bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effect of Sulfhydryl Blocking and Disulfide Cleaving Reagents on the Hemolytic Activity of C′2 and C′3 of Human Complement
Authors: Polley M. J. , Müller-Eberhard H. J.
Published: December 2022

The observation that treatment of C′2 with a critical concentration of iodine led to enhancement of the hemolytic activity of the component and to stabilization of the C′42 complex was previously postulated to be due to oxidation of one or more sulfhydryl groups within the C′2 molecule. Treatment of oxyC′2 with 10-3 M dithiothreitol fully reversed the iodine effect and subsequent treatment with iodine again caused enhancement of C′2 activity. These observations suggest the presence of two free sulfhydryl groups in the native C′2 molecule, which are situated in such a position as to be sterically capable of forming a disulfide bond subsequent to oxidation with iodine. Molecular weight determinations, utilizing s-rates obtained by sucrose density ultracentrifugation and diffusion coefficients, by Sephadex gel filtration, indicated that the molecular weight of the product of the reaction of C′1 esterase with C′2 (C′2i) possessed a molecular weight approximately 33,000 less than that of the native molecular (C′2 = 117,000; C′2i = 84,000).
Immunochemical Studies on Blood Groups
Authors: Lloyd Kenneth O. , Kabat Elvin A. , Beychok Sherman
Published: December 2022

The majority of blood group substances from hog stomach linings and one of two blood group substances from human stomach linings were able to precipitate with the plant lectin, concanavalin A. However, none of the blood group preparations from human ovarian cyst fluids tested were precipitable. Concanavalin A precipitates with branched polysaccharides having terminal α-D-glucopyranosyl or α-D-mannopyranosyl residues and has a fairly wide specificity directed toward any terminal non-reducing sugar residues having the D-arabino configuration at C-3, C-4 and C-6 and an α-configuration at C-1. The isolation, by degradation of hog gastric mucin with NaOH-NaBH4, of an inhibitory disaccharide: 3 or 4-O-α-(2-acetamido-2-deoxy-D-glucopyranosyl)-D-galactitol (α-D-GNAc-(1 → 3 or 4)-D-galactitol) indicates the basis for the reaction of concanavalin A with hog gastric blood group substances.

Two samples of hog gastric blood group substances were fractionated by concanavalin A on a preparative scale into two fractions: one had only blood group activity and was not precipitated by concanavalin A while the other fraction was precipitable by both anti-A and concanavalin A.
Antigen-Antibody Reactions in Agar
Authors: Hayden Alonza R. , Becker Elmer L.
Published: December 2022

Several conditions of storage of antisera to be used in quantitative determinations of antibody were investigated to determine their effect on the optical density measurements in gel precipitin techniques. Antisera fleshly stored at 4°C showed decreased O.D. after 3 days, Antisera stored at 25°C showed an initial decrease in O.D. followed by an increase which appeared maximal at 16 days, and, finally, a decrease in O.D. Antisera incubated at 56°C showed an initial decrease in O.D. followed by an increase in O.D. The initial decrease in O.D. following storage of antisera at 4°C or 25°C or heating of antisera at 56°C paralleled the loss of hemolytic complement activity of the antisera, the presence or absence of which was determined by complement fixation. The factor causing the initial O.D. decrease could be removed from the antisera by absorption with an immune precipitate. Also, preliminary experiments, not described here, show that the factor or factors responsible for the initial high O.D. found with fresh antisera could be removed by treating such antisera with either EDTA or zymosan. The increase in O.D. during storage at 25°C or incubation at 56°C appears to be due to denaturation or alteration of serum components in such a manner that they interact with the antigen antibody precipitation process to produce denser precipitates.

A statistical comparison of the results of analysis of nine antisera using both the classical precipitin technique and the agar gel precipitin techniques, by means of Student's t test, indicated no difference in the means at the 5% level, provided O.D. corrections of zero to 0.02 units were made in the gel precipitin measurements.

The agar gel precipitin technique offers considerable savings in both time and material, since an analysis can be obtained from O.D. measurements at a single concentration of antiserum after only 2 days of incubation at 25°C.

The data presented here indicate that for routine use of this technique, one should: a) inhibit the hemolytic complement of all antisera to obtain standard curves with zero intercepts, b) store such decomplemented antisera at subzero temperatures to prevent further changes which might influence O.D. values and c) keep the antiserum-agar mixture in the reaction bath at 56°C for the minimal time needed to charge triplicate reaction tubes to prevent abnormal O.D. increases, particularly those observable at higher antiserum concentrations.
Antigens of Normal and Atherosclerotic Intima
Authors: Intorp Hans W. , Milgrom Felix , Witebsky Ernest
Published: December 2022

Rabbit antisera to normal and atherosclerotic intima of human aorta were used to compare the antigenic composition of these tissues. The antigen characteristic for normal tissue was a thermolabile component with fast electrophoretic mobility. This component was considerably reduced in amount in atherosclerotic plaques. Atherosclerotic lesions contained two “novel” antigens, virtually absent from normal intima. These antigens were identified as fibrinogen and C-reactive protein, respectively.
Immunochemical Studies on Cobrotoxin
Authors: Chang C. C. , Yang C. C.
Published: December 2022

Cobrotoxin, the crystalline toxic protein obtained from Formosan cobra venom, is immunochemically homogeneous and was proved to be the main toxic protein in the venom.

One hundred per cent precipitable antibody was isolated immunospecifically by gel filtration on Sephadex G-100 after the antigen-antibody complexes had been dissociated with 0.53 M formic acid-0.15 M NaCl, pH 2.05. The specific neutralizing capacity of the purified antibody was 17.5 greater than that of the antisera.

The purified antibody was digested with papain and the univalent fragments formed soluble antigen-antibody complex with the homologous antigen. An estimate of the molecular weight of the soluble complex formed from cobrotoxin and Fragment I indicated that cobrotoxin consists of three antibody-combining sites/molecule.
Communications
Authors: Kubo Ralph T. , Benedict Albert A.
Published: December 2022

Enzymatic digestion of IgG immunoglobulins has been used extensively to study the relationship between function and structure of these proteins. Papain hydrolysis of rabbit IgG yields two kinds of fragments (1): Fab, which is composed of H- and L-chain determinants and is the structural site for antibody activity; and Fc, which contains only H-chain determinants and is crystallizable. The Fc fragment has been crystallized from rabbit IgG (1), human IgG (2), and guinea pig IgG (3). The conditions for formation of Fc crystals generally require dialysis against distilled water or low ionic strength buffers in the cold. Chicken IgG also is hydrolyzed by papain into Fab and Fc fragments (4–7), but crystalline like structures rarely have been seen in the Fc preparations. Evidence is given here that chicken Fc is easily crystallized under conditions quite unlike those used for the crystallization of Fc from mammalian IgG.
Genetic Factors in Leucocyte Responses to Endotoxin: Further Studies in Mice
Authors: Sultzer Barnet M.
Published: December 2022

Comparative studies of the leucocyte responses of two strains of highly inbred mice, which radically differ in their susceptibility to the lethal effects of endotoxin, indicate that the proportion of neutrophils to mononuclear cells in the peritoneal cavity after the intraperitoneal injection of endotoxin appears to be a function of the relative toxicity of the endotoxin. Therefore, the extravascular accumulation of leucocytes and mononuclears at the site of endotoxin injection may be a function of mechanisms that are influenced by the susceptibility of the host to the toxic effects of endotoxins. The degree of susceptibility appears to be determined in great measure by the genetic constitution of the host.
A Simplified Procedure for the Preparation of Immunoglobulin-Class-Specific Antisera
Authors: Henney Christopher S. , Ishizaka Kimishige
Published: December 2022

A method is described for preparing immunoglobulin-class-specific antisera which requires neither the isolation of homogeneous immunoglobulin preparations for immunization, nor the subsequent absorption of antisera to remove cross-reacting antibody. The method is based on the principle that guinea pigs rendered tolerant to one immunoglobulin by i.v. injection of 5 mg protein, and simultaneously challenged with another in complete Freund's adjuvant, make antibody only to the class-specific determinants on the challenging antigen. Guinea pigs were given 5 mg of human γG i.v., and simultaneously challenged with mixtures of immunoglobulins included in complete Freund's adjuvant. Antisera specific for α, µ, δ and ε chains were thus produced.
Cross-Reactions among Group A Streptococci
Authors: Wiley Grove G. , Bruno Pauline N.
Published: December 2022

C101/21 cocci, type 41, and strain G1, of uncertain type, shared a cross-protective relationship with each other and with C126/21 cocci, type 43. The relationship with type 43 was in one direction, i.e., type 41 antiserum was bactericidal for type 43 cocci but the reciprocal reaction was negative. Absorption experiments revealed bactericidal antigenic determinants shared with type 43 cocci in addition to others which were bactericidal for the homologous cocci only. The relationship of G1 cocci to type 43 was similar to that for type 41. In the reciprocal relationship between type 41 and G1 cocci absorption experiments demonstrated shared protective determinants plus others which were specific for the homologous cocci in each case. Agar gel tests with heterologously and reciprocally absorbed type 41 and G1 antisera showed lines of precipitate with each strain which corresponded to cross-protective and homologous bactericidal reactions. It is suggested that cross-protection among the types of group A streptococci appears to be common enough to play a part in streptococcal immunity if a corresponding protection occurs in vivo.
The Suppressive Effect of Rauscher Leukemia Virus on the Secondary Antibody Response of Spleen Cells Cultured in Cell-Impermeable Diffusion Chambers
Authors: Borella Luis
Published: December 2022

The effects of Rauscher leukemia virus (RLV) on the immune response to sheep red blood cells (SRBC) were studied in BALB/c mice and in isogeneic spleen cells cultured in cell-impermeable diffusion chambers implanted into irradiated hosts. Primary antibody formation to SRBC was depressed in mice inoculated with RLV. Mice injected with SRBC from 0 to 48 hr after virus inoculation had normal 19 S and depressed 7 S antibody production. Both 7 S and 19 S antibodies were depressed when SRBC were inoculated 3 to 15 days after RLV infection. There was a positive correlation between erythroblastic reaction and immunosuppression in two strains of mice: BALB/c was sensitive and C57BL partially resistant to the erythroblastic and immunosuppressive effects of RLV. Antibody production was not impaired in mice inoculated with RLV mixed with anti-RLV mouse antisera providing evidence for the viral specificity of immunosuppression.

Antibody production and plasmacytosis were demonstrated in SRBC-primed spleen cells cultured with SRBC in cell-impermeable diffusion chambers implanted into irradiated isogeneic hosts. RLV inhibited the secondary antibody response to SRBC in intact mice and in spleen cells cultured in diffusion chambers. Increase in blast cells, reduction in plasma cells, and depression of antibody formation were found both in primed spleen cells infected with RLV prior to culture and in primed spleen cells simultaneously exposed to RLV and SRBC in diffusion chamber cultures. This system provides a quantitative approach to the study of mechanisms of oncogenic virus-induced immunosuppression at cellular and subcellular levels.
Specificity of the Immunosuppression Caused by Passive Administration of Antibody
Authors: Cerottini Jean-Charles , McConahey Patricia J. , Dixon Frank J.
Published: December 2022

Antibody production against Fab and Fc determinants in rabbits immunized with varying doses of human immunoglobulin G (IgG) incorporated in incomplete Freund's adjuvant (ICFA) was determined using an antigen-binding capacity test. The kinetics of antibody formation and the amount of antibody produced against each group of determinants were usually comparable within the range of antigen doses used.

Intravenous administration of rabbit hyperimmune anti-Fab or/and anti-Fc antibodies 1 hr before and 7 days after injection of rabbits with IgG in ICFA specifically inhibited antibody production to the corresponding determinants without affecting the immune response to the unrelated determinants inasmuch as kinetics of antibody formation, serum antibody levels and relative avidity of antibodies were concerned. The results suggest that passive antibody may inhibit the antigenic stimulus at the level of antigenic determinants rather than at the level of the whole antigenic molecule.
Human Lymphocyte Membrane (HL-A) Alloantigens: Isolation, Purification and Properties
Authors: Mann Dean L. , Rogentine G. Nicholas , Fahey John L. , Nathenson Stanley
Published: December 2022

Human lymphocyte (HL-A) alloantigens have been isolated from human spleen and tissue culture cell sources. Solubilization was accomplished by digesting membrane extracts with papain. About 20% of all the alloantigens detected on the cell membrane were recovered in soluble form.

Purification of these materials was achieved by Sephadex chromatography and acrylamide gel electrophoresis. The final purified fraction contained similar levels of several alloantigen specificities. These materials lost alloantigenic activity at low pH, low NaCl concentration, and high temperatures.

Comparison of the amino acid composition of alloantigen preparations from two cell sources with different HL-A specificities revealed a general similarity in amino acid content. These proteins contained sialic acid and neutral sugars, but amino sugars were absent.

The analytic and quantitative findings indicate that the HL-A alloantigen-carrying molecules have a similar fundamental structure.
Interaction of Concanavalin A with Polysaccharides of Histoplasma Capsulatum
Authors: Markowitz Harold
Published: December 2022

Methods have been described for the preparation of serologically active polysaccharides from various strains of H. capsulatum. These polysaccharides were different by chemical analysis, carbohydrate composition, and serologic test by a quantitative C′-fixation technique. Polysaccharides from H. capsulatum cannot be differentiated by use of Con A in the immunodiffusion test. Some differences between these polysaccharides can be shown by quantitative “precipitin” and hemagglutination-inhibition tests. Whether this is due to the relative insensitivity of the reaction with Con A or to only slight differences in configuration among the active sites of the polysaccharides is not presently known.
Metabolic Studies of a Heavy Chain Disease Protein
Authors: Ein Daniel , Waldmann Thomas A.
Published: December 2022

The metabolic behavior of a heavy chain disease protein (γ HCD-JM) has been investigated in four patients with normal IgG levels, in the heavy chain disease patient himself, in a patient with a G-myeloma protein and in a patient with myotonic dystrophy. The protein was isolated from urine, labeled with 131I and administered once to each of the patients.

The γ HCD-JM belongs to the G3 subclass of IgG and its catabolic behavior is like that of intact proteins of the same G3 subclass. The survival half time of 6.6 days and the fractional catabolic rate of 39.5% per day is similar to that of intact G3 immunoglobulins. The γ HCD-JM resembles pooled normal IgG (predominantly composed of IgG1 molecules) in several other catabolic properties. Thirty-nine per cent is distributed in the intravascular pool, as compared with 44% for normal IgG. The survival half time and fractional catabolic rate of this protein is altered by the serum IgG level. Subjects with hypercatabolism of IgG associated with myotonic dystrophy or with high serum levels of IgG (multiple myeloma) also have shortened γ HCD survival and high fractional catabolic rate (FCR). The proteinuric rate is low, like that of IgG. Two per cent of the intravascular pool is lost as proteinuria daily. These findings support the concept that the Fc portion of the IgG molecule determines the catabolic rate of IgG.

Studies of the metabolism of the γ HCD-JM in the heavy chain disease patient himself show that the FCR is slightly elevated and the synthetic rate is high (0.42 g/kg/day compared to mean IgG synthesis of 0.034 g/kg/day in normal individuals). This indicates that the accumulation of H-chain in patients with heavy chain disease does not arise from defective catabolism of the protein but is due to excessive synthesis.
A Study of the Kinetics of the Anti-Dinitrophenyl Antibody Response at the Cellular Level
Authors: Havas H. Francis , Hraba Tomáš
Published: December 2022

An improved method for the detection of DNP-specific PFC was used for studying the kinetics of the immune response to the DNP-group in mice.

DNP-specific PFC were already detected in the spleens of immunized mice after the first challenge. Direct PFC predominated on days 4 and 6, but by the 8th day a conversion to predominantly indirect PFC had occurred.

A much stronger immune response was reached after the second challenge when the number of indirect PFC was consistently higher than that of the direct PFC. The immune response after the third challenge did not differ significantly from the response after the second challenge.

The number of background plaques against SRBC remained low following both the first and second challenge.

An attempt was made to induce tolerance to the DNP determinant by the injection of soluble DNP-MGG. However, the immune response in the pretreated group paralleled that of the controls, and the differences in the number of PFC or HA titers were not sufficient to warrant the conclusion that tolerance had been achieved.
Distribution Analysis of Brain-Localized Radioantibodies after Zonal Ultracentrifugation in a Sucrose Density-Gradient
Authors: Day Eugene D. , Rigsbee Lewis C.
Published: December 2022

Through the process of brain-localization in vivo after intravenous injection, of homogenization and zonal centrifugation in a sucrose density gradient and of multi-compartment distribution analysis, an antibrain radioantibody, paired with normal radioiodoglobulin, was shown to have localization properties in rat brain. Of the brain-localized immunoglobulin, 85% was shown to be antibody that had a distribution profile with four main density peaks and several lesser ones. In contrast, the 15% that was shown to be normal globulin had a profile with only one major density peak, that of the soluble protein compartment.
Insoluble Immunoadsorbents Containing Anti-IgE: Removal of Reaginic Activity and Subsequent Elution
Authors: Ito K. , Wicher K. , Arbesman C. E.
Published: December 2022

Reaginic antibody plays a significant role in immediate hypersensitivity of man. Ishizaka et al. (1) have reported that the human atopic reagin is primarily IgE protein, and these investigators have prepared rabbit (1) and guinea pig antisera (2) to human IgE by immunization with a purified reagin-rich fraction from ragweed sensitive patients. Johansson et al. (3) produced antiserum to human IgND (IgE), employing for immunization the protein from a patient with IgND (IgE) myeloma. Stanworth et al. (4) were able to block a Prausnitz-Küstner (PK) reaction with the Fc fragment of the IgND (IgE) myeloma protein, indicating that the reaginic antibody is related to the IgND (IgE). The antisera produced by Ishizaka and Johansson were shown to have identical serologic properties (5).

Through the courtesy of Dr. M. Ogawa, we obtained a small amount of plasma from a patient with IgE myeloma (for case description see (6)). With this material we were able to prepare rabbit antiserum to human IgE.
Studies on Heterologous Antilymphocyte and Antithymocyte Sera
Authors: Barth Rolf F.
Published: December 2022

Data on the effects of heterologous antilymphocyte sera (ALS) on the cellular kinetics of the immune response of mice to sheep erythrocytes were obtained using the hemolytic plaque technique as an assay system.

Mice given progressively increasing amounts of ALS had exponential decreases in the number of splenic plaque-forming cells (PFC) 4 days following primary immunization. There was a linear relationship between the logarithms of the numbers of PFC and the dilutions of ALS used.

Combined treatment with ALS and anti-sheep cell serum produced complete suppression of splenic PFC against sheep erythrocytes, whereas treatment with either serum alone did not. Since the mechanism of action of anti-sheep cell antibody is believed to involve the inactivation of immunogenic antigen at stages later than induction, it acts synergistically with ALS. The inability of ALS to suppress “background” PFC further supports the view that its primary effects are on inductive events of the immune response.

Increasing the antigenic stimulus only slightly enhanced the immune response of ALS-treated mice.

Infusion of ALS-treated mice with 5 × 107 syngeneic spleen or thymus cells resulted in an 8- to 10-fold increase in the number of splenic PFC. Removal of an adherent cell population markedly reduced the ability of these infused cells to restore immunocompetence.

These data support the view that ALS has a preferential effect on antigen-sensitive, thymus-derived cells.
Studies on the Immunochemical Properties of Aggregates and Subunits of Fe of Rabbit γG Globulins
Authors: Amiraian Kenneth , Ovary Zoltan
Published: December 2022

Fc, obtained by digestion of rabbit γG globulin with papain and by cation-exchange chromatography, yields at least six fractions upon lyophilization at low ionic strength and gel filtration. These fractions are comprised of Fc, its aggregates and subunits. A study of the antigenic, anticomplementary and skin-sensitizing properties of these fractions indicates that the structures necessary for these immunochemical properties are at variance with each other.
Experimental Murine Amyloid
Authors: Willerson James T. , Asofsky Richard , Barth Werner F.
Published: December 2022

The immunoglobulin response and the development of amyloidosis were studied in six strains of mice after repeated administration of casein. Strain differences were noted in the ease of induction and amount of amyloid deposited. Increases in immunoglobulins following administration of casein were predominantly in the 7Sγ1 class. There was no consistent relationship between serum levels of immunoglobulins and development of amyloid. Bence Jones proteinuria was not detected. Infusion of a 7Sγ1 myeloma protein to produce and maintain hyperglobulinemia did not induce amyloidosis. These studies do not support a direct relationship between elevation in serum immunoglobulin levels and experimental murine amyloidosis.
Effects of Antithymus Serum on Thymic Lymphopoiesis: A Radioautographic Study
Authors: Nagaya Hiroshi , Sieker Herbert O.
Published: December 2022

In an attempt to investigate immunosuppressive mechanisms of antithymus serum, especially its superiority to antilymphocyte serum as immunosuppressive agents, the effects of these antisera on thymic lymphopoiesis were studied by radioautography.

In rats treated with antithymus serum, thymidine incorporation by the thymic cortical lymphocytes was markedly suppressed although the number of thymidinelabeled lymphocytes appeared to be increased in the medulla and in the perivascular sheaths. In rats treated with antilymphocyte serum, on the other hand, there was increased thymidine incorporation by the cortical lymphocytes as indicated by the higher percentage of thymidine-labeled cells, the greater mean thymus weight and the histologic evidence. Despite lowered thymidine uptake, uridine uptake by the thymus cells of rats treated with antithymus serum was increased, suggesting an increased RNA metabolism in these cells. Histologically the majority of uridinelabeled cells were reticulo-epithelial cells and were mostly located in the medulla and in the perivascular sheaths, but some of them were seen in the subcapsular regions of the cortex showing positive periodic acid Schiff (PAS) stain.

These findings suggest that the greater immunosuppressive potency of antithymus serum than that of antilymphocyte serum is related to the former's ability to suppress proliferation of the thymic cortical lymphocytes, possibly by inactivating a thymic humoral factor, whereas the latter serum has no such suppressive effect. Increased uridine uptake by the thymus of the animals treated with antithymus serum may indicate a compensatory increase in the production of a thymic humoral factor by the reticulo-epithelial cells.
Studies of Cyclophosphamide Suppression of Experimental Allergic Encephalomyelitis in Wistar Rats
Authors: Paterson Philip Y. , Hanson Margaret A.
Published: December 2022

Experimental allergic encephalomyelitis (EAE) is completely inhibited in Wistar rats sensitized to spinal cord-adjuvant and treated with cyclophosphamide (CY) in doses of 5, 10 or 12 mg/kg from the day of sensitization through the 17th post-sensitization day. Such animals develop neither clinical neurologic signs nor histologic evidence of the disease. Furthermore, production of complement-fixing antibrain antibody is suppressed. Comparable inhibitory effects can be obtained by a single dose of CY of 100 mg/kg given 2 days after sensitization. Inhibition of EAE extends for at least 112 days after stopping CY. Limited “break-through” occurs in some rats re-sensitized to spinal cord by a different route after prior CY treatment. Evidence favoring the thesis that CY exerts its striking inhibitory action on this auto-immune disease through immunologic mechanism(s) is presented.
Coproantibodies to Milk: Identification by Radioimmunoelectrophoresis
Authors: Kletter B. , Freier S. , Davies A. M. , Gery I.
Published: December 2022

Precipitating antibodies against cow's milk, as well as high titers of hemagglutinins, have been found in the feces of subjects suffering from milk sensitivity or celiac disease (1, 2). Low titers of hemagglutinating, but not precipitating, coproantibodies were found in the majority of normal infants or patients with other diseases (2).

There is some direct and indirect evidence that coproantibodies to viruses consist mainly of immunoglobulin A (3–5). The data of Katz et al. (1, 6) suggest that this is also the case with coproantibodies to soluble proteins. The use of the radioimmunoelectrophoresis (RIEP) technique proved in the present study to be useful for direct identification of the immunoglobulin types which make up the coproantibodies to milk.

The sera and fecal supernatants examined in the present study were obtained from infants (aged 3 to 12 months) with gastrointestinal milk sensitivity. These infants suffered from diarrhea and vomiting that ceased after elimination of cow's milk from the diet and the diagnosis was confirmed by three challenges with milk.
Purification of the First Component of Complement by Zonal Ultracentrifugation
Authors: Colten Harvey R. , Bond Howard E. , Borsos Tibor , Rapp Herbert J.
Published: December 2022

The zonal centrifuge has been used to purify viruses, sub-cellular organelles and macromolecules of biologic interest (1, 2). In this report we describe a two-cycle preparative ultracentrifugation method for the separation from serum of chemically and functionally highly purified first component (C1) of guinea pig and human complement. The observation that the sedimentation rate of C1 varies as a function of ionic strength (3) made the development of this purification procedure possible.

Materials and Methods. The source and preparation of sheep erythrocytes (E), rabbit antibody to boiled stromata of sheep erythrocytes (A) and sensitized sheep erythrocytes (EA) have been described (4). Fresh frozen guinea pig and human sera were purchased from Suburban Serum Laboratories, Silver Spring, Md. EAC4 were prepared with guinea pig serum as described (5). C1 activity was determined by the method described in (6).
Comparison of the Effects of Divalent Ion Chelating Agents on Complement-Mediated Hemolysis in Rabbit and Human Serum
Authors: Marney Samuel R. , Des Prez Roger M.
Published: December 2022

The chelating agent, magnesium ethylene-glycol-diamino-ethyl-tetraacetic acid (MgEGTA), inhibits EAC′ hemolysis when human serum is used as a complement source, but does not inhibit rabbit platelet injury which occurs in the presence of antigen-antibody reactions unrelated to platelet antigens. This difference has been assumed to indicate that the mechanism of complement action in producing this type of immunologically-induced platelet injury differs from that subserving EAC′ hemolysis.

The present studies indicate that when rabbit serum rather than human serum is used as a source of complement, EAC′ hemolysis is not inhibited by MgEGTA. It is concluded that rabbit complement and human complement differ with respect to divalent ion requirement. The explanation for this difference may be that the concentration of calcium ion required for maintaining the rabbit C′1 molecule in an assembled state is lower than the concentration required for normal human C′1 activity.
Aerosol Immunization of Humans with Tetanus Toxoid
Authors: Wigley F. M. , Wood S. H. , Waldman R. H.
Published: December 2022

Soluble tetanus toxoid given as an aerosol to human volunteers led to an increase in serum antibody titers comparable to that resulting from conventional subcutaneous administration, thus suggesting that tetanus toxoid booster immunization can be administered by aerosol in man. The results also show that an antigen given by fine particle aerosol can stimulate the production of high levels of serum antibody. No increase in the titer of nasal antibody was demonstrated. Sputum antibody titers, however, were found to increase significantly in the persons who received aerosolized vaccine, which demonstrates that secretory antibody was produced in response to an antigen that normally does not enter the respiratory tract.
Inhibitor of Human C3 Associated with Preparations of C3 Dissociated from C3-Zymosan Complex
Authors: Blum Livia
Published: December 2022

Incubation of human serum with zymosan at 37°C, followed by removal of the zymosan residue by centrifugation, resulted in C3-zymosan complex. The C3-zymosan complex could be dissociated and C3 recovered in the eluate. Preparations of dissociated C3 did not contain the hemolytically active component of complement, C1 esterase, or fibrinolytic or caseinolytic activity.

Addition of dissociated C3 to EAC142 blocked the formation of EAC1423 or EAC14235 by partially purified C3,C5. The effect of the inhibitor was not limited to intermediate complexes, it also blocked the hemolytic activity of serum was restored by adding C2, but not C3. A positive immune adherence test was obtained by treating EAC142 with dissociated C3.

The data are discussed in relation to an hypothesis that the inhibitor in preparations of dissociated C3 is C3, which is hemolytically inactive, but participates in immune adherence and has greater affinity for SAC2 than the C3 molecule possessing functional integrity.
Heterogeneity of Mouse Myeloma γG Globulins as Revealed by Enzymatic Proteolysis
Authors: Gorini G. , Medgyesi G. A. , Doria G.
Published: December 2022

The sensitivity of mouse myeloma γG globulins to enzymatic proteolysis has been investigated under several experimental conditions. γG1 was found to be more resistant to papain or tryptic cleavage than γG2d and γG2b. Minor differences were found between the reactivities of the latter two proteins. γG2b showed extremely high sensitivity to the action of pepsin. The addition of cysteine to the incubation mixtures of globulins with papain or trypsin enhanced the degree of fragmentation, but some physical and immunochemical properties of these fragments were found to be different from those of fragments obtained in the absence of cysteine. The possible role of this reducing agent has been discussed.
On Mouse Complement: Genetic Variants
Authors: Rosenberg L. T. , Tachibana D. K.
Published: December 2022

There are a number of polymorphic genes in mice which control the expression of hemolytic complement levels. The levels of hemolytic complement vary greatly within a non-inbred strain, CF-1, and there is a characteristic level of hemolytic complement activity for each inbred strain. Studies of progeny of crosses between selected inbred strains suggest that the levels of hemolytic complement are controlled by relatively simple genetic mechanisms. The significance of recognizing these genetic variants in understanding the functions of the complement system is discussed.
Immunosuppression by Antibody: Localization of Site of Action
Authors: Ryder R. J. W. , Schwartz Robert S.
Published: December 2022

Immunosuppression by antibody was studied in mice passively immunized at birth and challenged with antigen 6 weeks later. At the time of antigen (sheep red blood cells (SRBC)) administration none of the antibody (anti-SRBC) given at birth was detectable by conventional serologic techniques. However, evidence for persistence of antibody was obtained by the demonstration that spleen cells from normal mice, when transferred to irradiated, passively immunized mice, were unable to mount a primary immune response. By contrast, spleen cells from mice primed with SRBC elaborated a normal anamnestic response when transferred to irradiated, passively immunized mice.

The locus of action of immunosuppressive antibody was studied in several ways. No evidence was obtained that the effect was due to binding of antibody to unproccessed antigen. SRBC retrieved from passively immunized mice were immunogenic in normal mice. Spleen cells from passively immunized mice responded normally when transferred to irradiated mice, thus excluding lymphoid cells as the site of action of immunosuppressive antibody. However, when macrophages from passively immunized mice were “fed” SRBC, they failed to induce an immune response in normal recipients. It was concluded that, in the system studied, antibody exerts its immunosuppressive effects at the level of the macrophage.
Human Lymphocyte Response to Phytomitogens in Vitro: Normal, Agammaglobulinemic and Paraproteinemic Individuals
Authors: Douglas Steven D. , Kamin Roberta M. , Fudenberg H. Hugh
Published: December 2022

Studies of the time course of thymidine incorporation during stimulation of lymphocytes from normal individuals with four plant mitogens over a 10-day period demonstrate several differences in response profiles. Whereas the peak responses for cultures stimulated with phytohemagglutinin (PHA), wax bean glycoprotein and concanavalin A (Con A) occurred at 3 or 5 days, the lower peak response to pokeweed mitogen (PWM) was observed at 5 or 7 days. Cultures stimulated with PHA, wax bean glycoprotein and PWM continued to show significant thymidine incorporation at 7 and 10 days, although the response to Con A had returned to baseline after 7 days.

Lymphocytes from eight adult “acquired” agammaglobulinemic patients and their four available parents had a diminished thymidine incorporation in 3-day cultures using each of the four mitogens (p < 0.001). A late response was observed with PHA or wax bean stimulation at 5 to 7 days, comparable to controls at 3 days. With Con A, lymphocytes from both the “acquired” agammaglobulinemic patients and their parents had a diminished incorporation throughout the 10-day period (p < 0.001). Stimulation with PWM distinguished the lymphocytes of the patients from their heterozygous parents; at 5 to 7 days the parents had a peak comparable to controls, whereas the “acquired” agammaglobulinemics had a diminished peak (p < 0.001) at 10 days.

Lymphocytes from patients with multiple myeloma and Waldenström's macroglobulinemia had diminished thymidine incorporation in response to Con A during a 10-day incubation period. These patients had normal or late responses comparable in magnitude to controls with PHA, wax bean glycoprotein and PWM.

These observations suggest that plant mitogens have selective and specific actions on lymphocyte subpopulations. Differences in the time course and magnitude of lymphocyte response to plant mitogens may reflect different circulating subpopulations of cells or alterations in the total number of cells responsive to mitogen. Detailed time studies using several mitogenic agents represent a possible approach to the assessment of lymphocyte function in immunologic diseases.
Mechanisms of Endotoxin Tolerance
Authors: Greisman Sheldon E. , Young Edward J.
Published: December 2022

Sieved spleen cells (3 × 109) from rabbits immunized with a Biovin preparation of Escherichia coli endotoxin were capable of transferring significant “anamnestic” endotoxin-tolerant responses to non-inbred normal recipient animals. Thus, whereas the initial febrile responses of the primed spleen cell recipients to a standard dose of E. coli endotoxin remained unaltered, upon retesting 48 hr later significant reductions in febrile responses occurred in comparison with recipients given comparable numbers of spleen cells from non-immunized donors. Simple carry-over of free endotoxin by the primed spleen cells could not explain such transfer of “anamnesis” since frozen and thawed primed spleen cells were ineffective. The transferred “anamnestic” tolerance was partially specific for the endotoxin employed in immunizing the spleen cell donors, i.e., spleen cells from donors immunized with a Boivin preparation of Salmonella typhimurium transferred significantly less “anamnestic” tolerance to the standard E. coli endotoxin test preparation. These latter observations support the concept that antibodies directed against the specific “O” terminal polysaccharide side chains contribute significantly to pyrogenic tolerance to bacterial endotoxins. Despite the specificity of the transferred “anamnestic” tolerance to endotoxin, increments in circulating anti-“O” antibodies were not generally observed at 48 hr, suggesting that “O” protective antibodies at this time may be primarily cell-fixed. The smaller but nevertheless significant transfer of “anamnestic” tolerance to E. coli endotoxin conveyed by spleen cells from donors immunized with S. typhimurium endotoxin provides evidence that antibodies with broader specificity also participate in pyrogenic tolerance to bacterial endotoxin.
Properties of Purified Mouse Homocytotropic Antibody
Authors: McVeigh Thomas A. , Voss Edward W.
Published: December 2022

Purified anti-DNP antibody was obtained from a pool of hyperimmune mouse sera. Purity was verified by radioimmunoelectrophoresis, disc gel electrophoresis and sedimentation equilibrium studies. A molecular weight of 141,500 was determined by the method of Yphantis. A corrected valence of 2 and an average intrinsic association constant of 6 × 105 liters/mol was measured by equilibrium dialysis.

The purified antibody elicited a positive PCA reaction in mice only after a 3-hr incubation period. The length of the incubation period, the electrophoretic mobility and the heat stability at 56°C indicated that this antibody belonged to the 7S γ1 class of mouse immunoglobulin.
Structural and Antigenic Relationships Between Avian Immunoglobulins
Authors: Leslie Gerrie A. , Benedict Albert A.
Published: December 2022

Ring-necked pheasants produced IgM hemagglutinating antibodies 3 days following a single injection of bovine serum albumin (BSA). Seven days following injection, IgG and IgM anti-BSA antibodies were detected by the method of radioimmunoelectrophoresis. Compared to chickens and pheasants, Japanese quail respond very poorly to injections of BSA.

Chicken IgG (C-IgG), pheasant IgG (P-IgG) and quail IgG (Q-IgG) were similar in several respects: their sedimentation coefficients, which ranged from 7.1S to 7.3S; their yields of heavy (H) and light (L) chains from reduced and acid-dissociated molecules; and the patterns formed by H- and L-chains in urea starch gel electrophoresis.

The most striking similarity between the IgGs of these avian species was that, unlike mammalian IgG, partial dissociation of the molecules occurred following reduction in the absence of a dispersing agent. In alkaline buffer, C-IgG and P-IgG each required 0.05 to 0.1 M 2-mercaptoethanol (ME) for release of L-chains, whereas Q-IgG required 0.2 M ME. About 35% of the C-IgG, P-IgG and Q-IgG each dissociated in the absence of a dissociating agent when reduced with 0.2 M ME. As with chicken IgM, reduction of pheasant IgM caused the release of some L-chains.
Fate of Antigen in Delayed Hypersensitivity and Arthus Reactions
Authors: Eisen Abraham H.
Published: December 2022

Hen egg albumin labeled with 125I was followed at the sites of delayed hypersensitivity and Arthus reactions, and in the lymph nodes draining the areas of both reactions. In the delayed hypersensitivity reaction, antigen localized preferentially in the cytoplasmic membrane of mononuclear cells. In the Arthus reaction, antigen localized in the polymorphonuclear leukocyte. The pattern of retention of antigen in the regional lymph nodes was the same in both delayed hypersensitivity and Arthus reactions and resembled that seen in the primed immune state. Antigen was not retained at the site of delayed hypersensitivity reactions. It is suggested that the in vivo observations correlate closely with the in vitro model of macrophage migration.
Studies on Rabbit Lymphocytes in Vitro
Authors: Sell Stewart , Lowe Jennifer A. , Gell P. G. H.
Published: December 2022

The addition of mixtures of anti-allotypic sera to cultures of lymphocytes obtained from allotypically heterozygous donors causes a greater amount of “blast” transformation than the sum of the maxima that can be obtained with each anti-allotypic serum added separately. This “superaddition” of anti-allotypically induced transformation implies that the expression of allotypic specificities by lymphocytes is not limited to one allele (allelic exclusion), as has been found for plasma cells. Therefore, the expression of genetically controlled immunoglobulin antigenic specificities by lymphocytes appears to be less restricted than that by plasma cells. Plasma cells may become restricted in regard to immunoglobulin expression during differentiation from precursor cells that are not so restricted.
Phylogenetic Insight into Evolution of Mammalian Fc Fragment of γG Globulin Using Staphylococcal Protein A
Authors: Kronvall Göran , Seal Ulysses S. , Finstad Joanne , Williams Ralph C.
Published: December 2022

The reaction between protein A of staphylococci and the Fc portion of γG globulins has been studied using representative sera and isolated γ globulins from species representing seven classes and 30 orders of living vertebrates. No reactions with immunoglobulins derived from either the primitive or later evolving fishes were noted. No reactions were recorded among Amphibia and Reptilia as well as all birds tested, except with serum from the primitive flightless bird Rhea americana. All members of the class Mammalia, including egg-laying monotremes, showed reactions with protein A except for the American oppossum. A comparison of patterns of reactivity between protein A and isolated human rheumatoid factors showed that in the majority of instances different specificities were involved. Protein A may provide a useful tool in the study of the evolution of γG globulin.
The Effect of Antigen Competition on Both the Primary and Secondary Immune Capacity in Mice
Authors: Hanna M. G. , Peters Leona C.
Published: December 2022

The primary and secondary immune capacity in mice was studied during antigen competition. Three antigens were used: sheep erythrocytes (SRBC), rat erythrocytes (RRBC), and human γ globulin (HGG). The results of these studies demonstrate the following. 1) In both competition systems, SRBC prior to RRBC or HGG, a marked suppression of the secondary immune capacity corresponds to a suppressed primary agglutinin response to the second antigen. 2) The degree of suppression of the secondary agglutinin capacity can be reduced by increasing the dose of the secondary challenge, which suggests that in the secondary immune response there may be participation of both reacting sensitized and uncommitted immunocompetent cells. 3) The entire level of the immune suppression during competition is dependent upon the dose of the first antigen. Further, the experimental results do not support the possibility that the competition can be attributed to either an increased or a decreased rate of antigen elimination. 4) Antigen competition cannot be attributed to a deficiency in the extracellular localization of antigen in reticular cells of splenic germinal centers. 5) The deficiency in a competition system is attributed to an impairment which ultimately affects the immunocompetent cell or unit. One reflection of this lesion on the antigen-sensitive unit is expressed in the competition between SRBC and HGG. A detectable increase in the latent phase of the primary HGG agglutinin response occurs when the HGG is injected at 1 or 3 days after SRBC. The recruitment of immunocompetent cells, and thus priming, is limited by the decreased level of preexisting tissue-associated and/or free antigen. On the other hand, in a condidition where there was limited immune progenitor cell recruitment to RRBC as a result of subsequent competition with SRBC the result was antigen-driven depletion of the existing sensitized immune cell compartment.
Enzyme-Activation by Antibody
Authors: Pelichová H. , Suzuki T. , Cinader B.
Published: December 2022

Antibodies that activate ribonuclease occur in fractions of rabbit IgG eluted at pH 8 from DEAE Sephadex A-50 between ionic strength 0.05 and 0.15. The fraction eluted at the lowest ionic strength (P1) activates the enzyme two- to threefold. Fractions eluted at higher ionic strength (P4 to P7) inhibit enzyme at relatively low antibody concentration, but become less inhibitory at high antibody concentrations. Activation and decreased inhibitory power at high antibody concentrations are observed with cyclic cytidylic and cyclic uridylic but not with ribonucleic acid as substrate. In fractions, such as P5, the zone of activation can be shifted when part of the antibody is removed by precipitation with ribonuclease. The resulting supernatants can activate ribonuclease (from 100% to 120%) at low concentrations of antibody.

The interaction of inhibiting and activating antibody was studied in terms of the neutralizing capacity of mixtures of enzyme and various proportions of activating and inhibiting antibodies. In antigen excess, the enhancing capacity of activating antibody was reduced by the admixture of inhibiting antibody. In antibody excess, the neutralizing capacity of inhibiting antibody was reduced by the admixture of activating antibody. The latter observation was not confined to experiments with cylic cytidylic acid but was also made when ribonucleic acid served as substrate. This was remarkable since enzymic hydrolysis of nucleic acid was not activated by isolated P1 or P5. These findings are analyzed in terms of relative concentrations and relative binding constants of inhibiting and activating antibodies and in terms of competition for overlapping determinants of the enzyme.