Passive Cutaneous Anaphylaxis Test Research Articles

Méthode radio-immunologique de détection des IgE spécifiques à l'alcuronium

A radio-immunoassay (RIA) was used to screen for specific IgE to myorelaxants. Alcuronium was coupled to epoxyactivated Sepharose. Sixteen patients with anaphylaxis to alcuronium (n=2), gallamine (n=2) or suxamethonium (n=12) were studied. The diagnosis was established by intradermal tests (ID), passive cutaneous anaphylaxis tests and human basophil degranulation tests. The amount of non specific label retained by Sepharose-ethanolamine (with sera of patients) and Sepharose-alcuronium (with sera of 11 control subjects) was estimated. The RIA was positive 10/16 (8/14 patients having reacted to a muscle relaxant other than alcuronium). The RIA seemed to be useful in the diagnosis of anaphylaxis to muscle relaxants. Drug-reactive antibodies were specific of the quaternary ammonium radical, which was the common allergenic determinant of all molecules of muscle relaxants. This test accounted for in vitro cross-reactivity, but had no predictive value for the clinical risk of crossed-anaphylaxis. This risk was best assessed by ID; it was positive in three cases. Although it was not possible to compare ID and RIA, the interpretation of which was different, both tests should be recommended for the detection of sensitivity to muscle relaxants.

Toxicity studies of VP 16-213 (VI)--Antigenicity in guinea pigs

VP 16-213 (etoposide, abbr. to VP), an oncostatic drug, and VP-protein conjugates were examined for their antigenic properties using male Hartley guinea pigs. The results of systemic anaphylaxis test, passive cutaneous anaphylaxis test, passive hemagglutination test and Schultz-Dale reaction revealed that VP possessed neither antigenic nor haptenic properties. The dose levels of VP employed in the present experiment were confirmed not to suppress immune reactions.

Immunological evidence of chemotherapeutic action of mebendazole against Ancylostoma ceylanicum (Looss, 1911) in hamsters (Mesocricetus auratus).

The immunological responses in hamsters during treatment with mebendazole against Ancylostoma ceylanicum are studied. Indirect haemagglutination (IHA) counterimmunoelectrophoresis (CIEP), passive cutaneous anaphylaxis (PCA), foot pad swelling for immediate (ITH) and delayed (DTH) types of hypersensitivities were employed for measuring the responses. Serum antibody which was 1:32 before treatment increased to the maximum of 1:512 (control 1:128) on the 10th day and it declined subsequently. The CIEP test was positive for 10 days and then became negative. The PCA test was positive throughout the observation period both in the treated and untreated groups. Foot pad swelling for ITH and DTH responses were comparatively more prominent than in the untreated control up to the 20th day and then both decreased simultaneously. The immunological responses remained prominent for a longer period and decreased more slowly in the untreated control group.

ChemInform Abstract: 1,2,4‐TRIAZOLO(4,3‐A)QUINOXALINE‐1,4‐DIONES AS ANTIALLERGIC AGENTS

AbstractTriazol‐Derivate (VIII) und (IX) erhält man auf dem im Formelschema beschriebenen Weg.

Reaginic and homocytotropic IgG antibodies in Schistosoma mansoni infected Mastomys natalensis: time courses in untreated infections and after chemoprophylactic and chemotherapeutic treatment.

Mastomys natalensis were infected percutaneously with 250 cercariae of Schistosoma mansoni and treated effectively with 5 X 50 mg amoscanate/kg body-weight 12-16 (I), 28-32 (II), 56-60 (III) and 101-105 days p.i. (IV) respectively. Levels of reaginic antibodies (RAb) and homocytotropic IgG antibodies (GAb) were assessed by passive cutaneous anaphylaxis tests (PCA) and compared with those in infected, untreated animals. In untreated animals RAb were first detected in the 6th week p.i. Maximum titres around 1:80 were reached after 100 days. High levels persisted until the end of the experiment 185 days p.i. RAb did not develop after treatment I. After treatment II only low PCA titres occurred. After treatment III the time course was similar to the controls but levels were reduced. Treatment IV did not interfere with the occurrence of specific RAb. In the case of GAb, independent of any treatment, after a first peak of PCA titres in the 5th week either a negative pattern or only very low titres were found in week 7. In untreated infected Mastomys GAb levels increased thereafter to maximum titres of about 1:80 at day 70. Later than 100 days p.i. relatively constant titres were observed. The second rise of antibody levels did not occur after early treatment. After treatment II only low transient titres developed. Animals treated in the early patency (III) showed reduced PCA titres and were negative at the end of the observation period. Treatment given 101 to 105 days p.i. rather enhanced the development of GAb.

Synthesis and antiallergic activity of some quinolinones and imidazoquinolinones.

A group of 1,4-dihydro-4-oxoquinoline-2- and -3-carboxylic acid esters with nitrogen functionality at the 8-position was synthesized, and 6-oxo-6H-imidazo[4,5,1-ij]quinoline-4- and -5-carboxylic acid esters were elaborated from these. Several of the compounds displayed activity in the rat passive cutaneous anaphylaxis (PCA) test for antiallergic activity. However, PCA activity in this series was accompanied by rat toxicity, as measured by a decrease in percent of normal weight gain over a 2-week period, following a single oral dose.

Alteration of Ovalbumin Immunogenic Activity by Glycosylation through Maillard Reaction

The immunogenic properties of glycosylated ovalbumin were investigated by measuring specific IgE and IgG antibody responses in mice. Ovalbumin was glycosylated with glucose through Maillard reaction. The glycosylated protein reacted well with the antibody to native ovalbumin. The glycosylated and native ovalbumins were injected into mice, and serum IgE and IgG responses were determined by the passive cutaneous anaphylaxis (PCA) test and a solid phase enzyme immunoassay (ELISA). The glycosylated ovalbumin induced rather lower responses of both IgE and IgG antibodies than native ovalbumin did, and the antibody directed against glycosylated ovalbumin reacted with native protein. These results suggested that immunogenic properties of proteins could be altered by the glycosylation through Maillard reaction.

IgE ANTIBODY AGAINST HEPATITIS B SURFACE ANTIGEN IN MICE

Hepatitis B surface antigen (HBs antigen) was examined for elicitation of IgE production by injection into mice. The Prausnitz-Küstner (PK)-type skin test in the rat was employed for detection of IgE antibody to HBs antigen, because no sufficient purified HBs antigen was available as the challenging antigen for the passive cutaneous anaphylaxis (PCA) test in rats. The positive PK test was considered to be due to IgE antibody, since the active principle was inactivated by heating the sera at 56 C for 30 min, did not bind to protein A and was eliminated by anti-mouse IgE antisera. These data indicate that the PK-type test in rats can be used for detection of mouse IgE antibody when the amount of a test sample is not sufficient for the PCA test in rats.

Cross reactivity studies with platinum group metal salts in platinum-sensitised rats.

Hooded Lister rats were sensitised to the halide salt of platinum ammonium tetrachloroplatinate(II)[(NH4)2PtCl4] in its conjugated form with ovalbumin. Sensitisation was achieved by intraperitoneal injection with Bordetella pertussis vaccine as adjuvant, followed 21 days later by a further injection in saline. The presence of specific anti-platinum IgE antibody was determined by passive cutaneous anaphylaxis (PCA) and radioallergosorbent test (RAST) using the platinum halide salt conjugated to a heterologous carrier. Sera exhibiting positive reactions were pooled and PCA tests performed on the titrated pooled sera with 3 conjugated platinum group metal salts, 5 platinum group metals in their free salt form and 6 platinum salts with differing ligands. PCA challenges with these compounds resulted in significant cross reactivity between ammonium tetrachloroplatinate(II), ammonium hexachloroplatinate(IV) and the conjugated tetrachloroplatinate. There was very limited cross reactivity with other platinum or platinum group metal salts in either free or conjugated forms. Furthermore, these results were confirmed by RAST inhibition studies.

Biologically active constituents of Centipeda minima: Isolation of a new plenolin ester and the antiallergy activity of sesquiterpene lactones.

Two pseudoguianolide sesquiterpenes were isolated as antiallergic constituents of Centipeda minima, which has been used as a folk medicine in southeast Asia. One of the sesquiterpenes, 6-O-senesioylplenolin (2), is a new pseudoguaianolide sesquiterpene lactone, which was isolated along with known pseudoguaianolide, arnicolide C (1). The two sesquiterpenes exhibited significant antiallergy activity in passive cutaneous anaphylaxis (PCA) tests. An amide isolated along with the sesquiterpenes was identified as aurantiamide acetate (3). Three flavonoids obtained from this plant also showed significant antiallergy activity.

Alteration of ovalbumin immunogenic activity by glycosylation through maillard reaction.

The immunogenic properties of glycosylated ovalbumin were investigated by measuring specific IgE and IgG antibody responses in mice. Ovalbumin was glycosylated with glucose through Maillard reaction. The glycosylated protein reacted well with the antibody to native ovalbumin. The glycosylated and native ovalbumins were injected into mice, and serum IgE and IgG responses were determined by the passive cutaneous anaphylaxis (PCA) test and a solid phase enzyme immunoassay (ELISA). The glycosylated ovalbumin induced rather lower responses of both IgE and IgG antibodies than native ovalbumin did, and the antibody directed against glycosylated ovalbumin reacted with native protein. These results suggested that immunogenic properties of proteins could be altered by the glycosylation through Maillard reaction.

Synthesis of the metabolites and degradation products of 2-amino-7-isopropyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3- carboxylic acid (Amoxanox).

The metabolites and degradation products of 2-amino-7-isopropyl-5-oxo-5H-[1] benzopyrano-[2, 3-b] pyridine-3-carboxylic acid (Amoxanox, AA-673, 1), a promising drug for the treatment of bronchial asthma, were synthesized to confirm the proposed structures and to determine their activity in the rat passive cutaneous anaphylaxis test.

Studies on Alternaria Allergens

Six extracts were prepared from <i>Alternaria tenuis </i>isolates ATCC 6663, ATCC 16086 and DAOM (Agriculture Canada) 183905 grown separately on two synthetic media, revised tobacco and Czapek’s, and their biochemical and immunological properties were examined. High-performance liquid chromatography revealed considerable variations in UV absorbance and carbohydrate profiles among extracts from the different isolates. These differences were less marked among samples of the same isolate cultured on different media. Enzyme screening showed that all extracts contained large amounts of phosphatases and glucosidases and moderate quantities of esterases. Only the <i>α</i>-galactosidase activity showed any correlation with allergenic activity. No significant variation was observed in isoelectric focusing patterns. Extensive antigenic cross-reactivity even between the different isolates was found in precipitin studies. In mouse IgE passive cutaneous anaphylaxis tests, all extracts gave reactions of similar intensity. In direct RAST and RAST inhibition assays, ATCC 16086 grown on revised tobacco medium was found to be the most potent and approached the activity of an extract from a commercial material (B-I). DAOM 183905 grown on either medium was next in potency while ATCC 6663 samples were the least potent. The results indicate that it is possible to obtain extracts of high allergenic potency for standardization purposes from growth of selected <i>A. tenuis </i>isolates on a chemically defined medium.

ChemInform Abstract: SYNTHESIS AND ANTIALLERGY ACTIVITY OF 10‐OXO‐10H‐PYRIDO(1,2‐A)THIENO(3,2‐D)PYRIMIDINES AND 10‐OXO‐10H‐PYRIDO(1,2‐A)THIENO(3,4‐D)PYRIMIDINES

AbstractTetrazolylsubstituierte Carbonsäureamide (III) und Heterocyclen (VI) erhält man direkt aus der Carbonsäure (II) oder über die Derivate (IV) und (V).

Trichinella spiralis: Immunization of rats with an antigen fraction enriched for allergenicity

Trichinella spiralis whole muscle larval extract was fractionated by gel filtration and anion-exchange chromatography, and the protein fractions were assayed for allergenicity by a footpad-swelling test in mice; IgE antibody levels in rats immunized with the fractions were determined by passive cutaneous anaphylaxis test in rats. By these methods, an allergenic fraction from T. spiralis was isolated. The fraction, F1-b, was shown to be monodisperse by analysis with SDS-PAGE, IEP, and isoelectric focusing, indicating that it is a single protein moiety with a molecular weight of approximately 45,000 and a p I of approximately 5.1. The Schiff-periodate test showed Fl-b to be a glycoprotein. Rats immunized with Fl-b had significantly fewer intestinal worms than did nonimmunized controls at 24 hr and 7 days after oral challenge with T. spiralis larvae.

Receptor-specific mediation by immunoglobulin E of antigen-induced contraction of tracheal and lung parenchymal strips isolated from the guinea pig.

The guinea pig is much like humans in the cells and mediators involved in immediate hypersensitivity reactions. However, the major anaphylactic antibody in this species is IgG1, not IgE. Recently, we have been successful in producing IgE antibody in guinea pigs. The current study examined whether guinea pig IgE antibody could mediate pulmonary smooth muscle contraction. IgE antibody to picryl and oxazolone determinants was induced by immunizing Hartley strain guinea pigs pretreated with cyclophosphamide. Hyperimmune serum from these animals was passed through a heavy chain-specific anti-IgG1 affinity column. The presence of IgE anti-hapten antibody in the filtrate fraction was verified by passive cutaneous anaphylaxis (PCA) testing with a 7-d period of local passive sensitization and by heat lability (56 degrees C X 4 h) of PCA activity. This IgE-rich fraction, and purified IgG1 anti-hapten antibody were transferred to normal guinea pigs. Both fractions sensitized trachea and pulmonary parenchyma for antigen-induced smooth muscle contraction. The IgG1-mediated antigen-induced contractile response was not affected by heat (56 degrees C X 4 h) and was inhibited in a dose-dependent fashion by IgG1 blocking antibody (anti-OA). The IgE-mediated antigen-induced contractile response was significantly decreased by heat and was not affected by the anti-OA blocking antibody even at a concentration of 100 mg/kg. Thus, two antigen-specific factors in guinea pig serum can mediate antigen-induced pulmonary smooth muscle contraction: IgG1 and IgE antibodies. Our data also suggests that these antibodies mediate the contractile response through separate receptors. The finding that guinea pig IgE can mediate pulmonary smooth muscle contraction suggests this species can be a model for IgE-mediated events in the lung.

IgG and IgE Antibodies to Hymenolepis nana Detected in Infected Mouse Sera by Gel Diffusion and Passive Cutaneous Anaphylaxis Tests

Specimens deposited: USNM Helminth. Coll. Nos. 77422-77427 (S. sagittifer). Four female specimens of an undetermined species of Neoechinorhynchus were collected from one of three Lisa macrolepis (Perciformes: Mugilidae) examined and one of 12 Pseudorhombus arsius (Pleuronectiformes: Paralichthyidae). They may represent a new species, but available material is not considered adequate for description. Body 7.80-13.20 (11.22) long by 0.48-0.88 (0.65) wide; with 6 dorsal giant subcuticular nuclei and 2 ventral ones. Proboscis 76-80 (79) long by 96106 (102) wide. Hooks in anterior ring rooted; i : el inth. Col . ( . itti er). lateral ones, 54-61 (58) long larger than dorsal or ventral 38-44 (42); hooks in median ring 2635 (32), in posterior ring 26-32 (28) long. Proboscis receptacle 352-420 (396) long by 112-140 (127) wide. Lemnisci 2,240 long by 140 wide. Eggs 26 long by 5 wide. These specimens most closely resemble N. doryphorus Van Cleave and Bangham, 1949, but have larger bodies and smaller proboscides, anterior proboscis hooks, and eggs. Specimens deposited: USNM Helminth. Coll. Nos. 77427, 77428 (Neoechinorhynchus sp.).

Studies on Alternaria Allergens

The biochemical and immunological characteristics of a series of commercial samples of <i>Alternaria tenuis</i> were examined, including samples from two different companies. There was a considerable variation in the biochemical parameters such as the gel filtration profile on Sephadex G-100, isoelectric focussing pattern and immunodiffusion precipitin band pattern. The content and proportion of two high molecular weight polysaccharides, one a galactomannan and the other a glucan, also varied greatly. In contrast, the allergenic properties of the samples were more consistent. In rat IgE passive cutaneous anaphylaxis tests, extensive cross-reactivity was observed among batches from the same company, but this cross-reactivity was weaker when batches from the two companies were compared. In direct radioallergosorbent tests (RAST) the samples showed similar reactivities, but RAST inhibition tests showed a 40-fold range in activity between samples from one company and 55-fold between batches from the two sources. It was found that the proportions of the major <i>A. tenuis</i> allergen and a related hypoallergenic antigen in the samples were relatively constant, hence RAST inhibition can be used as a measure of allergenic potency despite the presence of this cross-reacting antigen. The lack of correlation between allergenic and biochemical properties of the various samples suggests that a simple biochemical test is unlikely to be a satisfactory replacement for the RAST inhibition assay in standardizing <i>A. tenuis</i> extracts.

IgE antibody-mediated cutaneous basophil hypersensitivity reactions in guinea pigs.

Cutaneous basophil hypersensitivity (CBH) reactions are a heterogeneous group of delayed time course basophil-rich immune responses that can be mediated in the guinea pig by T cells, B cells, or IgG1 antibody. This study examined whether guinea pig IgE antibody could also mediate CBH reactions. IgE antibody to picryl or oxazolone determinants was induced by immunizing Hartley strain guinea pigs pretreated with cyclophosphamide. Hyperimmune serum from these animals was passed through a heavy chain-specific anti-IgG1 affinity column. The presence of IgE anti-hapten antibody in the filtrate fraction was verified by passive cutaneous anaphylaxis (PCA) testing with a 7-day period of local passive sensitization and by the heat lability (56 degrees C, 4 hr) of PCA activity. This IgE-rich fraction and the IgG1 fraction eluted from the column with base (0.2 M Na2CO3, pH 11.3) were transferred i.v. to separate groups of normal guinea pigs. Both fractions mediated delayed time course reactions that contained basophils. Macroscopic and microscopic reactions mediated by the IgE-rich fraction were abolished with heat (56 degrees C, 4 hr). Thus, two antigen-specific factors in guinea pig serum can mediate delayed time course basophil-containing reactions: IgG1 and IgE antibodies. IgE-mediated CBH reactions are similar to the late-phase reaction that follows IgE-dependent wheal-and-flare reactions in humans. The finding that guinea pig IgE can mediate a late reaction that contains basophils makes this a possible model for the human late-phase response, and suggests that some forms of CBH may play a role in human allergic disease.

ChemInform Abstract: (E)‐3‐(4‐OXO‐4H‐QUINAZOLIN‐3‐YL)‐2‐PROPENOIC ACIDS, A NEW SERIES OF ANTIALLERGY AGENTS

AbstractAuf den im Formelschema skizzierten Wegen werden die Acrylsäuren (V) sowie die Vorstufen (III) synthetisiert.