bluuebunny/crossref_metadata_2025 huggingface dataset:
Aspirin-Sensitive Asthma: Abnormal Platelet Response to Drugs Inducing Asthmatic Attacks
Authors: Ameisen Jean Claude , Capron André , Joseph Michel , Maclouf Jacques , Vorng Han , Pancré Véronique , Fournier Eric , Wallaert Benoît , Tonnel André Bernard
Published: August 2009

The pathogenesis of aspirin-sensitive asthma remains unknown. Using a new model of platelet activation, initially described as a response of platelets to IgE antibody-dependent stimuli, this study was designed to test the hypothesis of a possible involvement of platelets in aspirin-sensitive asthma. Washed platelets from 35 aspirin-sensitive asthmatics showed an abnormal in vitro response to cyclooxygenase inhibiting nonsteroidal anti-inflammatory drugs (NSAIDs) – aspirin, indomethacin or flurbiprofen – characterized by the generation of a cytocidal supernatant and (14 patients explored) a burst of chemiluminescence; these drugs had no similar effect on platelets from 31 controls (p <0.0001). It was shown that the abnormal platelet response to NSAIDs was not mediated by IgE. In contrast to platelets, aspirin-sensitive asthmatic leukocytes generated neither cytocidal factors nor chemiluminescence in the presence of NSAIDs. Sodium salicylate and salicylamide, which, though structurally similar to aspirin, do not inhibit cyclooxygenase and are well tolerated by aspirin-sensitive asthmatics, did not activate their platelets to release cytocidal factors. Moreover, preincubation of platelets with sodium salicylate, salicylamide or prostaglandin endoperoxide PGH2, highly prevented their abnormal response to NSAIDs (>80%; p <0.0001). Since several lipoxygenase inhibitors (NDGA, esculetin), including inhibitors of both cyclooxygenase and lipoxygenase (ETYA, BW755C), did not activate patient platelets and prevented the subsequent abnormal response to NSAIDs, it is suggested that the abnormal platelet activation by NSAIDs is not only the consequence of an inhibition of cyclooxygenase, but also involves generation of lipoxygenase metabolites of arachidonate. Besides, platelets from 4 aspirin-sensitive asthmatics undergoing aspirin desensitization were found to have completely lost their abnormal responsiveness to NSAIDs. These findings represent the first identification in aspirin-intolerant asthmatics of a specific abnormal cellular response to drugs inducing asthmatic attacks and open new perspectives into the pathogenesis, prevention and diagnosis of this disease. They also provide support to the concept of a role for platelets in asthma.
Synergism between Gammaglobulin Prophylaxis and Penicillin Treatment in Experimental Post-Splenectomy Sepsis in the Rat
Authors: Offenbartl Karsten S. , Christensen Poul , Gullstrand Per , Prellner Karin
Published: August 2009

In spite of long-term antibiotic prophylaxis and pneumococcal vaccination, there still exists a proportion of highly susceptible splenectomized or functionally hyposplenic patients at risk of contracting fatal overwhelming infections. We have studied the effect of gammaglobulin prophylaxis in experimental sepsis among splenectomized rats. Administration of 37.5 mg human gammaglobulin/kg body weight 24 h before challenge with 103 pneumococci resulted in the survival of 19 of 24 rats, in contrast to 1 of 24 controls. A dose of 19 mg/kg body weight was not protective (7 of 23 survived). However, treatment with penicillin 18 h after challenge in the gammaglobulin-pretreated group of animals saved 21 of 24 animals, although penicillin without gammaglobulin prophylaxis showed no effect. These data indicate that even relatively low circulating concentrations of specific antibody after gammaglobulin prophylaxis might nonetheless be adequate to render septic disease easier to treat.
Studies on Monovalent Anaphylactogens with Carbohydrate Auxiliary Groups: Exclusion of an Artefact
Authors: Guenin R. , Schneider C.H.
Published: August 2009

N1-DNCP-N6-lactobionoy1-1,6-hexanediamine is an elicitor of anaphylactic reactions in the guinea pig passively sensitized by antisera raised with a DNCP-bovine gammaglobulin conjugate. If the antisera contained anticarbohydrate antibodies against the carbohydrate residues of the globulin, significantly cross-reacting with the lactobionoyl moiety, anaphylactogenic cell triggering would be by classical hetero-specific bridging which involved the DNCP- and the lactobionoyl residues as ligands. This possibility was excluded by showing that a bis-lactobionoyl conjugate is unable to elicit anaphylactic reactions.
Human Antibodies against Formaldehyde-Human Serum Albumin Conjugates or Human Serum Albumin in Individuals Exposed to Formaldehyde
Authors: Patterson Roy , Pateras Vincent , Grammer Leslie C. , Harris Kathleen E.
Published: August 2009

Sera from patients undergoing hemodialysis with formaldehyde (F)-sterilized dialyzers were studied to determine if antibodies against F conjugated to human serum albumin (HSA) could be detected. F-human serum albumin (F-HSA) conjugates were prepared using ratios of F to HSA that did not precipitate the HSA. The F-HSA conjugates migrate differently electrophoretically than HSA with an increased negative charge of F-HSA as compared with HSA. The F-HSA was used in an ELISA. The results demonstrated that in certain sera, IgG, IgM, IgA and IgE antibodies against F-HSA could be measured. In the highest titered sera, it was shown that the IgG antibody was not directed against F alone or F-lysine but against an antigenic grouping of F-HSA. No correlation of either IgG or IgE antibodies with immune complex or allergic reactions was found in this series of dialysis patients. Some sera from dialysis patients had antibody activity against HSA. Sera from 2 physicians with rhinitis after F exposure had no antibody activity against F-HSA or HSA. Two nurses with a history of F-induced asthma had no IgG antibodies but did have IgE antibodies against F-HSA and HSA. This spectrum of immunologic responses is analogous to responses in dogs immunized with F or F dog albumin. We have not been able to identify anti HSA antibodies in patients reactive to other hapten-HSA compounds and it is suggested that anti HSA antibodies in F-exposed humans may relate to the F exposure.
Further Studies on the Ability of Different Metal Salts to Influence the DNA Synthesis of Human Lymphoid Cells
Authors: Nordlind K.
Published: August 2009

In a further study on the ability of different metal salts to influence the DNA synthesis of human lymphoid cells, aluminum chloride, beryllium chloride, cadmium chloride, cupric sulfate, ferric chloride, manganese chloride, palladium chloride, platinum chloride and silver nitrate, were tested regarding effect on thymocytes and peripheral blood lymphocytes in children. At certain concentrations in the range of 10––4–10––5M, all tested compounds but aluminum chloride and ferric chloride, were inhibitory, the latter compounds inhibited at 4.8 × 10––3M. A slight stimulation mainly on the thymocytes was obtained with beryllium chloride, cadmium chloride, palladium chloride, platinum chloride and silver nitrate, at certain concentrations in the range of 10––5–10––6M, while ferric chloride gave a slight stimulation at 1.2 × 10––3M. Thus, the tested metal salts should be suitable for use in lymphocyte transformation tests for diagnosis of contact allergy.
Modulation of Adjuvant-Enhanced Delayed-Type Hypersensitivity by the Interferon Inducers Poly I:C and Newcastle Disease Virus
Authors: Kraaijeveld C.A. , Kamphuis W. , Benaissa-Trouw B.J. , Harmsen M. , Snippe H.
Published: August 2009

The modulation by the interferon (IFN) inducers poly I:C and Newcastle disease virus (NDV) of the effector phase of adjuvant-enhanced delayed-type hypersensitivity (DH) was studied in mice. A strongly enhanced DH was induced in mice to ultraviolet (UV) light inactivated Semliki forest virus (SFV) by the use of the adjuvant dimethyldioctadecylammonium bromide. At day 6 after intracutaneous immunization, DH was elicited with SFV and measured 24 and 48 h later as increase in footpad thickness (footpad swelling test). Systemic, intravenous administration of either poly I:C, UV-inactivated NDV, or NDV-induced IFN prior to elicitation of DH with antigen resulted in a temporarily suppressed DH reaction. Both the poor swelling at 3 h and strong swelling at 24 h were suppressed, while the swelling at 48 h was enhanced. The model described provides a sensitive in vivo method to study modulating effects of drugs and microbial agents on the effector phase of DH.
IgE Responses of Wistar Rats to Schistosoma mansoni Infections
Authors: Reiner G. , Zahner H.
Published: August 2009

Using the Wistar rat as experimental animal for Schistosoma mansoni infections it was shown that reaginic antibody response to S. mansoni antigens depends on the infection dose in a way that low infection doses induce persistent titres whereas high doses provoke transient titres. Repeated infections strongly favoured the reagins. In contrast, total serum IgE levels measured by ELISA followed a rather constant time course, independent from the mode of infection. Antigen-specific IgE antibodies followed none of the time courses and reached maximum levels about 12–15 weeks post infectionem. The data allow to suppose that different types of IgE occurred possessing different capacities to sensitize mast cells.
Antigenic and Allergenic Determinants of Ovalbumin
Authors: Elsayed S. , Hammer A.S.E. , Kalvenes M.B. , Florvaag E. , Apold J. , Vik H.
Published: August 2009

The effects of enzymic cleavage and perturbing the conformation of the allergenic and antigenic determinants of hens egg white albumin (OA) were examined. Hens egg white extract of a total protein concentration 8.43 g/l was prepared. Isoelectric focusing in sodium docedyl sulfate and polyacrylamide gel peptide maps for the crude egg white extract showed 26 spots visualized by staining with Coomassie blue. The OA was purified using a TSK-2000 gel filtration chromatography column. The specific allergenic reactivity of the purified·A as measured by RAST inhibition and direct RAST was relatively high: 3 μg gave an inhibition of approximately 10%.The cleavage of OA with cyanogen bromide resulted in 4 fractions, all capable of binding specific IgE with the first peak showing the highest inhibition. Thermal denaturation of OA had no direct effect on the antigenic reactivity. RAST inhibition values for the denatured protein were similar to those of the native protein. Carboxymethylation of OA gave a product with only 20% of the inhibition reactivity. Further treatment with trypsin did not abolish the allergenic and antigenic reactivities as shown by RAST inhibition and by deflection of OA line in rocket line immunoelectrophoresis. On the other hand, limited pepsin hydrolysis destroyed the antigenic structure of the molecule. The reactivity of OA is thus relatively stable and could easily be retained making it possible to identify the allergenic determinants of enzymic hydrolysates used for elucidating the antigenic structure of the molecule.
The Effect of Some Immunomodulating Agents on the Sensitization Phase of Experimental Contact Allergic Reactions
Authors: Anderson Chris
Published: August 2009

In an experimental model which allows comparison of the macroscopic changes with the differentiated dermal inflammatory cell infiltrate of the allergic contact reaction to oxazolone in guinea pigs, the effects of single doses of the cytostatic agents cyclophosphamide, methotrexate and azathioprine and the anti-lymphocyte agent cyclosporin A administered prior to sensitization were studied. All the agents tested increased the degree of erythema and edema of reactions compared to controls. The dermal inflammatory mononuclear cell response showed little significant increase and would not seem to explain the enhancement of reactions. The number of basophils in reactions was not significantly different from controls, and there is no evidence for early arrival, degranulation and destruction of basophils prior to our first routine assessment at 24 h. The number of mast cells in the reactions was also unchanged. A role for the latter two cells involving degranulation without changes in the number of cells could provide a possible explanation for the augmentation of the vascular response. A mechanism for the effect of cyclophosphamide other than T-suppressor lymphocyte susceptibility based on selective cytostatic effects on precursor cells may need to be considered.
Assessment of Skin Reactivity and Specific Antibody to Egg and Milk Proteins in Patients with Atopic Dermatitis: Comparison with Responses to the House Dust Mite Antigen
Authors: Rowntree Stephen , Mitchell Bruce , Platts-Mills Thomas A.E.
Published: August 2009

Sixty patients with atopic dermatitis attending an allergy clinic were assessed for evidence of skin sensitivity and serum antibodies to egg and milk proteins. Prick skin test responses to egg were found in 23 patients and in 74% of these positive egg radioallergosorbent test (RAST) was demonstrable. Positive prick test for milk were present in 10 patients, but only 30% gave a positive milk RAST. Quantitative intradermal skin testing, RAST, and a double antibody antigen binding radioimmunoassay confirmed the presence of IgE antibody to egg proteins but indicated that the levels were very low when compared to those seen to the house dust mite antigen in sensitive patients. In contrast, IgG antibody to purified egg and milk proteins was present in large amount in most patients, the levels being significantly higher than in non-allergic controls.
Selective Enzyme Staining Procedures for Characterization of Mycobacterial Immunoprecipitates
Authors: Öhman Ronny , Ridell Malin
Published: August 2009

Selective staining procedures for four different enzymes (malate dehydrogenase, glutamic oxaloacetic transaminase, leucine aminopeptidase and glucose phosphate isomerase) in combination with two-dimensional immunoelectrophoresis were successfully applied to the analysis of antigen preparations from mycobacteria. Thus, a precipitate corresponding to malate dehydrogenase could e.g. be demonstrated in multilinear precipitation patterns of Mycobacterium avium, Mycobacteríum bovis BCG, Mycobacterium intracellulare, Mycobacterium kansasii, Mycobacterium marinum and Mycobacterium phlei. The analytical approach described can be advantageously used for identification of mycobacterial precipitinogens of enzyme character.
Cyclosporin A-Induced Suppression of Ongoing IgE Antibody Formation in the Mouse
Authors: Okudaira Hirokazu , Sakurai Yoshinori , Terada Kyoko , Terada Eiji , Ogita Tadaatsu , Miyamoto Terumasa
Published: August 2009

Persistent anti-ovalbumin (OA) IgE antibody formation in the mouse was suppressed by oral administration of cyclosporin A (Cy A). Spontaneous anti-OA IgE antibody formation in vitro was also suppressed by Cy A added to the culture. Anti-OA IgG antibody responses in vivo and in vitro were less affected by Cy A. Cy A-induced immunosuppression was T cell-dependent since removal of T cells from the immune spleen cell suspension abolished the Cy A-induced suppression of antibody formation. Supplementing normal spleen T cells resulted in recovery of Cy A-induced suppression of spontaneous antibody formation in vitro. Cy A-induced suppressor T cells carried both Lyt 1 and Lyt 2 surface markers.
P Cell Stimulating Factor Release: A Useful Assay of T Cell Activation in vitro
Authors: Fazekas de St. Groth Barbara , Thomas W.R. , McKimm-Breschkin Jennifer L. , Clark-Lewis Ian , Schrader John W. , Miller J.F.A.P.
Published: August 2009

The coordinate production of multiple lymphokines by activated T cells allows the investigator a choice of assays for antigen recognition in vitro. In this paper, we examine the production of P cell stimulating factor (PSF, a lymphokine identical to interleukin 3) by L3T4+ Lyt-2–– T cell clones. PSF production is antigen-specific, la-restricted and inhibited by a monoclonal antibody to L3T4. PSF, interferon-γ (IFN-γ) and interleukin 2 (IL 2) are all rapidly secreted after antigen or mitogen stimulation. The measurement of PSF avoids problems inherent in the IL 2 and IFN-γ assays, namely absorption of IL 2 by the producer T cells and failure of the interferon assay to distinguish between interferons of the α, β and γ classes.
Low Molecular Weight Eosinophil Chemotactic Factor (ECF) in the Serum of Murine Schistosomiasis Japonica
Authors: Owhashi Makoto , Horii Yoichro , Ishii Akira , Nawa Yukifumi
Published: August 2009

Eosinophil chemotactic activity was detected in the serum obtained at an acute stage of murine schistosomiasis japonica. Gel filtration of the dialyzable fraction of the serum on Sephadex G25 showed that the chemotactic component had an apparent molecular weight of less than 1,000. It was stable to heating, but was sensitive to pronase or carboxypeptidase A digestions, indicating its peptide nature. Eosinophil chemotactic activity of the dialysate of the serum from mast cell-deficient W/Wv mice infected with Schistosoma japonicum was far less than that from normal littermate +/+ mice, although the titers of specific IgE antibody to soluble egg antigen in the serum measured by passive cutaneous anaphylaxis was comparable between them. These results suggest that at least some part of low molecular weight ECF in the circulation seems to be a ECF-A derived from mast cells. Possible biological significance of circulating ECF in schistosomiasis has been discussed.
T Cell Subsets in Idiopathic Glomerulonephritis
Authors: Patel Raman , Connor Grace , Patel Derek R. , Soriano Agnes , Najera Daniel
Published: August 2009

Determination of T lymphocyte subsets using monoclonal antibodies revealed a deficiency of suppressor cells and an elevated helper:suppressor T cell ratio in 7 patients with untreated idiopathic immune complex glomerulonephritis and nephrotic syndrome. Possible pathogenetic mechanisms of immunoregulatory cell imbalance and their implications specifically in reference to idiopathic glomerulonephritis are discussed.
A Genetic-Epidemiologic Study of Human Immune Responsiveness to Allergens in an Industrial Population
Authors: Freidhoff Linda R. , Meyers Deborah A. , Marsh David G.
Published: August 2009

We quantitated puncture skin-test responses to 6 standardized, high-quality-inhalant allergen extracts in 320 adults – a stratified random sample, equally distributed between the sexes and among all decades between 20 and 60 years. For each subject, the ‘average’ IgE-mediated skin sensitivity to the entire panel of allergens (Allergy Index) was calculated. We evaluated correlations between log[total serum IgE] levels and Allergy Index versus quantitative measures of several environmental variables, including (a) infection in childhood, (b) residential history and (c) smoking history. Age, sex and month bled were also included in these analyses. Although many significant intercorrelations were found among these variables, subsequent analyses by stepwise multiple regression showed (a) that log[total IgE], age and smoking history were the best predictors of the Allergy Index and (b) that the Allergy Index and sex were the best predictors of log[total IgE]. The negative relationship between smoking and skin-test responsiveness may reflect a tendency for allergic people to avoid the irritative effects of smoking on respiration. Our analyses, which simultaneously evaluated the effects of age, sex, Allergy Index and smoking history on log[total IgE] levels, did not support other studies which concluded that smoking significantly increases total serum IgE levels.
Antigenic Competition in IgE Antibody Production
Authors: Soares Maria Fernanda M. , Soares M. , de Macedo Mahasti S. , Perini Adenir , Mota Ivan
Published: August 2009

An adoptive cell transfer system was utilized to evaluate the site of action of the suppressive mechanism involved in antigenic competition in IgE antibody production. Carrier-primed (OA) and hapten-primed (DNP-KLH) spleen cells were transferred to syngeneic irradiated recipients that were challenged with a heterologous conjugate (DNP-OA). To study the effect of antigenic competition on T and B cells, donor mice of one or the other cell type received in addition the competitor antigen (Asc) at immunization. The adoptive secondary IgE anti-DNP antibody response was suppressed in both situations. This effect could not be attributed to transfer of Asc-primed cells. Irradiation of donor mice before immunization with the two antigens abrogated the suppressive effect. These results indicate that both Th and B cells primed to the test antigen were affected by antigenic competition.
Characterisation of Cyclosporine-Induced Suppressor Cells in Murine Delayed-Type Hypersensitivity Responses
Authors: Mirisklavos A. , Mottram P.L. , Dumble L.J. , Clunie G.J.A.
Published: August 2009

The effect of Cyclosporine A (CsA) on T cell-mediated delayed-type hypersensitivity (DTH) reactions to murine alloantigens was analysed by transferring spleen cells from sensitised and suppressed mice into irradiated naive recipients. Selective removal of Thy1+, Ly1+ or Ly2+ cells prior to intravenous transfer of cells from alloantigen-sensitised mice showed that Ly1+ Thy1+ cells transferred sensitivity. Concomitant treatment of mice with alloantigen and CsA suppressed the DTH response to alloantigens, and spleen cells transferred from these suppressed mice abrogated the response of sensitised cells transferred at the same time. The suppressor cells were strain-restricted and antigen-specific with the surface phenotype Thy + , Ly1–– Ly2+.
Effects of H1 and H2 Receptor Agonists on Nonspecific Proliferative Response of Human Peripheral Blood Lymphocytes
Authors: Beaulieu Lucie , Beaudoin Jacqueline , Jobin Marie , Hébert Jacques
Published: August 2009

Histamine is known to modulate immune responses through the induction of suppressor cell subsets. The inhibition studies with antagonists suggest that the H2 agonist accounts for most of the suppression. This work studies the effects of various concentrations of 3-methyl histamine (as negative control), histamine, and pure H1 (2-methyl histamine, 2-pyridyl ethylamine) and H2 (4-methyl histamine, dimaprit) receptor agonists on the mitogenic (phytohemagglutinin A) proliferative response of normal human lymphocytes. At high concentrations of agonists (10––3, 10––4M) the suppression induced by the two types of agonists is comparable to that of histamine. At lower concentrations (10––6M) the suppression is seen only in the presence of the H2 agonist. The suppression induced by the two agonists is generally reversed in the presence of an H2 receptor antagonist. The H1 receptor antagonist did not abolish and even increased the suppression induced by histamine and the two agonists.
Effect of TMB-8 on Histamine Release from Isolated Rat Mast Cells
Authors: Grosman Nina
Published: August 2009

TMB-8 was capable of complete inhibition of the histamine release induced by antigen, compound 48/80, and the ionophore A23187 (2 μM). The effective concentrations for 50% inhibition (IC50) were 1.2 × 10––4M for antigen, 1.6 × 10––4M for compound 48/80 both in the absence and in the presence of calcium, and 0.7 × 10––4M for the ionophore. The inhibitory action was not affected by increased calcium concentration in the medium from 1 to 2 mM The presence of glucose in the medium counteracted the inhibition by TMB-8 and, for low concentrations of the ionophore (0.25 μM), TMB-8 caused a pronounced enhancement of the histamine release. Our results indicate that TMB-8 primarily exerts its effects by interference with oxidative metabolism rather than by affecting the intracellular calcium availability.
Bile and Milk from Cholera Toxin Treated Rats Contain a Hormone-Like Factor Which Inhibits Diarrhea Induced by the Toxin
Authors: Lange Stefan , Lönnroth Ivar
Published: August 2009

Protection against intestinal secretion induced by cholera toxin (CT) was studied in adult and suckling rats. Peroral CT treatment of lactating females protected their suckling offspring against diarrhea. This milk contained a protective antisecretory factor (ASF) as shown by passive peroral or intravenous transfer of the milk to adult, nontreated recipients in which the cholera response was tested in ligated jejunal loops. Bile from CT-treated rats also contained ASF, which was still present 100 days after the last treatment. Although milk and bile from untreated rats contained no ASF, bile from very old untreated rats did. Chemical characterization showed that ASF in bile and milk both had isoelectric points of about 5.0 and molecular weights of about 25,000. These data are similar to those previously obtained for ASF isolated from porcine pituitary glands, and show that the antisecretory activity is unrelated to immunoglobulins.
Limiting Dilution Analysis of Age- and Gender-Related Differences in Autoantibody Production against Bromelain-Modified RBC
Authors: Errington S.L. , Cox K.O.
Published: August 2009

Cultured peritoneal cells from untreated mice, after 3 days of in vitro culture, produce autoantibodies against bromelain-treated isologous erythrocytes. The autoantibody response varies with both age and gender. The effects of age and gender were demonstrated by culturing peritoneal cells using limiting dilution techniques. In neonatal mice there were no precursor cells that differentiated into autoantibody secretors. Cells from female mice gave higher responses than cells from males, and the effect was more pronounced in cells from older mice and cultures to which lipopolysaccharide/dextran sulfate (LPS/DS) had been added. Various cell separation and cell mixing experiments indicated that a non-B-cell, nonadherent cell was involved in the higher autoimmunity detected in the presence of LPS/DS in female and older mice. It is thus possible that low autoimmune responses are due to the absence or unresponsiveness of accessory cells rather than potentially autoimmune B-cells.
In vivo Reduction of Circulating C1q Binding Immune Complexes by Intravenous Gammaglobulin Administration
Authors: Lin Robert Y. , Racis Stan P.
Published: August 2009

Six patients with systemic lupus erythematosus were treated with high-dose intravenous gammaglobulin. Immunological parameters were studied and included solid-phase immune complex determinations, quantitative immunoglobulins G, A, and M, as well as C3 and C4 concentrations. Pretreatment values of circulating immune complex concentrations as measured by either C1q binding or anti-C3 binding assays were elevated in all patients. Posttreatment values showed reductions in all C1q binding immune complexes (p <0.01) and anti-C3 binding immune complexes also decreased in 5 out of 6 patients. These assays are described in detail and were also used to define in vitro interactions between the intravenous gammaglobulin preparation and heat-aggregated IgG or sera containing elevated circulating immune complexes. No reduction of immune complex levels were observed when IgG was incubated in vitro with either heat-aggregated IgG or sera with elevated immune complex concentrations. The duration of the in vivo effect and the patients’ clinical responses are described. These findings show that high-dose intravenous gammaglobulin administration can reduce certain types of immune complexes in patients with elevated levels of these substances.
Cyclophosphamide-Mediated Enhancement of Delayed Hypersensitivity Reactions in the Lung
Authors: Enander I. , Ahlstedt S. , Nygren H.
Published: August 2009

The appearance of mononuclear cells, mast cells and mucous cells in the airways was studied in Balb/c mice in relation to cell-mediated immunity reflected as delayed hypersensitivity (DH). The animals were pretreated with cyclophosphamide (Cy) to increase cell-mediated immunity and to decrease the influence of antibodies. Mice pretreated with Cy and epicutaneously sensitized with picrylchloride (PiCl) had stronger DH reactions as compared with sensitized mice not pretreated with Cy. The Cy treatment decreased the IgG antibody formation after sensitization. The Cy-pretreated, sensitized and challenged mice had increased numbers of mucus-producing cells and to some extent also mononuclear cells in their lungs compared with sensitized and challenged non-Cy-treated animals. However, the mucous cell numbers were also increased in Cy-treated, but nonsensitized mice in which most of the airway epithelium had differentiated into mucus-containing cells. The present results indicate that local cell-mediated immunity involves the appearance of mononuclear cells and mucus-producing cells in the lung. This inflammatory reaction is enhanced by Cy treatment, although mucous cell differentiation seems to be induced by exposure to Cy alone.
Effects of Cyclosporin A, Antilymphocyte Serum and Donor-Specific Transfusions on Murine Delayed-Type Hypersensitivity and Skin Graft Survival
Authors: Mottram P.L. , Mirisklavos A. , Dumble L.J. , Clunie G.J.A.
Published: August 2009

The effect of immunosuppressive reagents cyclosporin A (CsA) and rabbit anti-mouse antilymphocyte serum (ALS) on the response to alloantigens was studied in inbred mouse strains. Alloantigen was given either as a cell suspension which induced a delayed-type hypersensitivity reaction (DTH), or as a full-thickness skin graft. Dose-response studies showed that DTH reactions in CBA mice sensitised to BALB/c cells were reduced to background levels when recipient mice were treated with 100 mg/kg CsA on days 0, 4 and 6 after primary alloantigenic challenge. The response to a second challenge was significantly decreased by CsA treatment during primary or secondary exposure to alloantigen and CsA was as effective as ALS in abrogating both primary and secondary DTH reactions. Survival of full-thickness grafts of BALB/c skin on CBA mice was increased from 9 to 23 days by ALS treatment on days ––1 and +2, with grafts given on day 0. Long-term treatment with CsA, from day ––14 to +12, also prolonged graft survival from 9 to 18 days but donor-specific transfusions, with or without concomitant ALS or CsA treatment, decreased graft survival and often sensitised the recipients. This occurred with transfusions administered from ––63 to ––7 days and on the day of grafting. Thus, in H-2 mismatched mice, both CsA and ALS treatments produced a state of tolerance when administered during short-term exposure to alloantigen. With a stronger antigenic stimulus, in the form of a skin graft, ALS was only partially effective in overcoming the sensitising effects of donor-specific transfusions given prior to grafting and CsA could not prevent sensitisation of the recipient, but the continued presence of the drug delayed the onset of rejection in unsensitised recipients.
Induction of a Protective Immune IgE Response in Rats by Injection of Defined Antigens of Schistosomulum-Released Products: Immunochemical Properties of the Target Antigens
Authors: Damonneville Martine , Auriault Claude , Thorel Thérèse , Verwaerde Claudie , Pierce Annick , Strecker Gérard , Capron André
Published: August 2009

Brown Norway rats were injected without adjuvant with the soluble products liberated in a 16-hour culture by schistosomula (schistosomula-released products, SRP-A). A strong cytotoxic and protective IgE response was elicited, mainly directed against 22- and 26-kilodalton (kDa) SRP-A molecules. In the present study, we have attempted to characterize further those molecules. Metaperiodate denaturing treatment of the SRP-A glycans before injection into rats did not modify the immunogenicity of the SRP-A antigens. Results obtained by lectin affinity suggested that the 22- and 26-kDa molecules were glycoconjugates binding to ConA. Preparative sodium dodecylsulfate electrophoresis has allowed the separation of enriched fractions of 22- and 26-kDa molecules which have been injected separately into rats. The corresponding sera were tested in antibody-dependent cell cytotoxicity and displayed a significant cytotoxic IgE response (65 and 53%, respectively) towards the larvae. These results lend further support to the view that the 22- and 26-kDa antigens are the major targets of the protective IgE response and thus appear as potentially protective antigens.
Regulation of in vivo IgE Biosynthesis in Mice with Complete Freund’s Adjuvant
Authors: Smith W.G. , Butchko G.M.
Published: August 2009

Complete Freund’s adjuvant (CFA) administered before sensitization dampened the normal and cyclophosphamide-enhanced response of high and moderate IgE responder phenotype mice (CAF1 and C57Bl/6J, respectively). CFA-induced suppression of IgE biosynthesis was effective in reducing anaphylactic histamine release from ∼2,900 ng histamine per milliliter to background levels (<100 ng/ml). CFA-induced ascites fluid was able to reduce the cyclophosphamide-enhanced IgE response of low-responder phenotype SJL mice from 1:320 to <1:5 as determined by passive cutaneous anaphylaxis. Muramyl dipeptide, a mycobacterial cell wall component capable of eliciting effects similar to those seen with CFA, was shown to induce suppression of IgE production if incorporated in incomplete Freund’s adjuvant. Muramyl dipeptide administered in saline was ineffective, while incomplete Freund’s adjuvant alone had some immunoregulatory properties. Ongoing IgE responses were less susceptible to regulation. CFA administered to sensitized C57Bl/6J mice was ineffective in inducing IgE suppression when animals were challenged with antigen.
The Excretion of IgE with Feces from Healthy Individuals and from Others with Allergy and Diseases Affecting the Intestinal Tract
Authors: Kolmannskog Svein , Florholmen Jon , Flœgstad Trond , Kildebo Stein , Haneberg Bjørn
Published: August 2009

In this study we looked for the occurrence of immunoglobulin E (IgE) in feces from healthy individuals and determined the total daily excretion and day-to-day variation in IgE in feces from patients with allergy, as well as the correlation between concentrations of IgE in small samples of feces and the total amounts of IgE in feces collected over a longer period. Concentrations of IgE in extracts of small samples of dry feces correlated well with the total daily amounts of IgE in feces collected over a 3-day period. Thus, single small samples of feces can be used to measure the excretion of IgE with feces at that time. In 3 children, studied over a 5-week period, the IgE excretion varied somewhat from one day to another, but was largely within a certain range of concentrations. Addition of trypsin inhibitor to fresh feces had no influence on the IgE concentrations of the resulting fecal extracts. Less than 10% of 88 presumably healthy infants, children, and adults had detectable IgE in their feces, while 21 of 40 children with various kinds of allergy had measurable fecal IgE. Only 3 of individuals who were suffering from infectious acute gastroenteritis had IgE-positive fecal extracts. This was also the case for 6 of 25 adult patients in clinical remission of ulcerative colitis or Crohn’s disease. Seven of adult patients with chronic pancreatitis had measurable IgE in feces, and the concentrations were up to ten times the upper limit of IgE found in healthy individuals. Gel filtration experiments showed that fecal IgE from a patient with chronic pancreatitis mainly consisted of fragments corresponding to a molecular weight of approximately 40,000 daltons. Thus, the higher levels of IgE in feces from patients with chronic pancreatitis can only in part be explained by reduced destruction by pancreatic enzymes.
Cell Labeling and Sorting by Rosette Formation
Authors: Owen Charles S. , Rothong Carole L.
Published: August 2009

The binding of biotin by avidin has proven to be a useful adjunct to antigen-antibody reactions in a number of immunological labeling systems. This work has applied it to rosette formation, for identification and separation of selected lymphocytes. Three procedures were compared which incorporated the biotin-avidin reaction into a model system for the rosetting of murine T lymphocytes. The reaction was most effective in conjunction with antibodies binding to the red cell, instead of using red cells which were biotinated on endogenous cell surface proteins. Physical separation of these rosettes from unrosetted cells was demonstrated with magnetic filtration, in a preliminary experiment. One interpretation of differences in effectiveness of rosette formation in the three systems tested is in terms of the flexibility of the antibody molecule, as compared with avidin. The principal finding was that, if employed in a multiple-stage ‘sandwich’ labeling system, avidin-biotin binding was fully effective and provided new versatility in the design of such rosetting schemes.
Thymus Independence of Eosinophilia Induced by a Non-Parasite Antigen
Authors: Blomjous F.J.E.M. , Elgersma Anneke , Kruizinga W. , Ruitenberg E.J.
Published: August 2009

Eosinophilia is primarily a T-cell-dependent phenomenon, related to helminthic infections. Using a rat model (+/rnu), it has been shown that keyhole limpet hemocyanin (KLH) in complete Freund’s adjuvant (CFA) induced blood eosinophilia, but only after pretreatment with cyclophosphamide (CY). CY alone caused an eosinophilopenia which was reversed by CFA or KLH-CFA treatment. Eosinophilia was also induced in congenitally athymic rnu/rnu rats after CY pretreatment and KLH-CFA stimulation, the response being of a similar order as that in +/rnu animals. It is concluded that a non-parasite antigen can induce blood eosinophilia after pretreatment with CY, the response being thymus-independent. This model may be appropriate to study the biological significance of the eosinophil response.
Proliferative Responses by Immune Murine Popliteal Lymph Node Cells to High-Molecular-Weight Basic Allergen of Ryegrass Pollen and Its Cyanogen Bromide Fragments
Authors: Ekramoddoullah A.K.M. , Kisil F.T. , Cook R.T. , Sehon A.H.
Published: August 2009

High-molecular-weight basic allergen (HMBA) of ryegrass pollen and its fragmented form obtained by cleavage with cyanogen bromide (CNBr) (HMBA-frag) were evaluated for their immunogenicity in terms of their capacity to stimulate the in vitro proliferative responses of the in vivo antigen-primed popliteal lymph node (PLN) cells. For this purpose, mice (C3H/HeJ) were immunized in the hind foot pads with solutions containing 1–5 μg of HMBA or HMBA-frag emulsified in Freund’s complete adjuvant (FCA). Eleven days later, single-cell suspensions of the PLN were cultured in vitro for 4 days in the presence of either HMBA or HMBA-frag at concentrations ranging from 0.625 to 20 μg/ml. 3H-thymidine was added to the cultures 6 h prior to harvesting the cells. The degree of proliferation was assessed from the extent of intracellular incorporation of the 3H-thymidine by the PLN cells. Comparable degrees of lymphocyte proliferation were consistently obtained on stimulation with either HMBA or HMBA-frag of PLN cells from mice immunized with the allergen HMBA. A similar finding was made using PLN cells from mice immunized with the HMBA-frag. This study demonstrated that the CNBr-derived fragments of HMBA retained the immunogenicity of the parent molecule HMBA in terms of being able to (i) induce the in vivo priming of PLN cells and (ii) elicit the in vitro proliferative response of the antigen-primed PLN cells.
A Pro-Inflammatory Role of Lymphoid Cells in Acute Pleurisy in Rats
Authors: Bechara Gervásio H. , Bates Michael B. , Dunn Colin J. , Willoughby Derek A.
Published: August 2009

The effect of viable splenic lymphoid cells and their constituents (filtrate) on carrageenan-induced acute pleurisy was investigated in rats. Suspensions of lymphoid cells administered intravenously to recipients just prior to initiation of pleurisy enhance both the volume of exudate and cell accumulation in the pleural cavity 3 h after the irritation. Similar results were observed when filtrate of disrupted lymphoid cells was injected either 30 or 5 min before the carrageenan, but not when administered 30 min afterwards. Suspensions of bone marrow cells, on the contrary, were ineffective in producing an enhancement of the parameters studied. When administered into the pleural cavity together with carrageenan, the lymphoid cell filtrate augmented the inflammatory response to the irritant. Nevertheless, it was ineffective, per se, to elicit any local change. It is suggested that lymphoid cells may play a pro-inflammatory role in the initiation of the process by enhancing both the fluid and the cellular components of inflammation.
Complement Activation in Human Lymph: Modulation by the Contact Activation System and by Leukocytes
Authors: Vogt W. , Damerau B. , Lühmann B. , Hesse D. , Haller Y.
Published: August 2009

Complement components, their activation and the generation of C3a and C5a peptides were studied in human lymph used as a model of tissue fluid. Both, classical and alternative pathways could be activated by suitable agents such as immune aggregates or zymosan. C3 activation and C3a formation were marked while only 10–15% of the anyway low amount of C5 were converted during complement activation, yielding very low concentrations of C5a. Carboxypeptidase N activity was present in lymph and converted the peptides to their less (C5a) or not at all (C3a) active desArg derivatives. Contact activation of Hageman factor and kallikrein enhanced activation of the classical pathway up to C3 conversion. The search for additional processes apt to create efficient concentrations of C5a (desArg) in lymph led to the discovery that the presence of leukocytes in lymph greatly increases the release of C5a activity upon complement activation. This suggests a physiological role of leukocytes resident in tissues for the induction of inflammatory reactions.
Lack of Circulating Immune Complexes in Inflammatory Bowel Disease
Authors: Knoflach P. , Vladutiu A.O. , Swierczynska Z. , Weiser M.M. , Albini B.
Published: August 2009

Multi-organ involvement and especially extraintestinal manifestations have suggested an immune complex-mediated pathogenesis of ulcerative colitis and Crohn’s disease. Using various techniques controversial data have been reported on the incidence and levels of circulating immune complexes and their correlation to clinical presentation. Sera of 131 patients with inflammatory bowel disease (78 Crohn’s disease, 53 ulcerative colitis) representing a wide spectrum of disease activity, treatment and presence or absence of extraintestinal manifestations were tested for circulating immune complexes using Raji cell indirect immunofluorescence assay, Raji cell radioimmunoassay, C1q solid phase assay and polyethylene glycol precipitation coupled with measurements of optical density and subsequent immunoelectrophoresis or radial immunodiffusion. Circulating immune complexes in low concentrations were observed in a small number of patients with inflammatory bowel disease, the frequency and concentrations being slightly higher in patients with Crohn’s disease than in those with ulcerative colitis. No association of concentrations of circulating immune complexes with disease activity or presence of extraintestinal manifestations could be demonstrated. These data do not support the claim for a major role of circulating immune complexes in the pathogenesis of inflammatory bowel disease.
Comparative Studies on Tree Pollen Allergens
Authors: Vik H. , Elsayed S.
Published: August 2009

The previously isolated major allergen of birch pollen (fraction BV45), Int. Archs Allergy appl. Immun. 68: 70–78 (1982), was further purified by recycling chromatography. The purified preparation was run on a high-performance liquid chromatography (HPLC) TSK-G-2000 gel filtration chromatography column and, finally, on paper high-volt electrophoresis. The protein recovered met the homogeneity criteria required for performing the N-terminal sequence analysis. The allergenic and antigenic reactivities of the HPLC-purified protein, designated BV45B, was examined. A single homogeneous precipitation line in crossed immunoelectrophoresis (CIE) was shown. Specific IgE-inhibition tests and immuno-autoradiographic prints indicated that this allergen could bind reaginic IgE specificially and with good affinity. The homogeneity of BV45B was examined by isoelectric focusing (IEF). Several minor bands of pI differences of less than 0.1 units were visible, demonstrating the existence of some molecular variants of this protein. The N-terminal sequence analysis of the molecule was performed, and the following four amino acids were tentatively shown by sequential cleavage: NH2-Ala-Gly-Ile-Val-. The demonstration of one dominant N-terminal l-dimethyl-amino-5-naphthalene sulphonyl (DNS)-amino acid by polyamide thin-layer chromatography at each sequence step confirmed that (a) the N-terminal residue of the protein was not blocked; (b) the heterogeneity shown by the IEF system was merely due to the presence of several homologous polymorphic proteins with identical N-terminal amino acid, (c) the adequacy of the purification repertoire used.
Intraepithelial Migration of Nasal Mucosal Mast Cells in Hay Fever
Authors: Enerbäck Lennart , Pipkorn Ulf , Granerus Göran
Published: August 2009

Mast cells were studied by light microscopy in mucosal imprints and in biopsies of nasal mucosa of 12 birch pollen allergic individuals before and during the pollen season, using techniques optimized for the demonstration of mucosal mast cells. We also measured the histamine content of nasal mucosa, whole blood and plasma, and counted the numbers of circulating blood basophils. Before the pollen season the nasal mucosa was found to contain many mast cells located in the mucosal connective tissue stroma, and very few cells with basophilic and metachromatic granules were found in mucosal imprints. During the pollen season there was a redistribution of mast cells into the epithelium, many such cells now being recovered in mucosal imprints. The total number of mucosal mast cells counted in tissue sections did not change significantly with the onset of the pollen season, suggesting a redistribution of mucosal mast cells by migration. Judged by morphologic appearance and naphthol-AS-D chloroacetate esterase activity, the intraepithelial mast cells found in tissue sections had rather the properties of tissue mast cells than of blod basophils, and only a few of the basophilic cells of the imprints had a morphology compatible with blood basophils. The histamine content of the mucosa, as well as histamine levels of whole blood and plasma, and circulating blood basophil numbers did not change significantly in relation to the pollen season. These findings suggest that an intraepithelial migration of mucosal mast cells is part of the allergic mucosal response. This reaction resembles the nematode response of certain rodents and may explain how contact is established between mucosal allergens and effector cells when the allergic reaction is initiated.
Blocking of IgM-Mediated Single Radial Haemolysis by IgA and Non-Complement-Fixing IgM Anti-Type-III Pneumococcal Antibodies
Authors: Kearney Raymond
Published: August 2009

The results reported here provide information on the effects of mouse non-complement-fixing (NCF) IgM anti-type-III pneumococcal polysaccharide (SIII) in blocking single radial haemolysis (SRH) mediated by complement-fixing (CF) IgM or CF hybrid IgM/A anti-SIII antibodies. The results consistently showed that highly avid NCF IgM or IgA anti-SIII antibodies could prevent the detection of CF IgM or CF IgM/A anti-SIII antibodies by SRH. The altered haemolysis produced by NCF IgM and IgA antibodies in the primary response is similar to that reported when the SRH test was used to screen for primary rubella infection. The findings indicate that false-negative results can be due to the early production of NCF IgM antibodies which can block the detection of CF IgM antibodies particularly if the erythrocytes are coated suboptimally with antigen.