bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibitory Effect of Human Interferons on Lymphocyte Proliferation Induced by Phytomitogens in Cynomolgus Monkeys (Macaca fascicularis)
Authors: Tatsumi Masashi , Kohase Masayoshi , Yamazaki Shudo
Published: August 2009

In order to study the comparative cross-reactivity of human interferon (Hu-IFN) with cynomolgus monkeys, the effect of Hu-IFN on the immune responses of monkeys was assessed by measuring lymphocyte transformation induced by phytomitogens. The in vivo administration of Hu-IFN or its placebo to monkeys resulted in transient neutrophilia, but no significant alteration of the kinetics of T lymphocytes in the peripheral blood. Lymphocyte proliferation of Hu-IFN-injected monkeys showed a significant reduction depending on their serum Hu-IFN titers, whereas no significant decrease was observed in monkeys injected with placebo. The in vitro incubation of monkey lymphocytes with Hu-IFN before or simultaneously with the addition of phytomitogens caused a substantial decrease in the lymphocyte proliferative responses in a dose-dependent manner. However, the inhibitory effect was no longer observed when Hu-IFN was added 24 h after incubation of lymphocytes with phytomitogens. Hu-IFN also suppressed the blastogenic responses of lymphocytes pulsed with phytomitogens before the addition of Hu-IFN. Addition of peritoneal macrophages pretreated with Hu-IFN failed to exert any significantly different effect on the lymphocyte proliferation compared to untreated ones. Taken together, the inhibitory effect of Hu-IFN seemed to act directly upon the early phase of lymphocyte proliferation induced by phytomitogens. These findings indicate that cynomolgus monkeys were susceptible to Hu-IFN both in vitro and in vivo with respect to the inhibitory effect on lymphocyte proliferation and could serve as a useful experimental model to study the effect of Hu-IFN on the immune responses.
Changes of Beta-Adrenergic Receptor Number and Catecholamine-Sensitive Adenylate Cyclase in Guinea Pig Lung after Inhalation of Histamine Aerosol
Authors: Mita Haruhisa , Yui Yasuo , Shida Takao
Published: August 2009

After guinea pigs were exposed to histamine or acetylcholine aerosol for 1 week, alpha1- and beta-adrenergic and cholinergic muscarinic receptors on the lung membranes were measured by direct binding techniques using 3H-prazosin, l-3H-dihydroalprenolol, and l-3H-quinuclidinyl benzilate, respectively, and adenylate cyclase responses to /-isoproterenol (10––5 to 10––8M) and NaF (20 mM) were also examined on the membranes. After the inhalation, the number of beta-adrenergic receptors was decreased by about 25% without a significant change in binding affinity for l-3H-dihydroalprenolol, while the number and the affinity of alphar adrenergic and muscarinic receptors on the membranes were not significantly different from those in the control animals. When compared with the control animals, the lung membranes showed a reduced adenylate cyclase response to l-isoproterenol, and the response to 10––5M of l-isoproterenol was significantly decreased by 11 % in the lung membranes. The histamine inhalation showed no significant effect on basal adenylate cyclase activity and adenylate cyclase response to NaF.
Comparison of Posterior Capsular Opacification between a Combined Procedure and a Sequential Procedure of Pars Plana Vitrectomy and Cataract Surgery
Authors: Roh Joon Ho , Sohn Hee Jin , Lee Dae Yeong , Shyn Kyung Hwan , Nam Dong Heun
Published: August 2009

Purpose: To compare posterior capsular opacification (PCO) between a combined procedure and a sequential procedure of pars plana vitrectomy (PPV) and cataract surgery (CS). Methods: The medical records of 89 eyes of 85 patients who underwent PPV and CS were retrospectively reviewed. There were 56 eyes of 52 patients with a combined PPV and CS (the combined surgery group), and 33 eyes of 33 patients with CS in a previously vitrectomized eye (the sequential surgery group). The control group was comprised of 130 eyes of 102 patients who underwent CS alone. All patients were followed up for at least 1 year after CS. The major outcome measures were the PCO rate and the interval between CS and PCO formation. There were no significant differences in age and the incidence of diabetes among the 3 groups. Results: The PCO rate checked at 1 year after CS was 12.5% (7/56) in the combined surgery group, 24.2% (8/33) in the sequential surgery group, and 4.6% (6/130) in the control group. The differences in the PCO rate between the subgroups as well as among the 3 groups were statistically significant (p < 0.05). There were no significant differences in the interval between CS and PCO formation among the 3 groups. Conclusion: This study demonstrates that the PCO rate may be lower in patients who have a combined procedure of PPV and CS than in those who have a sequential procedure.
Colorimetric Analysis of Three Editions of the Velhagen-Broschmann Pseudoisochromatic Colour Plates
Authors: Kuchenbecker Jörn , Nicklas Sven , Behrens-Baumann Wolfgang
Published: August 2009

Purpose: Chromatic variations across different copies and different editions of pseudoisochromatic tests and violation of underlying principles of construction for individual plates can influence test results. Methods: We analysed the colorimetric characteristics of three different editions of Velhagen-Broschmann pseudoisochromatic plates (30th edition printed in 1995, 31st edition printed in 1997, 32nd edition printed in 2000). One hundred and twelve coloured dots of 18 plates were chosen from each edition. We measured RGB and CIE XYZ values using a spectrophotometer. Differences in lightness and chromaticity between corresponding dots of different editions were analysed in terms of ΔL* and Δu’v’, respectively. For each plate deviations from dichromatic confusion lines were analysed. Furthermore, we determined the relative luminance of a target compared to its background in terms of the Weber contrast. Results: The mean ΔL* across editions was 2.05 (±1.4) and the mean Δu’v’ was 0.0078 (±0.0029). For two plates the deviations of targets from dichromatic confusion lines exceeded suggested values. For a number of plates, the lightness contrast between the symbol and its background was high. Comparison with psychophysical data showed that these colour plates are easily detectable by colour-deficient observers. Conclusions: Lightness and chromatic variation across the three editions was moderate except for a small number of plates perhaps due to inaccuracies in the printing process. The design of several plates should be revised according to standard principles of construction of colour deficiency tests.
Genetic Interaction between Tau and the Apolipoprotein E Receptor LRP1 Increases Alzheimer’s Disease Risk
Authors: Vázquez-Higuera José Luis , Mateo Ignacio , Sánchez-Juan Pascual , Rodríguez-Rodríguez Eloy , Pozueta Ana , Infante Jon , Berciano José , Combarros Onofre
Published: August 2009

Abnormal tau hyperphosphorylation is one of the central events in the development of neurofibrillary tangles (NFTs) in Alzheimer’s disease (AD), and phosphorylation of tau is accelerated by the increase in the level of neuronal cholesterol. Apolipoprotein E (APOE) promotes the neuronal uptake of cholesterol via APOE receptors such as the low-density lipoprotein receptor-related protein 1 (LRP1), and the APOE ε4 allele is associated with an increase in NFT burden in AD brain. In a case-control study in 246 AD patients and 237 healthy controls, we examined whether the combined gene effects between tau (intron 9, rs2471738) polymorphism and LRP1 (exon 3, rs1799986) polymorphism might be responsible for susceptibility to AD, independently or in concert with the APOE ε4 allele. Subjects carrying both the tau (intron 9, rs2471738) T allele (CT and TT genotypes) and the LRP1 (exon 3, rs1799986) T allele (CT and TT genotypes) had a 6 times higher risk of developing AD than subjects without these risk genotypes (odds ration = 6.20, 95% confidence interval = 1.74–22.05, p = 0.005), and this genetic interaction was observed in either the presence or the absence of the APOE ε4 allele. These data suggest that the synergistic effects (epistasis) between tau and LRP1 might modify the risk of AD in an APOE ε4 allele-independent fashion.
Isolation and Characterization of an Allergen-Rich Fraction Derived from Cultures of Dermatophagoides pteronyssinus
Authors: Andersen Annette
Published: August 2009

An allergen-rich fraction derived from mature cultures of Dermatophagoides pteronyssinus was isolated. Results of radio-allergosorbent tests and the disclosure of IgE-binding proteins after their electrophoretic transfer from sodium dodecyl sulphate-polyacrylamide gel to nitrocellulose sheets revealed that both important groups of allergens, Der p I and Der p II, seem to be contained in the above-mentioned fraction. Analysis using crossed immuno-electrophoresis (CIE) disclosed 5–7 antigens and a corresponding enzymatic assay for the specificity of patient’s IgE in CIE revealed that 4 of the antigens also are allergens. Moreover, an allergen was detected which apparently was not discovered when using CIE. The fraction in question was fractionated further and subsequent analysis by CIE indicated the purification of 1 antigen.
Immunity to Ocular and Collagen Antigens in Childhood Arthritis and Uveitis
Authors: Petty Ross E. , Hunt David W.C.
Published: August 2009

Humoral and cellular immunity to ocular antigens (S antigen and alpha, beta heavy, beta light, and gamma crystallins) and connective tissue antigens (type I, II, III, and IV collagens) were studied in children with juvenile rheumatoid arthritis (JRA) with or without uveitis and in controls. There was no association between the presence of uveitis and antibody to any collagen type or of lymphocyte transformation to type II collagen. Antibodies to all crystallins were more common in children with JRA than in controls, and antibodies to BH crystallins were more frequent in children with JRA and uveitis than in those with JRA alone. Such reactivity did not appear to represent antibody cross-reactive with collagen antigens. Lymphocyte proliferation to alpha crystallin was increased in JRA with or without uveitis, whereas lymphocyte response to S antigen was associated with the presence of uveitis.
CD1-Positive Epidermal Langerhans Cells in Skin Reactions to Autologous Peripheral-Blood-Derived Mononuclear Cells in Leprosy Patients
Authors: Narayanan R.B. , Natarajan M. , Katoch K. , Sengupta U.
Published: August 2009

A comparison was made on the characteristics of the infiltrates, the number and distribution of CD 1-positive epidermal Langerhans cells (LC) at the sites of skin reaction induced by autologous peripheral-blood-derived mononuclear cells (PBMC) in leprosy patients. Clinically and histologically, the skin reaction was well expressed in tuberculoid patients as compared to lepromatous patients, erythema nodosum leprosum (ENL) patients and contacts. The quantum of lymphocytes in the infiltrates was maximal in the tuberculoid patients and it was minimal in lepromatous and ENL patients. The number and distribution of LC in the tuberculoid patients was significantly higher in the PBMC-inoculated sites as compared to control sites over 24 h. In contrast, no difference in the number and distribution of LC was noticed in the lepromatous and ENL patients. These observations indicate that the lymphocytes of tuberculoid patients in contrast to lepromatous leprosy patients are capable of sustenance in the local micro-environments of the skin and an effective interaction may be possible between LC and PBMC.
Selective IgG Subclass Antibody Response to Insulin in Diabetic Patients Receiving Animal Insulin Replacement Therapy
Authors: Siddiqi Mushtaq A. , Wangnoo Subash K.
Published: August 2009

Serum IgG subclass antibodies to insulin were semiquantitatively estimated using a highly sensitive and specific technique of enzyme-linked immunosorbent assay. The three groups of patients (categorised on the basis of duration and dose of exogenous insulin administered) showed a variable pattern. In spite of considerable variations from patient to patient in a group, IgG1 and IgG2 levels were higher in patients receiving a low dose of insulin for 9–18 and 30–72 months, respectively, while in the patients on high dose of therapeutic insulin, IgG4 well predominated over all other subclasses. It is postulated that IgG4 antibodies are directed against the biologically active site of insulin and explains the high requirement of insulin in these patients.
Antigen-Induced Bronchopulmonary Alterations in the Guinea Pig: A New Model of Passive Sensitization Mediated by Mouse IgE Antibodies
Authors: Desquand S. , Lefort J. , Liu F.T. , Mencia-Huerta J.M. , Vargaftig B.B.
Published: August 2009

A selective model to study the IgE-mediated anaphylactic bronchoconstriction (BC) in the guinea pig is needed, since human asthma involves mainly this class of antibody. However, most procedures presently available for passive homologous or active sensitization lead to responses which are mediated both by IgE and IgG antibodies. In this study, we developed an anaphylactic model in which guinea pigs are passively sensitized with mouse ascitic fluid containing dinitrophenol (DNP)-specific IgE antibodies. Challenge of sensitized animals with DNP coupled to bovine serum albumin evokes a bronchoconstrictor response that is maximal 5 h after sensitization. The resulting anaphylactic BC is not blocked by the H1 histamine antagonist mepyramine, by the peptido-leukotriene antagonist FLP 55712 nor by the platelet-activating factor antagonist BN 52021 alone. However, when the sensitized animals are pretreated with the three drugs in combination, significantly reduced BC was observed upon challenge with the antigen. This latter result indicates that IgE-dependent BC involves the participation of different mediators, a characteristic shared in common with allergic asthma in human.
Intestinal Permeability and Antigen Absorption in Rheumatoid Arthritis
Authors: Fagiolo U. , Paganelli R. , Ossi E. , Quinti I. , Cancian M. , D’Offizi G.P. , Fiocco U.
Published: August 2009

Intestinal permeability was measured using cow’s milk beta-lactoglobulin absorption (BLG) as a permeability marker in 14 patients with active and inactive rheumatoid arthritis (RA) under three different conditions: after a washout period, after treatment with acetylsalicylic acid (ASA) associated with disodium chromoglycate (DSCG), and with ASA only. No intolerance to cow’s milk was present and serum IgE levels were in the normal range in 12 of 14 patients. IgG anti-IgE were present in 7 of 13 patients tested. When off treatment the intestinal permeability to BLG in RA patients was not increased as compared to controls, but we found a significative difference between active and inactive RA. ASA administration strongly increased BLG absorption, not prevented by DSCG pretreatment. In normal controls treated with a single dose of ASA we obtained similar results. Our results suggest that prolonged treatment with nonsteroidal anti-inflammatory drugs induces an increase of food antigen absorption, apparently not related to anaphylaxis mediator release, with possible clinical effects.
Immunogenicity of Three Recombinant Treponema pallidum Antigens Examined in Guinea Pigs
Authors: Wicher Konrad , van Embden Jan D.A. , Schouls Leo M. , Zabek Jakub , Jakubowski Adam , Wicher Victoria
Published: August 2009

The immunogenicity of recombinant treponemal antigens TmpA, TmpB and TmpC incorporated in RIBI adjuvant and injected into inbred strain 2 guinea pigs has been examined. The immune status of these animals has been challenged by infection with Treponema pallidum, Nichols. The immune response evaluated by the fluorescent-antibody test, microhemagglutination test and ELISA demonstrated high titers of antibodies to the T. pallidum antigens. The immunoblot analysis proved that the antibodies were directed to the 43-(Tmp A) 34- (Tmp B) and 35-kdalton (Tmp C) polypeptides. Antibodies cross-reacting with Treponema phagedenis biotype Reiter were, however, also detected. In spite of high titers of antibodies the animals were not protected against challenging infection with 108 organisms of T. pallidum.
Biological and Immunological Properties of Sugi Basic Protein-Pullulan Conjugate
Authors: Taniguchi Yoshifumi , Ikegami Hakuo , Usui Mitsuko , Ando Shunsaku , Kurimoto Masashi , Sakaguchi Masahiro , Inouye Sakae , Matuhasi Tyoku
Published: August 2009

Sugi basic protein (SBP), a major allergen of Japanese cedar pollen, conjugated to pullulan acquires suppressive activities for IgE, but not for IgG antibody production, and for IgE-mediated PK reactions in a SBP-specific manner. When either normal or IgE anti-SBP antibody-producing mice were treated with the conjugate, IgE anti-SBP antibody levels scarcely increased even after immunization with SBP + alum. This suppression was shown to be SBP specific and IgE selective. The ability of SBP-pullulan conjugate to elicit PK reactions on passively sensitized rat skin was markedly reduced. Furthermore, the conjugate inhibited the reactions mediated by the IgE antibodies and native SBP when it was injected into the IgE antibody-sensitized sites 2 h prior to SBP challenge. The reduced ability of the conjugate to elicit PK reactions was not due to loss of antigenic determinants during the conjugation procedures because binding activity of the conjugate to anti-SBP antibodies was as strong as that of native SBP. These results indicate that SBP-pullulan conjugate may be useful for ‘desensitization therapy’ against the pollinosis.
Acute-Phase Proteins and the Serological Evaluation of Experimental Contact Sensitivity in the Mouse
Authors: Kimber Ian , Ward Rachel K. , Shepherd Christopher J. , Smith Michael N. , McAdam Keith P.W.J. , Raynes John G.
Published: August 2009

The evaluation of contact reactions in previously sensitized mice is assessed conventionally by measurement of increases in ear thickness following challenge. In an attempt to develop a serological method for the investigation of contact sensitization in mice, we have examined whether analysis of changes in the concentration of acute-phase proteins in response to challenge provides a reliable alternative means of evaluating elicitation reactions. Measurement of either the relative serum haptoglobin concentration, using radial immunodiffusion, or the absolute concentration of serum amyloid A, by an enzyme-linked immunosorbent assay, has been found to correlate well with induced increases in ear thickness following challenge. Changes in the concentration of acute-phase proteins proved to be of sufficient sensitivity to reflect the specificity of contact sensitization and its inhibition by antigenic competition.
Immunoglobulin E Response to Pertussis Toxin in Whooping Cough and after Immunization with a Whole-Cell and an Acellular Pertussis Vaccine
Authors: Hedenskog Staffan , Björkstén Bengt , Blennow Margareta , Granström Gunnar , Granström Marta
Published: August 2009

Immunoglobulin E antibodies to pertussis toxin (PT-IgE) were demonstrated in 15 of 23 (65%) patients with culture-confirmed pertussis. In 6 individuals there was a low-grade PT-IgE response after 6–9 weeks of disease and in 9 a rapid PT-IgE response, appearing 1–3 weeks after onset of symptoms. The PT-IgE antibody levels in immunized individuals were higher than in the non immunized. Following primary immunization of 23 children with a monovalent whole-cell pertussis vaccine (Burroughs-Wellcome, UK) or with an acellular pertussis vaccine (JNIH-6, Biken, Japan) a late low-grade PT-IgE response was found in 8 (35%). In 7/10 children previously immunized with the JNIH-6, a booster injection 16 months later with the same vaccine resulted in a rapidly appearing PT-IgE antibody response. In contrast, none of 13 children initially immunized with the monovalent whole-cell vaccine and then boostered with either this vaccine or JNIH-6 had detectable PT-IgE antibodies after the booster injection. The study shows that IgE-antibodies to pertussis toxin commonly appear in patients with whooping cough and that the kinetics and the magnitude of the response is influenced by previous exposure to the antigen. A PT-IgE response may also follow pertussis immunization.
Disposition and Immunogenicity of Penicillin in the Rabbit
Authors: Christie G. , Park B.K.
Published: August 2009

The immunogenicity, disposition and irreversible protein binding of benzylpenicillin (BP) were studied in male New Zealand White rabbits. There was an increase in IgG anti-benzylpenicilloyl (BPO) antibody activity, as detected by enzyme-linked immunosorbent assay (ELISA) following daily intramuscular administration (for 4 consecutive days) of BP (2.7 and 1.6 mmol/kg) freshly dissolved in 0.15 M NaCl. Antibody activity reached a maximum approximately 14 days after the last injection. There was a smaller immune response when a dose of 270 μmol/kg was administered. The specificity of the IgG antibody response for the BPO determinant was confirmed by inhibition of binding by BPO-aminocaproate. [3H]BP, administered intravenously to rabbits at a dose of 2.7 mmol/kg was rapidly cleared from plasma, and unchanged BP was not detected at 1 h. After 3 h, irreversible binding accounted for < 0.004% of the dose bound per milliliter of plasma, and this represented all the radioactivity present in plasma at this time. Covalent binding of BP to plasma proteins, in vitro, after 3 h was of the same magnitude for rabbit, rat and human plasma. Therefore, BP can induce a specific antibody response in the rabbit in contrast to the lack of immunogenicity observed previously in the rat.
Spontaneous Regression in Murine Hypersensitivity Pneumonitis: Lack of Immunological Tolerance
Authors: Takizawa Hajime , Suko Matsunobu , Kobayashi Nobuyuki , Ohta Ken , Shoji Shunsuke , Horiuchi Tadashi , Okudaira Hirokazu , Miyamoto Terumasa , Shiga Junji
Published: August 2009

We have reported a murine model of hypersensitivity pneumonitis (HP) by transnasal administration of Thermoactinomyces vulgaris (Tv) antigen and shown a potential usefulness in the study of the immunopathogenesis of HP. Since we observed spontaneous regression (SR) of the lung lesions with continuous administration of Tv antigen in our model of HP, attempts were made to study if SR is caused by immunological tolerance. The titers of anti-Tv IgG antibody in serum and bronchoalveolar lavage fluid (B ALF) did not show any decrement during the period of SR, at the 6th and 9th weeks after the first administration of the antigen. A delayed-type hypersensitivity reaction, i.e. footpad swelling with Tv antigen, did not change significantly during the SR period. Analysis of BALF cells disclosed that Lyt-2-positive cells, i.e. suppressor/cytotoxic T lymphocytes, did not increase to predominate Lyt-1 -positive cells. The lymphocyte proliferation test with Tv antigen showed that the lung lymphocytes reacted to Tv antigen as well as concanavalin A during the period of SR in a similar magnitude as in the development period (3 weeks after the first treatment with the antigen). Treatment with cyclophosphamide before the period of SR resulted in no delay of SR. Finally, the adoptive cell transfer of spleen T cells obtained from mice during the SR period did not manipulate the development of granulomatous pneumonitis in the recipient mice. We concluded that the events causing SR in our murine model of HP may not involve the induction of immunological tolerance in humoral or cellular immunity.
Unusual Precipitins in Pathological Human Sera
Authors: Kim D.S. , Anthone R. , Anthone S. , Milgrom F.
Published: August 2009

In conducting double diffusion gel precipitation tests, unexpected, strong precipitation reactions were noticed between a serum originating from a renal graft recipient and many pathological human sera. The highest frequency of positive reactions were produced by sera of patients with infectious mononucleosis (83%), chronic Epstein-Barr virus infections (72%), rheumatoid arthritis (57%), lepromatous leprosy (57%), and AIDS (44%). The precipitin in these pathological sera was identified as an antibody of IgM variety and the precipitinogen in the transplantation serum was shown to be a thermostable component with β-globulin mobility. No explanation of the nature of reactions observed can be given at present.
IgG Rheumatoid Factor in Human and Rabbit Transplantation Sera
Authors: Miyata Masayuki , Albini Boris , Kreis Henri , Milgrom Felix , Dausset Jean
Published: August 2009

Sera of human recipients of kidney allografts and rabbit recipients of skin allografts were tested by means of enzyme immunoassays (EIA) for IgG rheumatoid factor (RF) combining with human IgG (RF-H) or rabbit IgG (RF-R). In humans, the mean OD value in EIA for RF-H was significantly higher in 185 sera of allograft recipients than in 46 pretransplantation sera and 100 sera of normal subjects. Furthermore, the mean OD in patients who rejected their grafts was higher than in patients with satisfactory graft function. Interestingly, there was no difference in OD values for RF-R between sera of graft recipients and sera of normal subjects. Of 52 rabbit recipients of skin allografts, 24 were positive for both RF-R and RF-H, 4 were positive only for RF-R, another 4 only for RF-H, and 20 were negative for both. Renal allografts originating from the previous donors of skin grafts resulted, in most instances, in a significant decrease in the strength of the reaction for both RF-R and RF-H. The possible role of graft rejection in stimulation of RF formation as well as the possible role of RF in graft rejection should be considered.
Dendritic Cell Accumulation in Draining Lymph Nodes during the Induction Phase of Contact Allergy in Mice
Authors: Kinnaird A. , Peters S.W. , Foster J.R. , Kimber I.
Published: August 2009

Draining lymph node cells isolated from mice 24 h following topical exposure to a variety of contact-sensitizing chemicals, including the dinitrobenzene derivatives, 2,4-dinitrochlorobenzene and 2,4-dinitrothiocyanobenzene, contained increased numbers of dendritic cells (DCs). The increase in frequency of DCs was time-dependent and preceded significant changes in either lymph node cellularity or lymph node cell pro-liferative activity. The degree of DC accumulation was also influenced by the chemical used and the concentration employed for sensitization. In the context of contact allergy, the biological relevance of this phenomenon to the induction of hapten-specific responses is indicated by the fact that relatively small numbers of DC-enriched fractions of lymph node cells (comprising approximately 70% DCs), but not unfractionated or DC-depleted populations, transferred sensitization to naive animals. Moreover, using the skin-sensitizing fluorochrome, fluorescein isothiocyanate, it was observed that 24 h following exposure the majority of lymph node cells bearing high concentrations of antigen were within the DC-rich fraction.
Abrogation of Spleen Macrophage Suppressive Activity by 15-Deoxyspergualin
Authors: Yamamura Takashi , Tabira Takeshi
Published: August 2009

We studied the immunomodulatory mechanism of 15-deoxyspergualin (DSG), a novel antitumor agent. To assess this, we used a mixed culture of antigen-primed lymph node cells (LNC) and spleen cells (SPC). In normal or untreated conditions, SPC suppressed the proliferative response of LNC to the antigens. In contrast, SPC from DSG-treated animals did not show any suppressive activity. This suppression was mediated by adherent cells in SPC but the suppression mechanism was recovered when DSG-treated spleen macro-phages were reconstituted with control spleen nonadherent cells. These data suggest that DSG abrogates nonspecific suppressive activity of spleen cells by acting on both adherent and nonadherent cells.
Decreased Releasability of Basophils from Patients with Cold Urticaria after Cold Exposure
Authors: Hessler Hans-Jürgen , Pufahl Constanze , Christophers Enno
Published: August 2009

Histamine release from peripheral blood basophils challenged with C5a, f-met-peptides and calcium ionophore was studied in patients with cold urticaria before and after exposure to low environmental temperatures. Compared to healthy controls, stimulated mediator release before cold exposure was increased in 7 of 11 patients. When challenged by cold exposure mediator release from in vitro-stimulated basophils was decreased. This decrease was more pronounced after stimulation with receptor-mediated stimuli (e.g. C5a) as compared to receptor-unrelated stimuli, e.g. calcium inophore. In 4 of 11 patients stimulated mediator release before cold exposure was moderately increased. Also after cold exposure only a weak decrease of stimulated histamine release was seen. Levels of activated complement components (C3a) before and after cold exposure failed to provide evidence for complement activation in vivo. Also the number of circulating basophils as well as their cellular histamine content remained normal after cold exposure. The results show that in these patients release of histamine is altered before and after cold exposure. These changes in basophil responsiveness are not due to complement activation in vivo.
Protection against Experimental Allergic Encephalomyelitis with Complete Freund’s Adjuvant Is Unaffected by Prostaglandin Synthesis Inhibition
Authors: Weber Frank , Hempel Klaus
Published: August 2009

Pretreatment with complete Freund’s adjuvant blended with aluminium hydroxide (ALU-CFA) prevents the clinical as well as histological manifestation of experimental allergic encephalomyelitis (EAE) in Lewis rats. Suppression of prostaglandin biosynthesis with the cyclooxygenase inhibitor piroxicam (10 mg/kg body weight) from day 2 before to day 17 after EAE induction could not restore responsiveness in pretreated animals. In contrast piroxicam increased ALU-CFA-induced suppression of autoantibodies against myelin basic protein. In controls not pretreated with ALU-CFA, clinical signs of EAE were attenuated by piroxicam, whereas mononuclear infiltration of the brain remained unchanged. Therefore it is unlikely that prostaglandins exert an important function in the regulation of immune responses in this model of autoimmunity.
Characterization of Grass Pollen-Specific IgE, IgA, IgM Classes and IgG Subclasses in Allergic Patients
Authors: Desvaux F.X. , Peltre G. , David B.
Published: August 2009

Water-soluble grass pollen extracts (Dactylis glomerata) were separated by isoelectric focusing in a wide pH range (2–11) in agarose gel. After focusing, two successive gel prints were taken. The first one, on an ordinary nitrocellulose filter during 10 s, enabled the visualization of separated components of the pollen, after india ink staining. The second one, on a cyanogen bromide-activated nitrocellulose filter obtained after a 10-min transfer and saturation step, was incubated overnight with patient sera, and the specific antibodies bound to antigens or allergens were detected. The screening of different patient sera showed great variability in the antigen spectra. There was no obvious relationships between IgM and IgA antigen spectra as compared to IgE. Some association between IgE and IgG4 subclass was observed.
In vitro Sensitivity of Immature Human Mast Cells to Chemotherapeutic Agents
Authors: Butterfield Joseph H. , Weiler Deborah A.
Published: August 2009

Cells from the human immature mast cell line, HMC-1 were tested for their sensitivity to 11 chemotherapeutic agents by changes in viability and incorporation of tritiated thymidine ([3H]-thymidine) after 72 h of in vitro drug exposure. Doxorubicin hydrochloride and cytosine arabinoside were the most active agents against HMC-1 cells at the concentrations tested. Doxorubicin hydrochloride inhibited the incorporation of [3H]-thymidine and decreased the viability of HMC-1 cells by > 90% at a concentration of 0.06 μg/ml. Cytosine arabinoside inhibited the incorporation of [3H]-thymidine and decreased the viability by > 95% at a concentration of 0.1 μg/ml. A clone of HMC-1 cells, A4, with an enhanced percentage (70–80%) of metachromatically staining cells was equally sensitive to these two agents; however, A4 cells were more sensitive to vinblastine sulfate and less sensitive to methylprednisolone than was the parent cell line. Both HMC-1 cells and A4 cells showed approximately equal sensitivity to etoposide and to mitomycin. These results show that human mast cells are susceptible in vitro to a number of commonly used chemotherapeutic drugs.
Prolonged Exposure of Neutrophils to Saline or Bronchoalveolar Lavage Fluid Does Not Alter Superoxide Anion Generation
Authors: Nguyen Phong , Shusterman Richard D. , Barnett Ronald B. , Ishihara Yoko , Gottlieb Jonathan E. , Albertine Kurt H. , Gee Marlys H. , Peters Stephen P.
Published: August 2009

The role played by neutrophils (PMNs) in the genesis of lung injury in diverse clinical situations, such as bronchial asthma, idiopathic pulmonary fibrosis, and the adult respiratory distress syndrome, is an area of intensive investigation. Functional studies of PMNs, particularly those obtained from the alveoli by bronchoalveolar lavage, should shed light on their contribution to lung injury. However, it has not been demonstrated whether procedures used to harvest cells from the lung (bronchoalveolar lavage), particularly the potentially prolonged exposure to saline, commonly used to perform lavage, and other components of lavage fluid, can alter the functional characteristics of PMNs. In this report we demonstrate that a 2- to 3-hour exposure of neutrophils to saline from both humans and sheep in vitro does not alter the functional characteristics of PMNs as determined by superoxide anion generation after activation with phorbol myristate acetate (PMA; 6.96 ± 0.44 vs. 7.60 ± 0.32 nmol O2/250,000 PMNs for control and saline-treated human cells, respectively, after a 45-min incubation with 10––7M PMA, and 4.73 ± 0.30 vs. 4.50 ± 0.42 nmol O2/250,000 PMNs for control and saline-treated sheep cells). In a second series of experiments, we studied the effect of exposure of human PMNs to bronchoalveolar lavage fluid supernatants obtained from normal volunteers on superoxide anion generation by neutrophils. Again, spontaneous (1.27 ± 0.49 vs. 1.65 ± 0.71 nmol O2/200,000 PMNs for control and lavage fluid-treated cells, after a 45-min incubation), PMA-triggered (10––6M; 5.67 ± 0.27 vs. 5.18 ± 0.55 nmol O2/200,000 PMNs), and f-met peptide-triggered (10––6M; 3.34 ± 0.41 vs. 3.04 ± 0.64 nmol O2/200,000 PMNs) generation of superoxide anion was not affected. Therefore, we conclude that prolonged exposure of neutrophils to either saline or bronchoalveolar lavage fluid supernatants does not alter their capacity to generate superoxide anion and that biochemical studies of neutrophils obtained from the air spaces by bronchoalveolar lavage with saline should provide useful information about the functional characteristics of these cells.
Pharmacological Effect of KC-404 on Leukotriene Release from Human Leukocytes Induced by Ionophore A23187
Authors: Ohtsu Hiroshi , Fujimoto Yumiko , Yamauchi Kohei , Tamura Gen , Takishima Tamotsu
Published: August 2009

Release of leukotriene (LT) B4, LTC4 and histamine from human peripheral mixed leukocytes stimulated with calcium ionophore A23187 was evaluated. The amounts of LTB4 and LTC4 in supernatants were analyzed by high performance liquid chromatography and histamine was measured by an automated fluorometric technique. A time-course study demonstrated the characteristic patterns of the release of these mediators. LTB4 and LTC4 were more slowly released than histamine and the release of these LTs was inhibited by KC-404, a novel anti-asthmatic drug which has no effect on histamine release.
Effect of Reduction and Heat on the Detection of House Dust Mite (Dermatophagoides pteronyssinus) Allergen Der p I by Protein Blotting
Authors: Ford Stephen A. , Tovey Euan R. , Baldo Brian A.
Published: August 2009

Employment of reducing conditions during sample preparation alters the mobility of the house dust mite (Dermatophagoides pteronyssinus) allergen Der p I as determined by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) from an apparent MW of approximately 25 kD (unreduced) to an apparent MW of approximately 30 kD. Probing of nitrocellulose transfers with sera from subjects allergic to D. pteronyssinus showed that redcuction of Der p I was accompanied by a substantial loss of IgE-antibody-binding capacity by this allergen. An important consequence of the effect of reduction on Der p I is that the electrophoretic mobility of this protein becomes very similar to a closely spaced pair of protein-staining bands, probably Der p III, of MWs 30–31 kD. These bands bind IgE antibodies strongly and with high frequency and exhibit the same electrophoretic mobilities under both reducing and non-reducing conditions. Thus, for the clear resolution of allergen Der p I from other IgE-binding components in the same MW region, including Der p III, house dust mite samples for analysis by SDS-PAGE and blotting should not be reduced.
Characterization of the Proteases in the Crude Mite Extract
Authors: Ino Yoshitaka , Ando Thoru , Haida Michiko , Nakamura Kazunori , Iwaki Masahiro , Okudaira Hirokazu , Miyamoto Terumasa
Published: August 2009

Characterization of the proteases was performed in the crude mite extract fractionated by Sephacryl S-200 gel filtration. Three peaks of protease activities were detected in the fractions. From the results of substrate specificity and susceptibility to the inhibitors, PK.1 protease (about 60 kD) is suggested to be a trypsin-like protease of mites. From the results of susceptibility to various agents, PK.2 (about 30 kD) and PK.3 (about 20 kD) proteases may be cysteine proteases, e.g., papain and cathepsin B. PK.3 protease existed in the precipitate of 60% ammonium sulfate fractionation. The data in the present study suggest the possibility that Dermatophagoides farinae I allergen might be a cysteine protease probably derived from the gastrointestinal tract of the house dust mite.
Pigeon Breeder’s Disease: The Effect of Antigen-Specific Antibodies on in vitro T Cell Responses of Normal Blood Lymphocytes
Authors: Bouic P.J. , de Beer P.M. , Daniels J. , Joubert J.R.
Published: August 2009

The effect(s) of affinity-purified antibodies to pigeon serum immunoglobulins on normal T cell responses in vitro was measured. Our results show that the antibodies partially inhibited the concanavalin-A-induced proliferation of normal lymphocytes (32% at 10 μg/ml concentration; p < 0.001), whilst they had no significant effects at equivalent doses on the phytohemagglutinin-induced response. A study of the kinetics of this inhibition revealed that the antibodies exerted their effect(s) within the first 24 h of culture (p < 0.001); this is probably due to their interference in early events intimately involved in the de novo synthesis and expression of activation antigens such as HLA-DR and Tac: antibody-treated cultures expressed 51.6 and 41.4% less HLA-DR and Tac, respectively. Two-colour immunofluorescence analysis showed that the specific antibodies bind to a subset of CD8+ cells only.
An Immunopharmacological Analysis of Adrenaline-Induced Suppression of Human Natural Killer Cell Cytotoxicity
Authors: Hellstrand Kristoffer , Hermodsson Svante
Published: August 2009

The circulating catecholamine adrenaline effectively suppressed human natural killer cell cytotoxicity (NKCC) when added to mixtures of effector lymphocytes and 51Cr-labelled target cells in a 4-hour 51Cr release assay in vitro. The effect was mimicked by the β2-receptor agonist terbutaline but not by the β1-receptor agonist prenalterol or the α1/α2-receptor agonist clonidine. Adrenaline-induced NKCC suppression was completely and potently antagonized by the mixed β1/β2-receptor antagonist propranolol and the selective β2-receptor antagonist ICI 118,551 but not by the β1-selective antagonist metoprolol. By comparing the adrenaline sensitivity of high-density (HD) and low-density (LD) lymphocytes, fractionated by Percoll density gradient centrifugation, we found that HD cells appeared more sensitive to adrenaline-induced suppression than LD cells. In both types of effector cells, adrenaline significantly suppressed NKCC at a final concentration of 10–11 M. Pretreatment of LD effector cells with IFN-α reduced the NKCC suppression by subsequent adrenaline treatment. Pretreatment with recombinant IL-2 virtually abolished the response to adrenaline. This effect was noted also when IL-2 and adrenaline were incubated simultaneously during the 4-hour 51Cr release assay. Our data suggest a role for adrenaline, via lymphocyte β2-receptor activation, in the regulation of natural killer cells.
Allergenic Fragments of Ryegrass (Lolium perenne) Pollen Allergen Lol p IV
Authors: Jaggi K.S. , Ekramoddoullah A.K.M. , Kisil F.T.
Published: August 2009

To facilitate studies on establishing the nature of structure/function relationships of allergens, ryegrass pollen allergen, Lol p IV, was cleaved into smaller fragments by cyanogen bromide (CNBr) and the resulting peptides were further digested with trypsin. The resulting peptides were then fractionated by high performance liquid chromatography (HPLC) on a C-18 reverse phase column. The allergenic activity of the HPLC fractions was evaluated in terms of their ability to inhibit the binding of 125l-Lol p IV to serum IgE antibodies of a grass-allergic patient. Many of these fractions inhibited the binding between the native allergen and IgE antibodies in a dose-dependent manner. The inhibitions were specific, i.e., the fractions did not inhibit the binding between 125l-Lol p I (a group-I ryegrass pollen allergen) and the IgE antibodies present in the allergic human serum. The possibility that the allergenic peptide fractions were contaminated by the native undegraded allergen, which might have accounted for the observed inhibition, was ruled out by the fact that the native allergen could not be detected by SDS-PAGE and the elution profiles of allergenically active peptides did not coincide with that of native allergen. One of the allergenic sites recognized by monoclonal antibody (Mab) 90, i.e., site A, was located in HPLC fractions 90–100 while another allergenic site B (recognized by Mab 12) appeared to be lost following the sequential digestion of Lol p IV with CNBr and trypsin.
Production of Tumor Necrosis Factor/Cachectin by Peripheral Blood Mononuclear Cells in Patients with Systemic Lupus erythematosus
Authors: Malavé Inés , Searles R.P. , Montano Jean , Williams Jr. R.C.
Published: August 2009

Levels of tumor necrosis factor/cachectin (TNFα) assessed by ELISA were similar in the supernatant of cultures from peripheral blood mononuclear cells (PBMC) from either active or inactive systemic lupus erythematosus (SLE) patients and controls stimulated with mitogen alone. When PBMC were stimulated with mitogen plus phorbol 12-myristate-13-acetate (PMA), the amount of TNFα was significantly decreased in culture supernatants from active patients or from the entire group of SLE patients studied. However, spontaneous synthesis of TNFα in nonstimulated cultures was increased in the SLE patients. Interferon-γ (IFN-γ) enhanced TNFα synthesis in cultures from either SLE patients or controls stimulated by concanavalin A (Con A) alone, but depressed the high production of TNFα by normal PBMC stimulated with Con A plus PMA. IFN-γ enhanced TNFα production in response to Con A plus PMA in 2 of 3 SLE patients.
Actions of the Protease Inhibitor Phenylmethylsulfonyl Fluoride on Neutrophil Granule Enzyme Secretion and Superoxide Production Induced by fMet-Leu-Phe and Phorbol 12-Myristate-13-Acetate
Authors: Gomez-Cambronero Julian , Mege Jean-Louis , Molski Thaddeus F.P. , Naccache Paul H. , Becker Elmer L. , Sha’afi Ramadan I.
Published: August 2009

The protease inhibitor, phenylmethylsulfonyl fluoride inhibits granule enzyme release and, above 1 mM, superoxide production from rabbit peritoneal neutrophils induced by the chemotactic peptide, fMet-Leu-Phe. At concentrations below 1 mM, it enhances superoxide production. Superoxide generation stimulated by phorbol 12-myristate-13-acetate is increased by phenylmethylsulfonyl fluoride at all concentrations studied. Phenylmethylsulfonyl fluoride has no effect on the rise in intracellular calcium or the depolarization induced by fMet-Leu-Phe but does decrease the extent of repolarization and abolishes hyperpolarization. It depresses actin polymerization and abolishes cytoplasmic alkalinization caused by fMet-Leu-Phe. The increased phosphorylation induced by phorbol 12-myristate-13-acetate in four of the five proteins studied was not affected by phenylmethylsulfonyl fluoride, but the increased phosphorylation of the fifth, a 21-kD protein was enhanced. We conclude that phenylmethylsulfonyl fluoride acts on inhibitory and enhancing processes or steps induced by fMet-Leu-Phe which are subsequent to or independent of calcium mobilization and protein kinase C activity.
Recombinant Interleukin-1 Triggers the Increase of Circulating Fibronectin Level in Rats
Authors: Hagiwara Toshiyuki , Kono Ichiro , Nemoto Kazuo , Kashiwagi Heihachiro , Onozaki Kikuo
Published: August 2009

Interleukin-1 (IL-1) is one of the mediators responsible for the acute phase protein response during inflammation. We evaluated the effect of recombinant IL-1α given intra-abdominally on rat circulating fibronectin (Fn) level. Circulating Fn level, sampled between 3 h and 7 days after injection, was maximum at 24–48 h in Wistar rats treated with IL-1 at doses of 10 U and 100 U/100 g body weight (BW), respectively. Circulating Fn level in rats injected with 100 U/100 g BW IL-1 was higher than that of rats given 10 U/100 g BW IL-1. Circulating Fn level peaked at 24 h, whereas albumin levels decreased by 10% at this time. When frozen tissue sections of rat liver were examined using anti-Fn antibodies, the immuno stainings were denser in IL-1 -treated animals than in controls. These findings suggest that IL-1 triggers plasma Fn production by the liver.
In vivo Arming of Cutaneous Mast Cell Receptors by IgE Released from Macrophages
Authors: Holt P.G. , Bilyk N. , Vines J. , Turner K.J. , Thomas W.R.
Published: August 2009

Intracutaneous injection of purified peritoneal macrophages harvested from ovalbumin (OVA)-hypersensitive high-IgE-responder BN rats into naive animals sensitised the injection sites for subsequent OVA-specific passive cutaneous anaphylaxis (PCA) reactions. The underlying mechanism(s) were investigated using a macrophage cell line (WEHI 265.1), which exhibited comparable sensitising activity in rat or mouse skin, after initial pulsing in vitro with antiserum rich in OVA-specific IgE. Transfer of OVA-hypersensitivity by the cell line (1) was IgE-dependent and did not occur when the cells were pre-exposed to antiserum containing OVA-specific IgG alone, (2) was blockable by saturation of cell surface receptors in the recipient with myeloma IgE (but not myeloma IgG), and (3) did not occur in mast cell-deficient mice carrying the W/Wv mutation, in contrast to their normal heterozygous littermates which developed marked OVA-hypersensitivity at the injection site. These results are consistent with arming of IgE-receptors on cutaneous mast cells by IgE antibody released from macrophages, and hint at a possible role for phagocytes in amplifying IgE-mediated reactions in tissues.