Subject's Serum Research Articles

The binding affinity of Down's syndrome and control subject's serum IgG for group A streptococcal polysaccharide

Antibody binding data for naturally occurring anti-Group A streptococcal polysaccharide IgG, using the hapten, iodinated-GlcNAc, were determined by equilibrium dialysis for 22 institutionalizd subjects with Down's syndrome and for 23 non-Down's syndrome, institutionalized retardates. These data were compared to the binding data obtained from a group of 25 genetically normal, noninstitutionalized subjects. The number of antibody combining sites (N) was significantly higher in subjects with Down's syndrome than in either of the other groups. However, the binding affinity of Down's syndrome subject's anti-Group A streptococcal polysaccharide IgG was significantly lower than the binding affinities of the other groups of subjects. It is suggested that the lower binding affinity of IgG molecules of Down's syndrome subjects results in the commitment of more clones to the immune response; this may result in the increased serum levels of IgG seen in Down's syndrome subjects.

Statistical survey of “saturation analysis” calibration curve data for prednisolone, prednisone and digoxin

An extensive survey of radioimmunoassay calibration data for prednisolone, prednisone and digoxin indicated that the common practice of preparing calibration curves with individual subject's pre-dose plasma or serum, and using this to estimate unknown concentrations for the same subject, is not supported by statistical considerations. Preparation of calibration plots from pooled data is better because this introduces less bias in estimated concentrations. Such a method also saves a great deal of time, since it is not necessary to repeat the calibration procedure each time “unknowns” are being assayed. The data suggest that there is no optimum calibration plot for all radioimmunoassays. Rather, each antibody-drug combination should be investigated thoroughly to determine the best calibration plot for the particular combination. We found that the best calibration plots are; the logistic-logarithmic plot for prednisolone; nonlinear least squares fit to a polyexponential equation for prednisone; and a weighted least squares regression of normalized % bound versus concentration for digoxin. The error in the radioimmunoassay is usually concentration-dependent, and, in certain regions of the standard curve, is larger than the literature indicates, since, frequently, the error has been gauged from % bound values, but should be gauged from inversely-estimated concentrations.

Black—white differences in serum creatine phosphokinase (CPK) activity

The mean serum creatine phosphokinase activities of hospitalized black male and female, and white male and female psychiatric patients were established. The effects of both race and sex on mean serum creatine phosphokinase activities were both highly significant. Black males had significantly higher mean serum creatine phosphokinase activity than white males, black females and white females. White males and black females had significantly higher mean serum creatine phosphokinase levels than white females but were not significantly different from each other. Age, height and weight did not contribute to the difference between races. There was a trend for mean serum creatine phosphokinase activity to be greater for dark skinned black males than light skinned black males but this trend was not present in black females. The standard deviation of each subject's mean serum creatine phosphokinase activity was highly correlated with the mean. The levels of serum creatine phosphokinase activity found in these subjects were significantly greater than those previously observed with the method of Rosalki (J. Lab. Clin. Med., 69 (1967) 696).

61 Quantitative Aspects of Sensitivity in Allergic Children

Procedures were devised for quantitative chemical assay of histamine released by antigens from leukocytes separated from a 10-ml sample of blood. Nine concentrations of antigen can be utilized in each examination to find the amount required for maximum release of histamine or dose response. Studies have been conducted with a variety of antigens for comparison with clinical manifestations and wheal and flare dermal reactions to the antigens. Also the procedures have been employed to measure the capacity of the sera of sensitive persons to inhibit histamine release with specific antigens (presumably by antibodies) and to follow fluctuations in this capacity and in the sensitivity of leukocytes during injection therapy with antigenic extracts. Data have been accumulated from study of over 100 children, including 30 receiving injection therapy. Histamine is released from leukocytes of sensitive subjects by antigens specifically, in agreement with wheal and flare dermal reactions, and the leukocytes of normal non-allergic individuals are unaffected. Sera of normal persons enhance histamine release but the sera of allergic children inhibit histamine release by the antigens specifically involved. During injection therapy the sensitivity of leukocytes to release of histamine by antigen and the capacity of the patient's serum to inhibit histamine release may vary independently. The net effects are ascertained by determining the amounts of antigen required for release of 50 % of the total histamine in the cells in the presence of the subject's serum in contrast to normal serum. This comparison affords an objective index of any influence of injection therapy on sensitivity, and an objective means of grouping patients by immunochemical response before undertaking clinical appraisals. (APS)

Antigenic Variations in Herpes Simplex Virus Isolants from Successive Recurrences of Herpes Labialis

Summary Thirteen strains of herpes simplex virus (HSV) were isolated from recurrences of herpes labialis in four persons. Antigenic variations among these strains were defined by comparing the kinetics of their neutralization by rabbit antisera against a range of strains. Successive isolants from the same person frequently varied antigenically, but not in a regular manner. Variations could not be correlated with sequence or temporal proximity of recurrences, or with labial sites of lesions. The data substantiated a previously proposed classification of the majority of HSV strains into four serogroups, and indicated existence of two additional serogroups. Neutralization kinetic analysis of the reactions of eight strains from two of the subjects with five samples of their own sera likewise showed that successive isolants from the same individual are not necessarily identical. Relative rates of neutralization indicated that antibody specific for the HSV isolant associated with a given recurrence of herpes labialis is demonstrable in the subject's serum within 18 to 30 hr after the lesion appears. The respective serologic relationships, however, did not correlate with those established by rabbit antisera, and varied with different serum samples from the same person.

The Lack of Absorption of Ingested Bovine Antibody in Humans

Abstract In the past few years several articles in professional journals (1, 2, 3) and numerous reports in the public press have claimed that antibodies in cows' milk are absorbed in active form via the human gastrointestinal tract. Further, it has been suggested that by using milk from appropriately immunized cows, this method of passive immunization may be employed with benefit in the treatment of such human diseases as pollen allergies. So attractive is this hypothesis that clinical trials of milk from ragweed pollen immunized cows have been conducted in ragweed sensitive patients without, however, any apparent benefit (4). In these clinical trials the lack of a relatively pure antigen makes it impossible to determine quantitatively the amount of antibody in the milk or in the subject's serum after ingestion of milk, so that all results must be based on subjective observations.

Changes in the serum cholesterol and blood clotting time in men subjected to cyclic variation of occupational stress.

Accountants were selectively chosen as a self-controlled group for studying effects of cyclic occupational stress upon serum cholesterol and blood clotting time, since their routine work schedule is interrupted by urgent tax deadlines, associated with severe occupational stress. Forty male accountants (age 28 to 56) were bled biweekly for serum cholesterol and monthly for blood clotting time from January to June 1957. Complete records also were kept of weight, exercise, diet, relative work load, and any exposure to unusual avocational stress. When studied individually, each subject's highest serum cholesterol consistently occurred during severe occupational or other stress, and his lowest at times of minimal stress. The results could not be ascribed to any changes of weight, exercise, or diet. Marked acceleration of blood clotting time consistently occurred at the time of maximum occupational stress, in contrast to normal blood clotting during periods of respite. The possible implications of these results are discussed in relation to the problem of clinical coronary artery disease.

Some Investigations of Erythropoiesis in Human Bone‐Marrow Cultivated in Various Media

Summary.Experiments have been performed on human sternal marrow kept alive in vitro in Locke's solution with and without the addi‐tion of different antipernicious anemia preparationsor of serum from either the same or from another person. These materials have been shown to maintain erythropoietic activity a t various levels.The slight formation of red cells which takes place in Locke's solution comes to a standstill after 8–10 hours, but is resumed later when red cells are hemolyzed and presumably liberate nutri‐tive materials. Folk acid (0.1 yo) will increase the formation of red cells considerably and liver extract even more so. “Foreign” serum has about the same effect, but the subject's own serum causes most rapid erythropoiesis, thus pointing to the existence of a factor peculiar to each individual. A number of pathological cases were, investigated. The “spontaneous” production of the red cells was diminished in untreated pernicious anemia and leukemia, but was normal in various other conditions studied. I n most of the cases, the pathological inarrow responded quite as a normal marrow does when liver of folk acid is added to the nutrient fluid. Only in lymphogranulomatosis benigna was a significant decrease decrease below normal found in the product:on of red cells when these materials were used. Tyrosine increased the effect of liver extract in untreated pernicious anemia and in leukemia but had no added effect, on normal bone‐marrow.The production of red cells from the pathological marrow nour‐ ished by normal serum is slightly decreased below normal in some of the pathological cases, especially in leukemia. I n this disease and in pernicious anemia there was a decreased total function of bone‐marrow plus “native” serum. All pathological marrows, however, were considerably activated by the addition of normal serum to the nutrient fluid. The use of pathological serum with normal marrow results in a significant decrease in red cell produc‐tion over that found when normal marrow is activated by “foreign” normal serum.In patients in whom a decrease was observed in the activity of the bone marrow cells and of the serum, i. e. in pernicious anemia, effective therapy resulted in an increase in the amount of those substances n both cells and serum which activate the formation of red blood cells in the marrow cultures.These results considered together with the results found in investigations of the ripening of the reticulocytes in vitro show that there are certain active principles in the serum, some being necessary for the production of red cells and some for the ripening of the reticulocytes. It appears that certain of these principles are necessary for both processes.I extend my deep gratitude to the Carlsberg Foundation and to the King Christian the Tenth Pund for supporting these in‐vestigations.

Active Immunization of Human Beings with Tetanus Toxoid

Apparently the first attempt at active immunization of man was made by Louis Bazy working with Vallee. He gave iodized tetanus toxin obtained from Ramon at the Pasteur Institute to wounded soldiers. Three injections were given 5 days apart and 10 days after the last injection the men were bled and 1 cc. of the serum of each individual examined at the Pasteur Institute was found to contain from 10 to 100 “units” of tetanus antitoxin. In 1927 Ramon and Zoeller used tetanus anatoxin, toxin treated with formaldehyde, and then incubated. They injected several hundred subjects. The greatest immunity was obtained by giving 0.5 cc. of anatoxin, 1 month later 1 cc. and 2 weeks later 1.5 cc. The immunity acquired was apparently so great that 1 cc. of the subject's serum neutralized 500 to 1,000 minimum lethal doses for a guinea pig. Twelve to 18 months later the immunity was still higher. After 1 to 2 years a single injection of anatoxin, the so-called “injection de rappel”, resulted in a great increase in antitoxic power of the serum. 1 cc. of the serum neutralized 10,000 minimum lethal doses. The process of active immunization can not be hurried by shorter intervals between injections or large doses of anatoxin. In May, 1930, we assembled 18 volunteers, 5 adults 18 years and over, and the rest children varying from 5 years up, who were bled and no trace of tetanus antitoxin found in their blood sera. At weekly intervals 3 doses of tetanus toxoid, made by treating the toxin with 0.4% formalin and incubating at 38° for 10 days, were injected intramuscularly. The injections were painful for about 5 minutes, but no general reactions occurred and no local ones beyond moderately sore arms.