bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunomodulating Properties of Substances to Be Used in Combination with Liposomes
Authors: Zigterman Guy J.W.J. , Jansze Margriet , Snippe Harm , Willers Jan M.N.
Published: August 2009

Liposomes haptenated with tripeptide-enlarged dinitrophenyl (DNP) are known to act as thymus-independent antigens which induce a strong IgM response and only limited amounts of circulating IgG. When haptenated liposomes are used in vaccination studies, it is of practical importance to improve the immunogenicity of these complexes. Therefore, an evaluation was made of the potency of various substances to modulate the immune response in such a way that the total antibody production is increased, including a relative great increase of 2-merecaptoethanol (2-ME)-resistant antibodies and immunological memory is induced. The following substances were used: glycophorin A (GP-A), sialogangliosides (monosialo-, disialo- and trisialogan-glioside), 6–0-stearoyl-MDP (MDP-SA) and lipid A (lip A). Lip A incorporated into liposomes was the only substance inducing considerable increases of both total and 2-ME-resistant haemagglutination (HA) titre after immunization. Depending on the dose tested, the sialic-acid-containing protein GP-A had a small and varying influence on the serum antibody response. Sialogangliosides transiently decreased in a dose-dependent manner the total antibody titre in serum. In contrast to lip A, the lipophilic bacterial adjuvant MDP-SA did not influence HA titres significantly. The number of plaque-forming cells (PFC) in the spleen was enhanced considerably after both primary and secondary immunization with liposomes containing lip A. The other substances tested induced only minor differences of the number of PFC. To some extent, lip A induced immunological memory. In conclusion, it can be stated that of the agents tested, only lip A is a potent and consistent stimulator of the humoral immune response to liposomes haptenated with DNP groups. Sialogangliosides induced a transient decrease of the immune response to haptenated liposomes.
Enzyme-Linked Immunosorbent Assay (ELISA) Inhibition Method to Estimate the Level of Airborne Bovine Epidermal Antigen in Cowsheds
Authors: Virtanen T. , Louhelainen K. , Mäntyjärvi R.
Published: August 2009

A method based on inhibition of the enzyme-linked immunosorbent assay was developed for measuring airborne bovine epidermal antigen (BEA) in cowsheds. BEA extract served as the solid-phase reactant competing with the sample for binding of a rabbit immune serum. A sample of swine epithelial extract was used to ascertain the specificity of the reaction. Cowshed air samples were collected according to a standardized volumetric filter method. A specific inhibiting activity was detected in all samples tested corresponding to concentrations between 137ng/m3 and 19.8 μg/m3. The intertest coefficient of variation of cowshed samples was 9.5%.
Contact-Sensitizing and Tolerogenic Properties of 2,4-Dinitrothiocyanobenzene
Authors: Kimber Ian , Botham Philip A. , Rattray Nicola J. , Walsh Stuart T.
Published: August 2009

Epicutaneous application of 2,4-dinitrothiocyanobenzene (DNTB) is said to result in specific immunological hyporesponsiveness and fails to induce contact sensitization. However, we demonstrate that topical exposure to DNTB causes activation of the draining lymph node in mice and the induction of contact sensitization in both rodents and a single human volunteer. In mice and rats, pre-exposure to DNTB failed to impair subsequent responsiveness to the cross-reactive allergen 2,4-dinitrochlorobenzene. These data provide evidence that DNTB cannot be regarded as an exclusive tolerogen when applied epicutaneously and indicate that attempts to define the characteristics of tolerising chemicals from analysis of this agent may be misleading.
Enzyme Immunoassay with Cell Suspensions: Studies on Reactions between Serum Antibodies and Cell-Surface Antigens
Authors: Wasik M. , Muirhead D. , Rubocki R. , Milgrom F.
Published: August 2009

The following reactions of serum antibodies with cell surface antigens were studied by means of an enzyme immunoassay with cell suspensions: (1) Rh antiserum with human red blood cells; (2) H-2 antisera with murine red blood cells; (3) H-2 antisera with murine thymocytes and hepatocytes; (4) Thy-1 antiserum with murine thymocytes; (5) polyvalent and monovalent HLA alloantisera with human leukocytes and human mononuclear cells, and (6) antisera of murine origin with human and marmoset lymphoid cell lines. In all instances, with the exception of monovalent HLA antisera, the assay proved to be a sensitive and highly specific procedure.
Functional Evaluation of the Basophil/IgE System in the Cord Blood
Authors: Miadonna Antonio , Tedeschi Alberto , Leggieri Ennio , Motta Giuliano , Fabbri Carola , Froldi Marco , Agosti Massimo , Lorini Maurizio , Marini Antonio , Zanussi Carlo
Published: August 2009

Basophil releasability was studied in 24 cord blood samples from normal-term deliveries. The histamine content in cord blood basophils was similar to that of adult blood basophils. The response to IgE-independent degranulating stimuli such as calcium ionophore A23187 and zymosan-activated human serum was overlapping with that of normal adults. Conversely, a reduced releasability was observed after challenge with anti-IgE, even after sensitization with an IgE-rich serum. The IgE-dependent degranulation seems to be hampered by the low concentrations of circulating and cell-bound IgE antibodies. The number of IgE molecules bound to the specific receptors in cord blood basophils is significantly lower than in adult blood basophils.
Changes in IgG and IgE Antibody Levels to Bee Venom during Immunotherapy
Authors: Ferrante Antonio , Mocatta Frances , Goh David H.B.
Published: August 2009

IgE and IgG antibodies to bee venom were measured in sera of patients receiving bee venom immunotherapy. All patients selected for therapy had suffered severe reactions to bee stings. The results showed that within 2–3 months from the commencement of immunotherapy there was a marked rise in IgG antibodies and a slight but not significant rise in IgE antibodies. After this period, the IgE antibody level began to fall and was about one third of the pre-treatment level by the second to third year. The IgG antibody level began to decline from its increased level after 9–10 months but remained above the pre-treatment level even after 2 years. All of the patients who had subsequently been accidentally stung after reaching the maintenance dosage of bee venom allergen showed no severe reactions. A small group of non-treated patients of the same category as those receiving therapy did not show a rise but a decline in the level of IgG antibodies. However, like the treated group, they showed a significant decrease in IgE antibodies. Thus one of the main benefits of bee venom immunotherapy is the build-up of a high concentration of IgG antibodies, and this may be the critical factor in the protection against bee venom allergy.
Intraepithelial Migration of Mucosal Mast Cells in Hay Fever: Ultrastructural Observations
Authors: Enerbäck Lennart , Pipkorn Ulf , Olofsson Anita
Published: August 2009

Evidence has been presented suggesting that a migration of nasal mast cells from the mucosal connective tissue stroma into the epithelium is part of the mucosal response in birch pollen allergy. In a previous study, the identification of these intraepithelial cells as tissue mast cells rather than blood basophils was based on light microscopical morphology and histochemistry. We have now studied the ultrastructure of these cells in mucosal biopsies taken before and during the birch pollen season. Intraepithelial cells with basophil or metachromatic granules were only observed in biopsies taken during the season. Some of these cells had the ultrastructural appearance of tissue mast cells, including cytoplasmic lipid droplets and a granular substructure composed of multilamellar arrays and scrolls, serving to distinguish human mast cells from blood basophils. The ultrastructural traits of the remaining cells were heterogeneous, some reminiscent of human blood basophils, others of globule leucocytes of other species, but entirely typical blood basophils could not be identified. The results thus support our previous suggestion that a migration of mucosal mast cells from the connective tissue stroma into the epithelium is part of the human allergic mucosal response. It cannot be determined whether the ultrastructural heterogeneity of these cells is the result of an adaptation to the intraepithelial environment of one single mast cell type or to the existence of an ultrastructurally distinct mucosal mast cell.
The Immunocompetence of Children with Congenital Heart Disease
Authors: Radford D.J. , Lachman R. , Thong Y.H.
Published: August 2009

We studied the immunocompetence of 18 children with conotruncal malformations (13 with tetralogy of Fallot, 5 with truncus arteriosus) and 22 children with cardiac shunt lesions. There were reduced total T cell percentages and T helper cells in the conotruncal group but no T cell abnormality in the shunt group. Also, 7 of the 18 cases in the conotruncal group had facial dysmorphism reminiscent of the Di George syndrome. These results suggest that patients with conotruncal malformations fall into the wide spectrum of the Di George syndrome. There was some humoral deficiency in both groups with reduced levels of immunoglobulins IgG and IgA and low levels of complement C3 and C4. The clinical records showed a high frequency of infections. Hospital admissions for these episodes had occurred in 61 % of the conotruncal group and 32% of the shunt group. Thus, there is an increased susceptibility to infection in children with congenital heart disease, and the predilection to infection has an immunological basis.
Mucosal Mast Cells as a Component of the Inflammatory Response to Lower-Urinary Tract Infection
Authors: Cornish Julian , Vanderwee Mark A. , Ormrod Douglas J. , Miller Thomas E.
Published: August 2009

Globule cells have been observed in mucosae for many years. Recently, a subpopulation of these globule cells in the intestinal mucosa of man and rodents have been identified as unique mast cells. In this communication, intraepithelial globule cells and some lamina-propria mast cells found in the normal rat urinary-bladder wall have been characterized as mucosal mast cells, similar to intestinal mast cells, and differentiated morphologically and histochemically from rat peritoneal mast cells. The number of mast cells in the bladder wall increased significantly during various bladder manipulations, including mechanical trauma, parasitic infestation and bacterial infection. The origin and function of the mucosal mast cells remains unknown.
Effects of Thiazinamium Chloride on Histamine Release from Rat Peritoneal Mast Cells and on Phosphodiesterase Activity in Guinea Pig Lung
Authors: Carlson Richard P. , O’Neill-Davis Lynn , Chang Joseph , Lewis Alan J.
Published: August 2009

The effect of thiazinamium Cl (TCl) on histamine release from rat peritoneal mast cells (RPMC) was investigated. Although TCl inhibited compound 48/80-induced histamine release moderately (IC50 value 40 μM), the drug was a weaker inhibitor of ovalbumin-induced histamine release (100 μM, ––21 %). In contrast, promethazine HCl (PHCl) was more effective against antigen-induced histamine release (IC50 value 13 μM) than against compound 48/80-induced histamine release (100 μM, ––53%). Disodium cromoglycate (DSCG) was effective against both antigen and compound 48/80-induced release of histamine with IC50 values of 7 and 1 μM, respectively. Neither TCl nor DSCG at 1 mM increased spontaneous release of histamine from RPMC, whereas PHCl induced spontaneous release by over 50% at 1 mM. TCl did not inhibit phosphodiesterase (PDE) activity in guinea pig lung at 1 mM, whereas theophylline and DSCG inhibited PDE with IC50 values of 1.1 and 0.32 mM, respectively. These data suggest that high local concentrations of TCl may reduce histamine release during an asthmatic attack and improve its effectiveness as a bronchoprotectant.
Changes in the Coagulation System during Pseudoallergic Anaphylactoid Reactions to Drugs and Food Additives
Authors: Zimmermann Eberhard , Czarnetzki Beate M.
Published: August 2009

In 10 patients with known anaphylactoid reactions to drugs and food additives, oral provocation tests were performed with increasing doses of the implicated agents. Clinical reactions, vital signs, bleeding time, prothrombin time, partial thromboplastin time and platelet aggregation were measured before and 15 min after each oral dose. Six patients reacted clinically. In all 10 patients, platelet aggregation decreased with increasing challenging doses. All other parameters remained stable or changed without any correlation to the drug, the type or the severity of the reaction. Measurement of the bleeding time is, therefore, of no diagnostic value in anaphylactoid reactions, and major alterations in the coagulation system do not explain their pathogenesis.
Effect of Serotonin on T Lymphocyte Proliferation in vitro in Healthy Individuals
Authors: Khan I.A. , Bhardwaj G. , Malla N. , Wattal C. , Agarwal S.C.
Published: August 2009

The direct effect of different concentrations of serotonin on T lymphocyte proliferation in vitro was seen in 20 normal individuals. It has been observed that the neurotransmitter had a suppressive effect on mitogen-induced lymphocyte proliferation. This study shows that the effect of serotonin on the immune system is direct.
Distribution of a Guinea Pig Lymphocyte Cell Surface Antigen of Defined Function, as Detected by the Mouse Monoclonal Antibody MSgp 1
Authors: Healey D.G. , Parker Darien , Turk J.L.
Published: August 2009

A mouse anti-guinea pig monoclonal antibody, designated MSgp 1, was derived from a fusion between NS-1 myeloma cells and splenocytes hyperimmunised with guinea pig thymocytes. The MSgp 1 determinant is expressed by a subset of small thymocytes and lymph node T cells which participate in mixed leukocyte reactions. The determinant is modulated by antigen in vivo, and MSgp 1 antibody will prevent MHC class II driven proliferation in vitro. In addition, MSgp 1 reacts with a minor population of lymph node B cells, but not with a chronic B cell leukaemic cell line. Resident peritoneal macrophages express the MSgp 1 determinant, whereas chronic oil-induced peritoneal macrophages do not. The role of MSgp 1 defining guinea pig helper T cells is discussed by comparisons with other documented T helper cell reagents.
An Antigenic Determinant Common to Lymphocytes and Langerhans Cells of the Guinea Pig
Authors: Healey D.G. , Baker D. , Gschmeissner S.E. , Turk J.L.
Published: August 2009

A mouse anti-guinea pig monoclonal antibody, designated MSgp 2, was derived by the fusion of NS-1 myeloma cells with mouse splenocytes hyperimmunized with guinea pig B cells. MSgp 2 reacts with an antigen present on the majority of lymphocytes. An unexpected finding was the expression of the MSgp 2 antigen on epidermal Langerhans cells, as defined by cell morphology, expression of MHC class II antigen and presence of Birbeck granules in positive cells. It is suggested that the tissue distribution of MSgp 2 antigen on lymphocytes of the lymph node could indicate a role for this determinant in cell migration.
Clonal Analysis of T Lymphocytes Infiltrating the Thyroid Gland in Hashimoto’s Thyroiditis
Authors: Bagnasco Marcello , Ferrini Silvano , Venuti Daniela , Prigione Ignazia , Torre Giancarlo , Biassoni Roberto , Canonica Giorgio Walter
Published: August 2009

T cells isolated from thyroid tissue and peripheral blood of 2 patients with Hashimoto’s thyroiditis were studied by a high cloning efficiency microculture technique. Clonal efficiences of 37 and 24% were obtained from thyroid-derived T cell cultures, while 40 and 90% efficiencies resulted from peripheral-blood-derived cultures. A prevalence of T4––/T8+ T cell clones were found in thyroid infiltrates. The functional analysis of the clones demonstrated significantly higher proportions of clones with cytolytic activity in a lectin-dependent assay in thyroid-derived microcultures, as compared to peripheral blood-derived ones. The proportion of clones displaying natural-killer-like activity was increased in 1 patient only. Cytolytic activity was displayed not only by all T4––/T8+, but also by several T4+/T8–– intrathyroid clones. Remarkable proportions of cytolytic clones were also able to release interleukin-2 upon phytohemagglutinin stimulation. Finally, the proportion of T cell clones able to release gamma-interferon following mitogen stimulation was significantly higher in thyroid- vs. peripheral-blood-derived microcultures. These results provide further data about the possible pathogenetical role of both regulatory and effector T lymphocytes in human autoimmune thyroiditis.
Antibody Synthesis in the Rat Liver: An Association between Antibody-Forming Cells in the Liver and Biliary Antibodies following Intravenous Injection of Horse Erythrocytes
Authors: Carter Lisa , Barrington Patricia J. , Cooper G.N. , Jackson Graham D.F.
Published: August 2009

The transient appearance of IgM in the bile of rats injected intravenously with horse erythrocytes (HRBC) correlated with the occurrence of anti-HRBC IgM-secreting cells in the liver. Both responses were reduced in animals splenectomised at the time of immunisation. When rats were given 2 injections of HRBC spaced by 28 days, IgG, IgA and IgM antibody-forming cells (AFC) were detected in the liver along with IgM and non-IgM anti-HRBC antibody in bile. There was a 10-fold increase in the total numbers of mononuclear cells (MNC) retrievable from the liver at the height of the biliary antibody response, the majority of which were not AFC. These results suggest that antigen entering the spleen stimulates the release of a population of MNC which may localise in the liver and that a minor portion of these produces specific antibody which is secreted into bile.
Effect of Polymeric IgG on Human Monocyte Functions
Authors: Mannhalter Josef W. , Ahmad Rafi , Wolf Hermann M. , Eibl Martha M.
Published: August 2009

Fc and iC3b receptors are involved in various biological functions of phagocytic cells, such as immune adherence and phagocytosis of opsonized particles, degranulation and superoxide generation. In the present study we examined the expression of specific receptors for the Fc portion of IgG (FcR) and for iC3b (CR3), a cleavage product of the third complement component, on human monocytes following in vitro treatment with polymeric and monomeric IgG. Interaction of polymeric IgG (fluid phase) with the monocyte membrane led to a concomitant modulation of both Fc and iC3b receptors. Monomeric IgG, however, down modulated Fc receptor expression only if surface bound. Under these conditions, no concomitant modulation of the iC3b receptor could be observed. The down modulation of Fc and iC3b receptors induced by fluid-phase IgG polymers was also accompanied by a decrease in monocyte functions as expressed by reduced Fc receptor-mediated phagocytosis, decreased release of oxygen metabolites following stimulation by aggregated IgG and opsonized zymosan, as well as in impaired killing of bacteria. These data suggest that a down modulation of Fc and iC3b receptors might have important implications for host defense mechanisms, since interaction with these receptors is required for the proper elimination of many pathogens.
A Murine Anti-Sheep T8 Monoclonal Antibody, ST-8, that Defines the Cytotoxic T Lymphocyte Population
Authors: Ezaki Taichi , Miyasaka Masayuki , Beya Marie-Françoise , Dudler Lisbeth , Trnka Zdenek
Published: August 2009

A mouse monoclonal antibody (mAb), ST-8, reactive with a subpopulation of sheep T lymphocytes was investigated by tissue distribution analysis and functional studies of the antigen-bearing (ST-8+) cells. Two other mouse mAb, ST-1 and T-80 that recognize all sheep T cells and a T-cell subset, respectively, were also examined for comparison. The ST-8+ cells represented 61–69% of thymocytes and about 10–30% of peripheral T lymphocytes. Histologically, ST-8+ cells were found mainly in the thymic cortex, T-dependent areas of the peripheral lymphoid tissues and the splenic red pulp and marginal zone, but not in B-dependent areas such as germinal centers of lymph follicles of the Peyer’s patches. ST-8 mAb plus complement treatment completely abolished both the induction phase and the effector phase of alloreactive cytotoxic T lymphocytes but did not affect proliferative responses to T-dependent mitogens and allogeneic antigens. ST-8 mAb also blocked the cytotoxic T lymphocyte function of lysing specific targets in the absence of complement. ST-8 mAb immunoprecipitated an antigen from the surface of sheep T lymphocytes under reducing conditions with an apparent molecular weight of 33–35 kilodaltons. Therefore the antigen recognized by the ST-8 mAb showed striking similarities to human T8/Leu-2a and mouse Lyt-2 antigens not only in the tissue distribution and function of antigen-bearing cells but also the molecular weight. We conclude that the ST-8 antigen is the ovine homologue of the human T8 and mouse Lyt-2 antigens.
Production of an International Reference Standard Alternaria Extract
Authors: Helm R.M. , Squillace D.L. , Aukrust L. , Borch S.M. , Baer H. , Bush R.K. , Løwenstein H. , Znamirowski R. , Nitchuk W. , Yunginger J.W.
Published: August 2009

As part of a program to establish international standards of selected allergens, 6 coded extracts of Alternaria were assessed in 6 laboratories by immunochemical, biochemical and physicochemical procedures. Direct RAST, RAST inhibition, quantitative skin tests and leukocyte histamine release were used to assign relative orders of potency to the 6 extracts. The composition and major allergen content was tested by thin-layer isoelectric focusing and quantitative immunoelectrophoresis (crossed immunoelectrophoresis and crossed radioimmunoelectrophoresis). Three laboratories determined the quantity of purified allergens in each of the preparations. In addition, source materials were sent to an expert Alternaria taxonomist for independent identification. The results showed considerable variation with respect to total allergenic potency and content of individual allergens. Source materials could not be confirmed as Alternaria in some instances. Based on fulfillment of written specifications and assay results, extract No. 6 was recommended by the Alternaria Working Group as the candidate international standard to the Steering Committee of the Allergen Standardization Subcommittee of the International Union of Immunological Societies.
Further Studies of Patients with Both Honeybee- and Yellow-Jacket-Venom-Specific IgE
Authors: Reisman Robert E. , Wypych John I. , Lazell Mary I.
Published: August 2009

Twenty-five sera from patients with high titers of both honeybee- and yellow-jacket-venom-specific IgE were analyzed in RAST inhibition experiments, using each venom as the coupling and inhibiting antigen. Eight sera had unique antibody activity with no cross-reactivity between yellow-jacket- and honeybee-venom-specific IgE. In 5 sera, the IgE antibody activity was directed at a major allergen in yellow jacket venom cross-reacting with a minor allergen in honeybee venom. Honeybee venom inhibited only the honeybee venom RAST; yellow jacket venom inhibited both the honeybee and the yellow jacket venom RAST. Five sera showed the opposite pattern with IgE antibody directed at a major allergen in honeybee venom cross-reacting with a minor allergen in yellow jacket venom. The fourth newly observed pattern was that of extensive IgE antibody cross-reaction found with 7 sera. Both honeybee venom and yellow jacket venom inhibited both the honeybee and yellow jacket venom RASTs. There were no clinical features such as age, sex, atopy or type of anaphylactic symptoms which could distinguish patients in each group. All but one patient had a history of multiple sting exposures. These data suggest multiple allergens in honeybee and yellow jacket venom with differing patterns of cross-reactivity and have important implications for proper venom immunotherapy.
Analysis of Mast Cell Function in Biological Responses Not Involving IgE
Authors: Galli Stephen J. , Wershil Barry K. , Mekori Yoseph A.
Published: August 2009

Mast cells are critical for the expression of certain IgE-mediated responses, but the precise contributions of mast cells to biological processes not involving IgE are obscure. We have employed genetically mast cell-deficient WBB6F1-W/Wv and WCB6F1-Sl/Sld mice to investigate the roles of mast cells in several different biological responses. This work strongly suggests that mast cells are not required for the elicitation of contact sensitivity (CS) responses, suppressor T cell-dependent tolerance to CS, reserpine-induced inhibition of CS responses, or bleomycin-induced pulmonary fibrosis. By contrast, mast cells appear to contribute to the acute gastric injury induced by ethanol and the acute inflammation of the skin induced by croton oil.
Changes in Nasal Mast Cell Numbers In and Out of the Pollen Season
Authors: Viegas M. , Gomez E. , Brooks J. , Davies R.J.
Published: August 2009

Mast cells were counted in biopsies of the nasal mucous membrane from 8 patients with allergic rhinitis during (July 1985) and after (October 1985 and January 1986) the grass pollen season in both the superficial epithelium and the deeper lamina propria. There was an eightfold increase in the total number of mast cells in the nasal mucous membrane during the grass pollen season when compared to midwinter. In addition, there was a striking change in the number of mast cells present in the surface epithelial layer from almost total absence in biopsies taken in midwinter to counts of between 2,000 and 28,000/mm3 in all but 1 patient during the summer.
Seasonal Variation of Mast Cell Function in Human Adenoid Tissues
Authors: Schmutzler W. , Amandi M. , Aniol M. , Braam U. , Eichelberg D. , Greven T. , Kortemeier A. , Schmitz F.J. , Ziesche R.
Published: August 2009

The number of mast cells which can be recovered from human adenoid tissues varies characteristically in the course of the year: The yield was found to be highest in May and lowest in July. The average histamine content was above average in April, June, July and September. Although the spontaneous histamine release remained relatively constant, the concanavalin A-stimulated histamine release was significantly reduced from June to September.
Time-Course of IgE Binding to Rat Peritoneal Cells after Sensitization with Alum-Adsorbed Ovalbumin and Bordetella pertussis
Authors: Behrendt Heidrun
Published: August 2009

High IgE responder rats were sensitized intraperitoneally with alum-adsorbed ovalbumin and Bordetella pertussis organisms. At day 0, 7, 14, 21 and 28 following sensitization the peritoneal cells were harvested and further processed for light and immunoelectron microscopical investigations using a specific anti-rat IgE immunogold sandwich method. The results obtained show that the number of peritoneal mast cells increase significantly during the course of sensitization. There is a time-dependent increase in the amount of immunogold particles on mast cell surfaces together with a shift of particle distribution towards the surface folds. Sensitized mast cells exhibit altered releasability as is indicated by slightly degranulated cells at day 21 and day 28 postsensitization. Additionally, about 25% of small lymphocytes which had entered the peritoneal cavity between day 7 and day 14 are heavily stained with the immunogold complex between day 14 and day 28 postsensitization. This is not so for macrophages (≤0.2% positive cells) nor for eosinophils which do not show any staining activity at all despite they are present in high numbers.
The Role of Histamine in Allergen and Adenosine-Induced Bronchoconstriction
Authors: Rafferty Paul , Beasley Richard , Southgate Peter , Holgate Stephen
Published: August 2009

We have investigated the role of histamine in allergen and adenosine-5′-monophosphate (AMP)-induced bronchoconstriction in asthmatic subjects by performing inhalation challenge tests with histamine, AMP and allergen after treatment with placebo or the potent H1 histamine receptor antagonist, terfenadine. Single concentrations of each agonist which had previously been shown to produce a 30% fall in FEV1 were used. After placebo, AMP and histamine both produced rapid bronchoconstriction reaching a maximum within 5 min and returning to within 10% of baseline after 25 min. Terfenadine inhibited this reaction to histamine completely and to AMP by 86%. The response to allergen was slower in onset and was sustained over 45 min and was inhibited 50% by terfenadine. We interpret these results as reflecting the contribution of histamine to the various airway challenges, both histamine and the newly generated mediators comprise the response to allergen, whereas AMP selectively enhances mast cell degranulation without affecting the production of arachidonic acid derived mediators.
Airway Hyper-Reactivity and Eosinophilia in Rats Treated with Sephadex Particles
Authors: Laycock S.M. , Smith H. , Spicer B.A.
Published: August 2009

The intravenous injection of Sephadex particles (G200) into rats produced a specific increase in numbers of blood eosinophils, peaking 7 days later. A second injection, given on day 14 when the numbers of blood eosinophils had fallen to control levels, produced a dose-dependent increase in numbers, greater than the first, and peaking 5 days later. At this time, there was a dose-dependent increase in numbers of eosinophils, but not of other leucocytes, in broncho-alveolar lavage fluids and lung tissue, together with an increase in sensitivity of the rats to the respiratory effects produced by the intravenous injection of 5-hydroxytryptamine.
Structure and Function of Intestinal Mucin: Developmental Aspects
Authors: Snyder John D. , Walker Allan
Published: August 2009

Mucin glycoproteins are thought to play an important role in protecting the intestine from chemical or physical injury but the mechanisms of protection and the possible relationship between mucin structure and function are incompletely understood. Structurally, purified intestinal mucins are a heterogeneous and polydisperse group of large-molecular-weight glycoproteins which have regional and developmental differences in composition. Newborn mucin contains more protein and less carbohydrate than adult mucin and differs from adult mucin in buoyant density and mobility on electrophoresis. The primary function of mucin, protection of the intestine, appears to be dependent upon at least four factors: (1) the rate and quantity of mucin release; (2) the physical barrier of the viscous mucus blanket; (3) the provision of specific inhibitory binding sites to infectious agents and proteins, and (4) the inclusion of secretory immunoglobulins to provide a link to the immunologic component of the intestinal host defense system. Immune as well as infectious and chemical agents appear to play an important role in mucus release but developmental effects have not been studied. Preliminary studies indicate that developmental differences may exist in the provision of inhibitory binding sites and the inclusion of secretory immunoglobulins in mucin. These studies indicate that mucus may provide a link between the physical and immunological components of the intestinal host defense system and point to the need for further studies in this area.
Regulation of the Delayed Hypersensitivity Reaction in the Lung Reflected as Mononuclear, Mast Cell and Mucus Cell Appearance after T Helper Cell Depletion and Adoptive Transfer
Authors: Enander I. , Ulfgren A.-K. , Nygren H. , Holmdahl R. , Klareskog L. , Larsson P. , Ahlstedt S.
Published: August 2009

Epicutaneous sensitization with picrylchloride (PiCl) induced a strongly delayed hypersensitivity (DH) reaction in mice. Local challenge in the airways of these mice resulted in increased numbers of mononuclear cells, mast cells and mucus cells. Depletion of T helper cells in vivo by treatment with monoclonal antibody (GK 1.5) inhibited the DH reaction. This treatment also resulted in a decrease in the number of mononuclear and mucus cells in the lung after intranasal challenge. The DH reaction was transferred to recipients with immune lymph node cells and spleen cells from mice sensitized epicutaneously with PiCl. The recipient mice also showed a slight increase in the number of mononuclear cells in the lung after intranasal challenge. These results indicate that T cells are not only involved in the DH reaction but also in the accompanying lung reaction.
Inflammatory Mediators, Tachykinins and Enkephalinase in Airways
Authors: Lazarus S.C. , Borson D.B. , Gold W.M. , Nadel J.A.
Published: August 2009

Immunologic and nonimmunologic stimuli evoke responses in the airways by complex mechanisms, often involving mast cells and mast cell-derived mediators. These mechanisms can be investigated using isolated mast cell lines for biochemical and physiologic studies. Neuropeptides as well as the enzymes that degrade neuropeptides are present in airway tissues. In the case of substance P, metabolism in the airway has demonstrated physiologic significance. The physiologic response to a stimulus depends upon the interaction between locally released inflammatory mediators and neuropeptides and the enzymes which degrade them.
Effect of Capsaicin on Bronchial Reactivity and Inflammation in Sensitized Adult Rats
Authors: Alving K. , Ulfgren A.-K. , Lundberg J.M. , Ahlstedt S.
Published: August 2009

The involvement of tachykinins in the airway reactivity of ovalbumin (OA)-sensitized rats was studied by capsaicin (CAPS) treatment. In subcutaneously sensitized animals, the reactivity to both OA aerosol and serotonin given intravenously was decreased when CAPS was given after the sensitization period. No effects on the serum IgE and IgG antibody levels were seen in these animals. In contrast, when CAPS was given before the sensitization period, no effects were seen on the OA aerosol and serotonin reactivities. Immunohistochemical examination revealed that the la antigen expression in the bronchial epithelium was increased by both subcutaneous and aerosol sensitization. The CAPS treatment decreased this la antigen expression. Histological examination of mononuclear cells, mast cells and goblet cells revealed only small effects on the cell numbers by both the OA sensitization and the CAPS treatment. The results demonstrate a link between tachykinins, serotonin, the immune system and clinical lung reactivity. The mechanisms for this seem to be complex, since the timing of the CAPS treatment and antigen sensitization is crucial for the outcome.
Chemiluminescence Response of Polymorphonuclear Leukocytes in Atopic Dermatitis
Authors: Schöpf Erwin , Kapp Alexander
Published: August 2009

Atopic dermatitis (AD) is characterized by many signs of immunodeficiency. Our interest was to investigate if there are also alterations of the chemiluminescence (CL) response of polymorphonuclear leukocytes (PMN) as a measure of the release of toxic oxygen radicals. Isolated PMN of 13 patients with AD with mild to moderate disease activity were stimulated with a chemotactic peptide (f-met-phe), zymosan-activated serum (ZAS), zymosan particles and phorbolmyristate acetate. In the AD group, we found a significantly decreased response after stimulation with ZAS in comparison to the controls. With the other stimuli tested no significant difference was detected. The decreased response of PMN to stimulation with ZAS from patients with AD associated with a normal reactivity to the other stimuli could be due to specific desensitization of the PMN by C5a in vivo.
Regulation of the Human Allergic Response
Authors: Geha Raif S. , Leung Donald Y.M.
Published: August 2009

Over the last several years our laboratory has been engaged in the in vitro study of the regulation of IgE synthesis. We have found that polyclonal B cell activators fail to induce IgE synthesis in human B cells. Alloreactive T cell helper clones induced synthesis in B cells only under conditions of interaction and in B cells from allergic donors under cognate conditions of bystander stimulation as well. Isotype-specific regulation of the IgE response was mediated in the secretion of IgE-binding factors by T cells suppressing Fc receptors for IgE. Finally, IgE immune complexes in sera from patients with hyper-IgE states were shown to downregulate T cell proliferation to antigen and to stimulate monocytes to resorb 45Ca-labelled bone and to release prostaglandins. The implications of these in vitro findings for disease states in which IgE is elevated are discussed.
Seasonal Variations in in vitro Synthesis of Rye Pollen-Specific IgE by Human Peripheral Mononuclear Cells
Authors: Turner K.J. , Siemensma N.P. , Krska K. , Cameron K.J.
Published: August 2009

Peripheral blood mononuclear B cells from 6 rye pollen-allergic patients, with no consistent perennial symptoms, were isolated before (July), during (October) and after (February and May) the pollen season. The T-depleted cells were fractionated on a discontinuous percoll density gradient and the B cell fractions, together with unfractionated B cells, incubated in vitro for quantitation of spontaneous synthesis of rye pollen-specific IgE. Markedly higher levels of IgE were synthesised by the fractions, as opposed to unfractionated B cells. The low-density fraction (B5) contributed most towards synthesis in the pollen season and the denser B6 cells in the pre- and post-pollen season. All low-density B cell fractions (B1-3–B5) and some high-density fractions contained large but variable amounts of preformed specific IgE which was retained, even in the absence of de novo synthesis in vitro, during the post- and pre-pollen season. Since only a fraction of this preformed IgE escaped into culture supernatants the contribution of preformed IgE to in vitro IgE synthesis in general may require reappraisal.
Human T Hybridoma-Derived Immunoglobulin-Binding Factors
Authors: Stadler Beda M. , Gauchat Dominique , Hildbrand Marie-Louise , Yang Xiadong , de Weck Alain L.
Published: August 2009

A human T hybridoma was generated lacking demonstrable low-affinity IgE surface receptors. This cell line produces spontaneously immunoglobulin-binding factors. The factors have an affinity for human IgE and IgG as well as for IgG from other species. The binding factors were purified by concanavalin A affinity chromatography, an IgE immunoaffinity column and SDS-PAGE. This purification procedure resulted in five major proteins bands at 13, 15, 30, 60 and <150 kilodaltons. As judged by Western blot analysis, all bands were variably glycosilated and were capable of binding IgE. Biological activity resided even within the small molecular weight (13 kilodaltons) peptide in terms of inhibiting IgE synthesis of the U266 B cell line and net IgE synthesis of B cells from atopic blood donors. Additionally, semipurified binding factor preparations also inhibited the Pokeweed mitogen-induced IgG synthesis while stimulating IgE synthesis at the same time. At present it is not clear whether the different molecular entities are dimers or polymers of the same protein, or are all derived from one larger protein (proteolytically cleaved), or whether they are completely different peptides mediating similar immunoglobulin-binding capacities as well as similar biological activities. Glycosylation of binding factors is not responsible for binding to immunoglobulins, but might be important for its biological activity.
Eosinophil Response in Mast Cell-Deficient W/Wv Mice
Authors: Nawa Y. , Owhashi M. , Imai J. , Abe T.
Published: August 2009

The combination of cyclophosphamide treatment and Toxocara canis infection is known as an effective way of causing a high level of eosinophilia in mice. When this treatment was applied to congenitally anemic, mast cell-deficient W/Wv mice, eosinophil response was far less than that of their normal littermate +/+ mice. The degree of the defective eosinophil response in the peripheral blood of W/Wv mice was severer than that in the bone marrow. The defective eosinophil response of W/Wv mice was completely restored by bone marrow grafting 8 weeks prior to cyclophosphamide treatment and T. canis infection. The kinetics of the recovery of eosinophil response in the bone marrow of W/Wv mice after bone marrow grafting was faster than that in the peripheral blood. Chemotactic reactivity of eosinophils obtained from bone marrow or peritoneal cavity of W/Wv mice was essentially comparable to that of +/+ mice. These results suggest that, in addition to the production of eosinophils in the bone marrow, mast cell-derived factors play an important role in the mediation of peripheral blood eosinophilia.
Isolation of Interleukin-1 from Human Milk
Authors: Söder O.
Published: August 2009

Human milk was investigated in regard to the presence of lymphocyte growth factors. Precipitated and dialyzed protein from mature human milk was found to exhibit interleukin-1 (IL-1) activity in the murine thymocyte proliferation assay. A mitogenic effect of milk protein was also observed in cultures of human and guinea pig thymocytes, demonstrating that the effect was not species restricted. No interleukin-2 or thymocyte growth peptide activity was found. Gel filtration chromatography on Sephadex G 150 revealed three distinct peaks of IL-1 activity, with apparent molecular weights of 14, 31 and 60 kilodaltons, respectively. Chromatofocusing showed IL-1 activity in four peaks, the major peak eluting at an isoelectric point of 5.0. Similar molecular heterogeneity has previously been demonstrated for IL-1 isolated from macrophages and other sources. The physiological role of milk-derived IL-1 is unknown, but it is proposed that it might act as an immunostimulating factor in the mammary gland, and hypothetically also in the breast-fed infant. Since IL-1 is a potent pyrogen, it is also suggested that milk-derived IL-1 contributes to the well-known pyrogenic activity of milk.
Thermostable Ethanol-Insoluble Antigens of Human Heart Muscle
Authors: Kaise S. , Abeyounis C.J. , Kaplan M.H. , Milgrom F.
Published: August 2009

Organ-specific reactions were observed between thermostable ethanol-insoluble (BE) fraction of human heart muscle and its corresponding rabbit antiserum by means of tanned cell hemagglutination test, enzyme immunoassay, and gel precipitation test. Two heart-specific components could be identified using gel precipitation and immunoblotting procedures. Antibodies to human heart muscle BE combined also with heart BE preparations from mammals other than man; these interspecies cross-reactions included also rabbit heart BE pointing to the ‘auto’ nature of some of the antibodies under investigation. The heart antigens under study were distinct from the organ-specific antigen demonstrated by Espinosa and Kaplan in acid extract of human heart muscle.
Effects of Some Prostaglandins and Leukotrienes on Lymphocytes, Monocytes and Their Activity in vitro
Authors: Wasserman J. , Hammarström S. , Petrini B. , Blomgren H. , von Stedingk L.-V. , Vedin I.
Published: August 2009

The effect of some prostaglandins (PGs) and leukotrienes (LTs) on lymphocytes and monocytes was tested in vitro. Overnight incubation with PGD2 reduces the expression of Leu-2 antigen (suppressor/cytotoxic phenotype) and of Fc receptors for IgG. PGA2, PGD2 and to a lesser extent PGE2 inhibit PHA reactivity of lymphocytes. LTB4 does not have any inhibitory activity. Preincubation of lymphocytes with PGD2 and PGE2 decreases their NK activity. LTB4, but not the PGs tested, stimulates monocyte metabolism as assessed by chemiluminescence.
Immunogenicity and Tolerogenicity of a Major House Dust Mite Allergen, Der p l from Dermatophagoides pteronyssinus, in Mice and Rats
Authors: Stewart G.A. , Holt P.G.
Published: August 2009

Primary immunization of different inbred strains of mice with the house dust mite allergen Der p I (from Dermatophagoides pteronyssinus) adsorbed to adjuvant revealed differences in IgE but not IgG responses. Thus, the CBA and C57 Black strains were shown to be high IgE responders in that persistent IgE was induced which lasted for several months. In contrast, the C3H, AKR and Balb/c strains were judged to be poor responders. Subsequent experiments showed that such responses were adjuvant-independent, refractory to irradiation (850 rad) and that they could be adoptively transferred using immune spleen cells in a dose-dependent manner. At sub-optimal doses, adoptively transferred IgE responses were initiated by either intraperitoneal or intranasal challenge of recipients with allergen in the absence of adjuvant. Allergen-specific IgE in rats was also shown to be strain-dependent, with Brown Norway rats, in contrast to Wistar Furth (WF) and Lou/M rats, being the highest responder strain studied. Repeated intranasal administration of soluble allergen induced IgE tolerance in rats but not mice. However, tolerance was restricted to the low IgE responder phenotype strain, WF.
Purification, Identification and Elimination of a Natural Human Serum Antibody with Cytotoxic Effect on Guinea Pig Thymocytes
Authors: Sandberg G. , Rydlander L. , Stenvinkel C.
Published: August 2009

Normal human sera contain one or several factors cytotoxic for normal guinea pig thymocytes, and when serum is precipitated with ammonium sulphate (60% saturated) and the precipitate dissolved and dialyzed, the activity is preserved. Gel chromatography with Sephadex G-150 and Sepharose CL-6B indicated a molecular weight of approximately 900,000 daltons. The active fractions contained a high amount of IgM according to single radial immunodiffusion and two-dimensional gel electrophoresis. Qantitation of the IgM band in one-dimensional gel electrophoresis preparations by gel scanning indicated that IgM accounted for 65% of the eluted proteins in active fractions. Purified human IgM from myeloma patients eluted as the active factor during gel chromatography. Elimination of IgM from serum by affinity chromatography eliminated the cytotoxic activity. The serum could also be inactivated by heating. The mixing of IgM-depleted serum with either polyclonal IgM or heat-inactivated serum restored the activity. Thus, the cytotoxic activity is due to IgM antibodies plus a heat-labile component (presumably complement). The presence of the cytotoxic activity in autologous (guinea pig) serum was recently demonstrated. The possible functional role of these antibodies in the elimination process of a large number of cortical thymocytes is suggested.
A Sandwich Enzyme Immunoassay for the Detection of Human IgG Antibodies to Dog Allergens
Authors: Viander M. , Nieminen E. , Valovirta E. , Vanto T. , Ingeman L. , Koivikko A.
Published: August 2009

A double-sandwich enzyme immunoassay (DSEIA) was developed for the detection of human IgG antibodies to dog dander and hair (DH) allergens. Since DH allergens are immunogenic in rabbits, the gamma-globulin fraction of rabbit antiserum to DH (RGG-a-DH; 10 μg/ml) was used for coating of polystyrene microtiter plates. Dog allergens (1 μg/ml of DH extract) were bound to RGG-a-DH. Binding of human IgG Ab to nonimmune RGG (10 μg/ml) – used as a background control – was substracted from the DH specific one and nonspecific binding was further eliminated by using 0.5% of normal rabbit serum in the dilution buffer. Sera were diluted 1:10 in this buffer. Specificity of the assay was shown by absorption experiments: protein A, anti-IgE discs (Phadebas® PRIST) and dog or birch (e5, t3; Phadebas® RAST) allergen discs were used to remove total and allergen specific IgG and IgE, respectively. The results were expressed as net absorbances or as a percentage of a reference serum. A significant correlation (r = 0.65, p <0.01) between IgG (DSEIA) and IgE (RAST) antibodies to dog allergens was observed in 40 untreated dog allergic subjects. The DSEIA was found to be more sensitive than conventional ELISA in detecting IgG Ab in 15 asthmatic children during hyposensitization: a sinificant rise was observed in 14 compared to 12 with ELISA, while no significant increases were observed in the placebo-treated group.