bluuebunny/crossref_metadata_2025 huggingface dataset:
IgG-Class Insulin Autoantibodies in Autoimmune Thyroid Disease
Authors: Ng W.Y. , Thai A.C. , Lui K.F. , Yeo P.P.B. , Cheah J.S.
Published: August 2009

We studied the incidence of insulin autoantibodies (IAA) in 97 patients with autoimmune thyroid disease with an enzyme-linked immunosorbent assay. The sera were also tested for islet cell antibodies (ICA) and thyrotropin receptor antibodies (TRAb). IAA as assessed by a standard deviation score were present in 7 patients (7.2%). None of the patients were seropositive for ICA. Data from this random sampling and a retrospective study of 12 patients on antithyroid drug treatment indicated that the presence of IAA was not associated with TRAb.
Induction of HLA Class I Surface Expression Recruits Low-Affinity Cytolytic T Lymphocytes
Authors: Mentzer Steven J. , Burakoff Steven J. , Barbosa James A.
Published: August 2009

Recent studies indicate that the relative expression of HLA class I molecules can play an important role in regulating the cellular immune response. In this report, we used cloned human cytolytic T lymphocytes (CTL) and transfected HLA class I molecules to investigate the effect of HLA class I induction on CTL reactivity. CTL clones with identical antigen specificities but different antigen affinities were compared for their ability to lyse target cells with varying levels of HLA class I expression. The cytolytic reactivity of the high-affinity clone was independent of the level of HLA class I expression. In contrast, cytolytic reactivity of the low-affinity CTL clone was directly related to the level of HLA expression. These data suggest that the induction of HLA class I surface expression has a disproportionate effect on the reactivity of low-affinity CTL and may provide a mechanism for the recruitment of these cells in vivo.
Cytokine Control of Peripheral-Blood CD23 Expression and sCD23 Release: Differential Regulation by IL-2 and IL-4
Authors: Fischer A. , Pfeil T. , König W.
Published: August 2009

CD23 expression on peripheral-blood lymphocytes (PBL) was studied under the influence of cytokines. It is shown that IL-2 induced CD23 expression on human peripheral-blood B cells. Evidence is presented that the IL-2 induced CD23 expression and release of soluble CD23 (sCD23) are not mediated by IL-4. In comparison to IL-4, the IL-2-induced CD23 expression and sCD23 release revealed kinetic differences and were not inhibited by anti-IL-4. The prestimulation of PBL with Staphylococcus aureus strains Cowan 1 (SAC) led to a pronounced reduction in basal CD23 expression and to a change in the response of cytokines. Subsequent stimulation with IL-4 induced CD23 to the same extent as on unstimulated cells, whereas the IL-2-induced CD23 expression and sCD23 release were greatly reduced. Interferon γ showed no effects on the IL-4-stimulated CD23 expression of SAC-PBL, whereas the IL-2-induced CD23 expression was suppressed. Furthermore, we demonstrate that the stimulation with IL-2/IL-4 inhibits the effects of the individual cytokine; this inhibition is also seen for immunoglobulin (E, G, M) synthesis.
Manipulation of Macrophage Migration Inhibition/Stimulation Responses by Adjuvants and Interleukins
Authors: MacSween J.M. , Rajaraman R. , Rajaraman S.
Published: August 2009

C57 B1/6 mice were immunized with bovine serum albumin (BSA) and Freund’s complete and incomplete adjuvants in various concentrations. Spleen cells from these animals were subsequently stimulated with concanavalin A (Con A), purified protein derivative or BSA, and lymphokine responses were measured in one-stage migration assays. Con A consistently produced macrophage migration inhibition factor (MIF) responses in nonimmunized animals and those immunized with complete adjuvant. This was switched to migration stimulation factor (MStF) responses by prior immunization with incomplete adjuvant. Immunization with complete adjuvant and with BSA alone was followed by MIF responses to antigenic stimulation whereas incomplete adjuvant promoted MStF responses. The MStF responses to both mitogenic and antigenic stimulation were inhibited by the addition of affinity purified interleukin-1 to the spleen cells. Interleukin-1 also inhibited MStF responses and potentiated MIF responses to Con A stimulation by human mononuclear cells whereas interleukin-2 did not influence these responses
Rat Mast Cell Granules Reacting with a Mouse Monoclonal Antibody M6764 which Recognizes the Same Epitope of a Mouse Monoclonal Antibody HNK-1
Authors: Kurosawa Motohiro , Okayama Yoshimichi , Kobayashi Setsuo , Obata Kunihiko
Published: August 2009

Rat mast cell granules and plasma membrane fractions were obtained by homogenization of highly purified rat mast cells and isolation in a Percoll gradient and a sucrose gradient, respectively. Immunostaining of rat mast cells, granules and plasma membrane fractions was performed with mouse monoclonal antibody M6764 which was produced against the crude membrane fractions of the neural tubes. Rat mast cells and granules were immunostained with the monoclonal antibody, but not the plasma membrane fractions. The granules fixed with glutaraldehyde-paraformaldehyde showed ring-like forms. Chloroform-methanol treatment did not effect the staining of rat mast cells and granules with the monoclonal antibody. Western blotting analysis of rat mast cells and granules with the monoclonal antibody showed broad protein bands ranging from 100 to 250 kD.
Protection of Rats against Schistosoma mansoni Infection Induced by Platelets Stimulated with the Murine Recombinant Tumor Necrosis Factor Alpha
Authors: Damonneville Martine , Pancré Véronique , Capron André , Auriault Claude
Published: August 2009

Human recombinant tumor necrosis factor (rTNFα) and lymphotoxin (rTNFβ) have been shown to enhance the schistosomicidal activity of human platelets in vitro. In this report, we demonstrated that the murine rTNF was able to induce the in vitro cytotoxic activity of both murine and rat platelets towards the larvae of Schistosoma mansoni, whereas human rTNFs activated only the murine platelets. Passive transfer of platelets stimulated with murine rTNF significantly protected rats up to a 65% reduction of the worm burden. Thus, rTNF may constitute, via its interactions with effector cells, an important regulatory mechanism during the parasitic infection of rats.
Drug-Induced Fever: A Clinical Report and Challenge Test with Calcium Dobesilate
Authors: Puyana J. , Fraj J. , De La Hoz B. , Davila I. , Vergara C. , Cuesta J.
Published: August 2009

A 54-year-old man was receiving calcium dobesilate for retinopathy and after 8 days he presented fever of more than 39 °C, generalized myalgia, chills and headache. Other causes of fever were ruled out. A challenge test was done with a single therapeutic oral dose of calcium dobesilate and fever appeared 20 h later and lasted 8 h. Our patient fulfils Young’s stringent criteria for drug fever. To our knowledge drug-induced fever due to calcium dobesilate has not been reported previously.
Follicle Lysis in HIV-Free Lymphoid Tissues
Authors: Shiota Mami , Toyama Keisuke , Mori Shigeo
Published: August 2009

Follicle lysis (FL), the ‘moth-eaten’ degeneration of follicular dendritic cells (FDCs), previously regarded as specific for HIV-infected lymph follicles, was later claimed to exist in HIV-free patients. This study was performed to clarify the differences between HIV-free and HIV-induced FL. HIV-free and HIV-infected lymphoid tissues were analyzed immunohistologically. Most of HIV-free FLs were shown to lack the dark zones, the areas in germinal center composed mostly of much larger and clumped noncleaved cells, whereas HIV-induced FL in persistent generalized lymphadenopathy (PGL) lymph nodes included highly proliferative dark zones. Thus, FL in HIV-free lymphoid organs was suggested to be a physiological degenerative process with poor proliferative activity, while FL in PGL nodes includes two processes, FDC degeneration and B lymphocyte proliferation.
Possible Role for Platelet-Activating Factor in Neutrophil Infiltration in Allergic Inflammation in Rats
Authors: Watanabe Masako , Sugidachi Atsuhiro , Omata Mitsugu , Hirasawa Noriyasu , Mue Suetsugu , Tsurufuji Susumu , Ohuchi Kazuo
Published: August 2009

Allergic inflammation was induced by injecting an antigen solution into an air pouch made on the dorsum of immunized rats with the antigen azobenzene-arsonate-conjugated acetyl bovine serum albumin. In this model, leukocyte infiltration into the pouch fluid was prominent 4–8 h after the antigen challenge. Most of the infiltrated leukocytes were neutrophils. Administration of the platelet-activating factor (PAF) antagonists such as CV-3988 and L-652,731 into the air pouch 15 min before and at the time of the antigen challenge failed to suppress leukocyte infiltration at 8 h. However, when the PAF antagonist was injected into an air pouch 4 h after the antigen challenge, neutrophil infiltration at 8 h was suppressed in a dose-dependent manner. Combined treatment with the 5-lipoxygenase inhibitor AA861 and the PAF antagonist did not potentiate the effect of the PAF antagonist, suggesting that participation of leukotriene B4 in neutrophil infiltration in this model is negligible. Eosinophil infiltration was very weak at 8 h, and the PAF antagonist showed no significant effect. At 8 h, the PAF level in the serum of the immunized rats was significantly higher than that of the nonimmunized rats. Intravenous administration of the PAF antagonist 15 min before the antigen challenge suppressed leukocyte infiltration more effectively than local administration into the pouch. These results indicate that PAF plays a significant role in neutrophil infiltration in allergic inflammation.
Epitope-Specific Regulation of the Antibody Response against Alpha-Lactalbumins in the Mouse
Authors: Horiuchi Tadashi , Yagi Junji , Asano Yoshihiro , Tada Tomio
Published: August 2009

The immune responsiveness to human and bovine α-lactalbumin (HuALA and BoALA) was found to be under the control of immune response (Ir) gene(s) linked to the major histocompatibility complex. H-2k mice responded to both HuALA and BoALA, whereas H-2d, s, and f mice respond only to HuALA; H-2b mice were nonresponders to both HuALA and BoALA. A survey with B10.A recombinant mouse strains .enabled us to map the Ir gene in the I-A subregion. The responsiveness was shown to be dominant in F1 mice. The coimmunization of BoALA and HuALA resulted in the suppressed secondary antibody response to HuALA in B10.S (H-2S) and BALB/c (H-2d) but not in C3H (H-2k) suggesting that the low responsiveness against HuALA in these strains is due to an active suppression. The transfer of splenic T cells of B10.S mice primed with BoALA into syngeneic animals suppressed the response to HuALA. T cells specific for a particular epitope present on BoALA appeared to suppress the immune response to other epitopes on HuALA. Thus, the presence of epitope-specific suppressor T cells seems to account for this Ir-gene-controlled low responsiveness to ALA in H-2smice.
Susceptibility to Proteolipid Apoprotein and Its Encephalitogenic Determinants in Mice
Authors: Endoh Masumi , Kunishita Tatsuhide , Nihei Junko , Nishizawa Masatoyo , Tabira Takeshi
Published: August 2009

We investigated in mice strain differences in induction of experimental allergic encephalomyelitis by proteolipid apoprotein and studied encephalitogenic determinants. SJL/J, C3H/He, CBA/J and A/J mice were high responders, BALB/c and AKR/J mice were moderately susceptible, and DBA/2, B6 and congenic strains of B10 background were low responders. Synthetic peptide 136–150 was encephalitogenic for SJL/J mice, and 215–232 was encephalitogenic for C3H/He mice. These encephalitogenic derterminants are present in the extracellular portion of proteolipid apoprotein in myelin.
Mapping of Bet v I Epitopes by Using Murine Monoclonal Antibodies
Authors: Marc-Series I. , Boutin Y. , Vrancken E.R. , Hébert J.
Published: August 2009

The different determinants of birch pollen extracts, as shown by SDS-PAGE analysis, range from 10 to 94 kDa. These determinants were then electrotransferred on nitrocellulose strips and allowed to react with human IgE Ab from sensitive patients in order to identify the allergenic determinants. Several minor (43, 35, 28 and 21 kDa) and the major (17 kDa) allergenic determinants were identified. Murine monoclonal antibodies (mAb) were then produced against the major allergenic determinant (Bet v I) and their specificity confirmed by immunoblot. One of them, mAb 3F10, was used to affinity-purify the Bet v I. The purity of this material was confirmed by SDS-PAGE analysis and its reactivity on immunoblot against human IgE ensured its biological activity. These mAb were then gathered on four families based on their pattern of reactivity with Bet v I. Indeed four different epitopes on the molecule were identified. Binding inhibition studies using two of them (mAb 5F9 and 8F12) suggested that the epitopes of Bet v I recognized by these mAb are not overlapping. On another hand, the binding of 8H7 and 3F10 was partially inhibited by 5F9 and the binding of 3F10, by 8F12. These data suggest that those two latter epitopes are somewhat overlapping. Finally, the mAb 5F9 could inhibit the binding of human IgE on the affinity-purified Bet v I up to 40% and then shares a common idiotope with human specific IgE Ab of allergic patients.
Effect of Oral or Parenteral Sensitization to Cow’s Milk on Mucosal Permeability in Guinea Pigs
Authors: Heyman M. , Andriantsoa M. , Crain-Denoyelle A.M. , Desjeux J.F.
Published: August 2009

The systemic and local immune responses and the intestinal barrier function were examined in orally or parenterally milk-sensitized guinea pigs. Both types of sensitization led to positive passive cutaneous anaphylactic responses and high IgG titers against β-lactoglobulin (β-LG), especially in parenterally immunized animals. In Ussing chambers, sensitized jejunum had higher short-circuit current (Isc) than control jejunum, with and without β-LG challenge. The further increase in Isc induced by serosal β-LG treatment was higher in parenterally than orally sensitized animals. Barrier function was tested as the intestinal transport and degradation of horseradish peroxidase (HRP) in the presence and absence of β-LG. There was a 5-fold increase in degraded HRP transport in sensitized versus control animals, with and without β-LG challenge. Serosally applied β-LG enhanced the transport of intact HRP in sensitized but not in control animals. These results indicate that sensitization of guinea pigs to cow’s milk permanently increases endocytic and electrogenic activities. The challenge with β-LG induced a further transient rise in Isc and increased intact HRP transport.
Induction of Suppressor Cell Activity in Normal Peripheral Blood Mononuclear Cells by Sera from Predialytic Uremic Patients
Authors: Modai D. , Berman S. , Weissgarten J. , Klinovsky E. , Cohn M. , Averbukh Z.
Published: August 2009

Normal peripheral blood mononuclear cells (PBMC) were primed with sera from 22 predialytic uremic patients, normal sera, concanavalin A, or both. Their suppressive activity was subsequently tested on fresh phytohemagglutinin-stimulated allogeneic responders (i.e., genetically unrelated to either primed cell or sera donors). Uremic serum induced suppressor cell activity in the normal PBMC. No correlation was found between serum urea/creatinine levels and their effects on suppressor cell activity. The induced activity, expressed as percent suppression, was similar to that induced by concanavalin A. In PBMC primed with both concanavalin A and uremic serum, an additive suppressive effect was evident. The suppressor subset(s) induced by uremic serum proved to belong to adherent cell population. Addition of indomethacin or catalase to responder systems did not abolish the suppressive effects, thus suggesting a mechanism of action other than prostaglandin or hydrogen peroxide release.
Influence of IgE Fragments on IgE Determination
Authors: Vassella Claudia C. , de Weck Alain L. , Stadler Beda M.
Published: August 2009

IgE fragments were detected by polyclonal and monoclonal anti-IgE antibodies using Western blot techniques and immunoassays. Other antibodies detected only intact IgE molecules. In the presence of IgE fragments, higher IgE levels were estimated, depending on the epitope specificity of the antibodies used in the IgE determination assays. IgE fragments have been found together with intact IgE in isolated material from different human sera and also from culture supernatant of an IgE-producing cell line. Our data demonstrate that IgE fragments may be assessed and represent a potential pitfall in diagnostic IgE determinations.
Observations on NK Cells, K Cells and on Their Function a Long Time after Posttraumatic Splenectomy
Authors: Demeter J. , Pálóczi K. , Lehoczky D. , Benczur M.
Published: August 2009

Natural killer (NK) cell activity and antibody-dependent cellular cytotoxicity of 48 posttraumatic splenectomy patients was found to be normal despite the increase in the proportion and the absolute number of large granular lymphocytes (LGLs). The NK cell activity of 2 further posttraumatic splenectomy cases was severely decreased, despite elevated number of LGLs and normal number of CD 16+ peripheral blood mononuclear cells. Besides increased susceptibility to upper respiratory tract infections, 1 of these patients developed herpes genitalis recidivans. Two posttraumatic splenectomy patients were found to have developed chronic lymphocytic leukemia 5 and 31 years following splenectomy, respectively.
Allergens of Horse Epithelium
Authors: Franke D. , Maasch H.J , Wahl R. , Schultze-Werninghaus G. , Bretting H.
Published: August 2009

We investigated five horse epithelial allergen extracts prepared from different qualities of raw material by several biochemical and immunochemical methods. Horse serum albumin and horse serum were used to identify serum-related antigens. We found high similarities as well as marked differences between the extracts. There were strong differences in the protein contents, the protein patterns obtained by isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis, the total allergenic activities obtained by radioallergosorbent test inhibition assays and the amounts and numbers of serum-related proteins. The patterns of dander-related allergens in crossed radioimmunoelectrophoresis were similar but showed partly strong differences in allergen concentrations. Allergen compositions determined by blotting methods showed only minor differences between the investigated epithelial extracts. It appears that some of the differences, e.g. the content of serum-related proteins, depend on the way the different raw materials are prepared. Because serum-related extract components are not considered as major allergens, their content should be low in horse epithelial allergen extracts. For this reason whole skin cannot be recommended as starting material for horse epithelium allergen extract preparation.
Selective Alterations in Natural Killer Cell Subsets in Patients with Atopic Dermatitis
Authors: Wehrmann W. , Reinhold U. , Kukel S. , Franke N. , Uerlich M. , Kreysel H.W.
Published: August 2009

We examined the immunophenotypic characteristics of natural killer (NK) cell subsets in patients with severe atopic dermatitis (AD), rhinitis allergica (RA) and in healthy controls. Expression of CD16, CD56 and CD57 antigens on peripheral blood lymphocytes was evaluated by simultaneous double immunocytofluorometry. Our results showed significantly lower percentages of cells with CD 16, CD56 and CD57 surface antigens in patients with AD. Furthermore subdivision of the AD group into two subgroups, AD1 and AD0 (with and without antigen-specific IgE antibodies against potent inhalative allergens, i.e. mite, grass, rye, birch, cat) revealed that patients of subgroup AD1 showed a more prominent decrease compared to that of subgroup AD0. Moreover, we found a significant negative correlation between the percentage of CD56 + CD 16 + NK cells and total IgE levels in serum, which were significantly higher in patients of subgroup AD1 than in AD0. NK cell activity was deficient in patients with AD but there was no difference between both subgroups. These data indicate that considerable heterogeneity in immunologic regulation may exist in patients with AD with regard to their NK cell subsets.
Inhibition of 5-HETE, LTB4, and LTC4 Formation by Azelastine in Rat Mixed Peritoneal Cells
Authors: Chand Naresh , Pillar John , Nolan Kevin , Diamantis William , Sofia Duane
Published: August 2009

Azelastine produced a concentration-dependent inhibition of calcium ionophore A23187 (0.2 μM) stimulated generation of 5-HETE, leukotriene B4, and leukotriene C4 in rat mixed peritoneal cells, yielding IC50 values of 35.5,47.4, and 31.7 μM, respectively. Nordihydroguaiaretic acid (a potent 5-lipoxygenase inhibitor) also exerted a strong and concentration-dependent inhibition of 5-HETE and leukotriene B4 and C4 formation with IC50 values of 0.15, 0.09, and 0.1 μM, respectively. The inhibition of the formation of the products of the lipoxygenase pathway of arachidonic acid metabolism by azelastine may contribute to its overall antiallergic, antiasthmatic, and pulmonary anti-inflammatory activities.
Shared Allergenic Activity in Asian (Blattella asahinai), German (Blattella germanica), American (Periplaneta americana), and Oriental (Blatta orientalis) Cockroach Species
Authors: Helm Ricki M. , Squillace Diane L. , Jones Richard T. , Brenner Richard J.
Published: August 2009

Whole-body extracts of the feral and peridomestic Asian cockroach (Blattella asahinai) and the three domestic cockroach species, German (Blattella germanica), American (Periplaneta americana), and Oriental (Blatta orientalis), were compared allergenically using an IgE serum pool from 4 German cockroach sensitive individuals. In crossover radioallergosorbent inhibition analysis, the Asian cockroach shared allergenic activity primarily with the German cockroach polymer and to a lesser extent with either the American or Oriental cockroach polymers. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and thin-layer isoelectric focusing analysis of the extracts showed similar although varying intensities of Coomassie blue stained banding patterns among five extracts analyzed. Electroblotting analysis with 12.5% SDS-PAGE of the whole-body German cockroach extract and IgE serum from individuals sensitive to German cockroach revealed eight allergenic components with apparent molecular weights of 92, 80, 67, 48, 36, 27, 25 and 18 kD. Five components could be identified in the whole-body extract of the Asian cockroach corresponding to apparent molecular weights of 92, 67, 48, 40, and 32 kD. Analysis of individual serum by immunoblot analysis with each of the cockroach extracts showed considerable heterogenicity in the IgE-binding pattern. Although the Asian cockroach demonstrated considerable cross-reacting allergenic components to German and relatively fewer cross-reacting allergenic components to either the Oriental or American, it is too early to establish genus- or species-specific cockroach allergens. It is important to point out that German cockroach sensitive individuals should be made aware of the potential exposure of Asian cockroach aeroallergens both indoors and outdoors in areas with high infestations of Asian cockroaches.
K-Channel Blocking Drugs Induce Histamine Release and 45Ca Uptake in Isolated Mast Cells
Authors: Eleno N. , Botana L. , Espinosa J.
Published: August 2009

Histamine secretion and 45Ca uptake processes were studied in mast cells treated with four K+ channel blocking drugs in physiological saline and in media containing different ionic concentrations. Quinine, 4-aminopyridine and sparteine were effective as histamine-releasing agents when mast cells were incubated in physiologic saline solution. The dose-response profile obtained was in the range of 0.1–0.5 mM for quinine, 1–10 for 4-aminopyridine and 0.5–5 mM for sparteine and did not show significant differences between purified and unpurified mast cells. By contrast, tetraethylammonium (1–100 mM) did not induce histamine release. The presence of high K+ or Rb+ concentrations in the medium (Tris-K+ or Tris-Rb+, both at 150 mM) displaced the profile obtained to the right in cells stimulated with 4-aminopyridine or sparteine, but abolished histamine release induced by quinine. Additionally, all three K+ channel blockers increased 45Ca uptake in mast cells. The exact mechanism of the action of K+ channel blockers on mast cells is unknown. However, the fact that the drugs used were effective as histamine-releasing and 45Ca uptake promoters suggests both that mast cells might be endowed with a K+ channel activity and that the blockade of this should open certain calcium channels, leading to elevated intracellular Ca2+ levels which in turn activate mast cell secretion.
Identification of Soybean Allergens by Immunoblotting with Sera from Soy-Allergic Adults
Authors: Herian Anne M. , Taylor Steve L. , Bush Robert K.
Published: August 2009

Immunoblots were used to compare the IgE-binding proteins of various soybean products using sera from 7 soybean-allergic patients. Blotting results indicate at least two distinct individualistic patterns of IgE binding to soy proteins. Serum IgE from individuals allergic to both peanuts and soybeans binds to several proteins with molecular weights ranging from 50,000 to 60,000 daltons, while IgE from individuals allergic to soybeans, but not peanuts, binds strongly to a protein(s) with a molecular weight of 20,000 daltons. The soy products tested displayed varying amounts of IgE binding with sera from sensitive patients.
Comparison of Conidial and Mycelial Allergens of Alternaria alternata
Authors: Paris S. , Fitting C. , Latgé J.P. , Herman D. , Guinnepain M.T. , David B.
Published: August 2009

Alternaria allergens, separated by SDS polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane, were identified using sera from Alternaria-allergic patients. 30 Alternaria components of spore and mycelium, ranging in molecular weight from more than 150 to 12 kD, bound IgE antibodies. 15 were detected by more than 25% of the sera and only 4 (85, 56, 42, 31 kD) by more than 50% of the sera. Among these four major allergens, the 56-kD component was present mostly in the spore extract, the 85- and 42-kD fractions in the mycelium extract and the 31-kD in both extracts. The 31-kD component showed the highest IgE binding and the highest frequency of binding (95% with mycelium extract and 79% with spore extract). It was composed of two subunits.
Antigenic Analysis of Group I House Dust Mite Allergens Using Random Fragments of Der p I Expressed by Recombinant DNA Libraries
Authors: Greene W.K. , Chua K.Y. , Stewart G.A. , Thomas W.R.
Published: August 2009

Antigenic regions of a major house dust mite allergen, Der p I, were identified by a recombinant DNA strategy employing the technique of random fragmentation. Fragments of cDNA coding for Der p I were produced by sonication and used to construct lambda gt 11 expression libraries. Analyses of recombinant fragments reactive with a rabbit anti-Der p I antiserum showed that the B cell determinants expressed in Escherichia coli were limited, with the majority (86%) of antigenic clones isolated mapping to the region comprising amino acid sequence position 60–80. To define antigenic regions of Der p I more precisely, selected overlapping fragments were subcloned into the expression vector pGEX-1. Dot blot immunoassay and immunoabsorption studies using individual fusion proteins revealed five regions – 34–47, 60–72, 82–99, 112–140, and 166–194 – to contain B cell determinants responsible for the antigenicity of recombinant Der p I. Absorption of the anti-serum with Dermatophagoides pteronyssinus extract removed reactivity to all fragments, whereas absorption with an extract from the related mite Dermatophagoides farinae removed reactivity to peptides containing residues 34–47, 60–72, and 166–194, but not 82–99 and 112–140. Similarly, rabbit anti-D. farinae reacted strongly with peptides containing residues 34–47, 60–72, and 166–194, but not residues 82–99 and 112–140 which again showed antigenic differences in these residues between the group I allergens.
Isolation and Characterization of Rat Carcinoembryonic Antigen
Authors: Kim J.G. , Abeyounis C.J.
Published: August 2009

A rat tumor-associated antigen with properties similar to those of human carcinoembryonic antigen (CEA) has been detected with rabbit immune sera in extracts of transplantable rat colonic adenocarcinoma, RCA-1. This antigen, termed rat CEA, was also detectable by a monkey antihuman CEA serum and a rat monoclonal antibody to rat CEA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of perchloric acid extracts of RCA-1 tumor, followed by immunoblotting with the above-mentioned anti-CEA reagents, revealed that rat CEA activity resided in components with a molecular weight of approximately 350 kD. The glycoprotein nature of these components was indicated by positive staining with periodic acid-Schiff. Sephadex G-200 chromatography, as well as Sepharose 4B chromatography with and without sodium dodecyl sulfate indicated that the 350-kD components existed in the extracts as molecular aggregates. The 350-kD material, which had been purified by an affinity column containing rat monoclonal antibodies to rat CEA, reacted with the rabbit and monkey anti-CEA sera. This provided strong evidence that serological activity of rat CEA was confined to the 350-kD components.
Immunohistological Analysis of Nerve Granulomas in Neuritic Leprosy
Authors: Narayanan R.B. , Girdhar A. , Girdhar B.K. , Malaviya G.N.
Published: August 2009

Immunohistological analysis of infiltrates of nerves in patients with neuritic leprosy was carried out using monoclonal antibodies defining T cell subsets, Langerhans cells, HLA DR antigens, and indirect immunofluorescence. In all, eight nerves were analyzed. 2 of the 8 nerves showed epithelioid cell granulomas surrounded by large numbers of lymphocytes. The predominant lymphocytes in these granulomas were activated T cells expressing CD3 and HLA DR antigens. The proportion of CD3+ and CD4+ cells was higher than that of CD8+ cells. The ratio of CD4+/CD8+ cells in these two biopsy specimens was 5.6 and 1.5, respectively. In these nerves CD4+ cells were diffusely scattered into epithelioid cell granulomas, while CD8+ cells were localized at the periphery of the granuloma. The remaining six nerves showed macrophages containing numerous bacilli, and a few lymphocytes and plasma cells diffusely distributed into the granuloma. In these nerves, only occasional lymphocytes expressing CD3 or CD4 or CD8 and HLA DR antigens were noticed. In two of the biopsy specimens, a small proportion of CD8+ cells were visualized. Macrophages and Schwann cells were HLA DR+ in all nerves. CD1+ cells were not seen in the infiltrates of any of these nerves. A similar pattern and distribution of cells was noticed in the nerve granulomas of tuberculoid and lepromatous leprosy. These findings suggest that the mechanisms of nerve damage in the patients with neuritic leprosy could be either immunological or non-immunological, depending on the nature and characteristics of the infiltrates.
Mouse IgE Response against Exoantigens of Trypanosoma cruzi
Authors: Gruppi A. , Pistoresi-Palencia M.C. , Basso B. , Vottero-Cima E.
Published: August 2009

This work describes the occurrence of anaphylactic antibodies against exoantigens of Trypanosoma cruzi in infected or immunized mice. The results obtained show that the exoantigens induce an IgE response when injected into mice with complete Freund’s adjuvant or in animals infected with 102 trypomastigotes. Among the antigens recognized by anti-exoantigen IgE, two molecules of 27 and 45 kD were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by western blotting. Immunofluorescence analysis located some target antigens on the parasite surface.
Glomerular CR1 Express in situ Cofactor Activity for Degradation of C3b
Authors: Vedeler C.A. , Matre R. , Iversen B.M.
Published: August 2009

Adherence of sheep erythrocytes (E) sensitized with IgM antibodies (A) and C3b (EAC3b) to C3b/C4b receptors (CR1) in cryostat sections of human renal glomeruli was studied using the closed chamber technique. The adsorption was stable for at least 3 h at 37°C. In the presence of purified factor I, the indicator cells, however, detached from the sections after 30 min at 37°C. Factor H was not required. The release was not due to loss of CR1 activity in the tissue. The detached indicator cells were negative in the immune adherence test and were agglutinated by antibody to C3d, but not by antibody to C3c. Western blot of the detached indicator cells revealed the presence of C3d and C3c was found in the chamber fluid. Accordingly, detachment of the Indicator cells was due to degradation of C3b to C3d with the release of C3c into the chamber fluid. Protease inhibitors did not prevent the detachment of the indicator cells. EAC3b incubated with sections of renal glomeruli preincubated with anti-CR1 antibody were not degraded. The results therefore indicate that CR1 in situ in renal glomeruli can provide the necessary corfactor activity for factor I-mediated degradation of C3b to C3d and C3c.
Importance of Mast-Cell-Derived Eosinophil Chemotactic Factor A on Granuloma Formation in Murine Schistosomiasis japonica: Evaluation Using Mast-Cell-Deficient W/Wv Mice
Authors: Owhashi Makoto , Horii Yoichiro , Ikeda Teruaki , Maruyama Haruhiko , Abe Tatsuya , Nawa Yukifumi
Published: August 2009

The role of mast-cell-derived eosinophil chemotactic factor A (ECF-A) on eosinophil-rich periovular granuloma formation was examined by using mast-cell-deficient WBB6F1W/Wv mice infected with Schistosoma japonicum. The average size of granulomas formed around newly deposited eggs in the liver of W/Wv mice was significantly smaller than that observed in control +/+ mice. The number of adult worms recovered or specific IgE titers were comparable between W/Wv and +/+ mice. In contrast, immediate-type hypersensitivity response to specific antigen and dialyzable low-molecular weight ECF in the serum was detectable only in infected +/+ mice. When naive bone marrow eosinophils were incubated with the dialyzable fraction of infected +/+ mice sera, chemotactic reactivity of eosinophils to ECF derived from S. japonicum eggs was significantly enhanced, although that to synthetic ECF-A was depressed. Similar effects were observed when naive eosinophils were treated with synthetic ECF-A. The dialyzable fraction of infected W/Wv mice sera had no such modulating effect on the chemotactic reactivity of eosinophils. These results suggest that an immediate-type hypersensitivity reaction is important in the formation of eosinophilic granulomas around S. japonicum eggs, mainly through the modulating effect of mast-cell-derived ECF-A on the chemotactic reactivity of eosinophils.
Antiallergic and Anti-Inflammatory Action of Tioxamast in Rats I.
Authors: Tarayre J.P. , Aliaga M. , Barbara M. , Tisseyre N. , Caillol V. , Delhon A. , Bruniquel F. , N’Guyen X. , Puech L. , Tisné-Versailles J. , Couzinier J.P.
Published: August 2009

Tioxamast (F 1865) is an antiallergic drug that, administered systemically, reduces anaphylaxis in various models in rats. This action is due mainly to the inhibition ofthe synthesis and release of certain mediators. Orally or intraduodenally administered tioxamast inhibits IgE-dependent passive cutaneous anaphylaxis (ED50 = 0.8 mg/kg), IgE-dependent passive pulmonary anaphylaxis (ED50 = 0.5 mg/kg), and IgG-dependent passive cutaneous anaphylaxis (ED50 = 0.6 mg/kg). It has little or no effect on the increase of cutaneous capillary permeability induced by various mediators. In IgE-dependent passive peritoneal anaphylaxis in rats, tioxamast reduces the release of histamine (IC50 = 0.024 µg/ml) and of β-glucuronidase (IC50 = 0.102 µg/ml). Also, histamine release is inhibited in IgG-dependent peritoneal anaphylaxis (IC50 = 0.103 µg/ml). The antiallergic compound has less effect on the release of histamine induced by the compound 48/80 in the peritoneal cavity of rats (IC50 = 1.67 µg/ml). Tioxamast inhibits the synthesis in vitro of leukotriene B4 (LTB4) by peritoneal neutrophils from rats stimulated by A23187 (IC50 = 8.88 µg/ml). At higher tioxamast concentrations, metabolites ofthe cyclo-oxygenase pathway are inhibited at concentrations ofthe same order of magnitude as those that inhibit Naja naja phospholipase A2 (IC50 = 144 µg/ml). Tioxamast also reduces the production of free radicals by leukocytes from the pleural cavity of rats which had phagocytosed opsonized zymosan (IC50 = 5.21 µg/ml).
Antiallergic and Anti-Inflammatory Action of Tioxamast in Rats II.
Authors: Tarayre J.P. , Aliaga M. , Barbara M. , Tisseyre N. , Caillol V. , Delhon A. , Bruniquel F. , N’Guyen X. , Puech L. , Tisné-Versailles J. , Couzinier J.P.
Published: August 2009

Tioxamast is an antiallergic drug that inhibits anaphylaxis in various models in rats, and it inhibits the release and synthesis of certain mediators of inflammation [see Tarayre et al., this issue]. Here we report that the drug also has an anti-inflammatory effect in vivo in various nonimmunological models in rats. It reduces zymosan-induced inflammation in the paw and pleural cavity, starting at doses from 1.5625 to 3.125 mg/kg given intraperitoneally. In pleurisy, tioxamast lowers the concentration of leukotriene B4 (LTB4) in the exudate, at doses from 50 mg/kg i.p. Also, at doses from 12.5 mg/kg i.p., the compound reduces PAF-acether-induced pleurisy and the concentrations of LTB4 and peptidoleukotrienes in the exudate. An anti-inflammatory action against carrageenin-induced edema of the paw was seen only at doses of 50 mg/kg i.p. or more. The anti-inflammatory and antiallergic effect of tioxamast makes it a potentially useful drug in the treatment of allergies in humans.
Contact Hypersensitivity Induces Plasma Interleukin 6
Authors: Kimber Ian , Cumberbatch Marie , Humphreys Marie , Hopkins Stephen J.
Published: August 2009

Previous studies in this laboratory have revealed that elicitation reactions in contact-sensitized mice correlate closely with elevations in the serum concentration of acute-phase proteins. Regulation of the hepatic synthesis of acute-phase proteins is one of the many biological functions ascribed to interleukin 6 (IL-6). We now provide evidence that contact reactions are also associated with a rapid and substantial increase in the plasma concentration of IL-6. Quantitative assessment of plasma IL-6 may provide a useful serological method for the evaluation of experimental allergic contact dermatitis in mice, and possibly in other species.
Immunoglobulin G Subclass Deficiency in Children with High Levels of Immunoglobulin E and Infection Proneness
Authors: Loh R.K.S. , Thong Y.H. , Ferrante A.
Published: August 2009

Of 32 unrelated children with serum IgE > 1,000 U/ml, 17 were found to have infection proneness according to standard clinical criteria, and 15 were not infection prone. There were no statistical differences between these 2 groups of children with regard to age, sex, serum IgE levels or prevalence of asthma. However, the prevalence of eczema was significantly lower in the infection-prone group (p = 0.035). Of greater interest was the finding that 7 children in the infection-prone group had IgG subclass and/or IgA deficiency compared with none in the non-infection-prone group (p = 0.006). These results suggest that IgG subclass studies may be warranted in children with markedly elevated levels of serum IgE and proneness to infection.
Histamine Release from Cord Blood Basophils
Authors: Windelborg Nielsen Bent , Engberg Tine Margrethe , Herlin Troels , Bjerke Torbjørn , Schiøtz Peter Oluf
Published: August 2009

The histamine release (HR) after challenge with anti-IgE, concanavalin A, N-formyl-met-leu-phe and the calcium ionophore A23187 from 97 cord blood samples was determined by a microfiber-based assay. Maximum HR with anti-IgE showed great inter-individual variation (median: 20.5; range: 1–104 ng/ml blood), but was not significantly different from the results obtained in identically treated blood samples from 50 adults (median: 23; range: 1–93 ng/ml blood). Both the maximum HR and the sensitivity to anti-IgE were dependent on total plasma IgE content. Blood samples with plasma IgE < 0.5 IU/ml (n = 15) had significantly higher maximum HR than those with plasma IgE < 0.5 IU/ml (n = 82; median: 32 vs. 18 ng/ml blood; p < 0.01). Passive sensitization with IgE-rich atopic plasma increased the maximum HR with anti-IgE only in samples with a plasma IgE content of less than 0.5 IU/ml, although sensitivity to anti-IgE was universally increased. Preincubation with pharmacologic agents modulating the IgE-mediated HR produced effects generally similar to previous findings in adult blood. However, the effects of inhibiting the cyclooxygenase pathway in cord blood differed from our observations in adult blood, and may represent a maturational phenomenon. The familiy history of allergy was obtained by a questionnaire, and clinical observations were gathered from patient records. None of these parameters were found to influence HR with any secretagogue. However, HR stimulated by the calcium ionophore A 23187 was found to be highly dependent on the storage time of the EDTA-anticoagulated blood samples, which should be carefully controlled.
Regulation of the Expression of the Low-Affinity IgE Receptor (FcεRII) in the Human Monocyte-Like Cell Line U-937 by Phorbol Esters and IgE
Authors: Pucillo Carlo , Salzano Salvatore , Pepe Stefano , Vitale Mario , Formisano Salvatore , Rossi Guido
Published: August 2009

The expression of the low-affinity receptor for IgE FcεRII) in the human monocyte-like U-937 cell line can be upregulated by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) and by IgE. TPA induces terminal differentiation of U-937 cells and causes a four- to fivefold increase in the number of FcεRII. TPA also modulates the expression of several other membrane markers of U-937 cells. IgE alone has a modest effect on the expression of FcεRII (about a 10% increase), while simultaneous treatment of U-937 cells with TPA and IgE has a cooperative effect, causing an eightfold increase in the number of FcεRII. Cycloheximide strongly suppresses the expression of FcεRII, both in TPA-stimulated and unstimulated cells; this effect can be partly reversed by culturing the cells in the presence of IgE. These results suggest that TPA induces the expression of newly synthesized receptors, while IgE causes an accumulation of preformed receptors.
T-Cell Responsiveness towards Various Synthetic Peptides of the P28 Antigen in Rat and Mouse Models during Schistosoma mansoni Infection
Authors: Wolowczuk Isabelle , Auriault Claude , Gras-Masse Hélène , Mazingue Christine , Vendeville Catherine , Tartar André , Capron André
Published: August 2009

It has recently been demonstrated that the Schistosoma mansoni P28 antigen can induce a strong protective immunity after direct immunization in various experimental models. T lymphocytes from Fischer rats immunized with the recombinant P28 antigen were cultured in vitro in the presence of seven synthetic peptides derived from the amino acid sequence of the P28. The most significant and reproducible proliferation was obtained with the 24–43 and 115–131 synthetic peptides. In order to analyze whether these located determinants were also exposed to the host’s immune system during the natural S. mansoni infection or after immunization with crude antigenic extracts of various development stages of the parasite, the T-cell responsiveness of infected or immunized Fischer rats and BALB/c mice was tested towards these synthetic peptides. The results showed that, in both permissive (mouse) and non-permissive (rat) hosts, 24–43 and 115–131 synthetic peptides are recognized during the course of infection and that there is a dynamic variation of this recognition. These peptides are also recognized by T cells educated against crude antigenic extracts of different developmental stages of the parasite which contained the native form of the P28 molecule. Taken together, the results indicated that these synthetic peptides derived from the recombinant P28 antigen can activate T lymphocytes educated against the native P28 molecule during the development and maturation of the parasite in their hosts. Therefore, they might be useful for the construction of synthetic vaccines against schistosomiasis.
Selective Inhibition by Magnosalin and Magnoshinin, Compounds from ‘Shin-i’ (Flos magnoliae), of Adjuvant-Induced Angiogenesis and Granuloma Formation in the Mouse Pouch
Authors: Kimura Masayasu , Kobayashi Shinjiro , Luo Bao , Kimura Ikuko
Published: August 2009

Inhibitory effects of magnosalin and magnoshinin, compounds from the crude drug ‘Shin-I’ (Flos magnoliae), on angiogenesis and pouch granuloma formation induced by an adjuvant containing croton oil were investigated. Magnosalin inhibited angiogenesis 2.4-fold (intra-pouch) and 9.7-fold (intraperitoneal) more strongly than granuloma formation. The inhibition of angiogenesis by magnosalin was 5-fold (intra-pouch) and 21-fold (intraperitoneal) weaker than that by hydrocortisone. In contrast, intraperitoneal magnoshinin inhibited granuloma formation 2.5-fold more strongly than angiogenesis. The regression coefficients of anti-angiogenesis vs. the inhibition of granuloma formation were 1.79 for magnosalin, 1.11 for hydrocortisone, and 0.61 for magnoshinin. These results show that the anti-chronic inflammatory effect of ‘Shin-i’ was caused by selective inhibition of angiogenesis by magnosalin and of granuloma formation by magnoshinin.
Two-Dimensional Gel Electrophoretic Analyses of Kentucky Bluegrass and Rye Grass Pollen Allergens
Authors: Ekramoddoullah Abul K.M.
Published: August 2009

Allergenic extracts of Kentucky bluegrass (Poa pratensis) and rye grass (Lolium perenne) pollen were shown by 2-dimensional (2-D) gel electrophoresis to consist of several hundred protein components. The pollen extracts of the two related grasses had unique 2-D gel patterns. Two major grass pollen allergens, Poa p I and Lol p I, and their isoforms (i.e. isoallergens) were detected and localized on 2-D gels by immunoblotting with anti-Poa p I antibody mAb 60. Poa p I isoallergens were less acidic than Lol p I isoallergens. The relative proportion of four Poa p I isoallergens was (in decreasing pI): A, 13%; B, 37%; C, 35%; and D, 15%. The amino terminal amino acid sequences of the two major isoallergens of Poa p I were identical. However, the amino acid composition of these isoallergens showed enough differences to account for their charge differences. The amino terminal amino acid sequence of two major Poa p I isoallergens had a 70% homology in 20 amino acid overlap with the previously published amino terminal amino acid sequence of rye grass pollen allergen R-7.