Ragweed Pollen Extract Research Articles

Early eosinophilia induced in guinea pigs by intrapulmonary injection of antigenic determinants and antigens

Injections of antigens directly into the lungs of guinea pigs induced marked eosinophilia at the sites of injection within 72 hours. In contrast to generalized anaphylaxis, there was, at that time, no correlation between the eosinophilia in the lungs and the titer of circulating eosinophils, which remains stable. Antigen-induced eosinophilia was limited to the injected lung; no eosinophilia developed in the noninjected lung which served as a control. Neither homologous protein nor extract of ragweed pollen in aqueous solution or in Freund's adjuvant produced eosinophilia in the injected lung. Injection of arsanilic acid did not cause an increase in eosiniphils, but injection of arsanilic acid diazotized to homologous protein was followed by a marked increase in peribronchial eosinophils. p-Phenylene diamine, on the other hand, injected without conjugation in aqueous solution, produced a massive pulmonary eosinophilia.

Immunotherapy with antigen E

Ragweed-sensitive patients were treated for 4 years with antigen E, a purified ragweed fraction. The over-all results, including a double blind study and crossover studies, suggested that antigen E was not as clinically effective as whole ragweed pollen extract. Antigen E was well tolerated, allowing administration of relatively large doses with few local and systemic reactions. In vivo reagin neutralization studies also suggested that some allergens had been lost in the preparation of Antigen E. Antigen E was highly antigenic, stimulating high titers of hemagglutinating antibodies, and precipitins could be demonstrated in some sera. Antigen E appears to be an important component of ragweed pollen. However, other allergenic fractions may also be necessary in order to provide optimal clinical results.

Fine structural changes in livers of dogs in anaphylaxis

Reaginic serum was used in the passive transfer of ragweed pollen sensitivity to normal dogs. Anaphylaxis was induced by intravenous injection of a challenge antigen consisting of an aqueous extract of ragweed pollen, and pathologic changes in liver were studied in serial biopsies taken at intervals ranging from 15 seconds to 20 minutes. The results showed destruction of endothelial cells and hepatocytes as early as 15 seconds; later changes consisted of vacuolation of hepatocytes, continued cellular disruption, and filling of sinusoids by cell debris. These changes persisted throughout the period of observation.

Separation of New Skin Reactive Antigens from Dwarf Ragweed Pollens

Abstract The fractionation of dwarf ragweed pollen extracts has been investigated extensively (1–4) and the results of these studies have been reviewed (5). The principal products, isolated previously, contained one common antigen which was described by King and Norman (3) and which they called antigen E (AgE). King's product, IV-C (3) and Robbins' most recent products, IVRBB and IVRCC (6), contained AgE exclusively as determined by quantitative immunodiffusion methods. On the other hand, Goldfarb's product, pool C, contained 80% AgE. These three sets of products showed significant chemical and physicochemical dissimilarities. The amino acid analyses of IV-C, IVRBB and IVRCC were different in respect to methionine, half-cysteine, proline and tryptophan content. Pool C contained 11% arabinose whereas the other two products were reported to contain 1% or less. On the assumption that arabinose was an impurity, we attempted to extract AgE from pool C by a variety of methods, including the procedure used by King to separate IV-C.

Comparison of two dosages of ragweed extract in the treatment of pollenosis

A double-blind study was carried out to determine the effect of dosage in the treatment of ragweed hay fever with injections of aqueous ragweed pollen extract. Two concentrations of extract were used, one 20-fold smaller than the other. The volumes administered were such that the dose at the higher concentration was close to the maximum that it is practical to administer. During the ragweed season, the group of patients receiving the higher concentration fared significantly better than the group receiving the lower concentration. It is concluded that large dosages of pollen extract are more effective than small dosages in the treatment of hay fever. The findings indicate that the very low dosages recommended by some would have little influence on the course of pollenosis.

Comparison of Two Dosages of Ragweed Extract in the Treatment of Pollenosis

A double-blind study was carried out to determine the effect of dosage in the treatment of ragweed hay fever with injections of aqueous ragweed pollen extract. Two concentrations of extract were used, one 20-fold smaller than the other. The volumes administered were such that the dose at the higher concentration was close to the maximum that it is practical to administer. During the ragweed season, the group of patients receiving the higher concentration fared significantly better than the group receiving the lower concentration. It is concluded that large dosages of pollen extract are more effective than small dosages in the treatment of hay fever. The findings indicate that the very low dosages recommended by some would have little influence on the course of pollenosis.

The characterization of skin-sensitizing antibody induced in nonsensitive dogs

Skin-sensitizing antibody (SSA) to ragweed was induced in four nonatopic dogs following four weekly injections of ragweed emulsion. The characteristics of the SSA are similar to those occurring spontaneously in man and dog. A challenge of one of the dogs by insufflation and intravenous administration of ragweed pollen extract failed to produce any systemic symptoms. This suggested the presence of a high level of blocking antibody. One of the dogs developed ulceration and granuloma formation at each site of injection. Biopsies were taken during the stage of active ulceration and after complete healing.

Characterization of ragweed pollen extracts prepared by sonication and other methods

Extracts of ragweed pollen were prepared by stirring for 48 hours, homogenization, or sonication, and by various combinations of these methods. The extracts were compared with respect to protein yield and to degree and stability of skin-sensitizing activity in ragweed-sensitive patients. Compared with the usual method of extraction by stirring, both homogenization and sonication proved to be highly efficient, rapid methods with respect to all criteria evaluated. A decrease in allergenic activity was observed in all extracts during storage over a 20 day period. Following fractionation of extracts on Sephadex G-25 and G-100 columns, most of the skin-sensitizing activity was found in protein fractions with a molecular weight between 5,000 and 70,000, although some activity resided also in a fraction of low molecular weight.

Induction of immediate and delayed skin reactivity with emulsion of purified ragweed antigen

The induction of immediate or delayed skin reactivity in man following the injection of an emulsified whole ragweed pollen extract raised the question of the dependence of this activity on a nonantigenic component as an adjuvant and the possibility that a particular component might be responsible for a particular type of reactivity. Although we could not investigate all fractions of ragweed pollen, the availability of ragweed antigen E of King made possible a comparison of it with the extract of whole short ragweed pollen. Thirty-one nonatopic subjects were injected with an emulsion of 10,000 pollen units of the crude extract of short ragweed pollen, while 32 were given an injection of an emulsion of ragweed antigen E prepared from the same batch of pollen, the dose being comparable to the amount of E present in the crude extract. At approximately weekly intervals, intradermal tests were made with 0.05 ml. of 1:1,000 whole pollen extract and equivalent concentrations of antigen E. Several subjects who had immediate and delayed reactions were subjected to skin titrations with whole extract and antigen E, using serial dilutions of the material. The results indicate that emulsified antigen E is capable of inducing immediate or delayed sensitivity, and that the incidence and degree of this reactivity are not significantly different from that produced by whole extract emulsions. Antigen E and whole ragweed extract are equally capable of eliciting either an immediate or delayed reaction, regardless of the preparation employed in the emulsion. Exceptions to this generalization occur in individuals, and this may depend on other antigens than E in the crude preparation and/or trace heterologous antigens in the preparation of antigen E.

A familial tendency toward skin sensitivity to ragweed pollen

A study was undertaken to establish whether or not the ability to develop skin-sensitizing antibodies to ragweed pollen is familial. The frequency of positive intradermal skin tests to ragweed pollen extract was compared in the parents and sibs of children with ragweed hay fever and positive skin tests and the parents and sibs of children without ragweed hay fever and with negative skin tests, selected without reference to a family history of allergy. The frequency of positive skin tests to ragweed pollen extract was found to be higher in the sibs of children with positive skin tests and symptoms of ragweed hay fever (28.0 per cent) than it was in sibs of children with negative skin tests and no ragweed fever (4.8 per cent). The frequency of positive skin tests to ragweed pollen extract was higher in the sibs of children with positive skin tests and symptoms of ragweed hay fever when at least one parent had a positive skin test (48.5 per cent) than it was when neither parent had a positive skin test (14.3 per cent). These findings clearly demonstrate that the ability to form skin-sensitizing antibodies to ragweed pollen is familial and probably genetically determined. The ability to develop a positive skin test to grass pollen extract was also found to be familial.

Physicochemical properties of reaginic antibody: III. Further studies on the reaginic antibody in γA-globulin preparations

Three sera from ragweed-sensitive patients were fractionated by chromatography on DEAE cellulose columns and gel filtration. It was confirmed that the reaginic activity in the chromatographic fractions did not parallel γA-globulin concentration. The γA-globulin fractions from the same sera contained reaginic activity. When the γA-globulin preparations were further fractionated by chromatography on DEAE cellulosc columns, the reaginic activity was concentrated into one of the three subfractions. The precipitation of γA-globulin in the subfraction and the original γA-globulin preparation with rabbit anti-γA-globulin antibody was not accompanied by any detectable decrease of reaginic activity. The results indicated that the reaginic activity in the fractions was not associated with γA-globulin, but with impurities which were not detected by immunodiffusion methods. The subfractions of the γA-globulin preparation from a ragweed-sensitive patient blocked passive sensitization of normal human skin with reaginic antibody of a heterologous system. The blocking activity of the fractions paralleled the sensitizing activity against ragweed-pollen extract. It was also found that a fraction containing reagin but no detectable γA-globulin had a definite blocking activity, whereas γA-myeloma proteins did not. These findings suggest that both the blocking activity and reaginic activity are associated with the same protein other than γA-globulin.

Physico-Chemical Properties of Human Reaginic Antibody

Summary The reaginic activity in sera from ragweed-sensitive patients was inactivated after the precipitation of serum proteins with the rabbit antihuman globulin serum which did not contain any of the antibodies specific for γG-, γA-, γM-globulins and for light chains of immunoglobulins. The rabbit antiserum gave a γ1-globulin precipitin band with the reagin-containing fraction which contained γA but none of the γG-, γM- or γD-globulins. The specificity of the precipitin band was different from the γA-anti-γA band. By radioimmunoelectrophoresis and by radioimmunodiffusion, the antibody against ragweed pollen-extract (antigen E) was detected in the γ1-globulin, indicating the presence of a unique γ1-immunoglobulin. The protein was tentatively designated γE-globulin. Precipitation of γE-globulin anti-ragweed antibody in the reagincontaining fraction was accompanied by the loss of reaginic activity, whereas the precipitation of the γA-globulin antibody in the fraction did not result in any decrease of reaginic activity. The results highly suggest that the reaginic activity is associated with γE-globulin.

Some antigenic relationships between crude extracts and purified fractions of ragweed pollen

The capacities of two crude ragweed pollen extracts and of two purified fractions to inhibit the specific antibody binding of I 131-labeled ragweed pollen extract (I ∗RW) have been studied. Although the majority of antigenic groups capable of inhibiting the I ∗RW binding capacity had been removed from the purified fractions (Pool C and Fraction IV z), both probably contained at least trace amounts of most, if not all, of the antigens present in crude ragweed extracts. Pool C and Fraction IV z had similar but demonstrably different capacities to inhibit the I ∗RW-antiserum reaction. Whether the antigenic differences between Pool C and Fraction IV z were associated with skin-test reactive molecules is not known. The antigenic differences between Pool C and Fraction IV z were amplified by the use of related, but not identical, antibody populations in the inhibition test used for these studies. The useful role which antibody and antigen heterogeneity plays in the interpretation of the inhibition test has been discussed and is applicable to other I 131-labeled antigen-antibody inhibition test systems.

The bronchial provocation test: Its clinical evaluation and the course of induced asthma

Intracutaneous and bronchial provocation tests with an extract of short ragweed were performed on 33 asthmatic children. Nine of the highly sensitive (2 plus and 3 plus) patients were provoked with the defatted pollen suspension as well. Most of the 33 subjects were also tested with Alternaria and Hormodendrum extracts. The provocation was considered positive when the patients developed clinically apparent asthma (dyspnea, cough, wheezing) and a drop in vital capacity of 30 per cent or more. Negative skin tests (mean diameter of the wheal ≦ 6 mm. for the ragweed and < 4 mm. for the Hormodendrum by the intracutaneous method) were found to correlate well with negative provocation tests. The patients whose bronchial sensitivity to ragweed appeared to be of clinical significance had a mean wheal diameter ≧ 10 mm. The comparison between the provocative dosage and the theoretical natural exposure of the ragweed and evaluation of individual clinical history suggested that the provocation test can be regarded as negative if no asthma is provoked with an amount equivalent to the highest daily natural exposure. Thus, up to 24 inhalations of 1:500 ragweed pollen suspension or extract may be sufficient to detect clinically significant bronchial sensitivity. Late asthma occurred some hours after apparent spontaneous recovery from the immediate asthma in some cases showing a “biphasic” response to the inhalational challenge. In one patient who had late asthma consistently, steroids prevented this reaction. Further studies are necessary in order to confirm and evaluate the significance of the observations reported.

Biologic activities of soluble antigen-antibody complexes: X. Skin-reactive properties of allergen-reagin complexes

Mixtures of ragweed pollen extract with a fraction of an allergic serum containing reagin gave erythema-wheal reactions in normal individuals. Skin reactivity was not induced either with the fraction which had been heated at 56° C. for 4 hours or with the γ 2-globulin fraction of the same serum. The skin reactions with the allergen-reagin mixtures were inhibited by prior treatment of the skin sites with normal γ 1A-globulin, but not with γ 2- or γ 1M-globulin. After removal of free allergen in the mixture by gel filtration, no reaginic activity was recovered. A skin-reactive substance was eluted from a Sephadex G-200 column in the fractions containing γ 1A-globulin. The results definitely indicate that allergen-reagin complexes formed in vitro have an activity to induce erythema-wheal reactions in normal individuals.

Basophil degranulation tests in atopic allergic states: A pilot study of ragweed pollen sensitive patients

Of 22 patients with clinically active hay fever and positive skin tests to ragweed pollen extract, 27 per cent gave positive basophil tests with the use of the same extract. One half of these positive test reactors had received hyposensitization therapy One of 12 nonallergic controls gave a positive basophil test with the same extract. The basophil test, as performed by the methods described, does not lend itself to practical application in the detection of the ragweed-sensitive patient.

Skin-sensitizing antibody titer: III. Relationship of the skin-sensitizing antibody titer to the intracutaneous skin test, to the tolerance of injections of antigens, and to the effects of prolonged treatment with antigen

1. 1. The intracutaneous skin test tends to vary directly with the skin-sensitizing antibody titer. However, individual variations are so great that titer cannot be determined from the skin test. 2. 2. The doses of aqueous ragweed extract tolerated by patients vary inversely with the skin-sensitizing antibody titer. 3. 3. Development of blocking antibody increases the tolerance for injected extracts. 4. 4. Patients with high titers of skin-sensitizing antibody are more likely to have constitutional reactions to emulsions of ragweed pollen extract even with lower doses of antigen. The titer of skin-sensitizing antibody is a helpful guide in determining the dose of emulsified pollen extract to be used. 5. 5. Prolonged treatment with injections of aqueous ragweed extract is accompanied by a decrease in the titer of skin-sensitizing antibody. This decrease is greatest between the first and fifth year of treatment but continues up to 20 years. Comparative studies of untreated patients and of patients who, after 1 or 2 years of treatment, were untreated for an average of 9 years suggest that this decrease in titer of skin-sensitizing antibody results directly from the treatment and not from the increasing age of the patient or duration of hay fever.

Characterization of the dialyzable constituents of the aqueous extract of ragweed pollen

Four fractions of the dialyzable components of the aqueous extract of ragweed pollen were prepared and their physicochemical and immunologic properties were examined. All these fractions were skin active with respect to sera from both treated and nontreated individuals. All the antigenic and allergenic determinant groups of the dialysate fractions were found to be present in the nondialyzable components of the whole aqueous extract. Contrary to previous conclusions, it was demonstrated by cross-neutralization studies that new reagin(s) directed exclusively against components of the dialysate is not produced by hyposensitization treatment with the whole aqueous extract. Rather, any increase in reagin concentration caused by hyposensitization is considered to be the result of a rise in the concentration of reagin (s) already being produced by the allergic individual.

Local inhibitory effect of subcutaneously injected triamcinolone on the immediate and delayed types of skin reactions

One hundred sixty-six individuals were injected with a suspension of triamcinolone diacetate into the upper layer of the subcutaneous tissue to observe the effects on the immediate and delayed types of allergic skin reactions in the injected area. Eighty-one individuals were tested intracutaneously with Mantoux at various intervals of up to 7 days, after they were treated locally with subcutaneous injection of triamcinolone. Inhibition was present in 80 subjects. Of the 80 individuals tested intracutaneously with histamine up to 4 days after triamcinolone injection, 43 showed varying degrees of inhibition. Five allergic patients were tested with ragweed pollen extract up to 3 days after triamcinolone injection, and all showed inhibition. Age, sex, or the time interval between the triamcinolone injection and the intracutaneous testing had no significant influence on the inhibition.

Skin sensitizing antibody: I. Relationship of the skin sensitizing antibody titer to the occurrence of symptoms in untreated persons with a positive ragweed skin test

The titers of skin sensitizing antibody of the sera of untreated hay fever patients differ significantly from those of the sera of persons who react to intracutaneous tests with ragweed pollen extract but who do not have clinical hay fever.