bluuebunny/crossref_metadata_2025 huggingface dataset:
Reverse Enzyme Immunoassay for the Determination of Dermatophagoides pteronyssinus IgE Antibodies
Authors: Moneo Ignacio , Cuevas Manuela , Ureña Victoria , Alcover Roberto , Bootello Alfredo
Published: August 2009

Sera from patients with suspected mite sensitivity were assayed using both the radioallergosorbent test (RAST) and a reverse enzyme immunoassay for IgE antibodies (REIA). Of the 133 sera studied, the RAST gave positive results in 48 cases and the REIA in 59. Negative results for both assays were obtained in 72 of the sera. The immunoenzymatic assay described here, using micro-plates as solid phase, has proven to be IgE- and antigen-specific. The allergen conjugate has a prolonged shelf life and the method is easy to perform. The sensitivity of the REIA for Dermatophagoides pteronyssinus IgE antibodies is limited by the small amount of IgE bound by the well surface (less than 100 ng). However, we think that the method offers a reliable alternative for the diagnosis of D. pteronyssinus-allergic patients.
Trapidil, a Coronary Vasodilator, Inhibits IgE-Mediated Histamine Release from Human Basophils
Authors: Morita Yutaka , lnokuma Shigeko , Miyamoto Terumasa
Published: August 2009

The effect of trapidil, a coronary vasodilator, on histamine release from human basophils was studied. Trapidil inhibited IgE-mediated, anti-IgE-induced histamine release in a dose-dependent fashion (ID50:5.8 × 10––4M). In contrast, histamine release induced by calcium ionophore A23187, formyl-L-methionyl-L-leucyl-L-phenylalanine and polymyxin B was not affected by this agent. Two-stage experiments suggested that the effect of trapidil was exerted not only on the first Ca2+-independent activation stage, but also on the second Ca2+-dependent stage. These results, together with recent reports that trapidil has a bronchodilator action, suggest that trapidil could be used in the treatment of asthmatic patients.
Studies on Thymocyte Subpopulations in Guinea Pigs
Authors: Sandberg G.
Published: August 2009

Rosette-forming cells were counted in three thymocyte subpopulations separated by density gradient centrifugation with Percoll. A method for the electronic counting of rosettes is described which resulted in excellent agreement with results of visual counting. By both methods, significant differences in the rosette-forming ability of the studied subpopulations were detected. Thus, in population la, earlier shown to consist of large cells of low density and with high spontaneous proliferative activity, the rosette frequency was 54% as compared with 70% in the whole thymus population and 83% in population II, known to consist of small, immunologically incompetent cells. The third population studied (lb), consisting of intermediate density cells, had 74% rosette-forming cells.
Polymerization of Individual Species of Tree Pollen Allergens
Authors: Patterson Roy , Suszko Irena M. , Grammer Leslie C.
Published: August 2009

This report describes the preparation of individual tree pollen allergen polymers in contrast to our previous description of the polymerization of a mixture of tree pollens. These individual polymers may be used for immunotherapy as indicated by the IgE-mediated reactivity of each patient. A modified chromatographic fractionation process was used which results in tree polymers with skin reactivity 1,000- to 10,000-fold less from monomers and with a defined lower molecular weight limit. The modified preparative methodology results in polymer preparations as safe but with greater ease of production than polymers prepared with previous methodologies.
Effects of Anti-Idiotypic Immunizations on Autoimmune Uveitis Induced with Isologous Lens Proteins
Authors: Sunakawa M. , Seto A. , Ito Y. , Tsukahara I.
Published: August 2009

Autoimmune uveitis was induced in F1 hybrid rabbits by administering intramuscularly lens proteins in Freund’s complete adjuvant. Cellular infiltration in the anterior chamber of the eye appeared in a cyclic manner and anti-lens protein antibody in the serum was mostly of oligoclonal origin, as assessed by isoelectric focusing analysis. Immunizations of such rabbits with autologous or isologous anti-lens protein antibody affected little the appearance of cellular infiltration upon rechallenge with lens proteins, but anti-lens protein antibody was replaced by other antibodies with different isoelectric points. Serum from these rabbits contained antibody which inhibited the antibody activity of certain anti-lens protein antibody.
Cholera Toxin-Binding T Cells in the Human Peripheral Blood at Different Ages
Authors: Tsuru Sumiaki
Published: August 2009

The capacity of human T cells to bind cholera toxin was shown to decrease with age. In aged humans, the number of cells capable of binding high concentrations of cholera toxin was lower than that in young humans. The method presented in this paper may be useful as one of the indicators of aging of the immune system.
Phagocytosis, in vivo, of Immune Complexes by Dendritic Cells in the Lymph of Sheep
Authors: Hall J.G. , Robertson D.
Published: August 2009

Although the occurrence of dendritic, macrophage-like cells in the peripheral lymph of sheep, pigs, rabbits, and man is well documented, the exact nature of these cells has become controversial. Generally, such cells tend not to adhere firmly to glass or exhibit phagocytosis in in vitro systems. However, when immunized sheep were given a subcutaneous injection of specific antigen (horseradish peroxidase), most of the macrophage-like cells in the lymph from the injection site could be shown by transmission electron microscopy to have engulfed the immune complexes that had been formed in the subcuticulum.
Antigenic Variability of Different Strains of Aspergillus fumigatus
Authors: Wallenbeck Ingrid , Aukrust Lars , Einarsson Roland
Published: August 2009

10 strains (isolates) of Aspergillus fumigatus, grown and harvested under identical conditions, were compared with respect to biochemical and immunological properties. The total protein and carbohydrate content and isoelectric focusing pattern were determined for each of the individual strains. Their antigenic and allergenic activity was examined by a combination of crossed immunoelectrophoresis (CIE)/crossed radioimmunoelectrophoresis (CRIE) and RAST inhibition. Biochemical and immunochemical data indicated that considerable variation exists between the various isolates of A. fumigatus. 44 antigens were demonstrated in CIE of A. fumigatus, and 18 different allergens were identified in CRIE (2 major, 10 intermediate and 6 minor allergens).
Bone Marrow Origin of Mucosal Mast Cells
Authors: Crowle Patricia K. , Reed Norman D.
Published: August 2009

Infection with the intestinal parasite Nippostrongylus brasiliensis stimulates an accumulation of mucosal mast cells (MMC) in the villi of the small intestine of normal but not athymic or W/Wv anemic mice. W/Wv mice are congenitally deficient in both MMC and skin and connective tissue mast cells (CTMC). Athymic mice have normal or elevated numbers of CTMC but are severely deficient in MMC. CTMC derive from the bone marrow. To determine the origin of MMC, athymic and W/Wv mice were given various hematopoietic or lymphoid tissues from normal littermate or beige mice and the MMC response to N. brasiliensis infection was evaluated. The MMC defect in athymic mice was repaired by grafts of thymus cells, thymus gland, or spleen cells, but not by bone marrow cells or anti-Thy 1-treated bone marrow or spleen cells. The MMC and CTMC defects of W/Wv mice were repaired by grafts of bone marrow, spleen cells, or anti-Thy 1-treated bone marrow or spleen cells. Neither the MMC nor the CTMC defect in W/Wv mice was repaired by grafts of thymus cells or thymus glands. These results indicate the following. MMC, like CTMC, derive from the bone marrow and not from the thymus. MMC require a thymic influence for development. Athymic mice possess bone marrow precursors for both MMC and CTMC but lack a thymus-dependent component necessary for MMC development. W/Wv mice lack both MMC and CTMC mast cell precursors but possess the thymus-dependent component required for MMC development.
Allergens of Schistosoma mansoni
Authors: Damonneville Martine , Pierce Raymond J. , Verwaerde Claudie , Capron André
Published: August 2009

The interaction of Schistosoma mansoni crude soluble egg antigen (SEA) with IgE antibodies in sera from S. mansoni-infected mice, rats and humans has been studied by the radioallergosorbent test (RAST) and the Prausnitz-KÜstner (PK) technique. IgE antibodies recognizing egg antigens were present as early as day 21 after the infection in the mouse sera and day 28 in rat sera. IgE in sera of infected humans reacted with antigenic components in the Mr range 70,000–150,000 and focusing as a broak peak in the pH range 4.5–6.5 as measured by RAST. SDS-PAGE followed by western blotting showed the presence of major components at molecular weights of 117,000 and 35,000–43,000. In the PK test, using mouse sera, components focusing in the alkaline pH range also gave a positive reaction. Most of the allergenic activity was bound by concanavalin A-Sepharose and by wheat germ agglutinin-Ultrogel. IgE in serum from an infected non permissive host (the Fischer rat) apparently recognized egg-stage-specific allergen as indicated by differences in the time course of the IgE response to egg allergens compared to the adult material. When analyzed by SDS-PAGE and western blotting with day 45-infected rat serum, SEA showed some qualitative and quantitative differences to adult worm antigen. Molecules at molecular weights between 25,000 and 30,000 and at about 43,000 in SEA reacted with rat serum IgE and were absent from adult worm antigen. The allergenic similarities between egg and adult worm are discussed.
Effect of 15-Hydroxyeicosatetraenoic Acid (15-HETE) on Anti-Immunglobulin E- and Calcium Ionophore-Induced Histamine Release from Human Leukocytes
Authors: Dorsch Walter , Ring Johannes , Riepel Herbert
Published: August 2009

15-Hydroxyeicosatetraenoic acid (15-HETE) was prepared by soybean lipoxygenase-mediated oxygenation of arachidonic acid to 15-hydroperoxyeicosatetraenoic acid (15-HPETE) and subsequent reduction by NaBH4.15-HETE was identified, purified and proved as biologically active by thin layer chromatography, high pressure liquid chromatography, gas chromatography/mass spectrometry and biological experiments on horse thrombocytes and rabbit peritoneal leukocytes. 15-HETE (0.1–40 μM) was added to peripheral leukocytes of 48 human donors (33 atopies, 15 nonatopics) which were challenged with rabbit anti-human-IgE or calcium ionophore A 23187. Its effect was compared with the effects of nordihydroguaretic acid (NDGA) and eicosatetraynoic acid (ETYA). NDGA and ETYA markedly inhibited histamine release (17 μM: 74 ± 11 and 39 ± 14,3%, respectively), whereas 15-HETE neither stimulated nor inhibited spontaneous anti-IgE- or calcium ionophore A 23187-induced histamine release.
Serum Theophylline during Acute Asthmatic Attacks
Authors: Tønnesen Jan
Published: August 2009

To investigate serum theophylline levels during acute attacks of asthma, venous blood samples were taken from 41 consecutive patients. 36 had asthma and 5 suffered from chronic bronchitis. The patients were taking five different theophylline preparations. Blood samples were taken when the patients were admitted to the emergency unit because of serious deterioration in their pulmonary condition. Serum theophylline was analyzed by liquid column chromatography. 26 patients revealed subtherapeutic levels, 5 patients had levels above therapeutic range, and 10 patients had values within the therapeutic range. Thus, many patients taking oral theophylline derivatives are undertreated and a few are overmedicated.
Immune Inflammation in Newly Developing Facsimile Synovia
Authors: Sin Y.M. , Sedgwick A.D. , Willoughby D.A.
Published: August 2009

The injection of air into the subcutaneous tissue of the back in rodents has previously been shown to induce a lining which closely resembles synovial lining tissue. Air pouches of different ages represent the developmental stages of such tissue. We have found that pouches of 1 day in age induced in animals previously sensitised with bovine serum albumin (BSA) respond with low numbers of infiltrating leucocytes and a small fluid exudate volume 4 h after challenge into the pouch with BSA. In contrast, 3- and 6-day-old pouch tissue when challenged with antigen responds with a greater influx of leucocytes and a larger fluid exudate. These findings suggest that the constituent make-up of the air pouch-lining tissue is important for the full expression of the inflammatory response.
Staphylococcus aureus Protein A Antibody-Mediated Haemagglutination Assay
Authors: Jagannath C. , Sengupta D.N. , Kasinathan S.
Published: August 2009

A method has been described for the enhancement of the sensitivity of the passive haemagglutination (PHA) reaction through intact cells of Staphylococcus aureus (Cowan I) bearing protein A (SAPA) which bind Fc portion of IgG derived from many mammalian species. Interactions between Mycobacterium tuberculosis sonicate antigen-sensitized red cells and antimycobacterial antibodies derived from M. tuberculosis immunized goat, rabbit, guinea pig and mouse sera and sera from human tuberculosis patients have been used as model systems. SAPA cells were allowed to bind antibodies from sera and subsequently cross-react with antigen-sensitized red cells. Co-haemagglutination occurred only when specific antibodies were bound by SAPA cells. The sensitivity of this SAPA antibody-mediated haemagglutination assay (SAPA-AMHA) was found to be higher than that of PHA reaction and it depended upon protein A antibody affinity. The specificity of the interaction between SAPA cell-bound antibodies and red-cell-bound antigens suggests that these two reagents offer a simple and sensitive approach for the study of antigen-antibody reactions.
Essential Role of the hprK Gene in Ralstonia eutropha H16
Authors: Krauβe Désirée , Hunold Katja , Kusian Bernhard , Lenz Oliver , Stülke Jörg , Bowien Botho , Deutscher Josef
Published: August 2009

Ralstonia eutropha H16 possesses an incomplete phosphoenolpyruvate (PEP):sugar phosphotransferase system (PTS) composed of EI, HPr, EIIANtr (PtsN) and EIIAMan (PtsM). We could show that in vitro the incomplete PTS phosphorylation cascade is partially functional. HPr becomes phosphorylated by PEP and EI, and transfers the phosphoryl group to EIIANtr, but only extremely slowly to EIIAMan. Components of this system have previously been shown to regulate the metabolism of polyhydroxybutyrate. Downstream from ptsN this organism contains an hprK gene, which codes for a homologue of HPr kinase/phosphorylase. We show that this enzyme phosphorylates HPr using ATP as phosphoryl donor. Interestingly, hprK appeared to be essential in R. eutropha because this gene could not be deleted in the wild-type strain, but could be deleted in mutants lacking ptsH or ptsI. This suggests that an increase in the HPr and/or P∼His-HPr concentrations might be responsible for the growth defect. To test this hypothesis, various ptsH alleles were introduced into the ptsH hprK double mutant. Complementation of this mutant was possible only with the ptsH(His15Ala) allele, but not with the wild-type or ptsH(Ser46Ala) alleles. We conclude that elevated amounts of His-15-phosphorylated HPr, formed in the hprK mutant, are responsible for its growth defect.
Use of a Simple in vivo Technique to Investigate Components of the Immediate Hypersensitivity Reaction in Conscious Guinea Pigs
Authors: Hammerbeck D.M. , Swingle K.F.
Published: August 2009

A semiplethysmographic method was used to delineate the effects of aerosol antigen challenge on the respiratory pattern of conscious, sensitized guinea pigs. Chlorpheniramine pretreatment prevented death, delayed the onset of respiratory symptoms and decreased the severity of the hypersensitivity reaction. This reaction could be further diminished to varying degrees by the addition of FPL55712, atropine sulfate and/or imidazole to chlorpheniramine. Indomethacin and nordihydroguaiaretic acid (NDGA) with chlorpheniramine caused a potentiation in the severity of the hypersensitivity reaction. However, chlorpheniramine, NDGA and imidazole in combination blocked the hypersensitivity reaction. The technique appears to demonstrate the involvement of several suggested mediators in asthma and appears potentially useful as an indicator of effects on arachidonic acid metabolism and metabolites.
The Microvasculature of the Hamster Cheek Pouch as a Model for Studying Acute Immune-Complex-Induced Inflammatory Reactions
Authors: Björk Jakob , Smedegård Göran
Published: August 2009

The acute microvascular reactions induced by immune complexes were studied. Hamster cheek pouch preparations of ovalbumin (OA)-immunized animals were topically exposed to different doses of OA (0.1, 1, 10 or 100 μg/ml). Fluorescein-isothiocyanate-labelled dextran (FITC-dextran, molecular weight 150,000) was injected intravenously as a macromolecular tracer. The microvascular alterations observed upon exposure to OA ranged, in a dose dependent way, from a mild vasoconstriction and reversible vascular leakage from postcapillary venules exhibited by 0.1 μg/ml OA, to a severe long-lasting vasoconstriction, platelet aggregation, stasis, leukocyte accumulation in venules and prolonged vascular leakage from venules seen after exposure to 100 μg/ml OA. Exposure of immunized animals to 100 μg/ml fluorescein-labelled OA (FITC-OA) resulted in deposition of FITC-OA at the walls of small venules, indicating immune complex formation at these sites. A more diffuse pattern of FITC-OA localization at venules was seen with 10 μg/ml FITC-OA, which may possibly indicate formation of smaller more soluble immune complexes with lower doses of OA. The intravital model used in this study forms a basis for studying immunologically induced acute inflammatory reactions at the microvascular level.
Identification of Novel Proteins Secreted by Lactobacillus plantarum That Bind to Mucin and Fibronectin
Authors: Sánchez Borja , Schmitter Jean-Marie , Urdaci María C.
Published: August 2009

Lactobacillus plantarum is a lactic acid bacterium that can be isolated from a high variety of fermented foods, including dairy products. In the present work, eight novel proteins secreted by three L. plantarum strains have been identified by tandem mass spectrometry. Seven of them were predicted as extracellular proteins containing putative signal peptides. The sixth, identified as glyceraldehyde 3-phosphate dehydrogenase (GAPDH), is a cytoplasmic protein that has been detected on the surface of several microorganisms. Muramidase and GAPDH were secreted only by the L. plantarum BMCM12 strain. Two other bands present in this strain were not identified, in spite of their yielding good tryptic profiles, suggesting an absence of homolog sequences in the molecular databases. Four of these proteins, including GAPDH, bound to mucin and fibronectin. These proteins might play important roles in the physiology and ecology of this bacterium, notably in the interaction with the human host.
Occupational Allergy in Horticulture: Demonstration of Immediate-Type Allergic Reactivity to Freesia and Paprika Plants
Authors: van Toorenenbergen A.W. , Dieges P.H.
Published: August 2009

Patients A and M developed allergic symptoms when working in a greenhouse with paprika plants and freesia plants, respectively.The possible involvement of an IgE-mediated mechanism was investigated with the skin prick test, radioallergosorbent test (RAST) and the histamine release test (HRT). Paprika flower, leaf and stem extract released 58, 47 and 43% of the total amount of histamine from washed leukocytes of patient A. In serum A IgE antibodies against paprika leaves and flowers could be demonstrated by RAST (11% binding of 125I-anti-IgE added). Freesia flower and stem extract released 46 and 43% histamine, respectively, from washed leukocytes of patient M. In the RAST, specific IgE antibodies against freesia flowers and stems were found in serum M (37% binding of 125I-anti-IgE added).
Reduced Adenylate Cyclase Responsiveness to Histamine in Lymphocyte Membranes of Allergic Asthmatic Patients after Allergen Challenge
Authors: Meurs Herman , Koëter Gerard H. , Kauffman Henk F. , Timmermans Adiet , Folkers Betty , de Vries Klaas
Published: August 2009

The adenylate cyclase response to histamine was measured in lymphocyte membranes of allergic asthmatic patients and healthy control subjects, just before and 24 h after inhalation challenge with house dust mite allergen. In the non-acute phase before allergen challenge, the histamine response in the membranes of the patients was not significantly different from that in membranes of the control subjects. After the house dust mite-induced asthmatic reaction, however, the adenylate cyclase response to histamine in the membranes of the patients was significantly reduced by 47%, whereas allergen inhalation had no effect on the histamine response of the control membranes. The results suggest that allergic asthmatic reactions may cause reduced lymphocyte adenylate cyclase responsiveness to histamine. A similar mechanism in the airways might contribute to allergen-induced enhanced airway reactivity to histamine.
IgE-Binding Factors
Authors: Ishizaka Kimishige , Huff Thomas F. , Jardieu Paula , Moore Kevin W. , Martens Christine L.
Published: August 2009

Gene cloning of rodent IgE-binding factors was accomplished, using messenger RN A of a rat T cell hybridoma, 23B6, which produce IgE-suppressive factor. Transfection of cos 7 monkey kidney cells with a eDNA clone resulted in the formation of rodent IgE-potentiating factors. The results provided a definitive evidence that IgE-binding factors represent a single peptide chain, and that the IgE-potentiating factor and IgE-suppressive factor share a common structural gene. Nucleotide sequence of a cDN A provided predicted amino acid sequence of the 60K precursor molecules of IgE-potentiating factor. Human IgE-potentiating factors were obtaiend from a human T-T hybridoma. The factors have affinity not only for human IgE but also for rat IgE and selectively enhanced antigen-induced IgE response of rat lymphocytes. The same hybridoma could be switched to form IgE-suppressive factor by the addition of glycosylation-inhibiting factor (GIF) during its biosynthesis. Purified GIF has immunosuppressive activity in the mouse. This factor suppressed the primary IgE and IgG antibody responses in the mouse, and markedly suppressed ongoing IgE antibody formation.
Characterization of an Antigen from Rat Male Accessory Glands
Authors: Pesoa Susana A. , Galmarini Mirian , Pistoresi-Palencia Maria C. , Riera Clelia M.
Published: August 2009

An isolated soluble antigen involved in the autoimmune response against rat male accessory glands migrated as a double band when it was submitted to analytical disc polyacrylamide gel electrophoresis and as a homogeneous molecular species in SDS electrophoresis. The purified antigen had a molecular weight of approximately 78 K by SDS electrophoresis and 80 K by gel filtration chromatography. The antigen was identified as a protein and displays androgen dependency. The purified fraction was pathogenically active in microgram doses and induced humoral and delayed-type hypersensitivity responses. Besides, lesions were observed in the target organ. The cellular infiltration was located in prostate glands according to the localization of antigen by immunofluorescence findings.
Kinetic Study of Kurloff Cells in Guinea Pig Thymus
Authors: Sandberg G. , Hagelin Margareta
Published: August 2009

One single injection of estradiol to male guinea pigs resulted in the appearance of Kurloff cells in the thymus and spleen in a maximal number after 2–3 weeks. In both organs, Kurloff cells of three categories were observed with different size and number of the inclusion(s). In the thymus, cells with small and medium-sized inclusions were present almost exclusively among low-density thymocytes. Cells with one large inclusion were initially present among low-density cells, but with time an increasing proportion were found among high-density thymocytes. This indicates that the different categories of Kurloff cells represent maturational stages and follow the normal differentiation step of thymocytes from low to high density. According to electronic cell volume distribution analysis, estradiol treatment was associated with a shift in cell volume towards larger cells, and this shift was correlated in time with the appearance of Kurloff cells with large inclusions. In thymus sections there was evidence for a massive and selective migration of Kurloff cells via interlobular lymph vessels. No difference in localization of Kurloff cells was noted at various times after estradiol treatment. Attempts to induce Kurloff cell formation from bone marrow, spleen or thymus cells during 2 weeks in vitro were unsuccessful, also when culturing the cells in serum from estradiol-treated animals. This negative result, the long lag phase of estradiol-induced Kurloff cell formation in vivo, and the reported lack of estradiol receptor in Kurloff cells indicate that the Kurloff cell induction by estradiol may be indirect.
Prevalence of Specific IgE Antibodies against Piperazine in Employees of a Chemical Plant
Authors: Hagmar Lars , Welinder Hans
Published: August 2009

In 5 out of 72 (7%) workers exposed to the asthma-inducing amine piperazine, specific IgE antibodies against a conjugate between human serum albumin and piperazine were demonstrated with RAST and RAST inhibition techniques. No specific antibodies were found in 64 nonexposed workers at the same plant, and in 60 healthy reference subjects. Eight out of the 72 (11%) exposed employees had unequivocal histories of piperazine-associated asthma, but only 4 of them had specific IgE antibodies against piperazine. However, there was a statistically significant association between specific antibodies and disease, as 4 out of 5 RAST-positive subjects were asthmatics, as compared with 4 out of the 67 RAST-negatives (p = 0.0003). The 4 RAST-positive asthmatics had been exposed to piperazine for between 6 and 168 months before their asthma started, as compared with induction times of less than a month for the 4 RAST-negatives. This discrepancy may reflect different pathogeneses for piperazine-associated asthma; either type-I allergy, pseudo-allergic reactivity or nonspecific irritative effect on the bronchi. The RAST-positive subjects did not differ from other employees as regards smoking habits but had significantly elevated levels of total serum IgE (p = 0.004).
Human Splenic Fc Receptor Function and Dysfunction: Definition by Circulatory Clearance Rate Compared to Computerized Kinetic Scintigraphy
Authors: Lin Robert Y. , Racis Stan P. , Higginbotham-Ford Edith , Tanz Warren S.
Published: August 2009

In vivo reticuloendothelial Fc receptor function was studied in 14 individuals. To assess this function Tc-99m-labeled IgG-coated autologous erythrocytes were injected intravenously and circulatory clearance rates of radioactivity were determined. In 12 individuals [6 normals and 6 patients with systemic lupus erythematosus (SLE)], kinetic computerized scintigraphic imaging of various internal organs was compared to circulatory clearance rates. Both circulatory clearance rates and percentage splenic uptake (PSU) were significantly depressed in patients. The difference between patients and normals was more significant for PSU than for circulatory clearance. Splenic uptake curves showed significantly less linearity for patients, suggesting that splenic defects in SLE are not only quantitative, but also are qualitative. Understanding the nature of immune clearance defects in immune complex mediated diseases may allow for a more rational approach to treatment of patients with these disorders.
Plasminogen Activator Production from Murine Mastocytoma Cells. Dissociation of Plasminogen Activator and Histamine Production by Sodium Butyrate
Authors: Katayama Ichiro , Hashimoto Koji , Nishioka Kiyoshi
Published: August 2009

Plasminogen activator (PA) production from mouse mastocytoma P815 was investigated. A trace level of PA was detected in the culture supernatant of P815, but addition of Con A (5 μg/ml) stimulated the PA production to the 20- to 50-fold of that of the unstimulated culture. Time course studies suggest that PA is actively synthesized and secreted during the culture period with Con A. Furthermore, pretreatment of P815 with sodium butyrate, which is known to differentiate interleukin-3-dependent bone marrow mast cells or P815 cells, increased the PA production from P815 cells in the presence or absence of Con A.
Homologous Passive Cutaneous Anaphylaxis in Various Strains of Mice
Authors: Inagaki Naoki , Goto Shoichi , Nagai Hiroichi , Koda Akihide
Published: August 2009

Passive cutaneous anaphylaxis (PCA) was elicited both in the ear and in the dorsal skin of 13 strains of mice at the same time and assessed quantitatively by measuring the amount of extravasated dye. Body pigments of colored mice such as DBA/2 (chocolate), C3H/He (brown) and C57BL/6 (black) did not interfere with the measurement of dye. In the ear response, ICR was a higher responder, C57BL/6 and BALB/c-nu/nu were lower responder strains. In the dorsal skin response, however, ICR was a lower responder, BALB/c-nu/nu, Hairless and WBB6 F1- +/+ were higher responders. WBB6 F1-W/WV was a nonresponder in both responses. The ear response was highly reproducible and the dorsal skin response of each strain was 1/2–1/10 of its ear response except for BALB/c-nu/nu. The PCA bluing regions on the dorsal skin of BALB/c-nu/nu were clearly delineated and the response was almost comparable to its ear response.
Normal Chemotactic Migration of Polymorphonuclear Leukocytes Stimulated with Mononuclear-Derived Chemotactic Factor in Ulcerative Colitis
Authors: Hermanowicz Andrzej , Nawarska Zofia
Published: August 2009

The migration of polymorphonuclear leukocytes preincubated with autologous or heterologous serum was examined in 100 patients with untreated ulcerative colitis (UC) and in 100 age- and sex-matched healthy controls. The activity of complement-derived chemotactic factors and mononuclear-derived chemotactic factor (MDCF) was also investigated in the same group of patients. No significant difference was found in random and chemotactic migration of patients or control leukocytes preincubated in different concentrations (10, 50 and 100%) of autologous or heterologous serum. Defective chemotaxis of leukocytes stimulated with complement-derived chemotactic factors was found in UC and was more marked in patients in remission than with active UC independently of whether complement was activated by the alternative or the classical pathway. However, the random migration of neutrophils was enhanced in both groups of UC patients. The leukocytes of patients stimulated with MDCF (mononuclear cells were activated with lipopolysaccharide from Escherichia coli O55 :B5) show normal chemotaxis. Our data suggest an impairment of neutrophil receptors for complement-derived chemotactic factor in UC. The decreased response of neutrophils to these factors and normal response to MDCF suggest that the main way in which cells are attracted to the site of inflammation in UC may be a factor produced by stimulated mononuclear cells.
Effect of Drugs on the Increase in Cell Numbers in the Peritoneal Cavity of the Actively Sensitised Mouse after Intraperitoneal Challenge with Antigen
Authors: Spicer B.A. , Hatt P.A. , Laycock S.M. , Smith H.
Published: August 2009

The injection of antigen into the peritoneal cavity of actively sensitised mice produced an increase in the number of neutrophils in peritoneal washings collected 4 h later but after 1 day the numbers had returned to control levels. The increase in numbers of mononuclear cells and eosinophils in the peritoneal washings peaked at 2 days and persisted for at least 5 days. Dosing the mice with phenidone, a dual inhibitor of the cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism, potentiated the neutrophil infiltration at 4 h but had no significant effect upon the subsequent mononuclear cell and eosinophil infiltration. In contrast, treatment with the corticosteroid, dexamethasone, reduced the infiltration by all three types of cells, providing further evidence that the corticosteroids can inhibit immune-induced cellular infiltrations by mechanisms other than the inhibition of arachidonic acid metabolism. Isoprenaline, given to the mice before antigen challenge, had no effect on the subsequent neutrophil infiltration, but repeated doses did inhibit the mononuclear cell and eosinophil infiltration measured 4 days later. Aminophylline, disodium cromoglycate and cyproheptadine had no effect upon the cellular changes.
Effects of Muramyl Dipeptide and Trehalose Dimycolate on Resistance of Mice to Toxoplasma gondii and Acanthamoeba culbertsoni Infections
Authors: Masihi Noel , Bhaduri Ranjan , Werner Hans , Janitschke Klaus , Lange Werner
Published: August 2009

The effects of synthetic muramyl dipeptide (MDP) and natural trehalose dimycolate (TDM) against parasitic infections by intracellular Toxoplasma gondii and free-living Acanthamoeba culbertsoni were studied. Significant resistance against oral T. gondii infection was induced by intraperitoneal pretreatment with TDM but not with MDP. The protective effect of TDM against T. gondii was corroborated by a significant reduction in the number of cysts in brains of pretreated animals and elevated serum antibody levels. Partial protection against lethal intranasal A. culbertsoni infection was conferred by specific immunization with viable trophozoites of nonpathogenic Acanthamoeba lugdunensis. The nonspecific resistance induced by intravenous pretreatment with MDP was similar to, whereas that stimulated by TDM was lesser than the protection conferred by A. lugdunensis. The Fc receptor-mediated phagocytosis of 51Cr-labeled sheep red blood cells by alveolar macrophages was enhanced by MDP. The phagocytic activity of peritoneal macrophages was increased by lower doses of TDM.
Monkey Interleukin 2: Optimal Conditions for Production and Partial Characterization
Authors: Tatsumi Masashi , Yabe Mikiko
Published: August 2009

Cynomolgus monkey spleen cells and peripheral blood lymphocytes were tested for their ability to produce interleukin 2 (IL2) upon stimulation with phytomitogens. By assessing proliferation of murine IL 2-dependent cells, maximal release of IL 2 activity was found in the supernatants of nonadherent cells cultured for 48 h, after stimulation with 25 μg/ml concanavalin A. Addition of phorbol myristate acetate to the cultures resulted in enhancement of IL 2 production, but there was no significant promotion when indomethacin or human lymphoblastoid cell lines were added to the cultures. Adherent macrophages appeared to inhibit rather than enhance IL 2 production in this system. Monkey IL 2 exhibited a molecular weight in the range of 15,000–18,000 daltons in Sephadex G-100 gel chromatography.
Purification and Physicochemical Characterization of Schistosoma mansoni Egg Allergen Recognized by Mouse Sera Obtained at an Acute Stage of Infection
Authors: Owhashi M. , Horii Y. , Imai J. , Ishii A. , Nawa Y.
Published: August 2009

A major allergenic component recognized by mouse sera obtained at an acute stage of infection was purified from soluble egg antigen preparation (SEA) of Schistosoma mansoni by anion-exchange chromatography on DE52 and gel chromatography on Sephadex G-150. The purified allergen showed homogeneity by immunoelectrophoresis and by polyacrylamide gel electrophoresis. Its apparent molecular weight was 210,000 by gel chromatography on Sephadex G-200. This purified allergen could bind to Con A-Sepharose 4B, indicating its glycoprotein nature. After amino acid analysis, aspartic acid, glutamic acid, serine and theonine were found as the major amino acids. The allergenic activity was destroyed by heating at 100°C for 60 min and by pronase or periodate treatment. By double diffusion in agar gel, this purified allergen gave a strong single band against acute (8 w) stage serum, which fused to the major band formed by crude SEA. On the other hand, it showed a very faint band against chronic (22 w) stage serum, which is apparently different from the main band formed between crude SEA and the chronic stage serum. When specific IgE or IgG antibody titers in the serum of human schistosomiasis mansoni cases were measured by ELISA using this purified allergen, the results showed good correlation with those obtained by using crude SEA. Thus, this purified allergen is not only a major allergen in the acute stage of murine schistosomiasis but also an allergen in human schistosomiasis mansoni.
Differential Effect of Experimental Diabetes on the Early and Late Phase of Contact Sensitivity Reaction in Mice
Authors: Ptak W. , Rewicka Maria , Gryglewski A. , Bielecka Jolanta
Published: August 2009

Contact sensitization induces two different kinds of T cells (both Ly 1) that act in sequence to produce upon challenge with antigen a classical 24-hour local skin swelling reaction. One of these cells produces an antigen-specific factor. It has been suggested that it sensitizes mast cells, similar to IgE antibody, and causes them to release vasoactive amines in the presence of antigen. This results in an early (2-hour) swelling reaction. Increased vascular permeability facilitates the entry of the second, lymphokine-producing Ly 1 cell into the site of reaction to elicit the classical 24-hour delayed-type hypersensitivity reaction. In alloxan diabetic mice, contact sensitivity reactions are reduced significantly, and our experiments show that insulin deficiency affects only the activity of the late acting, lymphokine-producing cell and leaves the factor-producing cell responsible for the early swelling reaction unaffected. Our experiments demonstrate that insulin deficiency has different effects on distinct subpopulations of T lymphocytes.
Potentiation of the Cellular Immune Response by Adjuvants: A Limited Role for Adjuvant Induced Interferon
Authors: Kraaijeveld C.A. , Kamphuis W. , Benaissa-Trouw B.J. , van Haarlem H. , Harmsen M. , Snippe H.
Published: August 2009

Adjuvants differ in capacity to induce interferon (IFN). As a consequence IFN induction by adjuvants may influence their effectiveness in enhancement of delayed type hypersensitivity (DH) reactions. In this study, the lipophilic amine dimethyl dioctadecyl ammonium bromide (DDA), the synthetic double-stranded polynucleotide polyinosinic polycytidylic acid (poly I:C), liposomes and the polyols L 101 and L 121 were compared in BALB/c mice as inducers of IFN and also as adjuvants for DH to both lysozyme and inactivated Semliki Forest virus (SFV). The antigens were injected intracutaneously, alone or mixed with adjuvant. At day 6 after the immunization, DH was elicited and measured 24 h later as increase in footpad thickness. Negatively charged liposomes and polyol L 121 were unable to enhance DH to SFV and also lack the capacity to induce IFN. Polyol L 101 induced low levels of IFN and showed only slight adjuvanticity for DH to SFV. In contrast, DDA, a moderate IFN inducer, was shown to be a very effective adjuvant for induction of DH against both lysozyme and SFV. The excellent IFN inducer, poly I:C, at the tested dose range (0.03–3.0 mg/kg), displayed only a relatively weak adjuvant activity. But low doses of poly I:C (0.03 and 0.1 mg/kg) showed still adjuvanticity in contrast to equal doses of DDA. This might be related to sufficient induction of IFN by low doses of poly I:C but not by DDA. The discrepancy observed in the relation between IFN induction and a maximal DH induction suggests that IFN is not the only factor which enhances the effectiveness of adjuvants in induction of DH. Furthermore, various inbred mouse strains display considerable differences in DDA-induced enhancement of DH to both lysozyme and SFV, whereas no significant differences in DDA-induced IFN titers were detected. In conclusion, the capacity of an adjuvant to induce IFN is an important feature to achieve enhancement of DH reactions. The effectiveness of an adjuvant, however, is also determined by its other intrinsic properties and by factors determined by the host.
Local Intranasal Immunotherapy with High-Dose Polymerized Ragweed Extract
Authors: Georgitis J.W. , Nickelsen J.A. , Wypych J.I. , Barde S.H. , Clayton W.F. , Reisman R.E.
Published: August 2009

Thirty-one ragweed-allergic patients received preseasonal local intranasal immunotherapy (LNIT) with high doses of gluteraldehyde-polymerized ragweed extract (average total dose 544 μgantigen E). Minimal side effects were reported during treatment and did not interfere with the dosing schedule. During the ragweed pollen season, LNIT-treated patients had lower symptom scores for sneezing, rhinorrhea and nasal congestion than a comparable group of untreated ragweed-allergic patients. There was no difference in ragweed-induced eye symptoms between the two groups. Secretory ragweed-specific IgA and IgG rose following LNIT treatment. Absolute antibody titers and changes in titers did not correlate with clinical improvement. LNIT with the polymerized ragweed did not block the seasonal rise in serum ragweed-specific IgE. These results suggest that LNIT with high-dose polymerized ragweed extract is a safe, simple and effective form of immunotherapy.
Induction of Suppressor Cells in Normal Lymphocytes by Uremic Serum
Authors: Weissgarten Joshua , Modai David , Cohen Natan , Averbukh Zhan , Shaked Uri , Tieder Martin , Peller Shoshana , Kaufman Susana
Published: August 2009

Sera of patients on chronic hemodialysis induced suppressor cell activity (SCA) in normal peripheral blood mononuclear cells, which significantly impaired blastogenic response to PHA. This SCA is statistically not different from Con A induced SCA. Both SCAs are however additive. Speculations concerning the modes of action of this induced SCA are discussed.
Liposomes as Carrier for Antibiotics: A Comparative Study on the Immune Response against Liposome-Encapsulated Penicillin and Other Penicillin Preparations
Authors: de Haan P. , Claassen E. , van Rooijen N.
Published: August 2009

Liposomes do not stimulate a primary or secondary immune response against entrapped penicillin in mice.
Complement Activation by Female Protein, the Hamster Homologue of Human C-Reactive Protein
Authors: Etlinger Howard M. , Coe John E.
Published: August 2009

The capacity of Syrian hamster female protein (FP), a phosphorylcholine (PC)-binding pentraxin, to activate complement was tested in an in vitro system consisting of PC-coupled sheep red blood cells and guinea pig serum as the complement (C) source. FP was demonstrated to fix complement as reflected by hemolysis. Such hemolysis was eliminated by heat treatment (56°C, 30 min) of guinea pig serum and inhibited by PC chloride but not dinitrophenyl lysine. The inability of C4-deficient guinea pig serum to provide lytic activity indicated that lysis proceeded through the classical hemolytic pathway.
A Re-examination of Ryegrass (Lolium perenne) Pollen Allergens
Authors: Ford S.A. , Baldo B.A.
Published: August 2009

Electroblotting of crude ryegrass pollen extracts and purified group I, II and III allergens identified 14 IgE-binding components, 8 of which were previously unrecognized. In addition to allergen groups I, II and III, which are already regarded as clinically important, and on the basis of the frequencies and intensities of IgE binding with sera from 42 ryegrass pollen-allergic patients, proteins with molecular weights (MWs) of 60, 32, 30 and 28 kD were identified as allergens of possible major clinical importance. Six other pollen components with MWs ranging from 23 to 80 kD and which reacted with IgE antibodies in the sera of 33–50% of patients, should also be viewed as proteins with potential clinical relevance for at least a proportion of the patients. The electrophoretic separation patterns of ryegrass pollen extracts in both alkaline and acid gels and IgE-probed membrane transfers produced in this study should serve as useful reference patterns for standardization purposes. In addition, the identification of the complete allergen recognition pattern by individual patients will permit safer and more effective diagnosis and therapy of ryegrass pollen sensitivities.
Isolation and Characterization of Two Allergens from Dermatophagoides farinae
Authors: Yasueda Hiroshi , Mita Haruhisa , Yui Yasuo , Shida Takao
Published: August 2009

Two purified allergens, designated as DF1 and DF2, were isolated from the extract of the whole culture of Dermatophagoides farinae by a combination of ammonium sulfate precipitation and ion exchange, hydrophobic, chelate and gel chromatography. DF1 was isolated as a heat-sensitive acidic protein with an apparent molecular weight of 25,000 and an isoelectric point of 4.6–7.2. DF2 was isolated as a heat-stable basic protein with an apparent molecular weight of 15,000 and an isoelectric point of 7.8–8.3. No allergenic cross-reactivity was seen between DF1 and DF2. Both DF1 and DF2 were shown to be the major allergens of D. farinae by the results of radioallergosorbent test and histamine release assay.
Raised Serum IgG4 Levels in Patients with Atopy and Filariasis: Application of an Automated Particle-Counting Immunoassay Using Monoclonal Antibody
Authors: Magnusson C.G.M. , Cesbron J.Y. , Djurup R. , Capron A. , Johansson S.G.O.
Published: August 2009

We show here an automated (50 samples/h) assay for serum IgG4 having a throughput time of 40 min per sample and a sensitivity of 10 μg/ml. The assay procedure is based on the inhibition by sample of the agglutination reaction between monoclonal anti-IgG4 antibodies and latex particles to which IgG4 myeloma protein has been coupled. Assay reliability was ascertained by testing for linearity, analytical recovery (96.4%), interassay precision (≤8%), specificity and correlation between the results obtained with monoclonal and polyclonal anti-IgG4 antibodies (n = 84; rs = 0.97). Application of the assay to sera from various groups of patients indicated significantly (p < 0.00005) higher geometrical means (Gx) in patients suffering from atopy (n = 87; Gx = 617 μg/ml), atopic dermatitis (n = 28; Gx = 1,043 μg/ml), filariasis with Onchocerca volvulus (n = 48; Gx = 1,681 μg/ml) and Brugia malayi(n = 20; Gx = 1,078 μg/ml) as compared to nonatopic subjects (n = 103; Gx = 302 μg/ml) and randomized paired maternal/cord sera (n = 41; Gx = 276 and 296 μg/ml, respectively). IgG4 in the paired maternal/cord sera correlated (r = 0.98; p <0.00005). There was no significant influence of age or sex on the IgG4 levels either among the nonatopics or the atopies even though low IgG4 (≤30 μg/ml) was more common among women. The results suggest that IgG4 and IgE responses are somehow closely related in atopic and parasite-infested patients at the physiological, pathogenic or genetic level.