bluuebunny/crossref_metadata_2025 huggingface dataset:
Correlation of Three Potency Assay Methods for Smallpox Vaccines
Authors: Bartell Pasquale , Tint Howard
Published: January 2023

Comparisons were made between three methods for assaying the potency of smallpox vaccines. After the elimination of refractory animals, the rabbit scarification test proved to be as reproducible as titration in bovine embryonic skin tissue culture. Both these tests were more reproducible than infectivity titration in the egg.

Equations relating infectious units determined by the three tests were determined. An over-all correlation of 94% was obtained for the acceptance-rejection records of 126 vaccine samples when titration in the rabbit and in bovine embryonic skin were compared. Seventy-seven of the vaccines were acceptable on the basis of each test. Correlation between the egg and rabbit tests was 81%.

Stability studies on a lyophilized vaccine utilizing the several titration methods demonstrated, with remarkable consistency among tests, that the vaccine was stable for at least 18 months under vacuum or nitrogen.
Serum Properdin, Complement and Agglutinin Changes in Dogs with Staphylococcal Bacteremia
Authors: Baltch Aldona L. , Osborne Winifred , Canarile Leonard , Hassirdjian Anne , Bunn Paul A.
Published: January 2023

Serum properdin, complement and staphylococcal agglutinins were measured in 28 normal mongrel dogs, 16 operated dogs with an experimentally produced aorta inferior vena cava fistula and spontaneous or induced Staphylococcus aureus bacteremia, and seven normal mongrel dogs with induced S. aureus bacteremia.

Properdin levels in normal dogs ranged from 7.0 to 66.6 Ph N50/ml with a mean of 25.6 Ph N50/ml; complement levels ranged from 17.1 to 41.3 C1H2O/ml with a mean of 28.1 C1H50/ml; S. aureus agglutinin titers were very low (1:10 serum dilution) or absent. Rapid decline in the properdin and complement titers was seen in dogs surviving less than 24 hr following the first intravenous (i.v.) injection of S. aureus. In contrast, following the initial fall an increase in properdin and complement titers was seen in sera of dogs who survived at least 4 days following the first injection. A rather precipitous fall of both titers was seen in animals who succumbed after the 5th day of infection. The titer of agglutinins remained unchanged or increased slightly. If the animal survived the infection, the elevated properdin and complement titers returned to normal in 2 to 4 weeks. Agglutinin titers usually were found to be highest 9 to 28 days following the first i.v. injection of S. aureus. The height of the titer of serum properdin, complement and S. aureus agglutinins obtained before the experimental infection used in this study did not appear to influence the survival rate of this group of experimental animals. By altering the anatomical and physiological state of an animal, such as produced by an arteriovenous shunt, the animals were made very susceptible to bacterial infection, especially endocarditis, without any remarkable changes in serum properdin, complement or S. aureus agglutinins. When exposed to a bacterial agent, these operated animals responded with changes in the properdin, complement and agglutinin titers which were similar to those observed in physiologically intact animals.
Studies on the Production of Wassermann Reagin
Authors: Fowler Evelyn , Allen Rovelle H.
Published: January 2023

Antigens for the VDRL microflocculation test have been prepared containing: a) a synthetic substitute for cardiolipin, b) a synthetic substitute for natural lecithin, and c) synthetic substitutes for both cardiolipin and lecithin. Reagin-like antibodies were produced when the floccules formed by mixing each of these with syphilitic serum were injected into rabbits. Results are given for dimyristoyl glycerophosphoric acid as a substitute for cardiolipin and dioleoyl lecithin as a substitute for naturally occurring lecithin.

The relation of these findings to the formation of reagin in syphilitic infection is discussed.
Skin-Reactive Serum Factor in Rat Homotransplantation
Authors: Witebsky Ernest , Müller-Ruchholtz Wolfgang , Milgrom Felix
Published: January 2023

Large skin grafts were exchanged orthotopically between 29 pairs of unrelated female albino rats. The sera of these animals were tested for a skin reaction producing factor (SRPF) similar to that described in a previous paper on rat parabiosis. The sera of 17 rats out of 58 tested produced skin reactions when injected intradermally into the respective graft donor but always failed to produce a skin reaction in the animal itself. The sera which elicited a positive skin reaction in the graft donor also frequently produced positive results in other unrelated rats. In contrast, those sera which failed to elicit a skin reaction in the graft donor gave consistently negative results in other animals.

The skin reactions described in this paper were considered to be due to isoantibodies directed against the tissue constituents present in the skin of the graft donor but absent from the recipient of the graft.
Effect of Cations on Thermal Inactivation of Vaccinia, Herpes Simplex, and Adenoviruses
Authors: Wallis Craig , Yang Czau-Siung , Melnick Joseph L.
Published: January 2023

Three DNA viruses, vaccinia, herpes simplex, and adenoviruses, have been studied for thermal inactivation in the presence of monovalent and divalent cations. Na+ protected vaccinia and adenoviruses from inactivation at 50°C, but Mg++ and Ca++ markedly increased the rate of inactivation of all three agents. This confirms our earlier observation with another DNA virus (vacuolating SV40) (12) and is in contrast to the remarkable stabilizing effect exerted by divalent cations on the small RNA viruses (9, 10).
Observations on Incidence and Mechanism of Egg Insusceptibility to Trachoma Viruses
Authors: Jawetz E. , Chino S. , Hanna L.
Published: January 2023

In the fall of 1959 and the summer of 1960 there were periods of a few weeks when embryonated eggs failed to support the growth of trachoma and inclusion conjunctivitis viruses. Yolk sac material harvested from such insusceptible eggs and stored at -40°C for several months interfered with the lethal effect of trachoma virus challenge in susceptible eggs. Quantitation of these phenomena has been attempted and possible explanations proposed. During 1961 careful, frequent tests failed to reveal a period of insusceptibility. Eggs laid during and after a time of extreme heat appeared to be fully susceptible to psittacosis and trachoma viruses. Thus environmental heat and associated nutritional disturbances appear less promising as an explanation for egg insusceptibility than a transmissible, interfering agent.
Gel Double Diffusion Studies with Group B and Group A, Type 9 Coxsackie Viruses
Authors: Schmidt Nathalie J. , Lennette Edwin H.
Published: January 2023

The demonstration by the gel double diffusion technique of “specific” precipitating antibodies for the Coxsackie viruses suggests that the method offers considerable promise for use in the laboratory diagnosis of infections due to virus types A9, B1, B3, B4, B5 and possibly B6. Patients with Coxsackie B2 virus infections, however, could not be shown to develop “specific” precipitating antibody.

“Specific” precipitating antibodies for the infecting virus type were found in the sera of 97% of patients from whom a Coxsackie A9, B1, B3, B4, B5 or B6 virus was isolated and in 40 to 65% of these patients “specific” precipitating antibody developed over the course of the illness. However, since “specific” precipitating antibody disappears relatively rapidly during convalescence, its presence in any serum specimen may be evidence of a current or very recent infection. Heterotypic “specific” precipitating antibody was absent in 78% of the patients who yielded a Coxsackie virus, and when present would appear to be due to recent infection with the heterotypic virus, or to anamnestic response.
Studies on Mechanisms of Immunity in Listeriosis
Authors: Njoku-Obi Augustine N. , Osebold John W.
Published: January 2023

The interaction of peritoneal exudate cells of sheep with Listeria monocytogenes was studied in vitro in Kapral-type cell culture chambers. Four combinations of reactants were studied: immune cells + immune serum, immune cells + normal serum, normal cells + immune serum and normal cells + normal serum. Immune cells in immune serum resisted the destructive results of the parasitic interaction when compared with the other host cell systems. Samples taken at the 20th hr indicated that the exudate cell counts in the immune cell + immune serum chambers were 39% of the original, whereas the counts in the other systems had declined to less than 5% of their original values. Bacterial growth was suppressed in the immune cell + immune serum system. The increase in bacterial growth was greatest in flasks of media containing no host cells. However, bacteria also accumulated rapidly in chambers containing the three other combinations of reactants. Parasitic and phagocytic indices were comparable in normal and immune serum. Results are discussed regarding a cellular basis of immunity in listeriosis and relationships to other host-parasite studies.
A Kinetic Study on the Labeling of Serum Globulin with Fluorescein Isothiocyanate by Means of the Gel Filtration Technique
Authors: Tokumaru Tadasu
Published: January 2023

It was found that Sephadex gel filtration could be used for the quantitative analysis of reactants by rapid separation. Using simultaneous gel filtration of samples in a number of sister columns association constants and the number of sites were obtained for the kinetics of the labeling process of globulins with fluorescein. Reagents for protein precipitation were used to remove superficially adsorbed dye from fluorescent atibody. The results of such precipitation showed that the gel-filtered material contained smaller amounts of adsorbed dye than dialyzed material. The patterns of DEAE-cellulose and DEAE-Sephadex column chromatography were compared by the use of fluorescent antisera. It was shown that the complex of papain and fluorescent antibody could be separated from free papain satisfactorily on a Sephadex G-75 column.
Quantitative Studies on the Bis Diazotized Benzidine Method of Hemagglutination
Authors: Roberts Audrey N. , Haurowitz Felix
Published: January 2023

Quantitative studies on the BDB method of hemagglutination revealed that an average of 5 × 105 molecules of C14-anti-BSA rabbit globulin were fixed to each BSA sensitized rabbit erythrocyte which was agglutinated in antibody excess. Similar results were obtained in experiments using different suspensions of erythrocytes and freshly prepared BDB. There was a strictly linear relationship between the number of sensitized cells added and the number agglutinated in antibody excess. As the number of sensitized cells added per tube increased 6-fold, the quantity of homologous antibody bound per cell decreased by 50%. When the agglutinated red blood cells were hemolyzed with water, all of the bound C14 antibody was found in the stroma. Using H3-labeled BSA, it was demonstrated that an average of 1.3 × 106 molecules of antigen were coupled to each erythrocyte during the BDB sensitization procedure. The significance of these findings has been discussed.
Hyperreactivity to Endotoxin in Mice Infected with BCG
Authors: Suter Emanuel
Published: January 2023

The induction of hyperreactivity to bacterial endotoxin by BCG infection was studied in germfree mice and in mice chronically infected with Salmonella typhimurium. The results showed that infection of BCG in germfree and in chronically infected animals caused a similar increase in sensitivity to endotoxin. It is concluded from these experiments that concomitant infection is not a decisive factor in the emergence of endotoxin hyperreactivity during BCG infection. The experiments also indicated that germfree mice were almost equally resistent to BCG infection as were conventional animals.
Some Properties of an Inhibitor of Polyoma Virus Hemagglutination Isolated from Bovine Serum
Authors: Halperen S. , Schieble J. H. , Mori R. , Dinka S. , Ackermann W. W.
Published: January 2023

One of the several substances found in bovine serum which prevent agglutination of erythrocytes by polyoma virus has been isolated. The kinetics of its action indicate it reacts with the virus to form a product whose stability only at low temperature closely resembles the virus-erythrocyte complex. Like the red cell receptor, it is a mucoprotein. Some of its chemical and physical characteristics, as well as its visualization in the electron microscope, are described.
The Inhibitory Effect of Hydrocortisone on Lysis of Homologous Cells by Lymphocytes in Vitro
Authors: Rosenau Werner , Moon Henry D.
Published: January 2023

The effect of hydrocortisone was studied on the cytolytic activity of sensitized lymphocytes of BALB/c mice on homologous cells of the L strain in vitro. Hydrocortisone partially inhibited the destructive action of previously sensitized lymphocytes on homologous cells. There was no demonstrable lympholytic effect of hydrocortisone in vitro under the conditions of these studies. Recognition of the homologous cells by the sensitized lymphocytes was not altered by hydrocortisone. Therefore it is suggested that the inhibitory effect of hydrocortisone in vitro is due to blocking of the cytolytic activity of lymphocytes and not to the inhibition of the recognition of homologous cells by sensitized lymphocytes.
Two Different Anti-Insulin Antibodies in Guinea Pig Antisera
Authors: Yagi Yasuo , Maier Patricia , Pressman David
Published: January 2023

The antibodies in guinea pig anti-bovine insulin sera were fractionated by chromatography on DEAE cellulose. The anti-insulin antibody activity was found in two distinctly different fractions. Pooled antisera as well as antisera from individual animals contained both types of antibodies. However, the distribution of antibody activity in the two antibody fractions differed from serum to serum. The original antiserum and the two isolated antibody fractions showed very similar binding characteristics to crystalline bovine insulin. The antibodies differed in their electrophoretic mobility on starch: one migrated with the slow moving component of γ-globulin and the other with β2-globulin. Both antibodies were shown to have similar molecular size (probably of the order of 7 S globulins) by means of density gradient centrifugation.
Immunologic Identity of β-Lactoglobulins A and B
Authors: Gough Patricia , Jenness Robert
Published: January 2023

The quantitative precipitin test, analysis for cross - reactions, Ouchterlony double - diffusion plates, and micro-immunoelectrophoresis were used to show that the two types of β-lactoglobulin of bovine milk (β-lactoglobulins A and B) are serologically identical.
The Effect of Iodination on the Binding of Haptene to Arsonic Antibody
Authors: Koshland Marian Elliott , Englberger Frieda M. , Gaddone Suzanne M.
Published: January 2023

The effect of iodination on the binding capacity of purified antiphenylarsonic acid antibody was studied by means of the equilibrium dialysis technique. A direct correlation was found between the uptake of iodine and the loss in binding capacity. Since the presence of homologous haptene protected the binding sites from destruction by iodination, evidence was obtained that iodine reacts directly with groups at the active site. Furthermore, a comparison of the experimentally determined losses in binding capacity with the probable losses calculated from the assumption of one or two reactive groups at each site indicated that only one iodine reactive group is present at each site.
Enhancement by Endotoxin of the Primary Antibody Response to Bovine Serum Albumin in Chickens
Authors: Luecke Donald H. , Sibal Louis R.
Published: January 2023

Purified lipopolysaccharide of Salmonella abortus equi (endotoxin) was shown to enhance the antibody response of chickens to a single, intravenous injection of bovine serum albumin. Simultaneous injection of 5 or 10 µg of endotoxin/kg body weight and bovine serum albumin markedly potentiated all phases of antibody synthesis. The induction period was shortened, antibody levels were several-fold greater and detectable serum antibodies persisted for longer periods than in animals given antigen alone. Enhancement was demonstrated when endotoxin was given 24 hr after, but not 24 hr before, antigenic stimulation. The results are discussed with respect to the animal species involved as well as the time interval between endotoxin and antigen administration.
Plaques Produced by T2r Phage Particles Partly Neutralized by Normal and Immune Guinea Pig Sera
Authors: Cetin Enver Tali
Published: January 2023

Phage plated on agar medium produce plaques of a given size according to the properties of the phage and host strain. However, the size of the plaque depends upon the conditions of the environment, the composition and consistence of the medium. If these conditions are constant, phage particles of a given type produce plaques of the same size.

It has been observed that phage particles may, under certain conditions, give plaques that are smaller than normal. For instance, when phages active on a strain of Salmonella typhi Vi 4 were plated, they produced small plaques if exposed to considerable light (1). Moreover, it was demonstrated that T4 phage treated with immune serum taken early during immunization, in which period neutralized phage particles may reactivate, produced small plaques (2). In addition, following neutralization with high titer immune serum of T4, some of the active phage particles gave rise to small plaques (3).
The Relationship of Immediate Wheal Reactions to the Repeated Administration of Diphtheria and Tetanus Toxoid
Authors: Kuhns William J.
Published: January 2023

Immunization with single vs. repeated doses of diphtheria toxoid was carried out in human subjects to determine the effects of the mode and schedule of administering this antigen upon antitoxin titers and upon immediate wheal responses. A similar study was carried out using repeated doses of tetanus toxoid. Schick positive persons who remained Schick positive did not develop wheal reactions to toxoid. Schick negative persons who were immunized orally did not show appreciable increases in antitoxin titer or in immediate wheal reactions. All Schick negative persons who received repeated parenteral doses of toxoid showed considerable increases in antitoxin titer and immediate wheal reactions until after the time that toxoid was discontinued. The same amount of toxoid given in one dose usually caused a marked increase in antitoxin titer up to about 3 weeks, and this was followed by a rapid decrease in titer. Wheal reactions were not present as frequently or in as marked degree in persons who received one dose of toxoid as in persons who were repeatedly immunized. Delayed reactions occurred many times in most Schick negative persons who received repeated doses of parenteral toxoid, but did not occur in persons who received only one dose of toxoid. Delayed reactions following a dose of toxoid generally occurred prior to the onset of immediate wheal reactions. Evidence is presented for the concept that immediate wheal reactions, as inflammatory responses, are coincidentally within the framework of the immune response. The degree of coincidence may be influenced by modes and schedules of immunization. These factors may be important in complex formation, and complexes may in turn provide the potential for wheal reactions.
Antibody Responses Induced by Aqueous and Emulsified Influenza Virus Vaccines in Mice Exposed Daily to X-Rays
Authors: Berlin Byron S.
Published: January 2023

Hemagglutination-inhibiting (HI) antibody responses to aqueous and emulsified mineral oil adjuvant influenza virus vaccines were compared at 15, 27 and 43 days after vaccination in mice exposed to daily x-irradiation (except Sundays) beginning 1 day before vaccination. At each bleeding HI levels induced by the adjuvant were higher in similarly irradiated mice than those induced by an aqueous vaccine containing the same amount of antigen. Moreover, HI responses in the irradiated mice injected with emulsified vaccine were equal or higher at each bleeding than those in mice injected with aqueous vaccine but not irradiated. The findings were interpreted as indicating that the adjuvant effect of emulsified vaccine was reduced, but not abolished, by repeated exposures to x-ray.
Human Hypersensitiveness Induced by Very Small Amounts of Horse Serum
Authors: Hooker Sanford B.
Published: January 2023

Following toxin-antitoxin injections a noteworthy proportion of human subjects became hypersensitive to horse serum as shown by specific endermal reactions. The pertinence of this observation to serum therapy in general is discussed. In order to avert the widespread and disadvantageous influence which such induced hypersensitiveness is likely to exert upon the use of therapeutic horse sera, it is recommended that some other animal be chosen for the production of that antitoxin which is destined for use in toxin-antitoxin mixtures.
Desensitization to Local Action of Antigen (Arthus Phenomenon)
Authors: Opie Eugene L.
Published: January 2023

Sensitization to the local action of protein may be transferred to a normal animal by injecting the serum of an animal which has received repeated injections of the antigen. This observation may be explained by the assumption that the serum of the immunized animal contains an antibody upon which the local reaction (Arthus phenomenon) depends. Analogy with other reactions in which antigen and antibody are concerned suggests the possibility that saturation of the antibody with antigen may obliterate the phenomenon. Attempts have been made to desensitize animals which as the result of immunization exhibit acute inflammation and necrosis when injected with the antigen employed for immunization.

No desensitization of rabbits repeatedly injected with horse serum was observed by Knox, Moss and Brown (1) when they injected intravenously or into the peritoneum large quantities (5 to 10 cc.) of horse serum.
The Gradual Decrease of the Active Immunizing Value of Toxin-Antitoxin Mixtures Stored During One Year
Authors: Banzhaf Edwin J.
Published: January 2023

About two years ago when the active immunization from the 3 and 5 L+ toxin-antitoxin mixtures were compared the results showed that the 5 L+ did not give any greater percentage of active immunity in children than the 3 L+. The 5 L+ however, did give a higher percentage of local and constitutional reactions in children and adults. These were due to the greater amount, of protein substances in the mixture. The 5 L+ mixtures were made with the aged diphtheria toxin without any dilution other than that due to the addition of the proper amount of antitoxin, whereas the 3L+ mixtures were prepared from the 5 L+ toxin, diluted with water to contain 3 L+ and therefore had 40 per cent less of the reacting substances. Even with this lesser amount of reacting substances, the 3 L+ mixtures gave annoying reactions.
The Characteristics of Lipid-Adsorbed Virus Vaccines
Authors: Youngner J. S. , Axelrod Velma
Published: January 2023

A detailed study has been made of the properties of lipid-adsorbed virus vaccines. The influence of route of inoculation, suspending medium, and loading of lipid with antigen on antibody response were determined. Species differences in response to lipid-adsorbed vaccine were encountered; marked adjuvant action occurred in guinea pigs and mice but not in rabbits or chickens.

The toxicity and tissue reactions produced by lipid-adsorbed vaccines and the rate of disappearance of the antigen depot from the site of inoculation also were studied.

A series of water-insoluble lipids with different polar groups and sidechains were tested for their ability to act as immunologic adjuvants in order to determine the importance of available polar groups to the adsorbent and adjuvant properties of a lipid.
Specificity of Antisera to Mouse Ascites Tumor in Rabbits Immunologically Depressed with Mouse Red Cells
Authors: Reif Arnold E. , McVety Lora M. , Klein Esther R.
Published: January 2023

The concept tested is whether antisera showing increased specificity against a mouse ascites tumor can be obtained by immunizing with this tumor rabbits that were immunologically depressed by injection of mouse red cells when newborn.

One-half the young in three litters of rabbits (depressed) were injected with washed mouse red cells on the day of birth and several times thereafter until day 38 after birth. The other half (controls) were left uninjected. At 9 ½ to 12 months, one-half the rabbits in both depressed and control groups were immunized with mouse red cells, the other half with L4946 mouse tumor.

Antisera from depressed rabbits finally immunized with red cells had hemagglutination, hemolysin and cytolytic titers respectively 17, 6 and 6 times lower than their controls, indicating considerable immunologic depression. Antisera from depressed rabbits finally immunized with tumor cells had titers respectively 6, 2 and 2 times lower than their controls, indicating some cross-depression against tumor cells. The specificity of antisera against L4946 tumor was precisely the same in depressed rabbits and in nondepressed controls; absence of results for the latter control group in recent studies by Levi et al. and by Garb et al. throws serious doubt on a positive interpretation of their data.
The Effect of Excess Antigen on the Anaphylactic Reaction
Authors: Liacopoulos P. , Halpern B. N. , Frick O. L.
Published: January 2023

The following methods were used to determine the effect of large amounts of antigen in the eliciting of anaphylactic reactions: a) anaphylactic contraction of strips of ileum passively sensitized in vitro; b) liberation of histamine by slices of guinea pig lungs passively sensitized in vitro; and c) passive anaphylactic shock in mice. Rabbit anti-ovalbumin or anti-bovine serum albumin was used in the studies of isolated tissues, and mouse anti-ovalbumin was used for passive sensitization of mice. These three types of anaphylactic reaction could be specifically inhibited when a large quantity of antigen was used as the eliciting dose. The eliciting dose of antigen (ovalbumin, bovine serum albumin) was 1000 to 10,000 times higher than the minimal amount of antigen necessary to produce a maximal anaphylactic reaction or fatal anaphylactic shock. In this inhibition of anaphylactic reactions by excess of antigen, the larger the quantity of antibody used for passive sensitization, the greater the dose of the antigen necessary for inhibition.
The Temporal Relationship of Acquired Tolerance and the Immune Response Following Injection of Bovine Serum Albumin into Neonatal Mice
Authors: Sorem Gail L. , Terres Geronimo
Published: January 2023

In the present study the temporal relationship of acquired immune tolerance and the immune response was determined in mice injected at birth with BSA. It was found that both were finite in time and that their degree and duration were determined by the amount of BSA injected. The results also demonstrated that the acquired tolerance and the immune response that followed were due to the same antigen.
Antiglobulins in Normal Serum
Authors: Watson R. Glenn , Collins Nancy M.
Published: January 2023

The Waaler-Rose activity previously noted in normal rabbit sera was studied. The factor responsible for this activity was found in all normal rabbit sera tested in titers of 1:64 to 1:2048. This factor was primarily localized in the water-soluble fraction of rabbit serum, was heat-stable at 56°C, and could be absorbed from rabbit serum with sheep erythrocyte stroma highly sensitized with rabbit hemolysin. No detectable amount of the Waaler-Rose factor could be removed from the serum by absorption with latex particles coated with human Cohn Fraction II. The Waaler-Rose factor in rabbit sera was differentiated from the Forssman heterophile antibody and the rheumatoid factors and was not associated with conglutinin activity.

A latex-fixation factor was demonstrable in all normal rabbit sera tested in titers of 1:20 to 1:512. This factor differed from the rabbit Waaler-Rose factor in that it was primarily confined to the water-insoluble fraction of the sera and was completely inactivated when the sera were heated at 56°C for 14 min. The latexfixation factor could be completely absorbed from rabbit sera by latex particles coated with human Cohn Fraction II. However, the absorption of rabbit sera with sheep erythrocyte stroma highly sensitized with rabbit hemolysin did not remove all of this factor but did remove about 75% of it. The latex-fixation factor in rabbit sera was differentiated from the Forssman heterophile antibody and the rheumatoid factors and was not associated with conglutinin activity. The addition of small amounts of normal human serum to rabbit serum containing high titers of latex-fixation activity caused inhibition of latex-fixation activity. The inhibitory substance in human serum responsible for the inhibition could be inactivated by heating the serum at 56°C for 30 min.
Studies on an Endotoxin of a Group C Neisseria Meningitidis
Authors: Mergenhagen S. E. , Martin G. R. , Schiffmann E.
Published: January 2023

The lipopolysaccharide endotoxin of a Group C Neisseria meningitidis strain 1908 has been isolated by phenol-water extraction and further purified by differential ultracentrifugation of the water soluble lipopolysaccharide-nucleic acid complex. The lipopolysaccharide was found to contain 20% lipid (composed of cephalin, fatty acids and plasmalogen) with polysaccharide constituents identified as glucose, galactose, glucosamine and sialic acid. The endotoxin contained less than 1% protein, and less than 1% nucleic acid. Hemagglutination inhibition tests revealed that this meningococcal lipopolysaccharide was able to inhibit antibodies from agglutinating sheep erythrocytes sensitized with Group C specific capsular polysaccharide composed principally of units of sialic acid. The converse was not found, however. Analytical ultracentrifugation and the separation of lipopolysaccharide and of capsular polysaccharide by gel filtration provided evidence that the sialic acid formed an integral part of the biologically active lipopolysaccharide. “Crude” and “purified” lipids extracted from parent lipopolysaccharide failed to demonstrate lethal activity in high dosages in normal or pertussis-sensitized mice. They were found to be nonpyrogenic and nonpreparing for the local Shwartzman reaction in rabbits in comparison to the native lipopolysaccharide. In rather small dosages, however, they were capable of increasing nonspecific resistance in the mouse to a lethal E. coli infection.
Quantitative Studies of a Conglutinating System in Normal Bovine Serum
Authors: Sage Harvey J. , Ransby Arnold , Leon Myron A.
Published: January 2023

A conglutinating system present in normal bovine serum has been shown to consist of antipolysaccharide, conglutinin and complement. Conglutinated dextran yielded conglutinin, C′1 and C′4 on extraction with EDTA, and both high and low molecular weight antidextran on extraction with maltose. Details of the chromatographic purification of conglutinin and the high and low molecular weight antidextran are given. The isolated conglutinin was essentially homogeneous by free electrophoresis and by ultracentrifugation, but was heterogeneous immunochemically. Conglutinin migrated as a γ-globulin, had a sedimentation constant of 6.6 and a molecular weight of 650,000.
Recognition of Foreign Erythrocytes by the Liver in Mice
Authors: Nelson Eric L. , Buras Netty S.
Published: January 2023

An experimental model has been developed for the study of recognition by cells of the reticuloendothelial system (RES). A quantitative method employing Cr51 labeled cells has been devised for studying phagocytosis of homologous and heterologous erythrocytes. Homologous and heterologous red blood cells are phagocytized at different rates. The spleen primarily takes up homologous (mouse) red cells and the liver removes foreign (human, guinea pig, rabbit) erythrocytes. Genetic differences among strains of mice may be reflected in their ability to take up in the spleen red cells of nonrelated donor mice. The ratio of recovery of radioactivity (RR ratio) between the blood, spleen and liver may be used to characterize a system employing erythrocytes of a particular donor in any given recipient. A comparison of RR ratios obtained under varying experimental conditions may be expected to reflect responses in the recognition mechanism of the RES.
Studies on the Specificity of the Cellular Infiltrate in Delayed Hypersensitivity Reactions
Authors: McCluskey Robert T. , Benacerraf Baruj , McCluskey Jean W.
Published: January 2023

In actively immunized guinea pigs given H3 thymidine during the 2 days preceding skin testing, delayed hypersensitivity reactions contained a high percentage (87 to 92%) of labeled mononuclear cells.

Experiments were performed in which two noncross-reacting sensitivity systems were transferred by lymph node cells to single recipient guinea pigs, with the donors sensitized to one of the antigens having received H3 thymidine, whereas the donors to the other system supplied unlabeled cells. Delayed sensitivity reactions elicited by each of the antigens at separate sites in these recipients were found to contain comparable numbers of labeled cells. The average percentage of labeled mononuclear cells in the infiltrate was 4%.

Experiments were performed in which delayed sensitivity was transferred by unlabeled lymph node cells from actively immunized donors into nonimmunized guinea pigs which had received H3 thymidine during the 2 days preceding transfer. Delayed sensitivity reactions in these recipients contained a high percentage of labeled mononuclear cells (68 to 91%).

The observations indicate that the great majority of mononuclear cells which comprise the infiltrate in delayed sensitivity reactions are not specifically sensitized cells. In addition, the findings show that almost all of the mononuclear cells comprising the infiltrate are cells which have recently proliferated. The observations fail to provide evidence for the specific attraction of sensitized cells to the antigen. It is suggested that the delayed hypersensitivity reaction is initiated by the random arrival of a small number of specifically sensitized cells at the antigen site.
Development of Viral Antigen and Infectious Virus in HeLa Cells Infected with Reovirus
Authors: Spendlove Rex S. , Lennette Edwin H. , Knight Charles O. , Chin Jean N.
Published: January 2023

Development of viral antigen and infectious virus were followed in HeLa cells infected with type 1 reovirus. The antigen was found to go through a sequence of four stages determined by shape and intracellular location. Six to eight hours after infection, antigen appeared in particulate form in the cytoplasm. The particles coalesced to form a reticulum-like structure disseminated throughout the cytoplasm; the reticulum in most cases subsequently became perinuclear. In the final stage, the antigen lost much of its reticular structure and spread confluently throughout the cytoplasm. The cytoplasm began “bubbling” and cell processes containing viral antigen constricted and pinched off from the cell.

Cell-associated virus and viral antigen appeared simultaneously after 6 to 8 hr of infection and increased to a maximal concentration at 24 hr. The rate of virus release was slow and appeared to be related to the “bubbling” phenomenon. The effect of the location of antigen and its slow release from the cell may be related to the slow spread of virus in plaques.
Cellular Localization of Radioactive Antigen in Immunized and Nonimmunized Mice
Authors: Speirs R. S. , Speirs E. E.
Published: January 2023

Purified tritiated tetanus toxin was injected intraperitoneally into 42 mice previously immunized with tetanus toxoid. An equivalent amount of tritiated tetanus toxin neutralized with mouse antitoxin was injected intraperitoneally into 28 nonimmunized mice. Animals were autopsied at various intervals, and autoradiograms made of the peritoneal exudate, spleen, bone marrow, lymph nodes and thymus.

Neutrophils containing radioactive antigen were observed in all animals autopsied between 2 and 24 hr after injection, following which there was a marked reduction in number. Many of the neutrophils were phagocytized by macrophages. Lymphocytes and macrophages containing radioactive antigen were present in the inflammatory exudate 2 hr after injection. In the immunized animals injected with tritiated toxin, there was a progressive increase in the number of macrophages containing antigen, up to the 4th day. In the nonimmunized animals injected with neutralized toxin fewer labeled macrophages were observed, and there was a progressive decrease in their number after the 1st day of inflammation. Labeled eosinophils were observed between the 2nd and 8th days of inflammation. The eosinophils did not engulf antigen directly but appeared to receive it from disintegrating labeled mononuclear cells. Certain mononuclear cells became swollen, highly vacuolated, and acted as a center around which other cells became attached, forming a rosette of cells. The number and cellular composition of these rosettes varied at different stages of inflammation. Macrophages and basophilic mononuclear cells containing radioactive material were observed in the spleen, mediastinal, inguinal, axillary, mesenteric and cervical lymph nodes. These cells were also observed in the bone marrow and occasionally in the thymus. Consistently greater numbers of eosinophil and mononuclear cells containing radioactive material were found in the inflammatory exudate and lymphoid tissues of animals which had been immunized to tetanus toxoid prior to the injection of the radioactive toxin.

These results indicate that antigen localizes in different cells during various stages of an inflammatory response, but eventually becomes concentrated in macrophages. It was suggested that the role as well as the fate of the antigen in a macrophage varies depending upon the manner in which it reaches the macrophage.
The Opsonic Effect of Complement on the Phagocytosis of γ-Globulin-Coated Bentonite Particles
Authors: Rytel Michael W. , Stollerman Gene H.
Published: January 2023

Evidence has been presented which indicates that opsonization of γ-globulin-coated bentonite is diminished in sera lacking C′1, C′2 and C′4, but not in sera lacking C′3. These observations provide additional evidence for the similarity of γ-globulin-coated particles and specific antigen-antibody complexes in their interactions with the complement system.
Electrophoretic Boundary Disturbances as a Test for Antigen-Antibody Interaction and Some Observations on the Production of Nonspecific Boundary Disturbances
Authors: Farr Richard S. , Campbell Dan H. , Vinograd Jerome
Published: January 2023

The addition of antigen to the buffer over the ascending and descending limbs of a free boundary electrophoresis cell containing antiserum results in an immunologically specific disturbance of the γ-globulin boundary. This technique provides a sensitive method for detecting either antigen or antibody. The antigens studied have been bovine serum albumin, Pneumococcus polysaccharide (SSSII), Boivin-type antigen from E. coli and bovine serum globulin. The disturbance may develop in the ascending and/or descending limb depending upon whether the antigen migrates slower, faster, or at the same rate as the antibody component. The method detects as little as 0.1 µg of antigen/ml in the buffer and as little as 10 µg of precipitating antibody/ml in the serum compartment of the cell. The E. coli antigen differed from the others in that it induced γ-globulin boundary disturbances when tested against normal sera, providing further evidence for the presence of antibodies against E. coli in normal sera. The descending β-boundary disturbance frequently observed during routine free-boundary electrophoretic analysis can be eliminated by adding as little as 0.24 mg of albumin/ml to the buffer over the descending limb. Some nonspecific boundary disturbances which may be encountered when attempting to apply this method to detect antigen and antibody are discussed.
Factors Affecting Primary and Secondary Antibody Production by Splenic Tissues
Authors: Kong Yi-Chi M. , Johnson Arthur G.
Published: January 2023

The capacity of isolated rabbit splenic tissues to form antibody was investigated by using tissue culture and cell transfer procedures. Hemagglutinating antibodies to bovine γ-globulin (BGG) were regularly demonstrated when splenic tissues were cultured in vitro three or more days following a single injection of antigen. After a second injection of BGG, splenic tissues produced antibodies in vitro when removed two or three days after antigen administration. Bacterial endotoxin given simultaneously with antigen did not increase the primary or secondary antibody levels of splenic tissues cultured in vitro. The addition of BGG in vitro to splenic explants from normal or previously immunized rabbits failed to initiate a primary or a secondary antibody response.

The capacity of splenic tissues to form antibody after a single injection of BGG into rabbits was transferrable to x-irradiated recipients on days 1, 2, and 3 after antigenic stimulation. The enhancing effect of endotoxin, given either to the donor simultaneously with BGG or to the recipient immediately following cell transfer, was equivocal.

Fluorinated pyrimidines, 5-fluorouracil and 5-fluoro-2′-deoxyuridine, inhibited secondary antibody formation when added to splenic tissues obtained 2 days after a second injection of BGG. Preliminary experiments indicated that thymidine partially reversed the inhibitory effect of 5-fluoro-2′-deoxyuridine.
Competition of Soluble Antigens by Tissue Culture Assay
Authors: Cremer Natalie E.
Published: January 2023

Intravenous injection of 10 mg of hemocyanin (KLH) simultaneously with or 24 hr after intravenous injection of 10 mg of bovine serum albumin labeled with S35 (S35-BSA) into rabbits hyperimmunized to both antigens resulted in antibody formation to both antigens by spleen cells in vitro. However, intravenous injection of 10 mg of KLH 9 to 24 hr prior to S35-BSA into similar rabbits caused inhibition of BSA antibody production in vitro. The inhibitory effect was diminished if KLH was given 48 hr or 72 hr prior to S35-BSA and no inhibition occurred if KLH was given 6 days prior to S35-BSA.

The inhibition did not appear to be dependent upon excess concentration of S35-BSA in the inhibited cultures and could not be correlated with circulating serum titers just prior to challenge over the period tested or to depletion of nutrients in the medium. The inhibition was dosage-dependent, 10 µg of KLH being inadequate to cause inhibition.

The primary or secondary response to KLH was insufficient to cause inhibition of BSA antibody production in vitro by spleen cells from rabbits hyperimmunized to BSA only.
Studies on Immunity to Toxins of Clostridium Botulinum
Authors: Fiock Mary A. , Cardella Matteo A. , Gearinger Naomi F.
Published: January 2023

Purified toxins of Clostridium botulinum types A, B, C, D and E were converted to toxoids and adsorbed on aluminum phosphate; four preparations of pentavalent toxoid were investigated in groups of 15 to 30 volunteers. Each pentavalent product contained 1.7 Lf of type A, 0.54 Lf of type B, 50,000 LD50 equivalents of type C, 4 Lf of type D, and 0.5 Lf or 100,000 LD50 equivalents of type E toxoid/ml. All flocculation potencies are referred to standard antitoxins prepared by the Microbiological Research Establishment, Porton, England. Doses of the first preparation of pentavalent toxoid (0.5 ml) were administered subcutaneously at 0, 2 and 10 weeks; the other preparations were given in the same way except that a 0-2-12 week schedule was used. Booster doses of 0.5 ml were given 1 year after the initial injection. Serum antitoxin titers of individuals were determined 12 to 14 and 52 weeks after the initial injection, and again 8 weeks after the booster, by neutralization titrations in mice. After the initial series of injections a considerable proportion of the individuals developed antitoxin titers for all or most of the five toxins; the proportion of satisfactory titers varied with the type and also with the preparation. Good booster responses were obtained with all preparations, and antitoxin titers to each of the five types were found in 86 to 100% of the individuals immunized. Stability studies with one preparation showed that the toxoids retained considerable antigenicity for at least 16 months at 4°C.
Cross-Protection in Animals Infected with Group A Arboviruses
Authors: Hearn Henry J. , Rainey Cullen T.
Published: January 2023

Experiments were carried out to investigate the phenomenon of cross-protection among Venezuelan equine encephalomyelitis (VEE), eastern equine encephalomyelitis (EEE) and Semliki Forest (SF) viruses in a variety of laboratory animals after immunization by the intraperitoneal, subcutaneous or respiratory routes; the last was effected by exposing the animals to aerosols of virus. One injection of an attenuated strain of VEE virus (9t) protected guinea pigs against a lethal challenge dose of EEE or SF virus in guinea pigs and mice, respectively. Two injections of live SF virus protected guinea pigs against small doses of VEE or EEE virus. Mice vaccinated with strain 9t responded by demonstrating resistance mechanisms that appeared to operate in series. This consisted of, first, an early nonspecific interference phase, followed by a second, specific phase. The second phase also included a partially specific mechanism of resistance of unknown origin and of relatively long duration manifested as cross-protection in the group A viruses.
Immunologic Studies of Heart Tissue
Authors: Kaplan Melvin H. , Craig John M.
Published: January 2023

Rabbits given repeated injections of beef or rat heart homogenates in aluminum hydroxide gel adjuvant developed focal cardiac lesions associated with antibodies to heart. The lesions were characterized by myocardial cell necrosis or degeneration with infiltration of mononuclear and polymorphonuclear cells, among which eosinophils were frequently prominent. Myocardial lesions of varying severity were observed in 7 of 9 animals injected with beef heart and 6 of 12 animals injected with rat heart. An extensive myocardial degeneration and fibrosis was observed rarely. In 13 control rabbits injected with human γ-globulin in adjuvant, these scattered focal lesions were not observed.

The incidence and severity of cardiac lesions could not be related to the titer of antibodies to heart as measured by complement fixation or immunofluorescence, or necessarily, to presence of flocculating antibodies.

Bound γ-globulin was detected within cardiac myofibers in focal sites of myocardium in several animals of the experimental groups, but not in the control animals. The sparseness of such deposits of bound γ-globulin in myocardium of most of the experimental animals, despite the presence of circulating antibodies to heart, suggested that impermeability of myofibers to antibody may represent a limiting factor in the pathogenesis of these lesions.
A Factor Other than Streptococcal Nicotinamide Adenine Dinucleotidase Which Combines with Antibody to This Enzyme
Authors: Ayoub Elia M. , Wannamaker Lewis W.
Published: January 2023

An enzymatically inactive factor which binds with antibody to streptococcal nicotinamide adenine dinucleotidase (NADase), possibly an inactive form of NADase or an enzyme precursor, is produced in early streptococcal culture supernatant fluids. A similar factor is found in fully grown culture supernates in the absence of sufficient sulfhydryl compounds. The antibody-combining capacity of this inactive factor and of NADase chemically inactivated by sulfhydryl inhibitors is equivalent to that of active NADase.

These findings are of theoretical interest in that the site of enzymatic activity does not appear to be involved in the specificity of the antibody configuration. Of practical concern is the demonstration that the presence of inactive enzyme will result in spurious antibody titers.