bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigenic Analysis of Human Heart Tissue. Antigens with Restricted Organ Distribution in Acid Extracts of Human Myocardium
Authors: Espinosa Enrique , Kaplan Melvin H.
Published: December 2022

Antigens of human myocardium solbuilized by acid extraction at pH 2.5 were submitted to further analysis by double diffusion and immunoelectrophoresis using rabbit antisera to fractions of human heart acid extract obtained by DEAE-cellulose chromatography. Combined immunodiffusion and specific absorption tests demonstrated at least six antigens common to cardiac and skeletal muscle in acid extracts of myocardium. An additional heart-specific antigen was demonstrated in such acid extracts as well as in saline washes of myocardium. Of the antigens common to cardiac and skeletal muscle, four were found present in the different mammalian species tested, and two showed specificity for primates. The heart-specific antigen exhibited both species-specific and species-cross-reactive antigenic properties. Rabbit antisera to human heart fractions were reactive also with rabbit antigen common to heart and skeletal muscle and with rabbit heart-specific antigen. The antigens common to cardiac and skeletal muscle varied in susceptibility to proteolytic digestion by Pronase, trypsin and pepsin. The heart-specific antigen was susceptible to all three enzymes.
Immumochemical Studies of Foot-and-Mouth Disease
Authors: Cowan K. M. , Wagner G. G.
Published: December 2022

Guinea pig antiserum containing known quantities of antibody to foot-and-mouth disease virus (140S antigen) and virus protein subunit (12S antigen) was used as a standard in radial immunodiffusion (RID) analyses for determining the antibody content of other antisera. The antibody contents estimated by RID for the unknown sera were in close agreement with those subsequently established by quantitative precipitin analyses. Relatively small amounts of reagents are required and the procedure is simple to perform. Consequently, it provides a feasible procedure for the quantitative estimation of the antibody content of many antisera to the different antigenic components occurring in foot-and-mouth disease. The RID techniques used were approximately 10 times more sensitive for detecting either antibody or virus (140S antigen) than the Ouchterlony double diffusion method. The sensitivity of the procedure used was such that it approached that of complement fixation for the detection of 7S antibody. The greatest sensitivity for detecting either 140S antigen or antibody was obtained when the reactant sought was incorporated into the agar layer. Antibody was detected at a concentration of 1 µg/ml in agar, and 140S antigen was detected at a concentration of 1.25 µg/ml in agar by 19S antibody, and at 2.5 µg/ml by 7S antibody.
Complement Fixation and Immunodiffusion Tests for Assay of Hepatitis-Associated “Australia” Antigen and Antibodies
Authors: Schmidt Nathalie J. , Lennette Edwin H.
Published: December 2022

A serologic comparison was made of “Australia” (Au) antigens and antisera from different individuals using complement fixation (CF) and immunodiffusion tests. Titers of Au antigens varied widely in block CF titrations and in immunodiffusion tests against different antisera. CF titers of the antisera varied to a lesser extent in tests against different antigens. The importance of using optimal proportions of reagents for assay of Au antigen or antibody was demonstrated in both CF and immunodiffusion tests. In CF tests an excess of antigen produced an extensive “prozone” of inhibitory activity, and it was essential to examine a wide range of serum dilutions for antigen in order to avoid “false negative” results. An excess of either antigen or antibody was also shown to be inhibitory in immunodiffusion tests. The sensitivity of immunodiffusion assays for Au antigen or antibody could be enhanced by testing sera adjacent to wells containing positive reagents. Thus, of 204 sera from hepatitis patients which were positive for Au antigen by CF, 197 (approximately 98%) were positive by immunodiffusion; of 280 sera negative by CF, only 1 gave a positive immunodiffusion reaction. In contrast to findings reported by others, only a low proportion of sera from hepatitis patients in this study showed anti-complementary activity, and the activity was not reversed by an excess of Au antigen, although in some instances it was reduced by treatment with either antigen-containing or negative human sera. Sera from a few hepatitis patients in whom Au antigen had disappeared showed low titers of CF activity against their homologous antigens, which suggested that the convalescent-phase sera might contain very low levels of free Au antibody.
Staircase Rise in the Antibody-Forming Cell Population in Secondary Response
Authors: Sado Toshihiko , Perkins E. H. , Makinodan T.
Published: December 2022

Secondary anti-sheep RBC response by dispersed mouse spleen cells cultured in cell-impermeable diffusion chambers was assessed by the hemolytic plaque assay in combination with autoradiography and treatment with vinblastine. The results showed that, as in the spleen of mice undergoing a primary response, growth of the antibody-forming cell population occurred non-randomly in a staircase manner and recruitment of precursor cells into the antibody-forming cell population occurred repeatedly in a non-random manner. However, the size of the recruitment was significantly larger than that in the spleen of mice undergoing a primary response (i.e., a maximum of 40-fold vs 4-fold). Further studies on these precursor cells indicated that they transform into antibody-forming cells after a short but distinct lag following completion of mitosis. Another important observation was that the antigen-triggered proliferation of immunocompetent precursor cells could have occurred as early as 30 min after exposure of dispersed spleen cells to the test antigen, in contrast to a lag of more than 10 hr in the spleen of mice undergoing a primary response.
Detection of Immune Response Against Synthetic Polymers of Amino Acids Employing the Plaque-Forming Cell System
Authors: Egan Marianne , Maurer Paul H.
Published: December 2022

Incorporation of homologous polypeptide (10-5 M) into plaque assay plates containing anti-GLA30 (LLL) immune lymph node cells specifically inhibited these PFC. However, at lower levels of antigen incorporation an enhancement of PFC was noted. The enhancement was complement-dependent, did not occur with non-immune lymph node cells and was inhibited by specific anti-allotype serum. Cross-reaction studies suggested that the polypeptide did not enhance by merely optimally coating the erythrocytes. Serum titers were measured on modified localized hemolysis-in-gel plates. Titers were increased when certain concentrations of antigen were incorporated into these plates, thereby implicating the increased hemolytic efficiency of complement or antibody in the presence of excess antigen. Puromycin (5 to 25 µg/ml) did not inhibit the production of unenhanced plaques, but prevented the optimal enhancement of plaques by antigen. These same concentrations of puromycin inhibited (40% to 96%, respectively) the incorporation of 14 C-L-leucine into trichloroacetic acid precipitable protein. The inclusion of polypeptide in the incubation medium increased the above incorporation by 20% to 170%; puromycin in addition to polypeptide prevented this increase, although the radioactivity was above that obtained when puromycin alone was present. A similar effect was noted with chloramphenicol. The results suggest that enhancement of anti-polypeptide plaques by incorporation of polymer in the medium is a complex phenomenon and is probably due to at least two factors: the increased hemolytic efficiency of immunoglobulin or complement in the presence of excess antigen, and the stimulation of protein synthesis in immune lymph node cells in the presence of excess antigen.
Initiation of Antibody Responses by Different Classes of Lymphocytes
Authors: Strober Samuel
Published: December 2022

Thoracic duct cells and spleen cells from normal and immunized Lewis rats were injected into syngeneic recipients which had received 500 R x-irradiation 2 hr earlier. Tetanus and diphtheria toxoids were injected 24 hr after irradiation and the antibody response was measured for 2 to 3 weeks thereafter. Immunologically competent units were detected in normal spleen cells but not in normal thoracic duct cells. Immunization of the cell donors produced at least a 103- to 104-fold increase in units in the thoracic duct lymph and a much smaller increase in units in the spleen resulting in a change in the distribution (relative concentrations) of units in these lymphoid tissues.
The Evolution of the Immune Response
Authors: Pollara Bernard , Litman Gary W. , Finstad Joanne , Howell John , Good Robert A.
Published: December 2022

Lamprey produced agglutinins in high titer against Brucella organisms and the H antigens of human “O” cells. The antibody was shown to be labile at room temperature. The lamprey immunoglobulin was purified using Bio-Gel A 1.5 column and DE 52-cellulose column fractionation. Both purified immunoglobulin and antibody activity of serum were shown to have the electrophoretic mobility of an alpha globulin, a sedimentation velocity of 9S and a molecular weight of approximately 300,000. These characteristics indicate that the lamprey immunoglobulin is unique.
Studies on the Cytotoxic Immune Reaction
Authors: Manski W. , Ehrlich G. , Polack F. M.
Published: December 2022

Duck anti-rabbit cornea serum, which is cytotoxic to corneal epithelial or endothelial cells cultured in vitro, was found not to react with normal rabbit corneal tissue in vivo. No pathologic effects were observed after injecting such antiserum into the anterior chamber or intrastromally, or after repeated topical application to the corneal epithelium. Rabbit corneal tissues regenerating after injury were susceptible to damage by the anti-cornea serum. Corneal clouding developed after the antiserum was injected into the anterior chambers of eyes whose corneas had previously been injured by trauma or cold. In addition to pathologic changes in the endothelium, similar changes occurred in the epithelium after topical application of the antiserum. With the passage of time, the inflammatory response in the cornea gradually decreased in intensity, and the cornea became refractory to damage by antibody about 15 days after injury. The difference in the susceptibility to antibodies of regenerating as compared to normal tissue appears to be related to a difference in their cell membranes, which results in the accessibility of some antigens to cytotoxic antibodies during the active period of proliferation.
The Induction of Bacteriophage-Neutralizing Antibody by Mouse Spleen Immunogenic RNA
Authors: Herscowitz Herbert B. , Stelos Peter
Published: December 2022

RNA extracted from the spleens of Swiss Webster mice immunized with bacteriophage 81a, which infects Streptococcus faecalis var. zymogenes, induced the formation of specific phage-neutralizing antibody when added to nonimmune mouse spleen fragments in culture. Antibody formation could be elicited by the addition of immunogenic RNA derived from one inbred mouse strain to nonimmune spleen fragments from other inbred mouse strains. The activity of immunogenic RNA was inhibited by treatment with RNase, but not by DNase, trypsin, amylase or moderate amounts of pronase. The active RNA sedimented in the lightest fraction of a 5% to 20% linear sucrose gradient, and was estimated to have an average sedimentation coefficient of 8.2 S by analytical ultracentrifugation.
The Induction of Bacteriophage-Neutralizing Antibody by Mouse Spleen Immunogenic RNA
Authors: Herscowitz Herbert B. , Stelos Peter
Published: December 2022

Antigenic components derived from phage were detected in immunogenic RNA preparations extracted from immune mouse spleens. Injection of immunogenic RNA into previously immunized mice resulted in an anamnestic response, whereas challenge with non-immune RNA did not. Treatment of immunogenic RNA with mouse antiphage antibody reduced its ability to induce neutralizing antibody in cultures of normal spleen fragments. The extracted RNA was found to contain 14C-labeled material derived from phage. These findings indicate the presence of antigen in RNA extracted from the spleens of immunized mice.
Immunologic Tolerance to Escherichia Coli Somatic Antigen Assessed by Bacteriolytic and Hemolytic Plaque Assays
Authors: Allen Jerry L. , Friedman Herman
Published: December 2022

Immunologic tolerance to the somatic antigen of E. coli 0127:B8 was induced in neonatal mice by multiple injections with a lipopolysaccharide (LPS) antigen during the first weeks of life. The immune response of neonatally treated animals, as well as that of non-treated control mice, was assessed by two different kinds of immunoplaque assays: 1) direct bacteriolysis of viable E. coli in agar gel, and 2) hemolysis of sheep erythrocytes coated with specific antigen. Mice treated with differing amounts of LPS, and a variety of injection schedules, developed a marked tolerance as assessed by the indirect hemolytic plaque assay, as well as by serum hemolysin and hemagglutination tests. In contrast, significant numbers of plaque-forming cells were detected in spleens of neonatally treated mice when the direct bacteriolytic assay was used. Similarly, there were higher bacterial agglutinin titers in the sera of these animals. Larger numbers of antibody-forming cells were detected by the hemolytic assay in the spleens of control mice after challenge immunization than in neonatally treated animals. Significant numbers of bacteriolytic plaque-forming cells were present in the spleens of “tolerant” animals, in contrast to the marked suppression of hemolytic cells. These findings suggest that tolerance to bacterial antigens as complex as the somatic extracts of E. coli results in tolerance to “major” antigenic determinants and a concomitant immunity to some but not all of the “minor” antigens which fail to coat onto carrier erythrocytes.
Antigenicity of Polypeptides (Poly α Amino Acids): Immunogenicity of a Homopolymer Derivative
Authors: Maurer Paul H.
Published: December 2022

The γ-2-N ethyl morpholinyl derivative of the homopolymer polyglutamic acid (PAMEG) was immunogenic in rabbits but not in mice or humans. The specificity of the PAMEG-anti PAMEG reaction has been shown. The immunogenic behavior of PAMEG is analogous to polymers of D amino acids.
Herpes-Like Virus (EB Virus) in Continuous Lymphoblastoid Cell Lines from Patients with Viral Hepatitis
Authors: Stevens David P. , Barker Lewellys F. , Hopps Hope E. , Meyer Harry M.
Published: December 2022

In 1968, 21 continuous lymphoblastoid tissue culture cell lines were established in this laboratory from the peripheral blood of individuals with diagnosed acute infectious hepatitis (IH) (1). Concurrent attempts to establish similar cultures from normal subjects yielded a significantly lower number of continuous lines. These lines have been maintained in rapidly dividing suspension culture for 9 to 12 months.

Similar cell lines have been derived from individuals with lymphoma, leukemia (2, 3) infections mononucleosis (4), serum hepatitis (5) and other infectious diseases (6) as well as from presumably normal “healthy” donors (7, 8). Most have been shown to contain the Epstein-Barr virus (EBV), a herpes-like virus.

The present report summarizes studies employing immunofluorescence (IF) and electronmicroscopic techniques to determine whether EBV or any other viral agents were present in our cell lines which could be associated either with IH or with the lymphoproliferative phenomenon observed in vitro by leukocytes from patients with IH.
A Method for Typing Rabbit Sera for the A14 and A15 Allotypes with Cross-Linked Antisera
Authors: Tosi S. Landucci , Mage R. G.
Published: December 2022

The allotypes A14 and A15 are allelie speeificities on immunoglobulin-G (IgG) of rabbits (1, 2). They are located on the Fc portion of IgG and are genetically linked to specificities associated with the a locus which are localized on the Fd portion. Their assignment to a separate locus (e locus) awaits a definite solution of one of the crucial questions of immunogenetics, that is, whether the heavy polypeptide chain of a given immunoglobulin is the product of one or two genes. The A14 and A15 allotypes provide very useful markers. For example, by quantitating the distribution of al, a2, A14 and A15 among IgG molecules from heterozygous rabbits of known pedigree, we are able to evaluate whether genetic information from maternal and paternal chromosomes contributes to individual IgG molecules (3). In contrast to the a locus markers, the A14 and A15 markers are exclusively localized on IgG.
Biologic Properties of a Mouse IgG2a Myeloma Protein with Anti-Dinitrophenyl Activity
Authors: Warner Noel L. , Ovary Zoltan
Published: December 2022

An NZB-derived IgG2a myeloma protein with anti-dinitrophenol (DNP) activity was assayed for its ability to elicit complement fixation (CF) and passive cutaneous anaphylaxis (PCA) in guinea pigs after it had been combined with DNP antigens. The myeloma protein agglutinates difluorodinitrobenzene-coupled red cells and this agglutination can be inhibited by monovalent DNP amino acids, albeit with approximately 1000-fold more inhibitor than that needed to inhibit agglutination by a mouse anti-DNP serum. Despite the ability to agglutinate DNP red cells, no passive hemolysis occurred. The myeloma protein gave reverse PCA reactions in guinea pig skin with as little as 0.12 μg and challenge by anti-mouse globulin. However, even with 40 μg per skin site, no PCA reactions could be elicited on challenge with DNP conjugates. It is concluded that the complete biologically active sites on IgG2a molecules for CF and PCA reactions are created by the energy of binding of the molecule to its antigen. The myeloma anti-DNP protein does not create these sites after antigen binding because of its extremely low affinity.
Immunologic Detection of Single-Stranded DNA in Adeno-Associated Satellite Virions
Authors: Barnett E.V. , Torikai K. , Mayor H.D.
Published: December 2022

Double diffusion experiments with antibodies to single-stranded (s-s) DNA confirmed physicochemical data that adeno-associated satellite virions contain s-s DNA but the DNA extracted from such virions is double-stranded. Positive immunologic reactions of intact virions with sera from man and mouse with “autoimmune”disease and specifically immunized rabbits are consistent with the hypothetical role if viruses as immunogens or as the antigenic participants of immunecomplex vasculitis.
Cellular and Antibody Reactions to Streptococcal M Protein Types 1, 3, 6 and 12
Authors: Pachman Lauren M. , Fox Eugene N.
Published: December 2022

Immunization with purified M protein, a component of the group A streptococcal cell wall, induces the formation of circulating antibody and delayed cutaneous hypersensitivity (DCH). The specificity of these responses was evaluated in guinea pigs and man, using types 1, 3, 6 and 12 M protein. In a study of 10 human adults, delayed hypersensitivity and circulating antibody to more than one M protein was usually present. In several individuals, in vitro cellular reactivity to several M proteins was found in the absence of detectable DCH and antibody. Female Hartley guinea pigs sensitized with specific M proteins were reactive to homologous and, to a lesser extent, heterologous serotypes when DCH or spleen cell reactivity was determined. High titer homologous antisera reacted with heterologous M proteins measured by passive hemagglutination; only opsonic antibody was type-specific. Cellular responses to M proteins, assayed by spleen cell reactivity and inhibition of peritoneal exudate cell migration 6 months after sensitization when antibody was no longer detectable, were as great as when titers were high. Unimmunized controls, or those given Freund's adjuvant without M protein manifested negative skin tests, did not produce circulating antibody or respond with significant cellular proliferation when challenged with M proteins.
Hidden Rheumatoid Factors Reacting with “Non a” and other Antigens of Native Autologous IgG
Authors: Gaarder Per Ivar , Natvig Jacob B.
Published: December 2022

Several rheumatoid factor activities were revealed in 14 rheumatoid sera after the separation of IgM rheumatoid factors from inhibiting autologous IgG by gel filtration. These included a “new” specificity, anti-“non a.” Anti-Ga, found in 14 sera, anti-Gm(a), found in 10, and anti-“non a,” found in 6, were the most frequently detected rheumatoid factor specificities, while anti-Gm(g) was found in one serum. The anti-“non a” and anti-Ga rheumatoid factors reacted with antigens present in native autologous IgG. The original anti-Gm(p) and anti-Gm(e) rheumatoid sera contained anti-“non a” together with anti-Ga, and the respective original anti-Rh antibodies carried both the “non a” and Ga antigens. Gm(p) and Gm(e) appeared to be identical in part with “non a” and in part with Ga, and thus were not regular Gm types. The antigen for rheumatoid anti-“non a” was found to be located on the pFc' fragment of IgG. In contrast, the antigen for anti-Ga was present on Fc, but not on the pFc' fragment. Thus two distinct molecular regions of Fc were involved in the rheumatoid factor reactions.
Protein A from Staphylococcus Aureus
Authors: Stålenheim G. , Sjöquist J.
Published: December 2022

Myeloma IgG proteins become equally anticomplementary when aggregated with protein A from Staphylococcus aureus and thus contain equally efficient C-fixing sites. Furthermore, Fc fragments aggregated with protein A inactivate complement. Sheep erythrocytes stabilized with glutaric dialdehyde and coated with protein A were agglutinated by normal human IgG, by myeloma IgG, by fragment Fc from IgG, but not by fragment Fab. Thus protein A is a sensitizing substance for aldehyde-treated cells.
Biochemical Demonstration of the Activatable Esterase of the Rabbit Neutrophil Involved in the Chemotactic Response
Authors: Ward Peter A. , Becker Elmer L.
Published: December 2022

Direct biochemical evidence is provided that the rabbit blood neutrophil contains a precursor esterase involved in the cell response to the chemotactic stimulus C567, and that contact of the cell with chemotactic factor leads to activation of the proesterase. Rabbit blood leukocytes contain esterase 1, that is, an enzyme capable of hydrolyzing acetyl-dl-phenylalanine β-naphthyl ester. This activity can be ascribed largely, if not entirely, to the presence of neutrophils. Comparing blood leukocytes with peritoneal (exudate) neutrophils, the former cells contain greater esterase activity as well as greater chemotactic responsiveness. Treatment of blood leukocytes, but not peritoneal neutrophils, with chemotactic factor results in the acquisition of esterase activity (“esterase activation”) and chemotactic deactivation (loss of chemotactic responsiveness). By using highly purified complement and preparative (density gradient) ultracentrifugation, and employing agents that dissociate the macromolecular chemotactic factor, the phenomena of chemotactic activity, esterase-activating capacity and deactivating potential have been shown to be interrelated and clearly ascribable to C567. The esterase that appears in the blood leukocyte as a result of interaction with C567 can be identified as esterase 1, on the basis of exquisite sensitivity to inhibitory effects of phosphonate compounds at very low concentrations (10-8 M). These data provide direct biochemical support for the original hypothesis that the chemotactic response of the neutrophil to the complement chemotactic factor C567 involves an interaction which results in the conversion of the cell-bound proesterase to an active esterase. The biochemical role of this esterase in the kinetic behavior of the granulocyte is not known. This information may have special relevance to at least four other phenomena in which activation of a protesterase to an active esterase as the cause of an “allergic response” has been postulated on the basis of indirect evidence.
Abrogation of Immunologic Unresponsiveness by Whole Molecules and Fragments of Cross-Reacting Immunoglobulins
Authors: Scales R. W. , Cruse J. M.
Published: December 2022

A state of immunologic tolerance to a given antigen can be terminated by the injection of a cross-reacting antigen provided that the degree of cross-reactivity is within certain limits. The present investigation was designed to test the ability of human γG or its fragments to terminate tolerance to bovine γG or its fragments and vice versa. γG of both species was isolated by DEAE-cellulose chromatography and digested by mercuripapain. C57BL mice were injected with 1.0 mg of tolerogen and 20 days later were immunized with 0.2 mg of the cross-reacting antigen in Freund's incomplete adjuvant. Seven days after the last immunizing injection, 125I-tolerogen was injected intraperitoneally and the course of elimination followed by whole body counting. Sera were assayed for antibodies to the tolerogen by gel diffusion, immunoelectrophoresis and passive hemagglutination. Results of this investigation demonstrated that tolerance to bovine γG in mice may be terminated by injection of human γG or its fragments. Likewise, tolerance to human γG in mice may be terminated by injection of bovine γG or its fragments.
Immunologic Properties of the Main Encephalitogenic Peptide from the Basic Protein of Human Myelin
Authors: Lennon Vanda A. , Wilks Ann V. , Carnegie Patrick R.
Published: December 2022

The encephalitogenic basic protein from human brain was digested with pepsin and encephalitogenic peptides were isolated. Two overlapping peptides containing 15 and 22 amino acids had similar encephalitogenicity as the parent protein. Both peptides induced delayed hypersensitivity to the protein but no detectable circulating antibody in guinea pigs. There was a striking correlation between the occurrence of experimental autoimmune encephalomyelitis and low yields of peritoneal exudate cells. The main encephalitogenic determinant resides within the sequence Ser-Arg-Phe-Ser-Trp-Gly-Ala-Glu-Gly-Gln-. We postulate that the integrity of this region of the basic protein of myelin is vital to the normal functioning of the central nervous system. These small peptides should be of value in investigating the role of delayed hypersensitivity in autoimmune disease.
Homologous Serum Passive Cutaneous Anaphylaxis in Guinea Pigs Mediated by Two γ1 or γ1-Type Heat-Stable Globulins and a Non-γ1 Heat-Labile Reagin
Authors: Parish W. E.
Published: December 2022

Immunoglobulins with properties like those of human IgE (reaginic antibodies) occur in several laboratory animals, particularly the rat after antigenic injection (1, 2) or helminth infestation (3), the mouse after antigenic injection (4, 5) or helminth infestation (6), and the rabbit after antigenic injection (7, 8) and helminth infestation (9). Apart from a heat-labile antibody reported in guinea pigs infected with Trichinella spiralis (10), which conferred passive cutaneous anaphylactic (PCA) sensitivity on homologous skin for 21 days, reagin-like antibodies have not hitherto been described in guinea pigs, despite this animal's extreme susceptibility to anaphylaxis.

Benacerraf and Ovary and their associates (11, 12) described two 7S immunoglobulins in guinea pigs, a γ1 which sensitized homologous tissues anaphylactically, and a γ2 which did not. The investigations described below now reveal at least three antibodies in guinea pig sera capable of sensitizing homologous tissues anaphylactically.
Transfer of the Delayed Hypersensitivity Skin Reaction in the Guinea Pig Using Rna-Treated Lymphoid Cells
Authors: Jureziz R. E. , Thor D. E. , Dray S.
Published: December 2022

Ribonucleic acid extracts from the lymph nodes of guinea pigs sensitive to purified protein derivative, brain antigen, α, DNP-oligolysine and ε, DNP-oligolysine were shown to confer on syngeneic nonsensitive guinea pig lymphoid cells the ability to transfer for each antigen the delayed hypersensitivity skin reaction to nonsensitive guinea pigs. Successful transfers were obtained using intraperitoneal injections of approximately 109 nonsensitive lymphoid cells previously incubated for 15 to 30 min at 37°C with 120 to 300 µg/ml of RNA.
Specificity of Respiratory Secretion Antibody against Influenza Virus
Authors: Waldman R. H. , Wigley F. M. , Small P. A.
Published: December 2022

Serum, sputum and nasal secretion neutralizing antibody rises were measured against A2/Taiwan/64, A2/Hong Kong/68 and A/PR8 influenza viruses and parainfluenza type 1 virus in volunteers who had been immunized with A2/Taiwan/64 virus. There was approximately an eightfold rise in sputum, nasal wash and serum antibody to the homologous virus, and no rise to parainfluenza type 1 virus. There was no serum antibody rise to A/PR8 or A2/Hong Kong viruses, but in sputum and nasal washing there was approximately a three- to fourfold rise. Absorption studies with homologous and heterologous virus indicate that the heterologous response is due mainly to cross-reacting antibody. This could mean that secretory IgA is less specific than serum IgG. If these findings are substantiated for other viruses, it is possible that protection against a broad range of viruses may be inducible with fewer vaccines than would be expected on the basis of serologic studies.
Rosette Formation in Relation to Active Synthesis of Antibody
Authors: Moav Neomi , Harris T. N.
Published: December 2022

Studies were done on the requirement for an active synthetic process for the production of rosettes by mouse spleen cells, and on the relation of rosette-forming cells (RFC) to plaque-forming cells (PFC). Evidence for an active process was obtained by injection of mice with actinomycin D on the day of antigen injection. This prevented the appearance of both RFC and PFC. In other approaches, the inhibiting agents were applied in vitro to suspensions of cells producing rosettes. Rosette formation was inhibited on incubation at 37°C, in contrast with 4°C, by inhibitors of protein synthesis, cycloheximide and puromycin, and by anti-metabolites, iodoacetate and arsanilic acid.

The most direct evidence that rosettes and plaques can be produced by the same cells was obtained by differential density centrifugation. Following bovine plasma albumin density gradient centrifugation of original spleen cell suspensions, both rosettes and plaques were found to be produced by cells of the same, relatively low, density. When such suspensions were first incubated for rosette formation, however, the RFC, now in rosettes, were found in a fraction of higher density, intermediate between that of white blood cells and red blood cells. The number of plaques produced by this higher density fraction was found to be markedly increased over the analogous fraction from original spleen cell suspensions, in which rosettes had not been formed.

The nature of the receptors on the cell surface involved in rosette formation was examined in several ways. It was shown that rosette formation could be inhibited by exposure of the cells to an excess of antigen or to anti-IgG serum or its Fab fragments. The rosettes were partially sensitive to complement, indicating that IgM molecules as well as IgG molecules can be involved in rosette formation.
Bence Jones Proteins and Light Chains of Immunoglobulins
Authors: Solomon Alan , McLaughlin Carla L.
Published: December 2022

The immunochemical recognition of three classes of κ light chains, κI, κII, κIII, has been substantiated through immunochemical studies in which antisera prepared against several different κI, κII, κIII Bence Jones proteins were utilized. Heterogeneity within each κ-chain class has been demonstrated immunochemically and two distinct groups of proteins within each κ-chain class have been defined.

The availability of amino acid sequence data on Bence Jones proteins and light chains representative of the two groups of κI proteins made possible comparisons between proteins of these two groups. The ability to cleave selectively light chains, representative of the two groups of proteins within a κ-chain class, into their constituent variant halves and constant halves permitted the localization of the regions which were recognized by our antisera and immunochemically distinguished.
Antibody Inhibition of the Immune Response: Experimental Analysis of the Site of Action in Vivo
Authors: Eisenberg R. A. , Weissman I. L.
Published: December 2022

Analysis of inhibition of the immune hemolysin response was tested in vivo, utilizing sublethally irradiated mice injected with sheep red blood cells (SRBC) in order to separate antigen processing from central cellular events. The experiments demonstrate that passive anti-SRBC antibody, given to mice 2 days after SRBC and 3 days after irradiation with 600 rads, completely suppressed the response of incubated as well as non-incubated normal isologous spleen cells injected on the same day or 1 to 3 days after SRBC, but only partially suppressed a similar response in unirradiated mice when injected 2 days after SRBC. These experiments suggest that passive antibody can suppress the immune response by acting at some step subsequent to antigen processing.
Use of 51Cr in Studies on Cellular Immunity to Mycobacterium Tuberculosis, Brucella Melitensis and Poliovirus, Type I
Authors: Vickrey Herta M. , Elberg Sanford S.
Published: December 2022

An assay procedure was developed to assess resistance to cytopathic damage by virulent H37Rv in oil-induced peritoneal macrophages of normal and bacillus Calmette Guérin-immunized rabbits utilizing 51Cr. Immune serum was required for potentiation of maximal resistance. The radiolabeling procedure was standardized in terms of cultural and labeling conditions to obtain maximal radiochromate uptake with minimal nonspecific release, absence of re-utilization of released radiochro-mate and the low standard deviation of radiorelease in replicate cultures. Whereas functionally immune cells released only little more label than did their controls (sometimes less), nonimmune cells released appreciably greater amounts than did corresponding control cells. The radioassay demonstrated the importance of immune serum in potentiating cellular resistance to necrotization by tubercle bacilli, for effective immunity required the conjoint action of immune cells and immune serum. Immune macrophages in normal serum released significantly more isotope than in immune serum; normal cells in immune serum exhibited increased resistance by diminished label release. However, cells and sera reacted interdependently so that some immune sera enhanced the antimicrobial activity of certain normal cells nearly to the immune cell level; conversely, a number of immune cells in normal sera reacted in a manner comparable to that of normal cells. Further evidence that release of radiolabel reflects cellular damage was obtained through the use of an obligate intracellular parasite, poliovirus, type I, capable of rapid cytopathogenesis over a 48- to 72-hr period. When used to infect labeled monkey kidney cells, patterns of isotope release clearly indicated a close correlation between isotope escape and virus-initiated cytolysis.
Comparisons of the Specificities of Cell-Mediated and Humoral Immune Responses
Authors: Henney Christopher S. , Nordin Albert A.
Published: December 2022

The humoral and cell-mediated immune responses of guinea pigs to dinitrophenylated human IgG (DNP-HGG) have been concurrently assessed. Antibody-forming cells in draining lymph nodes and spleen were measured using a modified “plaque” assay and cell-mediated immunity measured on the same lymphocyte populations using a cytolytic technique, employing mouse mastocytoma cells coated with antigen as “target” cells.

The earliest plaque-forming cells (PFC) demonstrable in the spleens gave “plaques” only with DNP-HGG-coated SRC, although at the peak of the primary response the majority of the antibody-forming cells were producing antibody of antihapten specificity. The number of PFC formed against HGG-coated erythrocytes was always extremely low. Direct (γM) PFC of all specificities were negligible. PFC levels in the popliteal nodes were always higher than those observed in the spleen.

In contrast to the predominant anti-hapten specificity of the humoral response, cellmediated immunity was primarily directed against antigenic determinants of the carrier (HGG) molecule. The first cytolytic activity observable was specific for DNP-HGG-coated “target” cells, but by 14 days following antigenic stimulation DNP-HGG- and HGG-coated cells were lysed with equal efficiency. DNP-BSA-coated “target” cells were also effectively lysed, but cells coated with proteins immunochemically unrelated to the sensitizing antigen were not. The similar lysis rates observed with cells coated with the different antigens (DNP-HGG, HGG and DNP-BSA) suggested that a single lymphocyte population was involved in cytolysis, inferring that the “sensitized” lymphocytefr3 may be capable of recognizing more than one antigenic determinant of the immunizing antigen.
Corneal Hypersensitivity
Authors: Sery Theodore W. , Nagy Rose Marie , Winston Barber G.
Published: December 2022

The escape velocity of equine γ globulin or albumin introduced into a previously sensitized rabbit cornea is much greater than that found in the normal rabbit eye or in the systemically immunized animal. The loss of antigen from normal or immunized corneas follows first order kinetics. With equine γ globulin the normal cornea demonstrates an escape coefficient of -0.08/day. In contrast, the previously injected cornea which has passed through an immunologic response (Wessely phenomenon) and recovered will, upon reinjection of protein, demonstrate from the 1st day a rapid rate of escape (k constants averaging -0.44/day). This rapid rate is almost reached after the first corneal injection in systemically immunized animals, but it does not begin until after 5 to 6 days. Similar results were obtained with equine albumin. The rapid loss of antigen in previously sensitized corneas is believed to be caused by antibody in the tissues, and by a small population of immunocytes which are the residua of previous sensitization. In systemically immunized animals the longer period needed for removal of antigen is related to the time required for such cells to migrate into the corneal stroma. Other aspects concerning the development of an immunologic memory in the cornea and its significance for corneal disease are discussed.
Enhanced Antibody Response in the Offspring of Immunized Rats
Authors: Gill Thomas J. , Kunz Heinz W.
Published: December 2022

The inbred ACI strain of rats responds very well to immunization with poly Glu52 Lys33 Tyr15, whereas the F344 strain responds quite poorly; at least two genes are involved in the control of the antibody response (1). In the course of breeding the animals for these studies, we observed that the offspring of the F344 strain made considerably more antibody when the mothers were immunized with the antigen aggregated with methylated bovine serum albumin prior to becoming pregnant. It is important to define the factors involved in this effect, since they are presumably environmental and could interfere with the analysis of the genetic control of the antibody response.

In the studies reported here, the antibody responses of the offspring from unimmunized mothers and from immunized mothers are compared. Each of the immunized mothers received the antigen aggregated with methylated bovine serum albumin.
Two Complement Interaction Sites in Guinea Pig Immunoglobulins
Authors: Sandberg Ann L. , Oliveira Benedito , Osler Abraham G.
Published: December 2022

Numerous attempts have been made in recent years to identify and characterize the immunoglobulin site(s) associated with activation of the complement (C) system. Although there is considerable evidence that this activity can be attributed to the Fc fragment of rabbit antibody (1–8), other findings implicate the pepsin digestion products, i.e., the 5S or F(ab′)2 fragment (9–13). A possible basis for these divergent reports may be that multiple pathways exist for an immunologic attack on the C system. Recent studies in this laboratory with guinea pig immunoglobulins have strengthened this inference (14–17). These experiments disclosed that the hemolytic potency of C was diminished following interaction with either the γ-1 or γ-2 antibody classes, and that different pathways of complement utilization characterized these events. We now extend these observations in experiments whose results indicate that there are dual C interaction sites on the γ-2 guinea pig immunoglobulins.
Mouse Blood Leukocytes: in Vitro Primary and Secondary Responses to Two Synthetic Polypeptides
Authors: Tyan Marvin L. , Ness Dolly B.
Published: December 2022

The antibody responses of various strains of inbred mice to a series of multichain synthetic polypeptide antigens are quantitative traits under a dominant-determinant-specific type of genetic control (1). That is, a given strain of mouse may respond poorly, if at all, to one of the antigens while a second strain will respond quite well. These antigens are composed of a polylysine backbone with side chains of poly-dl-alanine and, in the case of the two antigens used in these experiments, terminating in short random sequences of either tyrosine and glutamic acid [(T,G)-A--L], or histidine and glutamic acid [(H,G)-A--L]. It has been shown that the genetic mechanism controlling these antibody responses is closely linked to the major histocompatibility locus (H-2) of the mouse (2). All strains of the same H-2 type (except H-2d) exhibit a similar pattern of immune response to these antigens regardless of the remainder of the strain's genetic background.
Immune Response to Alloantigens of Thymus Studied in Mice with Plaque Assay
Authors: Fuji Hiroshi , Zaleski Marek , Milgrom Felix
Published: December 2022

A plaque assay in which allogeneic thymus cells were used as target antigen has been developed. Inbred mice of various strains were stimulated with an intravenous injection of 4 × 107 allogeneic thymus cells. Whenever host and donor differed in ϑ antigens, plaque-forming cells (PFC) were detected in the spleen of the host. When host and donor carried identical ϑ antigen, PFC were detected only in certain strain combinations and only at the higher immunizing dose of 4 × 108 donor thymus cells.

Kinetics of the appearance of PFC was studied in the strain combinations differing in θ. In normal animals, the numbers of PFC were less than 10 per spleen. In the primary response, the peak numbers of PFC were observed in 4 to 7 days after the stimulations. In one strain combination, the mean number of PFC as high as 40,000 per spleen, was observed at the peak of the response. Marked differences in the magnitude of the PFC response to ϑ-AKR antigen were observed in various strains of mice which lack this antigen. In two strain combinations, the peak numbers of PFC were lower in the secondary than in the primary response. In one strain combination the reverse was true. Evidence was presented that PFC detected in this assay produce antibodies of IgM variety.
Rabbit Platelet Injury by Soluble Antigen and Antibody
Authors: Marney Samuel R.
Published: December 2022

Platelet aggregation induced by soluble antigen and antibody unrelated to the platelet membrane in heparinized plasma has been shown to be diphasic. The first wave of aggregation, which is complement- and divalent-cation-dependent, precedes the release of both vasoactive amines and adenosine diphosphate (ADP), is unaffected by the presence of 10 mM adenosine monophosphate (AMP), and coincides in onset with changes in the surface characteristics of the platelets detectable by retention of the platelets on glass bead columns. The second wave of aggregation is accompanied by release of platelet ADP and is totally inhibited by the presence of 10 mM AMP. Release of platelet serotonin occurs well after the second wave of aggregation. Citrate inhibits the second wave of aggregation and the release of both ADP and serotonin. It is postulated that the first wave is due to complement coating of the platelet membrane, which induces membrane injury and release of endogenous platelet materials in heparinized but not in citrated plasma. The second wave of aggregation is mediated by ADP released from the platelet. The first wave of platelet aggregation, which requires only the first four components of complement, clearly precedes and is separable from release of endogenous platelet materials, which requires at least six complement components.
C1q: Rapid Purification Method for Preparation of Monospecific Antisera and for Biochemical Studies
Authors: Yonemasu Kunio , Stroud Robert M.
Published: December 2022

C1q has been purified using precipitation in the presence of chelating agents at low ionic strength. The three times precipitated C1q was highly purified as shown by immunoelectrophoresis and analytic acrylamide disc electrophoresis. On immunoelectrophoresis, the purified C1q showed a single line in the slow γ-region against a potent anti-whole human antiserum, and the hemolytic activity and latex agglutination activity of C1q coincided with this region.

The final product showed hemolytic activity and latex agglutination activity but did not show any detectable C1r or C1s activity. The yield of C1q using these methods is about 40 to 60% and thus about 5 mg can be obtained from 100 ml of serum. The highly purified C1q obtained from 3.75% preparative acrylamide gels containing sodium dodecyl sulfate yields potent monospecific antisera when injected into rabbits. These antisera agglutinate EAC1q cells but not EA. The molecular weight of C1q as estimated by its relative mobility on acrylamide gels was 387,600 ± 10,790, and breakdown to smaller subunits has been demonstrated in gels containing urea.
Protein Biosynthesis in the Spleen
Authors: Selawry Helena S. , Starr Jason L.
Published: December 2022

The efficiency of spleen microsomes and ribosomes for the incorporation of phenylalanine after primary immunization was studied in an in vitro incorporation system developed for this purpose. Both microsomes and ribosomes were shown to become more efficient for the support of protein synthesis after antigenic stimulation. This change in amino acid-incorporating capacity appeared to be related to an intrinsic change which the ribosomes had undergone and was independent of the concentration of endogenous mRNA present in the system.
Development and Distribution of Immunoglobulin-Containing Cells in the Chicken
Authors: Kincade Paul W. , Cooper Max D.
Published: December 2022

The development and distribution of immunoglobulin-containing cells were studied in chickens using purified antibodies to µ, γ and light chains labeled with fluorochromes. Cells containing µ and light chains were found in lymphoid follicles of the bursa of Fabricius of 14-day embryos. IgG-containing cells were initially observed in bursal follicles at 21 days of incubation. Embryonic infusions of plasma from newly hatched and adult chickens had no effect on the appearance of immunoglobulin-containing cells within the bursa. The developmental sequence of immunoglobulin-containing cells within the bursa was later recapitulated in the spleen, tonsilla caecalis and thymus. Rapid expansion of IgM- and IgG-containing cells began in the spleen on the 3rd and 8th days, respectively, after hatching. The ratio of IgG- to IgM-containing cells was greatest in the intestinal wall of immunologically mature birds. Development of immunoglobulin-containing cells in extra-bursal sites was retarded in chickens hatched under germ-free conditions. In contrast, neither exclusion of exogenous antigens nor antigenic stimulation in ovo affected the time of appearance or proliferation of immunoglobulin-containing cells within the bursa. The medullary regions of individual bursal follicles frequently stained for both IgM and IgG. This pattern was not observed in lymphoid follicles of other tissues. Analysis of single cell suspensions revealed a high proportion of single bursal cells containing heavy chains of both IgM and IgG, wheras spleen cells rarely contained both classes of immunoglobulins. The results suggest that a developmental switch from IgM to IgG synthesis may occur within the bursa.