bluuebunny/crossref_metadata_2025 huggingface dataset:
Secretion of a Phytohemagglutinin-Binding Glycoprotein in Man
Authors: Strauchen James A. , Moldow Charles F. , Silber Robert
Published: December 2022

Phytohemagglutinin (PHA) is a glycoprotein purified from the red kidney bean, Phaseolus vulgaris, which binds to and agglutinates the erythrocytes of many species, including man (1). This reaction occurs without apparent blood group specificity. The finding that N-acetyl-d-galactosamine competitively inhibits agglutination by PHA suggests that, like the blood group isoagglutinins, PHA binds to a specific site on the cell surface (2). A PHA-binding material has recently been isolated and characterized (3, 4). There is also evidence that certain serum proteins react with PHA (5). This report describes the presence in saliva and ovarian cyst fluid of a glycoprotein which reacts with PHA.

Saliva was collected from normal donors and heated at 100°C for 10 min to inactivate salivary enzymes. After centrifugation at 1500 × G for 10 min, the opalescent supernatant fluid was tested for PHA hemagglutination-inhibition activity in the system described in Table I. Saliva from 30 individuals demonstrated hemagglutination-inhibition activity in dilutions up to 1:4. Inhibitory activity was independent of ABO group or secretor status. Ovarian cyst fluid demonstrated hemagglutination-inhibition activity in dilutions up to 1:16.
Cells Involved in the Immune Response
Authors: Abdou Nabih I. , Richter Maxwell
Published: December 2022

Antibody-forming cells (AFC) directed toward antigens on the sheep red blood cell have been localized to the sacculus rotundus of the normal unimmunized adult rabbit. These cells are either potential AFC or direct precursors of the AFC. The normal rabbit sacculus does not possess antigen-reactive cells (ARC) directed toward this or any other antigen. Only the rabbit bone marrow possesses the ARC. The significance of this finding with respect to accepted criteria for the definition of the primary immune response, both in vivo and in vitro is discussed.
Inv Allotypes of Cold Agglutinin Kappa Chains
Authors: Cooper Amiel G. , Steinberg Arthur G.
Published: December 2022

The normal globulins and the cold agglutinin kappa chains from 16 patients with the chronic cold hemagglutinin disease were tested for the Inv (1), Inv (2) and Inv (3) antigens. Ten of the patients had normal globulins with the Inv (-1, -2, 3) phenotype and this same phenotype was present on the cold agglutinin kappa chains tested. The remaining six patients had Inv (1, 2, 3) normal globulins, and of the cold agglutinin kappa chains from these patients, two had the Inv (-1, -2, 3) phenotype and four had the Inv (1, 2, -3) phenotype. Thus, the cold agglutinins of these six patients showed Inv allotypic exclusion compared with the normal immunoglobulins.
Specificity of Antibody Produced in the Bullfrog (Rana Catesbeiana)
Authors: Coe J. E.
Published: December 2022

The specificity of bullfrog (BF) (Rana catesbeiana) antibody to rabbit γ globulin (RGG) and hen egg albumin (HEA) was compared to the specificity of mammalian (guinea pig, mouse) antibody to these same proteins. Mammalian anti-RGG was relatively specific whereas BF anti-RGG produced a precipitin reaction with all mammalian sera tested. The BF anti-RGG contained antibody directed to both the Fc and Fab fragments of the RGG, although absorption experiments showed that antibody to the RGG Fc was primarily involved in the cross-reactions. Also, tests with fragments of the heterologous globulins showed that the BF anti-RGG was reacting with the Fc portion of the heterologous globulin. This BF antiserum to RGG detected similar antigenic determinants on the slow γ globulins from other rodents (7Sγ2 of guinea pig, hamster, Peromyscus) and man (IgG), and not on the electrophoretically faster or heavier immunoglobulins of rodents (7Sγ1, IgM) or man (IgA, IgM). A comparison of human IgG subclasses with BF anti-RGG showed that antigenic correspondence was greatest with γG1 and least with γG4. In contrast to the extensive cross-reactivity of BF anti-RGG, BF anti-HEA did not cross-react with other egg albumins (duck, goose) as did guinea pig anti-HEA.
The Localized Hemolysis-in-Gel Method Adapted to the Detection of Spleen Cells Releasing Virus-Specific Antibodies
Authors: Schwenk H.-U. , Lehmann-Grube F.
Published: December 2022

The localized hemolysis-in-gel method has been adapted to the detection of single cells releasing antibodies to influenza A virus. In mice infected with influenza A the number of cells releasing IgM antibodies specific for the inner component of the virus (RNP antigen) begins to rise between 24 and 36 hours after virus inoculation, reaches a maximum on the 4th day, and then decreases rapidly to the background level by day 20.
Target Cells of Human Ragweed-Binding Antibodies in Monkey Skin
Authors: Hubscher Thomas , Ian Watson J. , Goodfriend Lawrence
Published: December 2022

To determine the immunoglobulin nature of the allergen-binding antibodies which fix to monkey mast cells, sections from biopsies preincubated with sera of ragweed atopic or normal human individuals were stained by immunofluorescence techniques using antisera specific for human immunoglobulins. Evidence was obtained that one of the antibodies fixed to the mast cells was of the reaginic IgE type. By immunofluorescence and column chromatographic studies it was shown that non-reaginic ragweed-binding antibodies other than IgG, IgA, IgM or IgD present in the sera of hyposensitized individuals also fix to the monkey skin mast cells. The ability of both types of antibodies to render the mast cells fluorescent was lost after heating at 56°C for 4 hours.
Isolation, Chemical and Immunologic Characterization of κ- and λ-Type Light Chains from IgG of Normal Rabbits with b9 Allotype
Authors: Chersi Alberto , Mage Rose , Rejnek Jaroslav , Reisfeld Ralph
Published: December 2022

Immunoglobulin-G from the sera of b9 rabbits was subdivided into three fractions (I, II and III) by anion exchange chromatography so that the most basic fraction (I) contained essentially λ-type and the most acidic (III) κ-type light chains. Upon oxidative sulfitolysis and gel filtration of IgG I, λ-type light chains emerged with a Kd value of 0.35–0.38 (LI) whereas IgG III yielded κ-type chains which eluted with a Kd value of 0.48 (LII). The λ-type chains were characterized on the basis of immunologic reactivity, presence of C-terminal serine, absence of a free N-terminal amino acid and the presence of 5 half-cystine residues. The κ-type chains were characterized immunologically and by the presence of 7 half-cystine residues. The major b9 IgG fraction (II) yielded an LI peak considerably larger than LII (75:25) with about equal quantities of λ- and κ-type chains. This LI fraction contained an average of 6 half-cystine residues.
Immunoglobulins of the Hamster
Authors: Bienenstock John
Published: December 2022

An investigation of the distribution of immunoglobulins in the external secretions of the hamster revealed the predominance of γA. Only γA was found in saliva, whereas colostrum and urine both contained γ2 in addition to γA. Hamster γA was present in the serum at concentrations that were insufficient to allow its visualization by routine immunoelectrophoresis. No differences in electrophoretic mobility were found between serum and secretory γA regardless of derivation. The secretory γA from saliva, colostrum and urine had sedimentation characteristics intermediate between serum γA and 19S markers in sucrose density gradient ultracentrifugation studies. Similar results were obtained on Sephadex G-200 gel filtration. Isolated secretory γA had an s020,w of 15.2S. Mild reductive procedures did not appear to alter the sedimentation characteristics of this molecule. No antigenic differences were found between serum, salivary, colostral and urinary γA.
Anti-Pneumococcal Polysacharide Type III Hemolytic Plaques in Mice: Cellular Response in Immunity and Immunologic Paralysis
Authors: Kerman Ronald , Segre Diego
Published: December 2022

Sheep red blood cells sensitized with pneumococcal polysaccharide type III (SIII) react specifically with anti-SIII, and upon addition of complement are hemolyzed. This reaction provided the base for a hemolytic plaque procedure that was used to investigate the cellular response of mice to SIII given in amounts that induce either immunity or paralysis. Young offspring of immunologically paralyzed and normal mice showed differences in their response to SIII.
Communications
Authors: Iwasaki Tatsuo , Nozima Tokukichi
Published: December 2022

Antibodies to influenza virus have been shown to be present not only in serum, but also in secretions of the respiratory tract (1–4). Gamma A is the predominant immunoglobulin class in the secretions, whereas this immunoglobulin is a minor component in the serum. The γ A of the secretions differs chemically and antigenically from serum γ A. These observations suggest that antibodies associated with secretory γ A are produced locally, and are not simply the result of transudation from the serum. There is considerable indirect evidence for the synthesis of local antibodies in the respiratory tract. The problem remains to elucidate the mechanism of local antibody formation.

The lymphocytosis-promoting factor (LPF) obtained from Bordetella pertussis has been shown partially to suppress the formation of circulating antibody when given intravenously in mice (5). Therefore, the possibility of demonstrating a dissociation between the levels of antibody activity in the serum and in the secretions of mice infected with influenza virus was investigated.
Antigen-Induced Blastogenesis: The Human Cell Determining the Specificity of Response in Vitro
Authors: Levis William R. , Robbins Jay H.
Published: December 2022

Lymphocyte cultures devoid of phagocytic cells fail to undergo a blastogenic response to antigens. The addition of autologous or homologous macrophages to these cultures restores the response. While both lymphocytes and macrophages are required, the lymphocytes undergo blastogenesis and determine the immunologic specificity of the response.
Ontogenetic Development of Clq Synthesis in the Piglet
Authors: Day N. K. , Gewurz H. , Pickering R. J. , Good R. A.
Published: December 2022

The synthesis of C1q by tissues of pig embryos during gestation was studied using specific incorporation of 14C-labeled amino acids into C1q in vitro. C1q synthessi was first demonstrated in intestinal tissue (on the 48th gestational day) and this was the most active site of synthesis of C1q at all times. Significant synthesis of C1q subsequently was demonstrated in lymph nodes and spleen of older embryos (by gestational days 66 to 79) and at still later stages in tissues from other highly lymphoid organs such as liver and lung (gestational day 90 and later). We propose that there is a special relationship between development of synthesis of C1q and development of the lymphoid tissues, which suggests that the lymphoid cell is the site of C1q synthesis.
Immunochemical Relationships of Certain Capsular Polysaccharides of Klebsiella, Pneumococci and Rhizobia
Authors: Heidelberger Michael , Dudman William F. , Nimmich Wolfgang
Published: December 2022

Pyruvic acid is an immunologic determinant in the capsular polysaccharides of Klebsiella K32, pneumococcal types IV and XXVII, and eight strains of Rhizobia. All of these react heavily in anti-Pn XXVII. A cross-reaction between Pn IV and Pn XXVII is described and shown not to be due to pyruvic acid. Klebsiella K9 is characterized by non-reducing end-groups of D-glucuronic acid (or of 4-O-methyl-d-glucuronic acid) but differs in other respects from K2. Cross-reactions of K47 show it to be the only one of those tested with non-reducing end-groups of l-rhamnose. K52 reacts heavily in anti-Pn XXVIII. When depyruvylated, the rhizobial polysaccharides show much greater reactivity, and, occasionally, different reactivities in anti-Pn sera other than XXVII, in which depyruvylation virtually abolishes precipitation. The rhizobial polysaccharides may contain their pyruvic acid linked as a ketal to d-galactose; the linkage in S IV is more labile.
Studies on the Specificity and Affinity of α, DNP-Oligolysine Antibody: A Basis for Questioning the Role of Cell-Bound Antibody in Cellular Recognition of Antigen
Authors: Levin Howard A. , Levine Herbert , Schlossman Stuart F.
Published: December 2022

The specificity and affinity of serum antibody made to α, DNP-oligolysines was investigated by equilibrium dialysis, fluorescence quenching and inhibition of precipitation. The combining site of this antibody was found to have an upper limit in size complementary to α, DNP-Lys7. The increase in binding energy of α, DNP-Lys7 over α, DNP-Lys5–6 was found to be a relatively small fraction of the total binding energy. These results are contrasted to those of previous studies which showed that both immunologically competent or committed cells can readily distinguish between α, DNP-Lys5–6 and α, DNP-Lys7. The difference in specificity between the antibody-binding site and the proposed cellular receptor suggests that cell-bound antibody of the conventional type is not the mediator of antigen recognition by cells.
Some Observations on the Phylogeny of Serum Inhibitor of C1 Esterase
Authors: Donaldson Virginia H. , Pensky Jack
Published: December 2022

The antigenic properties of human serum inhibitor of C1 esterase were identified in serum from a number of primates, but were absent from serum of all lower species tested. Among primates the antigenic determinants in human and chimpanzee serum were identical with one another. In serum of the black ape, and rhesus monkey, antigenic determinants were also identical, but were deficient with respect to chimpanzee and human serum antigens. Although antigenic determinants of human serum inhibitor of C1 esterase apparently evolved among primates, functional serum inhibition was identified in the serum of some fish, fowl, rodents and mammals which were free of detectable antigen.
Effect of Physical State and Route of Inoculation of Diphtheria Toxoid on the Formation of Nasal Secretory and Serum Antibodies in Man
Authors: Butler William T. , Rossen Roger D. , Wende Reuben D.
Published: December 2022

To determine whether the physical state and route of inoculation of an antigen influence the appearance of IgA secretory antibody in nasal secretions, seven adult male volunteers were given fluid diphtheria toxoid and seven others alum-precipitated diptheria toxoid either intranasally or intramuscularly. Fluid diphtheria toxoid nose drops stimulated both nasal secretory and serum antibody production while nose drops containing alum-precipitated diphtheria toxoid failed to stimulate antibody production at either site. Intramuscular immunization with both types of toxoid stimulated serum antibody formation, but no secretory antibody formation followed the alum-precipitated toxoid. These results suggest that the physical state and route of administration of an antigen determine whether or not secretory IgA antibody is produced, and that a non-replicating antigen applied locally in the proper physical state causes a substantial local secretory antibody response.
A Comparison of PCA Reactive and Hemolytic Rabbit Antibodies to Sheep Red Blood Cells
Authors: Warner Noel L. , Ovary Zoltan
Published: December 2022

Rabbit antisera against intact sheep erythrocytes (SRBC) were assayed by hemolysis and PCA. In one serum, antibodies mediating both PCA and hemolysis were present, whereas another serum contained mainly hemolytic antibodies.

The PCA reaction could be developed by intravenous challenge with either fresh SRBC or their lysate, but not with stroma or formalin-fixed SRBC. Both stroma and formalin-fixed cells could, however, bind the hemolytic antibodies, which the lysate failed to do. The results indicate that one of the anti-SRBC serum contained antibodies against stromal antigens, which could be detected only by direct red cell binding assays (hemolysis or hemagglutination), and antibodies against a soluble internal antigen, possibly hemoglobin, which could be detected only by PCA reactions.
The Mixed Leukocyte Culture: A Three Component System
Authors: Rode Harold N. , Gordon Julius
Published: December 2022

The mixed leukocyte culture reaction is a proliferative reaction in which two cell types appear to perform three functions. The cell which proliferates requires the collaboration of a non-proliferating, supporting cell. The contribution of the latter is non-rate-limiting when added above a threshold concentration. Its role is probably nonspecific. The third function, i.e., the capacity to provoke a reaction, is shared by both cell types.
Complement Fixation by a Single Molecule of γG Hemolysin
Authors: Frank Michael M. , Gaither Thelma A.
Published: December 2022

Immunization of New Zealand White rabbits with boiled sheep erythrocyte stromata in complete Freund's adjuvant leads to the production of predominantly IgG hemolysin. Analysis of one such IgG hemolysin showed that it had unique biologic characteristics. A single molecule of this antibody on the surface of a sheep erythrocyte was capable of fixing C1̄. and initiating complement fixation. In many of its biologic activities the antibody, directed at the Forssman site on sheep erythrocytes, resembled a cross between IgG and IgM. This was true of the temperature sensitivity of C1̄ fixation by the antibody and the kinetics of Cl activation. There was almost no thermal effect on C1̄ fixation and the antibody activated Cl more rapidly than the usual IgG hemolysin and less rapidly than the usual IgM. Suggestive immunochemical evidence was obtained that the antibody may represent a unique subclass of immunoglobulin G.
Prevention of Immunologic Deficiency in Pituitary Dwarf Mice by Prolonged Nursing
Authors: Duquesnoy René J. , Good Robert A.
Published: December 2022

Baroni first demonstrated a deficiency of immunologic function in the Snell-Bagg pituitary dwarf mouse (1, 2). We have confirmed his observations and have also concluded that this deficiency appears largely to be limited to the thymus-dependent lymphoid system (3, 4). Another type of pituitary dwarf mouse, the Ames dwarf, has also been shown to be immunologically deficient (5). In both types of dwarfs the defect became particularly evident shortly after weaning at 21 days of age, as indicated by a sharp drop in the number of circulating lymphocytes.

We have studied the effect of prolonged nursing with foster mothers on the lymphoid system of dwarf mice. Three-week-old dwarf mice continued to have a strong suckling instinct for several weeks and consequently it was possible to foster-nurse them for a prolonged period beyond the usual time of weaning.
Relationship between Histocompatibility (H-2) Genotype and Immune Responsiveness to Low Doses of Ovalbumin in the Mouse
Authors: Vaz Nelson M. , Vaz Eda M. , Levine Bernard B.
Published: December 2022

A relationship has been recently described between the histocompatibility (H2) type of inbred strains of mice (1) and their immune responsiveness to low doses of hapten conjugates of bovine γ globulin (2,3). Several H-2k strains, as well as A/HeJ mice (H-2a) responded vigorously to immunization with repeated 0.1- to 1.0-µg doses of the conjugates, whereas H-2b, H-2d and H-2q strains failed to respond detectably. The present results indicate that the immune responsiveness to low doses of ovalbumin, or hapten conjugates of it, has nearly an opposite distribution. A single injection of 0.1 µg of ovalbumin in Al (OH)3 gel adjuvant induced intense and persistent primary antibody responses in H-2b and H-2q mice, whereas A/He mice (H-2a) and H-2k strains failed to respond.

Young adult female mice were used for immunization. Inbred animals from several different strains, described in the text, were obtained from the Jackson Laboratory, Bar Harbor, Maine.
The Immune Response to Heterologous Red Cells in Mice
Authors: Dukor Peter , Dietrich Felix M.
Published: December 2022

The effects of cyclophosphamide and cortisone acetate were examined in a model of antigenic competition between sequentially administered non-cross-reacting red cells from different species. In control mice receiving no drug, the relative immunogenic potency of heterologous red cells given with the first injection failed to determine the degree of unresponsiveness to subsequently injected unrelated cells. However, susceptibility to the suppressive action of previously administered erythrocytes was inversely related to the immunogenicity of the second antigen. Cyclophosphamide given 1 day after the initial red cell injection completely blocked competitive inhibition. On the other hand, impaired reactivity to the second of two sequentially administered antigens was further depressed by cortisone. The results are discussed in terms of targets of drug action and possible sites of antigenic interference.
Immunoglobulin Classes of Antibody-Forming Cells in Mice
Authors: Nordin Albert A. , Cosenza Humberto , Sell Stewart
Published: December 2022

The incorporation of tritiated thymidine (3HT) into hemolysin-in-agar plaque-forming cells (PFC) was determined for direct (IgM) and indirect (γ1) PFC following a single optimal injection of sheep red blood cells (SRC) into mice. After immunization with SRC, 3HT was injected into mice at various time intervals. The PFC of all mice tested were assayed at 5.5 days after immunization because γ1 PFC are not detectable in significant numbers until this time. The percentage of PFC labeled and the number of grains/labeled cell indicated that the onset, duration and extent of incorporation of 3HT were the same for both types of PFC. The potential γ1 PFC appear to be formed at the same time as the IgM PFC, but γ1 PFC do not synthesize or release γ1 antibody until 5 to 6 days after immunization, when proliferation of PFC has essentially stopped. IgM PFC appear 2 days after immunization and appear to synthesize DNA, divide and produce antibody simultaneously.
A Method for Preparing Monospecific Antisera to C1 Esterase (C1s): Microheterogeneity of Purified C1s
Authors: Nagaki K. , Stroud R. M.
Published: December 2022

Highly purified C1s has been obtained by immunization of rabbits using popliteal node injections of highly purified C1s followed by booster injections with gel segments containing C1s after disc-acrylamide gel electrophoresis. The antisera are monospecific and unexpectedly reveal electrophoretic microheterogeneity in highly purified C1s preparations. We have studied this electrophoretic heterogeneity in highly purified and in crude C1s preparations and hypothesize that it is due to two factors: 1) The partial proteolysis of C1s produced either by other serum enzymes or perhaps by autocatalysis. This can be demonstrated in highly purified C1s preparations. 2) The presence in crude preparations of another serum protein(s) which appears to influence the migration of C1s. These antisera are useful for quantitating C1s by single radial diffusion. We consider the heterogeneity of C1s to be an important feature of this molecule and future quantitative studies should take account of it.
Early Mast Cell Responses in Mouse Popliteal Lymph Nodes to Localized Primary Antigenic Stimuli
Authors: Roberts Audrey N.
Published: December 2022

Mast cell responses were quantitated in the regional draining lymph nodes of mice during the first 24 hr after primary footpad stimulation with three concentrations each of three soluble protein antigens. The time and intensity of response varied with the type and dosage of antigen. Immunization with 50 µg of bovine serum albumin (BSA) and ferritin induced the highest mast cell responses, with peak counts five times the normal mean at 5 and 3 hr, respectively. A low antigenic dose (10 µg) of BSA or ferritin stimulated peak responses at 16 hr, and the greatest response to 7.5 µg of sulfanilazo porcine γ globulin was observed at 3 hr. At doses of 100 to 150 µg, the three antigens yielded variable results and established no definite response patterns. During peak responses the mast cells accumulated in large clusters in the interfollicular cortex and medullary regions of the nodes. No degranulation or disruption of mast cells was observed, indicating the absence of “cytophilic antibodies” on the cell surfaces. Autoradiography was employed to study the incorporation of tritiated thymidine by responding mast cells at hourly intervals through the first 24 hr after immunization. No tritium was observed in mast cell nuclei, suggesting that the mast cell responses were not due to rapid cell proliferation.
Characteristics of the in Vitro Response of Guinea Pig Blood Lymphocytes to PHA and Antigen
Authors: Phillips S. Michael , Zweiman Burton
Published: December 2022

The in vitro response of guinea pig lymphocytes to phytohemagglutinin and tuberculin in tuberculin-sensitive and non-sensitive animals was studied. Measurements of tritiated thymidine incorporation, mitotic activity, and blast cell transformation were used to establish optimal conditions of culture and to investigate the mechanisms of in vitro interactions of antigen and cells. The pattern of response to phytohemagglutin (PHA) was found to be similar to that reported with human lymphocytes. Tuberculin-induced proliferative responses were limited to cells from sensitized animals, with a fairly consistent antigen dose-response and period of optimal incubation. Evidence was found to suggest a considerable dependence of the degree of in vitro responses to PHA or antigen upon the source of serum used in the culture medium. A high degree of correlation was found between isotope uptake and mitotic division in both PHA- and tuberculin-induced proliferative responses, with a less impressive correlation of these findings with the degree of blast cell transformation. When high concentrations of antigen were employed, there was an apparent dissociation of the blast cell transformation from DNA synthesis and mitotic activity in the lymphocytes of sensitized animals.
Migratory Response of Granulomatous Alveolar Cells from BCG-Sensitized Rabbits
Authors: Galindo Benito , Myrvik Quentin N.
Published: December 2022

Rabbits sensitized subcutaneously with bacillus Calmette Guérin (BCG) in oil routinely developed dermal tuberculin sensitivity but failed to develop pulmonary granulomas. Alveolar cells procured from such animals were not always inhibited by purified protein derivative (PPD) in the migration-inhibition test. In contrast, the majority of rabbits sensitized with the same dose of BCG by the intravenous route developed a pulmonary granulomatous response reflected by increased lung weight/body weight ratios and increased numbers of alveolar cells. The majority of such animals did not develop dermal tuberculin sensitivity. However, the granulomatous alveolar cells obtained from rabbits sensitized by the i.v. route were consistently inhibited by PPD in the migration-inhibition test. Alveolar cells procured from rabbits with extensive pulmonary granulomas exhibited inhibition of migration (auto-inhibition) in the absence of PPD. Granulomatous cells exhibiting auto-inhibition showed a tendency to escape both auto-inhibition and PPD-induced inhibition. Some BCG-immune sera potentiated PPD-induced inhibition, whereas other BCG-immune sera exhibited a tendency to relieve the auto-inhibitory response of granulomatous cells. Supernatant fluids obtained from granulomatous cells incubated with PPD inhibited the migration of normal alveolar cells. The possible role of the migration-inhibition response in granuloma formation is discussed.
Cells Involved in the Immune Response. XVII. The Elution of Antigen by Antigen-Reactive Cells from Antigen-Sensitized Glass Bead Columns. The High Affinity of these Cells for the Glass Bead-Adsorbed Antigen
Authors: Richter Maxwell
Published: December 2022

It was recently reported that antigen-reactive cells (ARC) of specific antigenic specificity in the bone marrow of normal rabbits (1–3) and antibody-forming cells (AFC) in the spleens of immune mice (4) can be isolated by the passage of these cells through an antigen-sensitized glass bead column. The cells which traverse the column possess no reactivity with respect to the antigen coating the glass beads; however, the cells which are eluted from the glass beads by vigorous shaking for several minutes react only with the antigen coating the beads and not with any other antigen (1, 2, 4). This highly specific antigenic reactivity of the eluted as compared with the effluent cells can be demonstrated by both in vitro incubation of the cells with the antigen (2, 3) or by transfer of the cells to irradiated immunoincompetent (800 R) rabbits (1) or mice (4).
Some Characteristics of Anti-Dinitrophenyl Antibody of the Gray Snapper
Authors: Russell Warren J. , Voss Edward W. , Sigel M. Michael
Published: December 2022

Gray snappers were immunized with dinitrophenylated hemocyanin and purified anti- bodies to the haptenic determinant (2,4-dinitrophenol) were obtained from two serum pools by by immune absorption. Examination of these antibodies revealed a single high molecular weight protein with an apparent valence of 5 and a relatively low binding constant.
Glycosyl Transferases for Mouse IgG
Authors: Schenkein Isaac , Uhr Jonathan W.
Published: December 2022

Cell-free fractions from a variety of mammalian tissues can catalyze the addition of carbohydrate to polypeptide (1–5). One of these reports describes the transfer of uridine diphosphate (UDP)-galactose to endogenously present IgG, mediated by the microsomal fraction of rabbit lymph node cells (6). The enzymatic activity appears membrane bound in all cases, and use of detergents (such as Triton X-100) has been necessary for its demonstration.

This paper describes initial studies on three distinct glycosyl transferases from a murine myeloma tumor and the partial purification of two of them: a galactosyl and an N-acetyl-glucosaminyl (NAG) transferase. In contrast to some reports by others, these latter two enzymes have been obtained in water-soluble form. An important aspect of our experiments was the use as substrate of mouse IgG from which a substantial portion of the carbohydrate had been removed by enzymatic digestion.
Evidence for Synthesis of Human Colostral γA as 11S Dimer
Authors: Bienenstock John , Strauss Hans
Published: December 2022

It is generally accepted that the secretory γA molecule in both rabbit and man is constituted by a serum type γA dimer plus an additional protein now termed secretory component (SC). It was first proposed by Tomasi et al. (1) that the secretory γA molecule was formed in plasma cells adjacent to the mucous surface and that combination with the secretory component, synthesized by glandular epithelial cells, occurred somewhere in transit of the γA molecule to the luminal surface. Examination of human colostral secretory γA isoagglutinin with anti-blood group A activity by Costea et al. (2) suggested to these investigators that the majority of the colostral secretory γA population was made up of molecules containing both κ and λ light chains. Thus, secretory γA appeared to be a hybrid molecule, implying random rather than specific combination of γA molecules.
Depletion of Plasma Complement in Vivo by a Protein of Cobra Venom: Its Effect on Various Immunologic Reactions
Authors: Cochrane Charles G. , Müller-Eberhard Hans J. , Aikin Barbara S.
Published: December 2022

The purified low molecular weight anticomplementary factor (CoF) in the venom of the cobra Naja naja was tested for its behavior in vivo and for its effect on various components of complement in several species of animals and the development of several immunologic reactions. CoF complexed with a plasma protein immediately after injection, the sedimentation rate of the complex being about 9S. The complex which inactivated purified C3 circulated with a half-life of 32 hr in rabbits. High blood levels of the CoF complex and minimal side effects were obtained using multiple injections of CoF over a 24-hr period. Under these conditions hemolytic complement levels fell to less than 10% of normal values in rabbits, rats and guinea pigs. C3 was markedly diminished in these species, and in guinea pigs C3 levels, as revealed in immune adherence, fell to less than 1% of normal. C3 and hemolytic complement levels remained low for 4 to 6 days. In two species tested, C5 protein was also lost during CoF treatment and in rabbits C6 activity diminished about half to one third of normal. The effect of CoF treatment in several immunologic diseases was assayed. Lesions depending upon neutrophil participation for full development were markedly inhibited. In acute nephrotoxic nephritis neutrophil accumulation in glomeruli and proteinuria were diminished. Arthus reactions, especially in the rat, were also inhibited. By contrast, immunologic reactions not requiring neutrophils, i.e., homologous cutaneous anaphylactic reactions in guinea pigs and contact (delayed) sensitivity were not affected by CoF treatment. Other possible effects of CoF in vivo were studied. The numbers of formed elements of the blood were not markedly altered. The most consistent finding was a modest elevation in neutrophils. Hemolysis was avoided by treatment of animals with multiple small doses of CoF. Neutrophils responded normally to immune adherence and chemotactic stimuli. Kininogen levels and clotting tests were unchanged.
Cytostasis of Mouse and Human Monolayer Cell Cultures by Normal, Unstimulated Lymphocytes
Authors: Jamieson Crawford W. , Wallace John H.
Published: December 2022

The effect of increasing concentrations of mouse and human lymphocytes on the growth of tumor cells, in vitro, has been studied by three independent methods. Growth was measured by sequential counts of the cells sheeted in predetermined microscopic fields of the culture flasks, by electronic counts of harvested cells and by thymidine uptake studies. All assay methods used indicate that inhibition of target cell growth is proportional to the ratio of lymphocytes to target cells. Control studies demonstrated that the effect is not due to exhaustion of the medium, the release of toxic breakdown products or elaboration of humoral antibody by the lymphocytes. Cell contact between the two cell types is essential, but there is no evidence that a cytotoxic factor is released as a result of this contact. No evidence for the accidental inclusion of a mitogenic agent was found.

The inhibitory capability is reduced by prior irradiation of the lymphocytes and is lost following their disruption. It is possessed, to a significantly lesser degree, by thymocytes but not by peritoneal macrophages. This cytostatic ability appears to represent a genuine capacity of apparently normal lymphocytes.
Streptococcal Group a Polysaccharide Haptens: Susceptibility to Degradation by β-N-Acetylglucosaminidases and Antibody-Combining Capacity of Haptens
Authors: Ayoub Elia M. , Schmidt Willard C.
Published: December 2022

Oligosaccharides of different sizes were prepared by acid hydrolysis of group A streptococcal carbohydrate. Separation of the oligosaccharides was performed by dialysis and chromatography on G-25 Sephadex. The activity of β-N-acetylglucosaminidase enzymes derived from the soil bacillus and from macrophage cells was assessed on the various haptens. The soil bacillus enzyme was equally active on the various sized haptens. However, the activity of the macrophage enzyme was strictly related to the size of the oligosaccharide, with hydrolytic activity increasing with decreasing size of the substrate. The capacity of the oligosaccharide fragments to inhibit the reaction between the group A polysaccharide and its antibody was studied using a radioimmune precipitation technique. The capacity of the haptens to inhibit antibody was directly related to the molecular weight of the oligosaccharides. Both the soil bacillus and macrophage β-N-acetylglucosaminidases were effective in eliminating most of the antibody inhibitory activity of the haptens, although this was achieved at lower concentrations of the soil bacillus enzyme. The amount of macrophage enzyme required to achieve a similar decrease in the antibody inhibitory activity of the haptens increased proportionately to the size of the haptens.
Antigen Binding and the Immune Response
Authors: Modabber Farrokh , Sercarz Eli
Published: December 2022

The cellular, immune response in the A/J mouse spleen to β-galactosidase (βGz) was examined by the A-H assay, which very sensitively measures the amount of antigen bound to cells. The early primary response can be subdivided into three phases: 1) a rise in activity which is completed by 12 hr, 2) a plateau from 12 to 48 hr, during which no substantial change can be discerned, and 3) a rapid increase from this time to the peak activity on days 4 to 6. There was no comparable latency in the secondary response. The existence of phase 2 supports the concept of a “true” preparative or latent period, not owing to difficulties in detection. The argument was favored that this first 48 hr represents splenic recruitment, cell-finding and interaction, followed by biochemical preparation for antibody synthesis.
The Cytophilic Activity of Human Immunoglobulins
Authors: Inchley Christopher , Grey Howard M. , Uhr Jonathan W.
Published: December 2022

The ability of human myeloma immunoglobulins of different subclasses to bind to the surface of guinea pig macrophages was investigated. Using 125I-labeled proteins, it was found that those belonging to the subclass IgG1 generally bound to a greater extent than did those in other subclasses, and that the cell-associated radioactivity remained largely on the cell surface during the short periods of observation employed. The Fc fragment of one such IgG1 protein was shown to contain virtually all of the cytophilic property of the parent molecule. Pre-incubation of guinea pig macrophages with IgG1 proteins, but not with representatives of other groups, resulted in a pronounced inhibition of the binding of cytophilic antibody, suggesting that the inhibiting proteins occupied the same site on the macrophage membrane involved in cytophilic binding.
Quantitative and Qualitative Studies of Antibody to Streptococcal M-Protein
Authors: Anthony Bascom F.
Published: December 2022

Rabbit antibody to group A streptococcal M-protein was measured by incubating dilutions of unabsorbed serum with partially purified and 131I-labeled M-protein from a type 12 strain (131I-M12) and coprecipitating the resulting complexes with anti-rabbit γ globulin. The method was found to be sensitive, quantitative and reproducible, but no antigen preparations, all of which were purified from acid extracts of cell walls, were found to be free of significant reaction with heterologous, unabsorbed antisera. This limitation was overcome by preincubating serum with an excess of unlabeled type 6 antigen; heterologous sera so treated subsequently reacted with 131I-M12 in a manner indistinguishable from normal rabbit serum (NRS). The specificity of the homologous reaction was indicated by its complete inhibition when serum was preincubated with unlabeled type 12 antigen and lack of inhibition with type 6 antigen. Significant binding of 131I-M12 was measurable with all type 12 sera examined and was present in early sera (20 days) which lacked detectable precipitating antibody and also in sera with insignificant or equivocal bactericidal activity. There was generally a linear relationship between the antigen-binding and precipitating capacity of different sera but a definite correlation between antigen-binding capacity and bactericidal activity of individual sera was not apparent. On indirect radio-immunoelectrophoresis, binding of 131I-M12 was observed only with IgG.
Development of Antibody-Forming Cells in Neonatal Mice: Stimulation and Inhibition by Calf Thymus Fractions
Authors: Hand Terry L. , Ceglowski Walter S. , Damrongsak Damri , Friedman Herman
Published: December 2022

Crude and purified saline extracts prepared from calf thymus glands, as well as two fractions prepared by disc electrophoresis from the purified fractions, were studied as to their effects on development of antibody-forming cells in neonatal mice. Mice injected at birth with microgram quantities of the purified extract or a stimulatory fraction, prepared by disc electrophoresis, had a 10- to 40-fold enhancement in the number of plaque-forming cells when challenged at 1 or 2 weeks of life, as compared with normal animals. There was also an enhancement of spleen size and weight. Another purified fraction, also prepared by disc electrophoresis and previously found to inhibit lymphocytopoesis, markedly suppressed development of antibody plaque-forming cells in the neonatal mice. The effect was relatively temporary, since older mice injected at birth with this fraction responded normally when challenged with the sheep red cells.
Dissociation of a Small Proportion of a Human IgM Population into Fcµ, Fabµ, and Free Light Chains Following Reduction
Authors: Morris James E. , Inman F. P.
Published: December 2022

Preparations of human IgM were treated with 0.1 M 2-mercaptoethanol and the reaction was terminated after 1 hr by adding neutralized iodoacetic acid. Dissociation into subunits, IgMs, was complete, but more slowly sedimenting material also was evident in the schlieren patterns. After gel filtration on Bio-Gel P-150 using a neutral salt buffer for solvent, several fractions were isolated in addition to the IgMs. The composition of the fractions was determined by ultracentrifugation, immunoelectrophoresis, and analysis of polypeptide chain contents. It was demonstrated that three fractions consisted primarily of IgMs, but that Fcµ and Fabµ also were present. One fraction comprised only Fcµ and Fabµ, and a fifth fraction consisted largely of free light chains.
Variation in the Size of Antibody Sites for the Poly-l-Aspartate Hapten during the Immune Response
Authors: Murphy P. Dianne , Sage Harvey J.
Published: December 2022

Rabbit antisera specific for poly-l-aspartate were prepared by immunization with poly-l-aspartyl-bovine serum albumin. Homogeneous l-aspartate oligopeptides were isolated from partially acid hydrolyzed commercial poly-l-aspartic acid by DEAE-cellulose chromatography. Hapten inhibition of the anti-poly-l-aspartylbovine serum albumin-poly-l-aspartyl-rabbit serum albumin reaction by l-aspartate oligomers was studied in a single rabbit with sera isolated after the first and second immunizations. The ability of oligopeptides to inhibit the poly-l-aspartate immune system increased with the size of the oligopeptide. With first course antisera an upper limit was reached with the triaspartate. With second course antisera the upper size limit of antibody-combining sites appeared to be larger than that found early in immunization.
Mechanisms of Release of Constituents from Rabbit Platelets by Antigen-Antibody Complexes and Complement
Authors: Henson Peter M.
Published: December 2022

The presence of neutrophils during the reaction of platelets with immune complexes increased the release of vasoactive amines. Two different neutrophil-dependent release mechanisms were involved, both requiring the complement system through C3, but not beyond C5. Soluble antigen, antibody in excess, C6-deficient plasma, platelets and neutrophils were used to demonstrate the first neutrophil-dependent mechanism. Release of vasoactive amines and nucleotides from the platelets followed the clumping together of the platelets, neutrophils and antigen-antibody-complement complexes. The release reaction required glucose and did not result from platelet lysis. Neutrophil procoagulant activity leading to thrombin activation in close proximity to the platelets in the clumps appeared to cause the vasoactive amine release. In the second process particulate antigens such as zymosan, which have fixed complement components, released increased histamine, serotonin and nucleotides from platelets when neutrophils were included in the reaction. The augmentation of release by neutrophils again required glucose and activated esterases and followed the aggregation of zymosan-complement, platelets and neutrophils.
Fetal Antigen Capable of Inducing Transplantation Immunity against SV40 Hamster Tumor Cells
Authors: Coggin J. H. , Ambrose K. R. , Anderson N. G.
Published: December 2022

Hamster, mouse and human cells transformed by simian virus 40 (SV40) share a common antigen capable of stimulating specific transplantation immunity against SV40-induced tumors in hamsters (1, 2). Cursory consideration of this observation might suggest that the viral genome must code for the common tumor-specific antigen in the different cell species. Knowledge acquired recently about the size of the SV40 genome limits the potential informational complement of this virus to seven to eight genes (3). Available data indicate that at least six genes are required for virus synthesis (3, 4), and radiation studies suggest that one or two different genes are required for transformation (5). It is becoming increasingly difficult to adhere to the concept that the virus codes directly for the tumor-specific transplantation antigen (TSTA) which is incorporated into the membrane of transformed cells unless the TSTA is a small protein and is a pleiotropic viral gene product.
Antigenicity of Polypeptides (PolyαAmino Acids): Calcium-Dependent and Independent Antibodies
Authors: Maurer Paul H. , Clark L. G. , Liberti P. A.
Published: December 2022

It has been shown that sheep antisera against negatively charged synthetic polymers of amino acids such as poly (L-glu60-L-ala30-L-tyr10)n of high glutamic acid content contain two populations of antibodies one of which requires calcium (“Ca++ dependent”) for reaction to occur. The Ca++-dependent antibodies are directed against immunogenic determinants created by the reaction of Ca++ with glutamyl residues which changes the conformation of the polymer and thereby creates the new determinants. The reaction of the specific polymer with this antibody is maximal at 0.008 M Ca++. Dissociation of the polymer-Ca++-antibody complex can be accomplished by removal of Ca++ with EDTA, citrate or oxalate or by washing the antigen-antibody precipitate with phosphate-buffered saline. Dilution of the antiserum in non-Ca++-containing diluents, which in effect lowers the Ca++ concentration, reduces precipitation of the antibody. This loss can be reversed by addition of Ca++ to the diluting buffer. The amounts of these two populations of antibodies vary from animal to animal within any species. No Ca++-dependent antibody was found with the polymers of slight negative or neutral charge or against several protein antigens. Peptide maps (finger prints) of these two kinds of purified antibody (both 7S IgG) revealed no significant differences.
Ribonucleic Acid Synthesis in Rabbit Lymph Node Cells
Authors: Donahue Helaine A. , Moore Virginia G. , Little J. Russell
Published: December 2022

Suspensions of lymph node cells were prepared from immunized and from unimmunized rabbits. In a totally synthetic medium at 37°C, these cells incorporated 3H-uridine into RNA at an initially rapid linear rate without lag. After 60 to 90 min of incubation, the rate of uridine incorporation changed to a second linear rate which was about 9% of the initial rate. Maximal initial rates of 3H-uridine incorporation were observed 5 days after immunization, though differences were small between cells from immunized and nonimmune animals. Sucrose gradient analyses of whole cell RNA revealed that the major classes of cellular RNA contained significant radio-activity and antigen-stimulated cells revealed predominant labeling in a 12 to 13S fraction. These experiments provide a reasonable approach to the preparation and isolation of high specific activity RNA for use in the RNA-DNA hybridization studies or in the functional characterization of various classes of RNA implicated in immune responses.