bluuebunny/crossref_metadata_2025 huggingface dataset:
The Surface Structure of Pseudomonas Aeruginosa
Authors: Homma J. Yuzuru , Suzuki Nachiko , Ito Fumiko
Published: January 2023

A “bacterial surface substance,” which contained cell wall and probably the cytoplasmic membrane, was prepared by disintegrating the intact cells of Pseudomonas aeruginosa with carborundum or by sonic oscillation. This substance was dissociated by various chemical and enzymatic treatments. The immunochemical and biologic properties of the fractions obtained were investigated and compared with the lipopolysaccharide-protein complex (Component I) isolated from the autolysate. The lipopolysaccharide of Component I was found in the nondialyzable, soluble fraction (LS) of lysozymedigested “bacterial surface substance”; however, it was not found in the protoplast. By treatment of the LS with n-propanol and lower phase solvent, a simple protein was isolated, which proved to be homogeneous by zone electrophoresis and ultracentrifugal analysis (s20,w, 1.7 S). By similar treatment a protein was isolated from the lipopolysaccharide-protein complex, which was shown to be homogeneous by zone electrophoretic and ultracentrifugal analyses (s20,w, 1.1 S). Cross-precipitin tests revealed that the two proteins were serologically identical. Biologically they had the same potencies in the Shwartzman phenomenon and pyrogenic reaction. Almost all of the ribonucleic acid present in the protoplast surface substance was liberated by lysozyme and Nagarse. Chemical studies on the substances dissociated by lysozyme and Nagarse treatment of “bacterial surface substance,” revealed the existence of a mucoprotein layer. By immunochemical, biologic and morphologic analyses the surface structure of the bacteria was divided into several components.
Classification of Trachoma Virus Strains by Protection of Mice from Toxic Death
Authors: Wang San-Pin , Grayston J. Thomas
Published: January 2023

Twenty-nine trachoma virus strains isolated on Taiwan and nine trachoma strains and two inclusion conjunctivitis strains obtained from other areas of the world have been tested in mice for cross-protection against intravenous toxicity. Crude yolk sac suspensions of these TRIC viruses were used both for protection and challenge. Protection was produced by intravenous injection of one-third and then two-thirds a minimum lethal toxic dose 14 and 7 days prior to challenge. All 29 of the Taiwan trachoma strains fell into one of two cross-protecting groups designated as the TW-1 and TW-3 groups. Three trachoma strains from New Delhi protected against each other and members of the TW-3 group. Two trachoma strains from Apache Indians in Arizona cross-protected with the TW-1 group. A Saudi Arabia and a Gambia trachoma strain protected against each other but not against the other strains tested. Two trachoma and two inclusion conjunctivitis strains failed to cross-protect with any strain. Therefore, seven different cross-protecting types were demonstrated. It is concluded that the total number of different groups of TRIC virus strains in the mouse toxicity prevention test are not yet known, but that in a given geographic area (as exemplified by Taiwan with two groups) the number of different groups is limited.
Demonstration of Human Measles Antibody by Mixed Agglutination
Authors: Barron Almen L. , Milgrom Felix , Karzon David T. , Witebsky Ernest
Published: January 2023

A method for the demonstration of human measles antibody by mixed agglutination was developed. The antibody attached to cells infected with measles virus and was detected by erythrocytes bound through an anti-γ-globulin antibody bridge. The mixed agglutination method was compared to hemagglutination-inhibition and complement-fixation tests and reasonable agreement was obtained. More positive results and higher antibody titers were obtained by mixed agglutination. Several types of studies were employed to evaluate the mixed agglutination method including: a) detection of measles antibody in acute and convalescent sera from patients with clinical measles, b) antibody response as a result of immunization with measles vaccine, and c) occurrence of measles antibody in different age groups.
The Use of Techniques of Absorption and Elution of Bovine Isoantibodies in the Study of Specificity of Antigen and Antibody and in the Development of Typing Reagents
Authors: Hatheway Charles , Ludwick Thomas M. , Weseli Donald F.
Published: January 2023

Some modifications of a heat elution technique are described. Heat elution was used for recovery of cattle erythrocyte antibodies absorbed from isoimmune cattle sera. The results of the use of eluates as typing reagents and for studying serologic relationships between antibodies and between antigenic factors are listed and discussed. The use of elution techniques in studying specific properties of biologic materials is suggested.
Studies on the Heterogeneity of the Rate of Combination of Antibody with Antigen
Authors: Grey Howard M.
Published: January 2023

By means of separating antisera into fractions with different rates of association, it has been demonstrated that rabbit anti-BSA is heterogeneous with respect to its rate of combination with antigen. This separation was accomplished by: a) fractionation according to total molecular charge; b) rapid, partial absorption with a particulate BSA preparation; c) partial in vivo clearance of anti-BSA with antigen. It was possible to select antibody molecules on the basis of their combination rate without a significant alteration in the dissociation rate, thereby indicating that these two processes are, at least in part, independent of each other.
Antibody Formation by Transplanted Bone Marrow, Spleen, Lymph Nodes and Thymus Cells in Irradiated Recipients
Authors: Stoner Richard D. , Bond Victor P.
Published: January 2023

Immunologic competence of immunized mouse bone marrow, spleen, lymph node and thymus cells was demonstrated when specific recall tetanus antitoxin responses were elicited after transfer of these cells to isologous irradiated recipients. Lesser amounts of antibody were obtained as the genetic strain distance was increased between the relationship of donor and host in the parental to F1 and in the homologous combination within the same species. It was not possible in the heterologous situation to elicit significant amounts of antibody from rat bone marrow and other lymphoid cells following their transplantation into irradiated mice. Minimal but not significant antibody responses were elicited from cells obtained from immunized rat spleen and thymus tissue. In a few experiments, it was possible to elicit antibody formation from a buffy coat suspension of circulating white cells following their transfer to irradiated recipients. Isologous nonimmunized bone marrow did not stimulate or hasten recovery of the ability to elicit secondary antibody responses in previously immunized irradiated mice. The capacity to elicit primary antibody responses to tetanus toxoid was depressed in parental bone marrow protected F1 mice when these chimeras exhibited varying degrees of secondary disease. The depression of primary antibody responses in irradiated F1 mice given parental bone marrow provides evidence for a donor mediated immunologic depression of antibody synthesis by host-lymphoid tissues.
The Effects of Treatment with the Emulsions of Ragweed Extract on Antibody Titers
Authors: Connell John T. , Sherman William B.
Published: January 2023

The skin-sensitizing and hemagglutinating antibody titers of most ragweed hay fever patients rise after an injection of emulsified ragweed extract reaching a peak at 3 to 6 weeks and showing a gradual decline by the end of 12 weeks. The least sensitive group of patients who received the highest dosage (10,000 PNU) showed no significant changes in skin-sensitizing antibody titer, but did respond with increased hemagglutinating and blocking antibody titers. All groups of patients showed a rise in blocking antibody after treatment reaching a peak at approximately 6 weeks. The group that received 10,000 PNU remained essentially unchanged from the 6th to 12th weeks whereas those that received 1000, 2500, or 5000 PNU showed a decrease in titer. Patients who received two injections of emulsified ragweed extract showed further increases of all antibody titers after the second injection.
Effect of Formalin and Periodic Acid on Red Cell Rho (D) and Enzyme-Exposed Rho (D) Antigen
Authors: Kaufman B. , Masouredis S. P.
Published: January 2023

The effect of formalin and periodic acid on the Rho (D) red cell antigen at 37°C for 30 min was studied with incomplete I131 anti-Rho (D). Pretreatment of the Rho (D) positive red cell with formalin resulted in a decrease in the amount of I131 bound to the red cell which was directly proportional to the formol concentration up to 0.66 M. Periodic acid to a concentration of 0.0008 M produced a marked decrease in the ability of red cells to bind anti-Rho (D) which was not directly proportional to the concentration of periodic acid. The antibody able to bind to the formalin-treated cells was stable to hemolysis, whereas the small quantity of antibody able to bind to the periodic acid-treated cell was bound poorly as judged by the inability to recover the cell bound I131 in the stroma.

The Rho (D) antigen exposed by papain was unaffected by pretreatment of the red cell with formalin. Periodate treatment of the intact red cell produced losses of from 20 to 90% of the Rho (D) which would normally be exposed by papain. The susceptibility of the enzyme-exposed Rho (D) antigen to inactivation with formalin was similar to that of the Rh antigen on the untreated red cell, whereas the enzyme-exposed antigen was more resistant than the Rho (D) antigen on the untreated cell to periodic acid inactivation. Quantitative absorption studies indicate that the I131 bound to the red cells following the various treatments represents specific anti-Rho (D).

The results are discussed in terms of the mechanism of action of formalin and periodic acid on the Rho (D) antigen and in terms of the nature of the Rho (D) antigen exposed by proteolysis.
The Molecular Properties of Antibodies Synthesized in Response to Simple Haptenes
Authors: Bauer D. C.
Published: January 2023

Rabbit hemagglutinating antibodies to sulfanilic acid and p-aminobenzoic acid were found to be heterogenous with respect to their molecular size. These differences in the physical properties of the anti-haptene globulins were determined by zone electrophoresis, density gradient ultracentrifugation, ion exchange chromatography and susceptibility to reduction by mercaptoethanol. Examination of the anti-haptene globulins by these techniques indicated that the antibody activity was of both the γ-1, 19 S and γ-2, 7 S types.

The proportion of the two types of antibody present in a particular antiserum was found to change with the extent of immunization. After an initial series of antigen injections the antihaptene activity was predominantly the macroglobulin type. With continued immunization the antibody activity shifted to the 7 S type.

The distribution of anti-haptene activity between the two globulin fractions changed independently of the physical properties of the antibodies to the carrier protein. Early in the course of immunization the anti-protein globulin was primarily the 7 S type in contrast to the macroglobulin nature of the haptene antibody. These changes in the proportion of 19 S to 7 S antibodies in various antisera do not necessarily represent the same distribution of antibody on a molar basis, since the efficiency of the hemagglutination test in detecting the two antibodies may not be the same.

The origin of the two types of antibody is discussed in relation to the inductive phase of antibody formation.
Fetal Response to Antigenic Stimulus
Authors: Silverstein Arthur M. , Thorbecke G. J. , Kraner Keith L. , Lukes Robert J.
Published: January 2023

In addition to “β2A-” and β2M-globulins, the serum of adult sheep contains 7 S γ-globulins composed of two cross-reacting components, an electrophoretically fast and a more slowly migrating component. The appearance of each of these components in the sera of stimulated fetal lambs at different times suggests that they may be functionally distinct.

The normal fetal lamb during the latter half of gestation has small amounts of β2M and 7 S γ-globulin in its serum. Tissue culture results indicate that the β2M-protein is produced predominantly in the spleen, with some production also evident in the lymph nodes. This technique has also indicated the production by normal fetal lymph nodes of a slowly migrating γ-globulin which appears to be distinct from the fast and slow γ-globulins that appear after stimulation.

In response to antigenic stimulus, a heightened production of β2M protein is induced. In the absence of complete adjuvant, little 7 S γ-globulin is formed. The presence of adjuvant in the stimulus elicits the production of relatively great amounts of fast γ-globulin, and only much later of slow γ-globulin and of β2A-globulin. Large amounts of 7 S γ-globulin may be present in the fetal serum even in the absence of detectable circulating antibody, or in cases in which the antibody that is produced is predominantly associated with β2M-globulin. These observations are discussed in relation to the possibility that some of the γ-globulin formed by the fetal lamb may lack specific antibody function.
Immunologic Studies of Anthrax
Authors: Klein Frederick , Haines Bertram W. , Mahlandt Bill G. , Lincoln Ralph E.
Published: January 2023

The effects of anthrax immunization on the serum antibody titer and the immunity index were compared for guinea pigs and rats immunized by two protocols. An association between the two measures could not be detected in either species of animals immunized at any one level; however, when data on animals immunized at several levels were combined, a weak but significant association became apparent. The results of the agar diffusion test were more variable than those of the immunity index, and at relatively low or intermediate levels of protection the agar diffusion test produced a much larger portion of false negatives. This weakness did not carry through to animals protected to a high level of resistance, although at nearly all levels of protection the immunity index was more precise than the serologic test. From these observations on animals we conclude that with human beings serum titers of less than 1:32 may be unreliable as an index of protection against anthrax. Titers of 1:64 or greater probably indicate some significant degree of resistance although the same titer observed for two individuals may indicate different degrees of resistance to the disease.
Studies of Pasteurella Pestis
Authors: Eisler Daniel M. , Von Metz Eleanor , Heckly Robert J.
Published: January 2023

Fraction IB (FIB) from Pasteurella pestis induced immediate protection of mice concomitantly infected with plague. Moreover, the serum withdrawn from mice 1 or 2 days after treatment with FIB passively protected mice against P. pestis. Both the serum β-globulin and leukocyte concentrations increased as protection increased during this time. Subsequently the concentrations of both β-globulin and leukocytes decreased. About 6 days after treatment there was a second period of increased resistance which was accompanied by an increase in serum γ-globulin. Leukocytosis was not accompanied by increased phagocytosis. An increase in β-globulin concentration was also induced by inoculating mice with protein from Escherichia coli, but these mice were not protected against plague. Either the β-globulin contained antibodies responsible for the early protection or the early increase in β-globulin was coincidental.
Studies on Immune Human Hemolysis
Authors: Hinz Carl F. , Mollner Anna Marie
Published: January 2023

The 11 S component of normal human serum is required in the initial cold phase of the Donath-Landsteiner reaction, an autoimmune hemolytic reaction entirely of human origin. The 11 S component is present in crude preparations of C′1-esterase, and removal of 11 S renders C′1-esterase inactive in the cold phase of the reaction. The 11 S component precedes C′1-esterase in the initiation of the hemolytic reaction, and, in contrast to C′1-esterase, is not inhibited by Na3HEDTA or serum inhibitor of C′1-esterase.
The Effect of Bursectomy at Different Ages on Precipitin and Natural Hemagglutinin Production in the Chicken
Authors: Graetzer M. A. , Cote W. P. , Wolfe H. R.
Published: January 2023

White Rock and White Leghorn chickens bursectomized between 1 and 5 weeks of age were challenged with bovine serum albumin at 6 weeks of age. The effect of bursectomy on precipitin production gradually became less during this period. There appeared to be a breed difference in response, as bursectomy at 4 and 5 weeks of age significantly lowered the response of White Rocks whereas bursectomy at 4 weeks did not affect the response of White Leghorns.

The development of natural antirabbit hemagglutinins was also affected by bursectomy. White Leghorns bursectomized at 1 week of age did not exhibit hemagglutinin titers until 9 weeks of age, and those bursectomized at 10 days did not begin to show titers until 6 weeks of age. All control birds had titers by 5 weeks of age. Groups bursectomized between 2 and 4 weeks of age were also delayed in attaining the control titer but did so by 9 or 12 weeks of age. These hemagglutinins may be produced in response to some unidentified antigen.

The observed reduction in antibody production following bursectomy may have been influenced by the induction of at least a degree of unresponsiveness in some birds.
Alteration of the Specificity of Lectin by Chemical Modification
Authors: Matsubara Shoji , Boyd William C.
Published: January 2023

What is thought to be the first instance of the modification of the specificity of a lectin by chemi-treatment is reported. Coupling diazotized p-aminophenyl - N - acetyl - D - galactosaminide to anti-A lima bean lectin greatly increased the agglutinating power of the lectin for group B cells without significantly altering its agglutinative activity for A1, A2, or O cells. The question is raised; do these observations possibly point to a role of carbohydrates in the specificity of lectins (and possibly of antibodies)?
Antigenic Analysis of Herpes Simplex Virus by Neutralization Kinetics
Authors: Ashe Warren K. , Scherp Henry W.
Published: January 2023

Antigenic differences among 15 strains of herpes simplex virus (HSV) were defined by the kinetics of their neutralization by 11 HSV antisera, using rabbit kidney cell monolayers for assaying virus by the plaque technique. A strain of HSV can be uniquely characterized by the rate at which it is neutralized by its homologous antiserum; heterologous strains are neutralized at slower rates. By converting absolute rate constants (K) to normalized rate constants (NK), the rates at which a strain of virus is neutralized by several antisera can be compared. NK values obtained in reciprocal neutralization tests divided these 15 strains into four distinct serogroups and one heterogenous group. Two strains of HSV isolated from the same individual at an interval of two years were antigenically distinguishable. Multiple passages of HSV in rabbit kidney cell cultures had no effect upon its antigenic specificity. These 15 strains cannot be antigenically correlated on the basis of the year of isolation, the clinical manifestation, or the methods used in isolating them.
Destruction of Immune Lymphoid Cells in Transplantation Immunity
Authors: McKhann Charles F.
Published: January 2023

The rapid transfer of H3-thymidine from labeled immune lymphoid cells to the cells of a rapidly growing ascites tumor from a foreign strain of mice against which the immunity is directed gives indirect evidence that immune cells may be selectively destroyed while reacting against the homograft.
The Effect of Adjuvant Amounts of Endotoxins on the Serum Hemolytic Complement Activity in Rabbits
Authors: Pearlman David S. , Sauers James B. , Talmage David W.
Published: January 2023

An attempt was made to correlate the adjuvant effects of bacterial endotoxins with a decline in serum complement activity. Rabbits were injected intravenously with sheep red cell stromas with or without endotoxins, and their serum complement activity was followed at various intervals. Serum complement activity was found to decline over a period of hours after the injection of adjuvant amounts of endotoxin, the most marked depression occurring 6 hr after injection. Possible mechanisms behind this effect of endotoxin on serum complement activity have been discussed.
The Common Protein Agglutinogen of Staphylococcus Aureus
Authors: Lenhart N. A. , Mudd S. , Yoshida A. , Li I. W.
Published: January 2023

A protein agglutinogen common to the internationally recognized serotypes of Staphylococcus aureus is described. Sera of all human subjects studied, healthy or infected, possess a high and fairly uniform level of antibodies against this agglutinogen. The agglutinin is concentrated in commercial human γ-globulin and is deficient in the blood of agammaglobulinemic children. No immunizing agent has been found which will regularly increase agglutination titers in man to a significant degree, although some increase has been induced in rabbits. Serum absorbed with the pure protein agglutinogen shows decreased agglutination titer against all serotypes but is not deprived of its phagocytosis promoting power.
The Common Protein Agglutinogen of Staphylococcus Aureus
Authors: Yoshida A. , Mudd S. , Lenhart N. A.
Published: January 2023

Staphylococcal agglutinogen was extracted from undigested cell walls and purified through carboxymethyl (CM)-cellulose column chroma tography and molecular sieving by Sephadex G-75. The sedimentation patterns of ultracentrifugation showed a high degree of homogeneity of the purified agglutinogen. The sedimentation constant is 1.6 S, the diffusion constant is 11.6 × 10-7 cm2/sec, and the molecular weight is 13,200. It showed single line precipitation reactions with human and rabbit antisera in agar gel diffusion and immunoelectrophoresis. From the nitrogen content, ultraviolet absorption, and sensitivity against digestion by proteolytic enzymes, it was shown that the agglutinogen is a protein. Jensen's antigen A contains the agglutinogen.
Phylogeny of the Immune Response
Authors: Grey Howard M.
Published: January 2023

The immune response of turtles to giant keyhole limpet hemocyanin was studied by means of the precipitation of I*-hemocyanin by antiserum. The pertinent findings were: The 18 S protein constitutes 13 to 26% of the total normal serum protein in the turtle.In an antibody response rapidly sedimenting antibody predominates for the first month and is gradually replaced by slowly sedimenting antibody.The slowly sedimenting antibody activity as well as that of the rapidly sedimenting antibody activity is almost entirely destroyed by 2-mercaptoethanol treatment.No anamnestic response occurred.The precipitin reaction between turtle antibody and antigen remains, throughout the course of immunization, similar to a mammalian primary response in that it is greatly effected by dilution of the test system.
Chromatographic Separation of the First Component of Complement and Its Assay on a Molecular Basis
Authors: Borsos Tibor , Rapp Herbert J.
Published: January 2023

A rapid and simple procedure for separating the first component (C′1a) of guinea pig complement in stable form is described as well as a quantitatative method for measuring C′1a activity on a molecular basis.
Demonstration of Two Types of Low Molecular Weight γ-Globulins in Normal Human Urine
Authors: Takatsuki Kiyoshi , Osserman Elliott F.
Published: January 2023

Low molecular weight γ-globulins in normal human urine (γ-u) are antigenically related to the S fragment of papain-digested γ2-globulin and have thermosolubility properties similar to those of Bence Jones proteins.

By immunodiffusion techniques, it has been established that there are at least two distinct populations of γ-u, designated γ-uA and γ-uB, which are respectively related to the Korngold and Lipari type A (= Mannik and Kunkel group 2) and Korngold and Lipari type B (= Mannik and Kunkel group 1) Bence Jones proteins.

The low molecular weight γ-u-globulins are increased in patients with various diseases associated with hypergammaglobulinemia and diminished in patients with hypogammaglobulinemia.
Studies of Acute Phase Protein
Authors: Kushner Irving , Kaplan Melvin H.
Published: January 2023

Following the injection of I131-labeled BSA into rabbits, CxRP was found to appear in the serum of one half of the animals demonstrating an immune response. The time of appearance of CxRP indicated that the inflammation began coincident with or immediately following the appearance of immune complexes in the circulation. It was not possible to differentiate on the basis of the level of circulating antigen-antibody complexes or the day of onset of immune elimination between animals with and those without CxRP responses. It was considered probable that the CxRP response was related to an inflammatory effect of circulating antigen-antibody complexes.

The serum level of CxRP, measured by a quantitative immunochemical procedure, was not related to the level of circulating antigen-antibody complexes, as measured by the Farr technique. Quantitative levels of CxRP stimulated by antigen-antibody complexes were in the low or moderate range as compared with responses to other inflammatory stimuli, such as typhoid vaccine or croton oil.
Evidence for Immunological Specificity of the Properdin System
Authors: Blum Livia
Published: January 2023

Treatment of serum with zymosan at 0°C, followed by removal of the zymosan-residue by centrifugation resulted in a supernatant serum in which C′3 could not be completely inactivated by fresh zymosan at 37°C. Although properdin could not be measured in the supernatant serum (0°-supernatant) by the zymosan assay, its presence could be demonstrated by direct isolation. These phenomena have been shown to be related to the interaction of zymosan with a previously unrecognized serum factor which is required for inactivation of C′3 in human serum by zymosan at 37°C. This factor (0°-factor) is distinct from other known constituents of the properdin system, from components of complement and from zymosan agglutinating antibody. Zymosan forms a complex with 0°-factor in serum at 0°C (0°-residue) which inactivates C′3 completely in 0°-supernatant at 37°C. Some of the kinetic and physicochemical characteristics of the interaction of zymosan with 0°-factor have been investigated.

0°-Residue could be dissociated and 0°-factor activity recovered in the eluate. Optimal conditions for elution of 0°-factor from 0°-residue were determined. An assay for measurement of 0°-factor was devised employing 0°-supernatant as reagent.

Experiments indicated that RP (serum reagent deficient in properdin) was limited in 0°-factor as well as in properdin. This hypothesis was confirmed by demonstration of the potentiating effect of partially purified 0°-factor on the properdin titers of serum and partially purified properdin.

Preliminary evidence was obtained for immunologic specificity of 0°-factor by adsorption experiments employing dextrans and levans as well as zymosan. The data are discussed in relation to an hypothesis that properdin is a nonspecific factor functioning as part of an immune system requiring antibody.
Immunoelectrophoretic Characterization of Human Serum Proteins with Primate Antisera
Authors: Lichter Edward A. , Dray Sheldon
Published: January 2023

Monkeys produce precipitating antibodies to at least 25 human serum proteins identified by immunoelectrophoresis. Some of the proteins with which the antibodies react were further characterized by modified histochemical techniques utilizing the specific chemical reactive sites on the serum proteins fixed in agar by the precipitating antibodies. In this way, two lipoproteins and five serum esterases were identified. At least three γ-globulins (γ-A, γ-B and γ-C) of different electrophoretic mobilities were identified with the monkey antisera. The appearance of parallel precipitin arcs for β2A, β2M and γ-C with monkey antisera suggests that these proteins each exist in at least two antigenically different forms in normal human serum. Chimpanzees produce precipitating antibodies to lipoprotein and to 7 S γ-globulin.
Immunochemical Studies on Rabbit Anti-Antibody
Authors: Fudenberg H. H. , Goodman J. W. , Milgrom F.
Published: January 2023

Anti-antibody in rabbits has been found to be a 19 S globulin with specificity for rabbit 7 S γ-globulin altered either by combination with antigen or by certain nonspecific methods. The anti-antibody combines with antigenic determinants on Fragments I and II, but not III, of the papain-digested 7 S γ-globulin molecule. The implications of these findings are discussed.
Doubling Time of Mouse Spleen Cells during the Latent and Log Phases of Primary Antibody Response
Authors: Capalbo Elmo E. , Makinodan T.
Published: January 2023

Cellular events were followed during the latent and log phases of primary antibody response by culturing spleen cells in 0.1-µ diffusion chambers. The results show that the decrease in T2 of the total cells and the increase in the number of mitotic cells after antigenic stimulation are reflections mainly of proliferative activities of blast and plasma cells. Estimates of the ratio, S:GT, indicate that after antigenic stimulation a significant increase was detected only among the immature plasma cells and lymphocytes. This would indicate that either the S period is prolonged or the GT decreased. Of interest also was the fact that the ratio for the reticulum-blast cells was 0.9 (68/75). This would indicate that the G1 period of these cells is very short. These results indicate that one is confronted with at least three variables when he is dealing with an intermediate cell compartment of a differentiation process in an immune response, i.e., the variable cell cycle dependent upon the stimulant and the input and output rates. Taking into consideration the cell compartment size, the T2, T1/2, and S:GT of each cell compartment, we can conclude that the terminal plasma cells are the end state of differentiation of reticulum-blast cells, suggesting that if other unique somatic cells can give rise to terminal plasma cells they must do so by feeding into the primary cell compartment.
Evaluation of the Diffusion Chamber Culture Technique for Study of the Morphological and Functional Characteristics of Lymphoid Cells during Antibody Production
Authors: Capalbo Elmo E. , Albright Joseph F. , Bennett William E.
Published: January 2023

Diffusion chambers constructed of 0.1 µ porosity filters and implanted into the peritoneal cavities of irradiated recipient mice have been found most useful for quantitatively evaluating cultures of isologous spleen cells during antibody response. During the first 3 days of culture there is an 88% decrease in the total number of cells which can be recovered; thereafter, the number of cells remains constant for at least 12 days. The production of hemagglutinin follows the typical time-course of antibody production by cells cultured freely in the peritoneal cavity of recipients. Synthesis of new antibody by cells cultured in chambers continues for at least 14 days and during the stationary phase of the response is presumably balanced by an equivalent rate of decay of existing antibody. During the over-all decline in total cell number during the first 3 days of cultures undergoing primary antibody response, certain types of cells undergo a relative increase. These are reticulum, blast and plasma cells and phagocytes which are induced to selectively proliferate by antigen-stimulation. Other types of cells, viz., granulocytes and normoblasts decrease disproportionately.
Toxicity, Pathogenicity and Antigenicity of Inhibitor-Sensitive and Nonsensitive Substrains of Influenza A2 Isolates
Authors: Lief Florence S.
Published: January 2023

Inhibitor nonsensitive (S-) and inhibitor sensitive (S+) substrains were recovered from a series of apparently sensitive A2 influenza viruses. The selection of S- substrains was facilitated by an initial passage in de-embryonated eggs of high concentrations of the parent strains in the presence of horse serum. Depending on the ratio of S- to S+ particles originally present in the mixed population, S- progeny appeared either immediately in the de-embryonated egg fluids or on a further passage in ovo of de-embryonated egg fluids in which hemagglutinins were nondetectable or of very low titers. Pure lines of S- virus were then established by passage of the S- progeny at limiting dilutions beyond the 50% infectivity level in the absence of horse serum. Pure populations of S+ particles were obtained by similar passage without horse serum at limiting dilutions of the parent virus.

It was found that S- particles were slightly more toxic than S+ particles when tested in mice by the intravenous or intracerebral routes. However, neither appeared pathogenic for the same host when given by the respiratory route. Similarly, no differences could be demonstrated with regard to their lethality for the chick embryo for both of them caused death to the same degree.

On examination of the two kinds of substrains by the strain-specific complement-fixation test, it became apparent that they could be distinguished serologically. S+ particles recovered from a given A2 virus appeared to be antigenically identical with S+ particles recovered from other A2 viruses of the same year, but distinct from S- particles and vice versa. Avidity for antibody played no role in this phenomenon. However, it was noted that on primary immunization, S- particles were capable of provoking more substantial homologous antibody titers than S+ particles.

The implication of these findings are discussed.
Antibody Formation by Rabbit Lymph Node Cells
Authors: Attardi Giuseppe , Cohn Melvin , Horibata Kengo , Lennox Edwin S.
Published: January 2023

To determine whether one cell can make two antibodies requires a series of controls that attempt to detect artifacts of technique, e.g., scoring of cells, and artifacts due to passive absorption of antibody by a cell. In addition, evidence that inactivation of bacteriophage is actually due to antibody as well as data on the relative amounts of the two different antibodies in single cells is presented. The results of the two previous articles (1, 3) in this series are discussed.
Direct Immunoelectrophoresis
Authors: Wilson Armine T.
Published: January 2023

A technique called direct immunoelectrophoresis is described, by which it is possible to visualize the size, shape, position and homogeneity of an entire antigen spot after electrophoresis in agar, by flooding the surface of the agar with antibody. The appearance of the visualized spot is similar to that seen when a protein substance is stained directly with dyes in simple agar electrophoresis. Protein antigens in too low concentration to be demonstrated by staining may be seen by direct immunoelectrophoresis, as may unstainable nonprotein antigens. Physicochemical heterogeneity of immunologically related antigenic substances can, in some cases at least, be demonstrated better by direct than by standard immunoelectrophoresis.
Effect of Corticosterone on the Skin Homograft Reaction and on Precipitin and Hemagglutinin Production in Thymectomized and Bursectomized Chickens
Authors: Meyer Roland K. , Aspinall Richard L. , Graetzer Mary Alice , Wolfe Harold R.
Published: January 2023

Neonatally thymectomized and/or bursectomized chicks treated daily with corticosterone were studied in terms of their abilities in producing precipitins and natural hemagglutinins and in rejecting skin homografts. Corticosterone treatment maintained a high percentage of homografts in the thymectomized, bursectomized as well as intact groups. This treatment, however, did not appreciably affect circulating antibody production. It was suggested that corticosterone may be acting either through the thymus and/or through its activity as an anti-inflammatory agent in maintaining skin homografts.
Studies on Human C′1-Esterase
Authors: Haines Anne L. , Lepow Irwin H.
Published: January 2023

Partially purified human C′1-esterase is antigenic in the rabbit. The immunizing antigen contained seven components by immunoelectrophoresis, only two of which were demonstrable in highly purified preparations of C′1-esterase. Antigenic similarity of C′1s and C′1-esterase was observed both by Ouchterlony gel diffusion and immunoelectrophoresis, providing additional evidence for the identification of C′1s as C′1-proesterase.

Anti-C′1-esterase in suitable concentrations completely inhibited the inactivation of C′4 by C′1-esterase, but only partially inhibited hydrolysis of ATE, a low molecular weight substrate. C′1-esterase completely inhibited by DFP was still capable of precipitating with anti-C′1-esterase. These data suggested that the antigenic and catalytic sites of C′1-esterase are distinct. The mechanism of inhibition by antibody is probably referable to steric hindrance or distortion of the catalytic site.

Anti-C′1-esterase inhibited hemolytic function of whole complement. Studies with the intermediate complexes EAC′1, EAC′1,4, and EAC′4 indicated that most of the antibody activity was directed toward C′1 and further implied the functional participation of C′1-esterase in immune hemolysis. At high concentrations of antibody, some agglutination of complexes which contained C′4 occurred, probably as a result of low titers of anti-C′4 in the antiserum. Anti-C′1-esterase did not inhibit the activity of the complex EAC′1,4,2, a finding consistent with previous work indicating that C′1 was not necessary for hemolysis once C′4 and C′2 had attached to the sensitized cell.
Characterization of Antibodies Produced by Guinea Pigs Inoculated with Inactivated Foot-and-Mouth Disease Antigen
Authors: Graves J. H. , Cowan K. M. , Trautman R.
Published: January 2023

Two types of neutralizing antibodies with markedly different physical-chemical and complement-fixing properties are found after inoculation of guinea pigs with inactivated foot-and-mouth disease virus. One, present during the 1st and 2nd week after inoculation, is of 19 S class sedimentation, of high adsorption to DEAE-cellulose, of β-globulin electrophoretic mobility, noncomplement fixing, and precipitation with viral antigen in agar gel. The other, appearing during the 2nd week after inoculation and continuing in excess of 30 days, is of 7 S class sedimentation, is poorly adsorbed on DEAE-cellulose, is of γ-globulin electrophoretic mobility, fixes complement readily and also precipitates with viral antigen in agar gels. Because of this marked change in their properties, it was concluded that quantitative studies of neutralizing antibody to assay antigens could not be done during the first three postinoculation weeks.
Neutralization of Bacteriophage ΦX174 by Specific Antiserum
Authors: Bowman Bernard U. , Patnode Robert A.
Published: January 2023

Three lots of bacteriophage ΦX174 were prepared by different techniques. Lot I was concentrated by dialysis against Carbowax 4000. Further concentration and purification were performed by differential centrifugation. Lot II was purified and concentrated by (NH4)2SO4 precipitation, followed by differential centrifugation. Lot III phage was treated exactly as the lot II phage except that it was further purified by RbC1 equilibrium density gradient ultracentrifugation. Analytical ultracentrifugation studies revealed that the lot II phage was composed of 114 S and 76 S particles and that the lot III phage contained only 113 S virions.

The antibody levels of sera from rabbits injected with lot I ΦX174 showed a typical primary response. The levels increase more rapidly and to a greater extent following the second injection of phage. Serum obtained from one rabbit on the 6th day after the second injection of phage was designated S24 and was used for all subsequent neutralization studies. Serum from rabbits injected with filtrate material from a broth culture of the host organism, Escherichia coli strain C, failed to neutralize phage.

In neutralization reactions with S24 diluted 2.8 × 104 times and lot I ΦX174 at an initial phage concentration (P0) of between 108 and 105 plaque-formers ml-1, the over-all rate of change of the survivor fractions was not constant for every P0/serum concentration ratio. This fact could not be attributed to reversible neutralization nor to the strain of host bacterium used for assay of surviving phage.

With P0 between 6 × 108 and 6 × 105 plaqueformers ml-1, neutralization of lot I ΦX174 by S24 diluted 6 × 104 times obeyed the percentage law, but the neutralization curves were not exponential. When the highly purified lot III ΦX174 was used in neutralization experiments with S24 diluted 6 × 104 times, the percentage law was obeyed when P0 was between 4.08 × 108 and 4.08 × 105 plaque-formers ml-1. In addition, all of the neutralization curves were exponential.

The specific rate constant of the ΦX174-S24 system was dependent on the temperature of incubation. The Arrhenius constant was approximately 9600 calories mole-1, corresponding to a Q10 of about 1.8.
γ-Globulin Metabolism in Germfree Guinea Pigs
Authors: Sell Stewart
Published: January 2023

Germfree guinea pigs have serum γ-globulin levels one-half to one-sixth those of non-germfree guinea pigs. The fractional rate of catabolism of homologous γ-globulin is the same in germfree and control guinea pigs, and there is no significant loss of γ-globulin into the intestinal tract of the germfree animal. Therefore the low levels of serum γ-globulin in germfree animals are due to lower rates of synthesis. The calculated absolute rates of synthesis are 2 to 5 times lower in germfree than in non-germfree guinea pigs. The catabolic rates of γ-globulin from germfree and from normal guinea pigs are the same in germfree and non-germfree guinea pigs.
A New Member of the Herpesvirus Group Isolated from South American Marmosets
Authors: Melnick Joseph L. , Midulla Mario , Wimberly Ira , Barrera-Oro Julio G. , Levy Barnet M.
Published: January 2023

A herpesvirus was isolated from two sick marmoset monkeys (Tamarinus nigricollis), that were believed to be infected with a helminth. The same virus was isolated directly from the kidneys and lungs of two different monkeys. It produced an effect in tissue culture cells resembling that of herpesvirus. The virus produced rapid, fatal infection in marmosets and in suckling mice; and it produced pocks on the chorioallantoic membrane.

Antigenically, this virus of new world monkeys was found to be different from classical herpes simplex virus and from B virus of old world monkeys, with no detectable cross reactivity. Its size and structure also show that the virus belongs in the herpes group, which may be briefly characterized as viruses containing DNA cores and possessing icosahedral symmetry with 162 hollow capsomeres. The behavior of the marmoset agent in tissue culture places it in herpesviruses of subgroup A (high titer of free virus) as against those in subgroup B (free virus uncommon).

By using primary kidney cell cultures of rabbit and of green and rhesus monkeys, it is possible to distinguish among the primate herpesviruses of subgroup A on the basis of the fact that the rabbit cultures are susceptible to all three, and green monkey cultures to both, of the monkey viruses (old world and new world), but not to the virus of man, whereas rhesus cultures are susceptible only to the old world monkey virus.
A Single Gene Controlling Hemolytic Complement and a Serum Antigen in the Mouse
Authors: Erickson Robert P. , Tachibana Dora K. , Herzenberg Leonard A. , Rosenberg Leon T.
Published: January 2023

Mice of strains deficient in hemolytic complement have been immunized with sera of strains positive for hemolytic complement activity. The resulting antisera react only with sera from mice with hemolytic complement activity. Typing of a variety of individuals for both the serum antigen and complement activity indicate that the same locus, Hc, determines the two. An absorbed rabbit antiserum prepared against the partially purified antigen also reacts only with serum from mice positive for hemolytic complement activity. This suggests that the allele at the complement locus, previously described as Hc0, determines no recognizable product.
Effects of Active Immunization and of Total Body X-Irradiation upon the Humoral Bactericidal System of the Guinea Pig as Measured with Strains of Enteric Bacilli
Authors: Steward J. P. , Collis L. R. , Roantree R. J.
Published: January 2023

For a given strain of E. coli sensitive to serum, a characteristic number is killed when placed in intraperitoneal diffusion chambers in normal guinea pigs. A serum-resistant strain of E. coli multiplied within the chambers in a normal guinea pig whereas a serum-sensitive mutant strain derived from it was killed. Immunization of the guinea pigs did not result in significant increases in the number of bacteria killed within the diffusion chambers when a serumsensitive mutant or a naturally serum-sensitive strain were used as test organisms. There was no evidence of a significant depression of complement-dependent humoral bactericidal activity after exposure of guinea pigs to total body x-irradiation (LD50). Contrary to some recent reports in the literature, no fall in the titer of naturally-occurring bactericidins for an E. coli and for an S. typhosa was found within 10 days after x-irradiation (LD50). Neither was there a decrease of bactericidal complement. A small and unexplained decrease in the ability of the guinea pig to kill E. coli in diffusion chambers was noted at 6 or 7 days after exposure to x-irradiation, but this was not associated with corresponding changes in the titer of bactericidal antibody or complement. Infections in guinea pigs following total body x-irradiation (LD50) are predominantly due to Gram-positive organisms. This is in contrast to the mouse, in which it has been well established that a high percentage of postirradiation infections are due to Gram-negative enteric bacilli.
Acquired Tolerance to Skin Homografts in Mice
Authors: Argyris Bertie F.
Published: January 2023

A quantitative analysis was carried out on the induction and maintenance of homograft tolerance in the CBA → C3H strain combination of mice. These strains share the H2-histocompatability locus. The results indicate that the incidence of tolerance, as measured by the ability of neonatally injected C3H mice to accept CBA skin grafts for 2 months or longer, decreases proportionally with spleen cell doses of less than 2 million. One-eighth million CBA spleen cells will induce 22% tolerance. CBA lymph node cells are slightly less capable in inducing tolerance than spleen cells. F1 hybrid lymph node cells are equally as effective in inducing tolerance as parental homologous lymph node cells. When the duration of tolerance is determined the results suggest that the length of the tolerant period cannot be correlated to the initial size of the donor cell population. The duration of tolerance is variable, even within groups of mice injected with equal cell doses of homologous spleen cells. Tracing the fate of donor cells by radioautography during the early phases of tolerance and by the “chimera” test during the later stages, indicates that the injected donor cells settle primarily in the spleen and lymph nodes of the tolerant host. Donor cells can be detected by chimera analysis, during all stages of graft acceptance, graft contraction, and even shortly after graft rejection. The results are discussed in the light of the hypothesis that in the CBA → C3H strain combination of mice, induction and maintenance of homograft tolerance is dependent on a critical balance between the immune potential of the host and the size of the donor lymphoid cell population.