Antitoxin Content Research Articles

Mechanism of Antibody Formation

Summary and Conclusions A study has been made of the secondary antibody response in guinea pigs immunized with purified diphtheria toxoid. The antitoxin content of the alum granuloma at the injection site, the draining lymph node and the blood was measured by the rabbit intracutaneous test and, whenever possible, by the quantitative precipitin technique. From the observed time-concentration curves it is postulated that lymph antibody is a measure of the rate of antibody production and that serum antibody represents the accumulation of antibody liberated into the blood reservoir. The antibody pattern in the alum granuloma appears to reflect both the processes of synthesis and accumulation. A theoretical curve for serum antibody calculated from the above assumptions agrees well with the experimental data and strongly supports the hypothesis. These findings confirm the results of a previous study on the kinetics of cholera antibody formation in guinea pigs (1, 4). Furthermore, since similar rates of antibody production were obtained with either a soluble or particulate antigen, with different routes of inoculation and at different sampling sites, it is concluded that a general measure of secondary antibody synthesis has been demonstrated for one species of experimental animal. A correlation of the rate of diphtheria antitoxin formation with the determinations of toxoid decay in the lymph node indicates that the anamnestic production of antibody proceeds in at least two steps. The current theories of antibody formation which postulate various intermediates are considered in the light of the kinetic evidence.

Enhanced Production of Antibodies by Local Irradiation

Summary Local irradiation with 1000 r of the site of injection of alum precipitated diphtheria toxoid results in an enhanced production of circulating antitoxin. There is an even larger increase apparent in the antitoxin content of the irradiated injected skin site.

Concentration of diphtheria antitoxin in the blood of young adults.

Summary1. The diphtheria antitoxin titres of several thousand samples of cord blood have been analysed with relation to the histories of the mothers.2. Among mothers without histories of diphtheria or of immunization those born in 1933 or later showed a higher degree of natural immunity.3. The distribution of antitoxin titres among mothers who had suffered from diphtheria was similar to the average distribution among the unimmunized.4. The differences in proportions with any antitoxin in different groups of unimmunized mothers were almost entirely made up of those with titres above 0·04 and below 1·0 unit per ml.5. Among the artificially immunized mothers there were many more with low titres, i.e. below 0·04, and a few more with high titres, i.e. over 1·0, than among the unimmunized.6. Mothers immunized between the ages of 9 and 14 with the peak at 11 or 12 years showed the highest immunity rate.7. The greatest number of high titres occurred among recovered cases subsequently immunized.

Selection of antitoxins by the foetal membranes of rabbits.

Antibodies pass from the uterine lumen of pregnant rabbits into the foetal body fluids. It has been shown that antibodies formed in rabbits enter the foetal circulation by this route much more readily than those formed in horses or cows. It is the intention of this paper to investigate further this selective admission to the foetal circulation, to determine if there is a differential selection of heterologous antibodies according to the species in which they were produced and if this selectivity changes with foetal age. Antitoxins were employed throughout and were prepared in rabbit, man, guinea-pig, dog, horse and cow, against diphtheria or tetanus toxin or Clostridium welchii α-toxin. Mixtures of sera from two or three of these species, each serum containing a different antitoxin, were administered so far as possible. The antitoxin concentrations observed in the experimental samples were corrected to unit concentration in the serum administered, the term concentration quotient (c.Q.) being used for this quantity. Measured in this way the entry of antitoxin to the foetal serum varied with the species in which the antitoxin was produced; rabbit, man, guinea-pig, dog, horse and cow forming a descending series in that order. The entry of antitoxin into the amniotic fluid, or the foetal stomach contents derived from it, does not vary with the species in which the antitoxin was produced. This is so also for the maternal serum. Thus the species in which an antitoxin is produced affects the concentration at which it will appear in the foetal serum but not that at which it will appear in the other foetal body fluids or in the maternal serum. The entry of antitoxin prepared in rabbits to the foetal circulation increases with foetal age from 20 to 26 days after copulation and decreases thereafter.

On the Lack of Agreement of the Constant Toxin and Constant Antitoxin Flocculation Reactions of Diphtheria Toxin and Equine Antitoxin

Summary and Conclusions Six equine antitoxins and 4 toxins and toxoids were investigated, with respect to their flocculating properties. The earlier, graphical presentation of Boyd demonstrating the existence of distinct optima for constant toxin and constant antitoxin procedures was fully confirmed in all cases. The flocculation reaction was equated with respect to total concentration and ratio AT/T with the following general conclusions: Log time of flocculation bears an inverse linear relationship to log concentration of toxin plus antitoxin. Log time of flocculation bears a third-order parabolic relationship to ratio AT/T. The overall speed of reaction and the magnitude of relationships a. and b. are dependent upon both the toxin and antitoxin in question. Evidence is offered to indicate that toxin-antitoxin pairs showing different optima by routine flocculation procedures can be made to give identical optima in constant toxin and constant antitoxin titrations by using procedures which eliminate all pertinent variables except the ratio AT/T. On the basis of these findings the authors suggest that failure to obtain identical results by the constant toxin and constant antitoxin methods is primarily because of failure to control the total concentration of reagents and, of lesser importance, to establish comparable AT/T intervals. For routine purposes, either the constant toxin or the constant antitoxin method can be made most nearly ideal (in the sense of measuring the single variable, AT/T ratio) by using a toxin of about 3.5 times the titer of the antitoxin and ignoring any difference in volume from tube to tube. A more exact procedure would depend upon knowing the specifically precipitable N of the antitoxins and applying this information to adjusting the total concentration (of antitoxin plus toxin) in each tube to a constant value. The latter would be most readily accomplished by diluting the antitoxin to contain the same concentration of specifically precipitable N as the toxin against which it is to be titrated and to proceed with the titration without regard for the final total volume.

The estimation in vitro of small amounts of diphtheria antitoxin by means of a haemagglutination technique.

1. Treatment of washed sheep erythrocytes with dilute solutions of tannic acid rendered them capable of adsorbing diphtheria toxin from solution. Diphtheria toxoid was not adsorbed under similar conditions.2. Cells thus sensitized with diphtheria toxin were agglutinable by a proportion of children's sera and human cord sera; sera of rabbits, mice and horses, immunized with soluble or adsorbed diphtheria toxoid, also agglutinated sensitized cells, as did also the sera of children suffering from diphtheria and treated with antitoxic serum. Clear-cut results were usually obtained, but the end-point of titrations on horse sera was, in some cases, difficult to determine.3. Using intradermal toxin neutralization tests, a highly significant linear relationship was demonstrated between the haemagglutinating titres of children's sera for the sensitized red cells, and the antitoxin content of their sera. The antitoxin content corresponding to the smallest detectable amount of haemagglutinating antibody was 0·008 unit per ml. serum.4. The haemagglutinating titre was an accurate measure of the mean antitoxin level of a group of sera and a close measure of the antitoxin content of most specimens in individual tests.

The effect of peptic digestion on the properties of diphtheria antitoxin.

Diphtheria antitoxin prepared in the horse and refined by peptic digestion when injected in very large doses into women in an advanced stage of pregnancy did not pass to the infant. In pregnant guinea-pigs diphtheria antitoxin (naturalserum, ex -guinea-pig) passed to the young in abundance; but, after peptic-digestion, this homologous antitoxin failed entirely to pass the placenta, the young being devoid of antitoxin at birth. The passage was not affected by the treatment of the natural serum with ammonium sulphate as used in the Gibson-Banzhaf (1910) process for the concentration of antitoxin. Diphtheria antitoxin (natural serum ex -horse) passed from pregnant guinea-pigs to their off spring in smaller amounts and much less readily than homologous antitoxin, and the quantity of antitoxin( ex -horse) so passing was reduced even further and very considerably as a result of peptic digestion. Even under the most favourable conditions homologous antitoxin takes sometime (2 or 3 days) to attain the same concentration in the young as in the mother; but once this concentration has been attained it is preserve data high level for long periods. Passive anaphylactics ensitization of guinea-pigs, either of the whole animal or the isolated uterus, is easily effected, in vivo or in vitro , by small quantities of diphtheria antitoxin (either natural serum or ammonium sulphate concentrated, ex -guinea-pig), but this property is completely lost when the homologous antitoxin is subjected to peptic digestion. It is not possible to sensitize anaphylactically guinea-pigs, in vivo or in vitro , by means of diphtheria antitoxin, ex -horse, whether the antibody is presented either in the form of natural serum, or concentrated by means of ammonium sulphate; and the result is the same when pepsin-refined diphtheria antitoxin ex -horse is used. When 5 or 10 units of diphtheria antitoxin ex -horse, whether as natural serum, ammonium-sulphate concentrated or pepsin-refined, are injected subcutaneously into guinea-pigs, the animals are rendered Schick-negative in a few hours. These antitoxins are eliminated in about a week, after which time the injected guinea-pigs are found to be Schick-positive again. If, however, the same amounts of antitoxin made in guinea-pigs are injected into guinea-pigs the result is different; the animals also become Schick-negative, but this condition is maintained for a month or longer. That is, homologous antitoxin is eliminated much more slowly; but if this natural serum antitoxin from the guinea-pig is subjected to peptic digestion it is eliminated as quickly as diphtheria antitoxin made in the horse. When diphtheria toxin is injected intracutaneously into guinea-pigs, a quantity of diphtheria antitoxin 50,000 times as large as that required to neutralize it in vitro is required for neutralization in the animal, and then only if injected intravenously within 1hr.; but little or no neutralization in vivo occurs if the intravenous injection is longer delayed, whatever type of homologous or heterologous antitoxin is administered.

Book Reviews: MEDICAL SCIENCES & RESEARCH

Book Reviews: MEDICAL SCIENCES & RESEARCH Get access A Study of Diphtheria in Two Areas of Great Britain with Special Reference to the Antitoxin Concentration of the Serum of Inoculated and Non-Inoculated Patients and Other Persons; and the Relation of this to the Incidence, Type, and Severity of the Disease Percival Hartley et al. London : His Majesty's Stationery Office , 1950 . ((Medical Research Council Special Report Series No. 272.) viii + 162 . pages. 24 × 15 cm. 4s. [£0.2] E. James E. James Search for other works by this author on: Oxford Academic PubMed Google Scholar British Medical Bulletin, Volume 7, Issue 4, 1951, Page 360, https://doi.org/10.1093/oxfordjournals.bmb.a073983 Published: 01 April 1951

Book Reviews: MEDICAL SCIENCES & RESEARCH

Journal Article Book Reviews: MEDICAL SCIENCES & RESEARCH Get access A Study of Diphtheria in Two Areas of Great Britain with Special Reference to the Antitoxin Concentration of the Serum of Inoculated and Non-Inoculated Patients and Other Persons; and the Relation of this to the Incidence, Type, and Severity of the Disease Percival Hartley et al. London : His Majesty's Stationery Office , 1950 . ((Medical Research Council Special Report Series No. 272.) viii + 162 . pages. 24 × 15 cm. 4s. [£0.2] E. James E. James Search for other works by this author on: Oxford Academic PubMed Google Scholar British Medical Bulletin, Volume 7, Issue 4, 1951, Page 360, https://doi.org/10.1093/oxfordjournals.bmb.a073983 Published: 01 April 1951

The borderline Schick test

<h2>Summary</h2> The Schick test should be so standardized that the scale of reactions between no inflammation and definite erythema should fall on either side of a detinite threshold reaction, occurring in all persons with a threshold concentration of antitoxin. Using the dosage of 1/50 of a minimum lethal dose, even circumscribed by other standards, different lots of Schick test product cannot produce constant results in the many borderline reactors. Careful interpretation of borderline reactions is indicated, based on an understanding of the present status. The use of a fraction of the L+ dose of each toxin for the Schick test, would be practically possible and preferable to the use of a fraction of the minimum lethal dose.

Immunological Studies of Three Antitoxin Containing Globulin Fractions of the Horse

Summary The γ- and T-globulin fractions from tetanus toxin antiserum in the horse are shown to be closely related immunologically although some differences are apparent when the quantitative precipitin technique is employed. A third fraction, IV-C2, also rich in antitoxin content, was less closely related to the other two fractions. The IV-C2 antigen consists largely of β-globulin and contains essentially none of the other two components.

Influence of Type and Concentration of Antitoxin on the in vitro Toxigenicity Test for C. diphtheriae

Influence of Type and Concentration of Antitoxin on the in vitro Toxigenicity Test for C. diphtheriae

Studies on Diphtheria. III. The duration of immunity against diphtheria following vaccination as compared with that following clinical disease and carrier state

SUMMARYA comparison is made between the antitoxin values found in the following groups of subjects: I. Subjects vaccinated two years and one year ago; II and III. Cases of clinical diphtheria 2 and 20 years ago; IV and V. Diphtheria carriers 2 and 20 years ago and VI. Non‐immunized control subjects. The results are condensed in table 6. The following facts should be stressed: The vaccinated subjects are by far better off, as judged from the antitoxin concentration, than the former clinical cases or carriers. The former carriers have on the whole somewhat better antitoxin values than the former clinical cases, when examined after 2 and 20 years but the difference is not very striking.The results do not give support to the view often held, namely, that the diphtheria cases represent a selection of poor reactors.

Combined active and passive immunization against diphtheria

Combined active and passive immunization against diphtheria

Quantitative Studies on the Neutralization of Pathogenic Agents in Tissues by Circulating Antibodies

Summary The effect of substances augmenting capillary permeability, on the in vivo reactions between toxins and antitoxins, has been studied. The capillary effect can be demonstrated by the indirect intracutaneous test with diphtheric toxin and the indirect intramuscular test with tetanal toxin. It cannot be demonstrated by the indirect intramuscular test with diphtheric toxin. These findings have been explained on the basis of the nerve carriage theory of Meyer and Ransom. Nutrient broth, Difco peptone, Savita broth without peptone, histamine, the blood group substances, saliva, testicular extract and leech extract were shown to have a strong capillary effect. It is undecided whether or not the capillary effect of the blood group substances is due to impurities. The capillary factor in testicular extract is distinct from the spreading factor. Most substances affecting capillary permeability increase capillary permeability to such an extent that the concentrations of antitoxin in blood plasma and tissue fluid become identical. The indirect test provides a convenient method for studying the effects of substances affecting capillary permeability quantitatively. Some practical implications of our findings are discussed.

STUDIES ON THE DENATURATION OF ANTIBODY

Diphtheria antitoxin has been shown to be inactivated in the presence of urea. In 7.5 molar area at pH 7.82 nearly half the activity is destroyed in 24 hours, as measured by the neutralizing power of toxin. The reaction appears to be first order with respect to antitoxin, since the initial rate is independent of the antitoxin concentration, but the rate decreases rapidly with time. Higher or lower pH of the presence of M/10 sodium sulfite increases the antitoxin inactivation. The meaning of the results is discussed.

VARIATION IN DIPHTHERIA ANTITOXIN TITERS OF HUMAN SERUM

The literature records few determinations at frequent intervals of the diphtheria antitoxin titer of the blood serum of the same person over a period of several years. In 1925 Hamburger 1 reported a study of 6 subjects on whose blood serum antitoxin determinations were done on several occasions. No more than five determinations were done on any 1 subject, and the longest interval between the initial and the final determination of the antitoxin titer was one and one-half years. In 1 subject there was an increase in the antitoxin titer from 0.01 to 0.75 unit per cubic centimeter within two and one-half months without any apparent stimulus. In the other subjects there was considerably less variation. In 1929 Fitzgerald 2 published a paper on variations in the antitoxin content of serums of 6 adults over periods of from one to six years. In this article the amount of antitoxin in

A Simple Method for the Concentration of Blood Plasma and Serum

In the course of the preparation of plasma for human use it was observed that on the thawing of frozen plasma, separation into a number of layers takes place; the lowermost deep-yellow layer is rich in proteins, while the uppermost, nearly colorless layer, contains only a small amount of protein. On repeated freezing and thawing, the separation becomes more distinct. A more rapid and sharp separation of plasma into layers is obtained by centrifuging the frozen material.It is interesting that Bujwid1 used the freezing method for the concentration of diphtheria and tetanus anti-toxin in 1897. His results were confirmed by Ernst, Coolidge and Cook.2 Rossi3 used the freezing and centrifuging procedures for fractionating plasma and other colloidal solutions, and Hati4 used this technic for the concentration of complement, opsonins and anti-sera. In view of the above observations, the question arose as to whether these procedures might be employed for the concentration of plasma without excessive loss of solids...

THE BODY BUILD ASPECT OF IMMUNITY TO DIPHTHERIA

The anthropometric studies that have been made in various diseases, such as tuberculosis,¹allergy and poliomyelitis,²may be considered an indirect approach toward correlating body build with susceptibility and resistance to disease. However, not until there was some means of measuring an immune substance could immunity be subjected to anthropometric studies and the results of these studies, in turn, correlated with body build. The diphtheria antitoxin in human serum is at present the only readily measurable immune substance. The antitoxin does not occur in measurable amounts in the blood of most children unless, or until, its production is stimulated by administration of diphtheria toxin or toxoid. A study of the antitoxin response to such stimulation has been previously reported in one cross section of the child population.³The diphtheria antitoxin content of the blood serum was determined by the method of Jensen.⁴The distribution of the

BULLOUS SCHICK REACTIONS

Positive Schick reactions become bullous have been described in the medical literature. Schick 1 stated that such reactions were seen in very susceptible who had no diphtheria antitoxin in the blood. Dudley 2 mentioned a rare type of Schick reaction which vesiculates and produces a shallow ulcer of the skin. Griffith and Mitchell's textbook 3 has an illustration of a bullous reaction which, it is stated, occurs occasionally in individuals who have practically no However, no studies of the antitoxin content of the blood of patients having these bullous reactions have been recorded. Young and his co-workers, 4 who studied Schick reactions in adults, mentioned necrosis and vesiculation but were unable to discover the significance of such tissue reactions or to correlate them with an unusually low titer of antitoxin. Cooke and Sharma, 5 who performed over 600 Schick tests on newborn infants and their mothers,