Ragweed Pollen Extract Research Articles

Influence of Electrolytes on the Cutaneous Reactivity of Seitz-Filtered Ragweed Solutions

Summary Methods for the preparation of “electrolyte-free” ragweed pollen extracts (i.e. having an electrical conductivity less than 20 × 10−6 reciprocal ohms) are presented. The reduction of the electrolyte-content of a ragweed pollen extract per se did not significantly diminish its capacity to produce intracutaneous reactions in ragweed pollen-sensitive subjects. If “electrolyte-free” ragweed pollen extracts of specified concentrations were passed through Seitz filters, the filtrates did not produce intracutaneous reactions in ragweed pollen-sensitive subjects. The specific activity of Seitz-filtered “electrolyte-free” ragweed pollen extracts could be restored or enhanced by the addition of electrolytes in proper concentration. A definite relationship between the specific activity of Seitz-filtered ragweed pollen extracts and the electrolytes contained therein has been demonstrated. This relationship was found to be dependent upon: (a) a certain alteration of unknown character produced by filtration through a Seitz filter-pad. (b) the concentration and type of contained electrolytes.

Ultracentrifugation of ragweed pollen extracts: Edward B. Sanigar

Ultracentrifugation of ragweed pollen extracts: Edward B. Sanigar

Observations on Sensitization of Guinea Pigs with Alum-Precipitated Ragweed Extract

Alum-precipitated ragweed pollen extract is a potent antigen for producing anaphylactic sensitization in guinea pigs,1-4 whereas the watery extract is by comparison a poor one. Alum-precipitated extracts have been successfully applied in sensitizing guinea pigs to house dust5-8 and cotton linters5, 6, 9 and in sensitizing dogs to horse serum.10 The enhanced antigenic response of the alum-precipitated extract is generally attributed to two factors: first, retention of the antigenic substance by the precipitate so that it is only slowly liberated into the system, thus producing a continuous stimulation for the production of antibodies; and second, the local tissue response to the precipitate at the site of injection is said to delay absorption of the antigen. Thus diphtheria toxoid has been detected at the site of injection from a few days to several weeks after administration of alum-precipitated toxoid.11, 12 The experiments recorded here were designed to determine whether antigenic constituents of ragwee...

Studies With Antigens, VIII. Preparation of Purified Extracts of Ragweed Pollen

Summary1. Fractionation of aqueous ragweed pollen extracts with water-miscible organic liquids produces purification and concentration of the allergens of these extracts. 2. Fractionation of extracts, purified by the above treatment, with high concentrations of sulphate salts produces further purification and concentration of the allergenic factors.

Experimental and clinical study of fresh and modified pollen extracts

1. 1. Fresh extracts of pollen were modified by formolization, radiation with ultraviolet light, heating, acctylation, and alum-precipitation and were compared in clinical and experimental studies with fresh and regular extracts. 2. 2. Treatment of undialyzed extracts of pollen with formaldehyde resulted in an increase in nitrogen precipitated with protein by phosphotungstic acid. It was shown that after formaldehyde treatment part of the dialyzable nonprotein nitrogen was precipitated with protein, and experiments suggested that protein split products, such as amino acids, might be precipitated under such conditions. Heating to 70° C. caused a precipitation and loss of protein nitrogen in ragweed extracts, but none in timothy extract. 3. 3. The protein in pollen extracts was stabilized by treatment with formaldehyde, heating, or radiation with ultraviolet light. In alumprecipitated ragweed extract there was a loss of protein on ageing, similar to the loss in regular extract. 4. 4. The gelatin liquefying enzymes in ragweed pollen extract were inactivated by formolization, heating to 70° C. for an hour, acetylation or radiation with ultraviolet light, and subsequent heating to 40° C. 5. 5. In neutralization of sensitive serum, the extracts modified by formolization, heating, radiation with ultraviolet light, and acetylation were reduced in activity in comparison with fresh extract. Regular extract one year old showed a loss of neutralizing capacity out of proportion to the decrease of protein content. 6. 6. The serologic response in hay fever patients treated with acetylated, formolized, and heated extract was less than that in patients treated with regular, fresh, or alum-precipitated extracts. The fresh extract gave the greatest response. 7. 7. Modification of the pollen extracts did not eliminate constitutional reactions. Since the various phenomena produced by the modified extracts required different degrees of specificity, no single test could be considered a measure of all types of allergic activity. 8. 8. The clinical results were best in the group of hay fever patients treated with fresh extract. Alum-precipitated and formolized extracts gave results which seemed fairly comparable to those obtained by regular extract. Heated and acetylated extracts gave the least satisfactory results. 9. 9. The antigenic relationship of the fresh, formolized, and heated extracts was studied by means of the Dale test. The formolized and heated extracts showed a modification of specificity in that they failed to neutralize uteri of pigs sensitized with regular extract.

Isolation of an Unpigmented Skin Reactive Constituent from Extracts of Ragweed Pollen by Electrophoresis.

In 1938 experiments were undertaken, using a conventional moving boundary technique, to separate the fractions and to determine the electrical charge of the constituents of giant ragweed pollen extracts. At that time it was observed that the pigments in dialyzed ragweed extracts at pH 7.0 were negatively charged. The results during that summer and the succeeding winter, however, were contradictory, because the relation of the pigments to the biologically active constituents was uncertain.Using the Tiselius1 cell for the study of the moving boundaries of protein solutions and the Philpot-Svensson2 technic to analyze the quantity and nature of the constituents, further progress may now be reported. Figures la and 1 b are illustrations of the type of curves given by the major constituent. This major constituent is negatively charged, slow moving, unpigmented, and highly skin reactive in persons sensitive to ragweed. A minor constituent, about 1/5 that of the unpigmented major constituent, appearing in the pi...

TREATMENT OF RAGWEED DERMATITIS

The first treatment of ragweed (contact) dermatitis was made in 1919. Hannah1reported good results in a single instance with coseasonal injections of an aqueous extract of ragweed pollen. In the same year, Sutton2reviewed 4 cases of the same condition, in 3 of which the patients were treated with a glycerin-phenol-saline solution extract of the pollen and showed decided improvement. However, none of these persons was given patch tests either with the oleoresin, which has since been proved to be the causative factor, or with ragweed leaves, which contain an abundance of the potent product. In 1930 Sulzberger and Wise3noted improvement in 1 person following intradermal injections of water-soluble ragweed pollen antigen. In 1931 Brown, Milford and Coca4showed that ragweed (contact) dermatitis is due to the fat-soluble fraction of the plant or pollen rather than to the water-soluble fraction, which causes hay fever

The thermolability of ragweed pollen extract and its corresponding reagin

Ragweed pollen extracts have been found to be highly thermolabile. Approximately 75 per cent of their reagin-neutralizing activity is destroyed within fifteen minutes at 56° C., 90 per cent within one hour, and over 95 per cent in four hours. The reagin-neutralizing activity of four similar ragweed pollen extracts prepared in 1939, 1937, 1933, and 1929 and since stored at 4° to 8° C. were found to be as 100 :16 :4.5 :1.2, respectively, indicative of a fairly rapid deterioration. Despite the widely differing biologic activities of these four extracts, their protein nitrogen contents, and the ratios of protein nitrogen total nitrogen , were of the same order of magnitude. This does not necessarily exclude the possibility that the reactive constituent is a protein. Short ragweed pollen reagin has also been found to be highly thermolabile. On heating at 56° C. approximately 50 per cent of the skin-sensitizing activity was destroyed within one-half hour, approximately 90 per cent within two hours, and approximately 99 per cent within four hours.

On the presence in rabbit antisera vs. ragweed pollen of skin-sensitizing antibodies passively transferable to man: Winkenwerder, W. E., Eagle, H., and Arbesman, C. E.: J. Immunol. 36: 435, 1939

Sixteen of 29 antisera produced in rabbits by the injection of various extracts of short ragweed pollen contained ragweed-specific skin sensitizing antibodies passively transferable to man. There was no regular correlation between the precipitincontent of the serum and its ability to sensitize the human skin. Defatting the pollen, or grinding it in a ball-mill had no apparent effect on its ability to excite the production of skin-sensitizing antibody. Although there was some indication that antisera of animals injected intravenously may contain less of this antibody than those injected subcutaneously or intraabdominally, further work will be necessary to establish this point.

The assay of ragweed pollen extracts

As suggested by Cooke and his co-workers, 1 the relative activity of ragweed pollen extracts may be measured by the determination of the minimal concentration of extract which is necessary to neutralize a serum containing the homologous reagin. It is essential to carry out the comparative assays simultaneously, in the same recipient and with the same serum. The details of the technique, the various factors which affect the assay, the degree of accuracy which may be expected, and the limitations of the method are discussed in the text.

Antigenic studies by the dale test: I. The antigenic relationship of certain pollens

1. 1. The protein fraction of low ragweed pollen extract produced muscular contraction and completely neutralized the uterine antibodies of guinea pigs sensitized by whole low ragweed pollen extract, as evidenced by the Dale test. Whereas, dialyzed low ragweed pollen extract also produced muscular contraction and neutralized the uterine antibodies to the whole extract, the dialysate completely failed to cause any contraction. The standardization of whole pollen extracts on the basis of their protein content was a measure of the reactive substance in whole pollen extract. 2. 2. Specific pollen extract always neutralized to the related pollen extract. 3. 3. A related pollen extract in every instance produced a contraction of the uterine horn sensitized by a specific pollen. 4. 4. The related pollen extract neutralized to the specific in 21 instances and did not neutralize in 17 experiments. 5. 5. These experiments indicate an antigenic relationship but not an identity of the pollens used in this study.

On the Presence in Rabbit Antisera vs. Ragweed Pollen of Skin-Sensitizing Antibodies Passively Transferable to Man

Summary Sixteen of 29 antisera produced in rabbits by the injection of various extracts of short ragweed pollen contained ragweed-specific skin sensitizing antibodies passively transferable to man. There was no regular correlation between the precipitincontent of the serum and its ability to sensitize the human skin. Defatting the pollen, or grinding it in a ball-mill had no apparent effect on its ability to excite the production of skin-sensitizing antibody. Although there was some indication that antisera of animals injected intravenously may contain less of this antibody than those injected subcutaneously or intraäbdominally, further work will be necessary to establish this point.

Electrokinetic phenomena : XIV. The inactivation of ragweed pollen extracts by adsorption and the electric charge of the resultant surface

1. 1. The electrical properties of quartz particles covered with material adsorbed from ragweed extracts are those typical of ampholytes. 2. 2. The isoelectric point of these particles covered with a fresh extract was at pH 3.9. On standing this shifted to pH 4.3 where it was stabilized for some time. 3. 3. Experiments with paraffin oil and collodion particles showed that these particles acquired similar amphoteric surface properties in ragweed extracts, corresponding exactly with those found when the adsorbent was quartz. 4. 4. The titration curves of ragweed extracts were typical of solutions of proteins with a maximum acid-binding capacity near pH 2.6. 5. 5. The data obtained are evidence supporting the point of view that the skin reactive constituent in ragweed extracts is protein-like in nature.

Sensitization of Guinea Pigs to Cotton Linters and House Dust Extracts

Since Cooke first directed attention to the importance of house dust as a specific excitant of asthma, there have been many attempts to characterize a hypothetical dust allergen. Its existence, as an entity, was recently questioned by Walzer on the basis of critical examination of available clinical data pertaining to dust sensitiveness. He found among these data evidence of reactions which he attributed to non-specific irritative principles in extracts of house dust or other dusty substances. Noted also was the fact that no one had succeeded in sensitizing experimental animals to house dust nor to suspected parent substances, cotton linters, kapok fiber dust, or mold(Aspergillus fumigatus). Adaptation of the alum precipitation technic devised by Harrison and employed by Caulfeild, Brown and Waters in sensitizing guinea pigs to ragweed pollen extracts, has proved effective for establishing anaphylactic sensitiveness to water extracts of linters and house dust, respectively. Comparing anaphylactogenic properties of these extracts led to recognition of a non-specific factor in house dust extracts which induced anaphylactoid symptoms in unsensitized guinea pigs. Two samples of clean, unused cotton linters were extracted with distilled water saturated with toluene. A 100 g sample was divided into 5 equal portions. To one 20 g portion sufficient solvent (100 ml) was added to saturate the fibers. After agitating for 48 hours, in a jar fixed on a rocking platform, the mixture was transferred to a Buchner funnel. The solution, recovered by suction and mechanical pressure, was made up to 100 ml with water and used in the same manner to extract in succession the 4 remaining portions of the sample.

Further studies on the allergenic activity of protein and nonprotein nitrogen fractions of ragweed pollen extract

1. 1. Protein prepared either by dialysis of ragweed extract, ammonium sulfate precipitation, or precipitation by alcohol and ether at −10° C., neutralized the skin sensitizing antibody of ragweed sensitive serum at approximately the same strength as whole extract when all were diluted to contain the same amount of protein nitrogen. This is further evidence that the activity of ragweed pollen in producing allergic reactions is associated with protein. 2. 2. The dialyzable nitrogen of ragweed pollen extract is shown to be inactive in neutralization of the skin sensitizing antibody. A small amount of this fraction is precipitated by phosphotungstic acid. 3. 3. Dried protein precipitated from ragweed extract at −10° C. with alcohol and ether when redissolved after ten months showed almost the same activity in neutralization as fresh extract. 4. 4. Ragweed extract at the end of ten months showed a loss of protein, and the remaining protein showed a marked loss of neutralizing capacity.

Activation of ultrafiltrates of ragweed pollen extracts by means of colloidal substances

Activation of ultrafiltrates of ragweed pollen extracts by means of colloidal substances

STUDIES ON IMMUNITY IN A TYPE OF HUMAN ALLERGY (HAY FEVER): SEROLOGIC RESPONSE OF NON-SENSITIVE INDIVIDUALS TO POLLEN INJECTIONS

Large injections of ragweed pollen extract into normal non-sensitive volunteers did not produce a sensitization to ragweed. Group 1 volunteers in whose skin many reactions were induced by injections of ragweed extract mixed with ragweed sensitive serum failed to show any serological changes. The theory that the immune substance found in the serum of treated ragweed sensitive cases was due to the reaction or to some substance created by it and not to the ragweed per se was not upheld. On the contrary in group 2, volunteers who received larger amounts of ragweed but no sensitive serum, serological changes were induced resembling those previously observed to occur in ragweed sensitive patients after treatment. They were demonstrable by an inhibition of the immediate reaction and by interference with the neutralization of sensitive serum by its antigen. These serological changes are therefore independent of the specific reaction characteristic of this type of allergy. The inhibiting factor was found to be related to the pseudoglobulin fraction of the serum and was shown to be specific.

Reactions to the Alcohol-Insoluble Fraction of Ragweed Pollen

It was shown by Coca and Grove1 that tryptic digestion of pollen did not destroy its ability to elicit a positive reaction in allergic individuals, although after dialysis no trace of its original nitrogen-content remained. Stull, Cooke and Chobot2 found that the active principle of ragweed pollen is present in that fraction which is insoluble in 90-95% alcohol. Black3 by repeated precipitation of ragweed pollen extract in 60% alcohol and redissolving of the precipitate in acid saline, obtained an extract which contained very little nitrogen, and which gave many of the chemical reactions characteristic of polysaccharides. This fraction also gave typical allergic reactions in 43 ragweed-sensitive patients, and no reactions in 50 normals. The highest dilution giving a reaction when injected intradermally was 1:240,000. Unger, Moore, Cromwell and Seeber,4 in desensi-tization experiments using both purified protein derivatives and purified polysaccharide fractions, concluded that the specifically active subst...

Studies on pollen and pollen extracts: XIV. The effects of varying hydrogen ion concentration upon the stability of pollen extracts

It has been shown that the presence of buffer salts in dextrose menstruum does not increase the potency of ragweed pollen extracts prepared with such solutions. Ragweed pollen extracts in isotonic dextrose buffered at either a slightly acid (hydrogen ion concentration 6.1) or slightly alkaline (hydrogen ion concentration 7.5) reaction have been shown to be at least as stable as similar unbuffered extracts. There is some indication that such buffering may increase stability. Alkaline buffer is probably to be preferred, as it prevents precipitation of the glucoside and thus prevents the development of a cloudy appearance on standing. Therapeutically, the same results were obtained with the extract buffered with sodium bicarbonate as with a similar unbuffered extract.

Suitability of the Monkey (Macacus rhesus) as a Recipient for the Prausnitz Kustner Reaction

The skin of the monkey (Macacus rhesus) is suitable for local passive sensitization to ragweed pollen extract by the intradermal injection of either serum from a sensitized guinea pig or serum from a human case of ragweed hay fever.