Ragweed Pollen Extract Research Articles

Electrophoretic studies on the chemical fractionation of ragweed pollen extracts

1. 1. An electrophoretic study has been made of fractions separated from extracts of low ragweed pollen, prepared first by extraction with concentrated potassium phosphate buffers at pH 7.0; second, by extraction in the cold with ethyl alcohol of various strengths; and third, by precipitation of aqueous extracts by ammonium sulfate at one-half and at full saturation at pH 4. 2. 2. All fractions, except those soluble in 80 per cent alcohol (by volume) or saturated ammonium sulfate, were active by scratch test on sensitive persons. 3. 3. A peak in the electrophoretic patterns, which represents a substance or substances having no net charge at pH 4.0 or 7.0, was found in every fraction studied. This is the material which was described by Abramson, Moore and Gettner as “USD” or “artefolin.” It is most prominent in the fractions low in nitrogen and high in carbohydrate. This peak cannot be used as a measure of the skin test-active substance nor is it related to carbohydrate. 4. 4. Material extracted in 60 per cent alcohol at minus 15° C., and precipitated by 80 per cent alcohol was found to be the most nearly pure of all fractions, being composed largely of uncharged substance with two minor components. This fraction corresponds with Stull's substance “A.” 5. 5. All the fractions showed several components, but the electrophoretic patterns spread and overlapped to such an extent that it has not been possible to distinguish them clearly. 6. 6. None of the different chemical means of fractionation of the pollen extracts have yielded any fraction which is even approximately a pure chemical substance.

A Comparison of the Collodion-Particle Technic with Other Methods of Measuring Antibody

Summary The collodion-particle technic described by Cannon and Marshall was compared with the gelatin-interface test and the complement-fixation test. The comparisons were made using rabbit antisera for ovalbumin, ragweed-pollen extract and horse-serum. The collodion-particle technic consistently permitted the greatest dilution of serum without loss of the ability to give an easily visible antigen-antibody reaction. The titers obtained with the collodion-particle technic paralleled those obtained with the gelatin-interface and the complement-fixation tests.

Studies of tryptic and peptic digestion of extracts of giant ragweed pollen

1. 1. Giant ragweed pollen extract was subjected to digestion by freshly assayed pancreatin in calibrated dialyzing sacs. Digestion was also attempted with a highly active pepsin. In some cases as many as nine daily additions of enzyme were made to insure complete proteolysis. The digestion was followed by prolonged dialysis. 2. 2. All digests showed a marked loss of activity which was not accounted for by dialysis alone. Nitrogen analysis indicated that digestion was complete in the case of tryptic digestion. 3. 3. We cannot be certain that it was complete in the case of peptic digestion, because the membranes were not sufficiently permeable to permit all of the products of peptic activity to dialyze away. 4. 4. There is a small but constant amount of activity in this pollen which is unaffected by proteolytic digestion.

Reactions of ragweed-sensitive individuals to skin tests with nucleic acids and related compounds

1. 1. Eleven of thirty ragweed-sensitive patients reacted to test with nucleic acid from yeast, but only three of these reactions were + + or + + +, and none were comparable to the reactions of the same patients to ragweed pollen extract. 2. 2. Four of twelve patients reacted to thymus nucleic acid, and a few patients reacted slightly to guanine hydrochloride, adenine sulfate, histadine monohydrochloride, allantoin, proline, and xanthine. 3. 3. In several cases, the reactions to nucleic acid were also demonstrated by the method of passive transfer. 4. 4. The significance of these reactions was not apparent. 5. 5. None of sixteen normal controls reacted to any of these compounds on direct skin test.

Skin reactions to electrophoretic fractions of timothy pollen extract

1. 1. Timothy pollen extract has been fractionated electrophoretically. 2. 2. As in the case of ragweed pollen extract, there is present a major, colorless, slow-moving component which is electrophoretically homogeneous at pH 7.4. 3. 3. In addition, there are also present six or more negatively charged, pigmented components which migrate more rapidly than the colorless fraction. 4. 4. On direct skin tests, the slow-moving, colorless component, the fast pigments, and the intermediate pigments showed about the same skin reactivity. 5. 5. On passive transfer, both the pigments and the colorless fraction also showed positive reactions. 6. 6. Preliminary data obtained by means of the ultracentrifuge and diffusion techniques indicate that all of the constituents under discussion are of small molecular size compared with ordinary proteins. 7. 7. The diffusion rate of the colorless fraction of timothy is very close to the analogous component of ragweed. 8. 8. Questions arising connected with the immunologic specificity of the various fractions are briefly discussed.

Antigenic fractions in ragweed pollen: II. Further purification of the water-soluble fractions and a study of alkali-soluble fractions and an estimation of these fractions in pollen extract

1. 1. Various alkali-soluble protein fractions separated from low ragweed pollen differed chemically and immunologically from Fraction 1 and Fraction 2. They resembled each other chemically and gave positive skin tests on certain ragweed sensitive cases and on skin sites passively sensitized with the sera of certain ragweed-sensitive patients (passive transfer). Therefore, they have been designated Fraction 3. 2. 2. When diluted to contain the same protein nitrogen content, different preparations of Fraction 1 were alike in allergic activity. Similarly the different preparations of Fraction 2 or the different preparations designated Fraction 3 when compared on their protein nitrogen content were of equal allergic activity. 3. 3. The three fractions showed marked differences in their capacity to neutralize ragweed-sensitive sera and their relative activity varied with different sera. 4. 4. Dale tests on sixteen animals sensitized to whole low ragweed extract and on six animals sensitized with preparation of Fraction 3 failed to show sensitivity to Fraction 3 although many sensitizations to Fraction 1 and Fraction 2 resulted. 5. 5. A method is given for the estimation of these three fractions in ragweed extract. In a typical extract of low ragweed pollen, Fraction 1 and Fraction 3 were each approximately one-fourth of the total protein nitrogen while Fraction 2 was approximately one-half.

A quantitative study of anaphylaxis in the guinea pig: Kabat, E. A., and Landow, H.: J. Immunol. 43: 69, 1942

An immunosorbent prepared by attaching covalently the constituents of the aqueous extract of ragweed pollen to aminocellulose was used to remove reagins from a fraction of allergic serum enriched with respect to skin-sensitizing activity. The adsorbed antibodies were eluted with a solution of 2M NaI, pH 9.1, or of glycine-HCl, pH 2.5. The presence of ragweed-binding IgE antibodies in these eluates was revealed by radioimmunoelectrophoresis and radio-Ouchterlony analysis. Guinea pigs, which had been rendered tolerant to normal human cord serum proteins, were immunised with these eluates and produced anti-IgE antibodies, as demonstrated by radioimmunodiffusion and by reversed PCA in monkeys. These results confirm that human reagins elicited to the allergens of ragweed pollen belong to the IgE class of immunoglobulins.

Studies with antigens. XIII. The significance of reactions to intracutaneous tests performed with solutions of purified extracts of ragweed pollens: Efron, B. G., and Boatner, C. H.: J. Invest. Dermat. 5: 49, 1942

Studies with antigens. XIII. The significance of reactions to intracutaneous tests performed with solutions of purified extracts of ragweed pollens: Efron, B. G., and Boatner, C. H.: J. Invest. Dermat. 5: 49, 1942

Electrophoretic and ultracentrifugal analysis of hay-fever-producing component of ragweed pollen extract: Abramson, H. A., Moore, D. H., and Gettner, H. H.: J. Physical Chem. 46: 192, 1942

Electrophoretic and ultracentrifugal analysis of hay-fever-producing component of ragweed pollen extract: Abramson, H. A., Moore, D. H., and Gettner, H. H.: J. Physical Chem. 46: 192, 1942

Studies with Antigens: XIII. The Significance of Reactions to Intracutaneous Tests Performed with Solutions of Purified Extracts of Ragweed Pollens

Studies with Antigens: XIII. The Significance of Reactions to Intracutaneous Tests Performed with Solutions of Purified Extracts of Ragweed Pollens

A comparison of ball-mill and Coca extracts of ragweed pollen in the treatment of hay-fever: Winkenwerder, W. L., Arbesman, C., Gay, L. N., and Eagle, H.: Bull. Johns Hopkins Hosp. 69: 287, 1941

A comparison of ball-mill and Coca extracts of ragweed pollen in the treatment of hay-fever: Winkenwerder, W. L., Arbesman, C., Gay, L. N., and Eagle, H.: Bull. Johns Hopkins Hosp. 69: 287, 1941

Studies on the immunology of ragweed pollen proteins: II. Anaphylactic experiments

1. 1. Short ragweed pollen extracts as usually prepared contain two main types of molecular aggregates roughly classified as large and small. 2. 2. These may be separated by means of heat at the isoelectric point. 3. 3. The small molecular aggregate has a capacity to elicit skin reactions in patients sensitive to ragweed pollen. The large molecular aggregate as prepared in the present study has this capacity but to a less marked degree. 4. 4. The small molecular aggregate does not give precipitin reactions with ragweed precipitin antiserum. The large molecular aggregate gives positive precipitin tests with such antiserum. 5. 5. The small molecular aggregate is incapable of eliciting anaphylactic shock in guinea pigs sensitized to whole ragweed pollen. The large molecular aggregate is capable of eliciting anaphylactic shock in guinea pigs so sensitized.

Electrophoretic and Ultracentrifugal Analysis of Hay-fever-producing Component of Ragweed Pollen Extract.

Electrophoretic and Ultracentrifugal Analysis of Hay-fever-producing Component of Ragweed Pollen Extract.

Investigations on the presence of Forssman antigen in ragweed pollen

1. 1. Rabbits immunized with ragweed pollen extract produce ragweed antibodies. 2. 2. Ragweed antibodies are easily demonstrable by precipitation and complement fixation tests. 3. 3. The increase in titer of Forssman antibodies in such rabbit sera is not significant. There is no evidence for the assumption that Forssman antigen is present in ragweed pollen.

The stability of the reagin-neutralizing substance in ragweed pollen at various hydrogenion concentrations

The effect of pH on the inactivation of ragweed pollen extracts at 37° C. has been determined. After four hours, more than 75 per cent of the activity is destroyed in solutions the pH of which lies outside the approximate limits of 4.5 and 7.5. The pH region of maximum stability has been found to be approximately 5.5 to 6.5. It is tentatively suggested that the inactivation may be due to the denaturation of an active protein component of ragweed pollen.

Studies with Antigens: XI. The Significance of Scratch Test Reactions to Purified Extracts of Ragweed Pollens

Non-irritant concentrations of ragweed pollen extracts are those which produce significantly fewer positive skin reactions in control cases than in patients whose symptoms are attributable to ragweed pollen. Positive reactions produced by such extracts are presumptive evidence of sensitization to ragweed pollens (1, 2). The purpose of this investigation is to evaluate the significance of scratch test reactions produced by extracts of short and giant ragweed pollens purified according to our technic (3).

A slowly absorbed gelatin-pollen extract for the treatment of hay fever

1. 1. Gelatin, a slow absorption medium, was mixed with ragweed pollen extracts in an attempt to overcome difficulties encountered with the standard aqueous extracts. 2. 2. A delay in absorption of the active ragweed substance of the gelatin-ragweed extract as compared to the aqueous ragweed extract was noted following subcutaneous injections at passive transfer sites in a series of normal nonsensitive adults. 3. 3. A series of ninety-five adult patients suffering from ragweed hay fever symptoms were treated preseasonally with the gelatin-ragweed extracts. Another group of ninety-five adult patients were treated with the standard aqueous extracts during the same period. Twenty-two patients of the first group continued their treatment on a monthly basis during 1940 after having completed a course of preseasonal injections. 4. 4. A greater tolerance for the ragweed-gelatin extract was noted; higher maximum dosages (often twice the amount attained with the aqueous extracts) were given with safety and with half the number of injections. 5. 5. Constitutional reactions were milder and occurred infrequently. 6. 6. More effective clinical relief was obtained following treatment with the gelatin-ragweed extracts. 7. 7. An average dosage schedule for preseasonal treatment with gelatin-pollen extracts was determined. It was modified to suit the individual's tolerance. 8. 8. The gelatin extract must be prepared according to the technique outlined if standard and satisfactory results are to be obtained. 9. 9. At present the chief value of the gelatin extracts is in the treatment of hay fever patients so sensitive as to accept poorly the usual aqueous pollen therapy. 10. 10. Further studies are in progress to determine whether the use of the more slowly absorbed gelatin extracts will result in a more sustained and lasting immunity.

Changes in Serological Reactions and Tissue-Sensitivity in Hay-Fever Patients during the Early Months of Treatment

Summary Thirteen typical cases of ragweed hay-fever were studied with intracutaneous, conjunctival, and serological tests before and after the first three months of treatment with injections of pollen-extract. In 12 of the 13 patients the reaction to intracutaneous tests with ragweed pollen-extract, which had been preserved by the cryochem-process, was less after treatment. In 11 of 13 patients the reaction to conjunctival tests with ragweed pollen-extract was less after treatment. The sera of 12 of the 13 patients showed more skin-sensitizing antibody, as measured by the dilution-test, after treatment. All of the sera required more ragweed pollen-extract to neutralize the skin-sensitizing antibody after treatment. All post-treatment sera inactivated one of the antigenic fractions (fraction 1) of low-ragweed pollen-extract. In only one case did the serum inactivate the second antigenic fraction (fraction 2) of the pollen-extract. None of the sera taken before treatment inactivated ragweed pollen-antigen. Patients who were sensitive to both antigenic fractions of ragweed pollen but who developed antibody inactivating only one fraction did not develop tolerance for large doses of ragweed pollen-extract and had more constitutional reactions than those who were sensitive to only one fraction and developed antibody inactivating that fraction. Sera that inactivated only the more active fraction (fraction 1) of the ragweed pollen-extract showed blocking activity when tested by the neutralization-method. When such sera were tested by the dilution-method, the results obtained depended on the way in which the serum used to sensitize cutaneous sites reacted to the two antigenic fractions of the ragweed pollen.

Antigenic fractions in ragweed pollen: I. Water-soluble fractions

Two water-soluble fractions have been separated by fractional precipitation from low ragweed pollen extract. Each of these had certain characteristics of proteins, but showed marked differences in their nitrogen content and in chemical reactions. The two fractions gave positive skin reactions in direct tests on ragweed-sensitive patients and on testing sites prepared with the serum of ragweed-sensitive patients (passive transfer). The relative activity of these two fractions varied greatly in different cases. The two fractions also showed marked differences in their capacity to neutralize sensitive sera. Here again, their relative activities varied with different sera. Studies by the Dale test showed that each of the two fractions had a certain antigenic specificity as well as cross relationship.

Skin reactions: X. Preseasonal treatment of hay fever by electrophoresis of ragweed pollen extracts into the skin: Preliminary report

The electrophoretic method of introducing certain allergens into the skin is a useful reproducible technique for transporting these allergens into the dermis. Giant ragweed, dwarf ragweed, and timothy extracts have been employed in this way. An electrophoretic unit of pollen dosage is defined. Giant and dwarf ragweed extracts have been used satisfactorily in the preseasonal treatment of a series of seven cases of hay fever by electrophoresis of the skin-reactive constituents. Analysis of undialyzed ragweed extract, using both the Philpot-Svensson and Longsworth techniques to estimate the amount of skin-reactive, unpigmented constituent present, has revealed that a very large amount of the unpigmented fraction is present in undialyzed extract of giant ragwee. We have, 7 therefore, been using undialyzed extracts in treatment experimentally since they are much simpler to prepare and less expensive. It is possible to introduce large quantities of skin-reactive material from undialyzed extracts by electrophoresis. Several patients treated preseasonally in this way in New York City have thus far (Aug. 27, 1940) had a satisfactory season.