matched controls. IgG2 and IgG3 were the predominant subclasses of antibodies in TB and SBE, whereas in ARF IgG1, IgG2 and IgG3 antibodies were detected. IgG4 antibodies were detected only infrequently. Antibodies of the IgG subclasses were detected in a greater proportion of sera when the PG-PS antigen was further purified by extraction with sodium dodecyl sulphate.
Western Immunoblotting of Cereal Proteins with Monoclonal Antibodies to Wheat Gliadin to Investigate Coeliac Disease
Authors: Freedman A.R. , Galfre G. , Gal E. , Ellis H.J. , Ciclitira P.J.
Published: August 2009

Western immunoblotting was used to investigate the binding of two monoclonal antibodies raised against unfractionated wheat gliadin to different cereal protein fractions separated by SDS-PAGE. Our results confirm the presence of considerable epitope sharing between the gliadin subfractions as well as barley and rye prolamins; however, there was less binding of these antibodies to bands present in oat avenins and maize zeins. The pattern of binding of one of these two antibodies to different cereal prolamins as well as to Frazer’s fraction III corresponds closely to the known toxicity of these proteins to patients with coeliac disease.