bluuebunny/crossref_metadata_2025 huggingface dataset:
Amino Acid Sequence Studies of a Complement-Fixing Immunoglobulin Fragment
Authors: Kehoe J. M. , Capra J. D. , Fougereau M.
Published: December 2022

Previous studies have established the complement-fixing capacity of a 62 amino acid cyanogen bromide fragment derived from the amino terminal area of Fc of the Ig 2a molecule produced by the Potter murine myeloma MOPC-173. Amino acid sequence studies allowed a comparison of this region with the published sequences of the corresponding region of two human IgG proteins, one (Eu) belonging to the complement-fixing γG1 subclass and the other (Vin) to the noncomplement-fixing γG4 subclass. Within the NH2 terminal 45 amino acids, a tyrosine in position 296 (Eu numbering) was the only residue common to the two complement-fixing molecules which was replaced (by a phenylalanine) in the non-complement-fixing immunoglobulin. Other substitutions were observed that are apparently not correlated with this particular biologic activity, but a strong mouse-man sequence homology was apparent for the region.
Enzymatic Activation of C3 by a Complex Derived from a Purified Cobra Venom Factor and a Serum Globulin
Authors: Bitter-Suermann D. , Dierich M. , König W. , Hadding U.
Published: December 2022

A cobra venom factor (VF) and a serum factor (SF) form a complex that is able to activate C3. The complex formation has been studied using preparations of purified VF and SF, which on polyacrylamide gel electrophoresis showed one single homogeneous band. The SF and all complement components have been purified from guinea pig serum. VF and SF did not react with each other except when Mg++ was present. Ca++, Zn++, Mn++, Cu++, Fe++ did not lead to complex formation. It could be demonstrated that the amount of formed complex is a function of the concentration of free Mg++ ions. The formation of an active complex is inhibited by EDTA. Reversal of inhibition can be achieved with Mg++. The complex formation is temperature dependent and starts at 37°C without lag phase. Applying the reaction velocity of the C3-turnover as a parameter for the complex formation it is shown that ca. 90% of VF and SF have interacted after 45 min at 37°C.
Elastase of Pseudomonas aeruginosa: Inactivation of Human Complement Components
Authors: Schultz D. R. , Miller K. D.
Published: December 2022

Pseudomonas infections are characterized by hemorrhagic and ulcerative necrosis with little or no inflammation. An elastolytic protease, implicated in the necrotic process, was isolated from cell cultures and was purified to homogeneity as shown by disc electrophoresis. Adding elastase to fresh human serum destroys the functional hemolytic activity of the nine components of complement (C1–9). Pretreating sheep erythrocytes (E) or sensitized erythrocytes with elastase affects neither the antibody-combining sites nor the combining sites for the sequential reactions of C. Of the nine isolated functionally pure C components tested individually, only C4 and C7 are not inactivated by elastase. The functional hemolytic activity of C3 is completely destroyed by elastase treatment, and highly purified β1C is converted to β1A as judged by immunoelectrophoresis. Exposure of human serum or a mixture of nine functionally pure C components to elastase did not cause hemolysis of unsensitized E.
Presence in Rabbit Colostrum of IgA Autoantibodies to a Gastric Antigen
Authors: Wise Ronald D. , Lichter Edward A. , Dray Sheldon
Published: December 2022

Rabbits had IgA antibodies in their colostrum that precipitated an organ-specific antigen in their own stomach tissue as well as that of their progeny and of unrelated rabbits. Colostrum from a pseudopregnant virgin doe also precipitated this gastric antigen. Precipitin activity was not found in serum during pregnancy or post partum. Electrophoretically, the gastric antigen migrated faster than albumin to the rho region, and according to gel filtration the molecular weight resembled that of albumin.
Recessive Inheritance of Rapid Anti-Allotype Antibody Production
Authors: Kindred Berenice , Weiler Eberhardt
Published: December 2022

BALB/c mice made antibody against an allotype determinant of C57 immunoglobulin after only three injections of non-agglutinated serum. Evidence was obtained indicating that the determinant was carried by the γG2a globulin class. C3H and CBA mice did not produce anti-allotype antibody when submitted to the injection schedule to which BALB/c animals responded. The results of treating F1 and backcross mice demonstrated that the strain difference was under polygenic control.
The Influence of Transfusions from Unrelated Donors upon Marrow Grafts Between Histocompatible Canine Siblings
Authors: Storb Rainer , Rudolph Robert H. , Graham Theodore C. , Thomas E. Donnall
Published: December 2022

Previous studies indicated that marrow grafts between canine siblings matched by serologic histocompatibility typing and for non-reactivity in mixed leukocyte culture uniformly resulted in prompt and sustained engraftment. In contrast, marrow rejection was the rule when the recipient was given a single prior blood transfusion from the matched sibling marrow donor. The present study investigated the effect of multiple prior blood transfusions from randomly selected unrelated dogs on the outcome of marrow grafts between histocompatible siblings. Recipients were given three transfusions on each of three occasions, 24, 17 and 10 days before grafting. Recipients were prepared for grafting by exposure to 1500 R whole body irradiation followed within 4 hr by infusion of marrow from the matched sibling. Seven of the 11 recipients had prompt and sustained marrow engraftment and survived beyond 70 days. Four recipients died between 9 and 40 days, three of marrow rejection and one with a functioning graft and graft-vs-host disease. The data indicated that multiple prior transfusions from unrelated dogs can in some cases immunize a recipient and prevent subsequent successful marrow engraftment from a histocompatible canine sibling.
The Third Cell Type Required for the Immune Response of Spleen Cells in vitro
Authors: Cosenza Humberto , Leserman Lee D. , Rowley Donald A.
Published: December 2022

Cells that adhere to glass or plastic surfaces, obtained from spleens of nor mal and immunized mice, were compared for the ability to interact synergistically with nonadherent cells in the in vitro response to sheep erythrocytes. Adherent cells obtained from normal and immunized mice were equally x-ray resistant at several levels of x-irradiation and equally effective in enhancing the response of nonadherent cells. The plaque-forming cell response of cultures that contained dilutions of adherent cells from immunized mice were equivalent to responses of cultures containing corresponding dilutions of normal adherent cells. Adherent cells, therefore, did not appear to increase in number or change in function following in vivo antigenic stimulation. Adherent cells, whether derived from normal or immunized mice, were required for the in vitro response of non-adherent cells obtained from immunized animals. Spleen cell cultures obtained from immunized mice, however, required higher dilutions of non-adherent cells to interact synergistically with adherent cells. The enhanced responsiveness of non-adherent cells from immunized mice was consistent with an increase in number or change in function of non-adherent cells as a result of antigenic stimulation in vivo.
In Vitro Studies of the Response of Hamster Lymphocytes to Phytohemagglutinin and Antigenic Stimulation
Authors: Fernald Gerald W. , Metzgar Richard S.
Published: December 2022

A semi-micromethod of in vitro culture of hamster lymphocytes was developed for studies of the immune response to cellular and soluble antigens and to microbial pathogens. The number of cells necessary for optimal stimulation in the 0.2-ml volume employed was 0.5 × 106 to 1.0 × 106; 20 to 60 cultures could be prepared from a single donor lymph node under these conditions. The response to phytohemagglutinin did not differ significantly from that described for several other laboratory species. A mixed lymphocyte reaction to human and other xenogeneic cells was demonstrated but allogeneic mixtures of Golden Syrian and MHA strains did not stimulate DNA synthesis. Responses to tuberculin-purified protein derivative and keyhole limpet hemocyanin were elicited in cultures from animals immunized with these antigens. Although cells from the mesenteric lymph node were employed for most experiments, similar reactivity was obtainable with peripheral blood lymphocytes.
The Role of Complement in the Phagocytosis of Candida Albicans by Mouse Peripheral Blood Leukocytes
Authors: Morelli Remo , Rosenberg Leon T.
Published: December 2022

The phagocytosis of Candida albicans by peripheral blood leukocytes obtained from complement-deficient (C-) CF-1 female mice was tested in the presence of specific mouse antiserum, complement-positive (C+) or C- normal mouse serum. Phagocytosis was also tested with leukocytes collected from C+ B10.D2/Sn “new line” × B10.D2/Sn “old line” F1 hybrid mice and C-F1 hybrid × old line backcross mice in the presence of either C+ or C- normal mouse serum. The results indicated that phagocytosis of Candida by mouse blood leukocytes was rapid and was enhanced by a complete hemolytic C system. Also, phagocytosis of the yeast was enhanced by specific antiserum. The extent of intracellular killing of Candida measured with mouse peritoneal cells was not influenced by the presence of antibody, or of a complete C system, or the C status of the animals providing the cells.
The Immunologic Response to Xenografts
Authors: Sachs David H. , Winn Henry J. , Russell Paul S.
Published: December 2022

The humoral response of rats to mouse xenografts included production of antibodies that detected the major histocompatibility alloantigens of donor mice with remarkable specificity. Although these antibodies were present in high titer, their effects were generally masked by “species-specific” activity until after appropriate absorptions. The absorption process produced significant anticomplementary activity in the absorbed sera, which may explain difficulties previously experienced in recognizing allospecificity in xenoantisera. The fact that histocompatibility alloantigens, unlike other alloantigens, are recognized readily by a xenogeneic host may indicate greater homology between major histocompatibility antigens of different species than was previously recognized.
Alterations in the Kinetics of Hemagglutinin Formation Following Embryonic Bursectomy
Authors: Moticka Edward J. , Van Alten Pierson J.
Published: December 2022

Surgical removal of the bursa of Fabricius 2 days before hatching caused a significant decrease in the number of cells capable of rosette formation and in the amount of antibody produced to sheep erythrocytes. This was accompanied by an alteration in the kinetics of the response and production of predominantly 2-mercaptoethanol sensitive, high molecular weight antibody. Bursectomy at 19 days of incubation appears selectively to eliminate a population of immunologically active cells. The remaining immunocompetent cells are incapable of developing a normal antibody response to sheep erythrocytes, perhaps because of a decreased population size and/or a derangement in the normal homeostatic mechanism responsible for the control of antibody synthesis.
Tumor Immunity in Hamsters Immunized with Fetal Tissues
Authors: Coggin J. H. , Ambrose K. R. , Bellomy B. B. , Anderson N. G.
Published: December 2022

Hamster and mouse fetal cells were shown to contain antigen cross-reactive with simian virus 40 (SV40)-induced tumor specific transplantation antigen in stimulating a surface reactive, cytostatic (C) antibody against SV40 tumor target cells in diffusion chambers. The antibody was synthesized when 10-day, but not 14-day, irradiated fetal cells were injected into adult, syngeneic hamsters, and these animals were subsequently found to exhibit immunity to SV40 tumor cell challenge. The spectrum of fetal antigens present in hamster fetus was extended to adenovirus 31-stimulated tumors, and the experimental data afford an explanation of the variability previously noted in the status of immunity to tumor challenge in hamsters immunized with syngeneic fetal tissues. The failure of unirradiated fetal cells to induce transplantation immunity was correlated in this work with the development of embryomas or teratomas at the site of immunization. Males responded to fetal immunization better than did female animals although multiparous, pregnant hamsters developed C antibody during pregnancy, and lost it post-partum. The role of fetal antigenic expression in cancer and cancer immunology is considered in relation to these findings.
Renal Tubular Disease and Autoantibodies against Tubular Basement Membrane Induced in Guinea Pigs
Authors: Steblay Raymond W. , Rudofsky Ulrich
Published: December 2022

Guinea pigs injected with heterologous renal basement membranes (RBM) and complete Freund's adjuvant (FA) develop a new renal cortical tubular disease and form antibodies that react with renal tubular basement membrane (TBM) (1–3). We now document the characteristics and immunopathology of this renal tubular disease and the presence of anti-TBM antibodies in the serum and along the cortical TBM. Finally, we demonstrate a positive correlation between the incidence of severe tubular disease and large amounts of anti-cortical TBM autoantibodies.

Materials and Methods. RBM, rich in TBM, were prepared from frozen New Zealand White rabbit kidneys3 by a modification of Krakower and Greenspon's method (4). Briefly, rabbit cortex was buttered through a 100-mesh metal screen. The screenings were washed by centrifugation in saline to remove free cells. By repeated shaking and washing of the residue, glomeruli settled faster than tubules. Supernatants rich in tubules were examined microscopically (4, 5).
Localization of Immunoglobulins in Human Prostatic Tissue
Authors: Ablin R. J. , Gonder M. J. , Soanes W. A.
Published: December 2022

Prompted by recent reports of the successful therapeutic use of cryosurgery for destroying benign and malignant human prostatic tissue (1) and of the reported occurrence of an “immunologic response” accompanied by remission in cases of metastatic adenocarcinoma of the prostate following cryoprostatectomy (2), we have endeavored to study the immunologic and immunochemical properties of the prostate in man and in experimental animals (3).

In view of the identification of rabbit IgG in coagulo-prostatic fluid, a secretory product of the coagulating and prostate glands of the male rabbit (4), and of the recent observation of the in vivo binding of rabbit IgG to the stroma in a serpentine manner within the folds of each acinus and to accumulations of secretory granules lying within the lumen of the rabbit prostate (5), we though it would be of interest to examine human prostatic tissue for the presence of in vivo bound immunoglobulins.
The Role of Cyclic AMP in the Cytolytic Activity of Lymphocytes
Authors: Henney Christopher S. , Lichtenstein Lawrence M.
Published: December 2022

The cytolytic activity of lymphocyte populations toward cells bearing antigenic determinants against which the lymphocytes are specifically sensitized has received increasing attention recently, principally on account of the relevancy of this phenomenon to allograft immunity. Although it is established that such cytolysis is immunologically specific (1) and requires intimate contact between the “sensitized” lymphocytes and the antigen-bearing (or “target”) cell (2), presumably via the intermediary of a specific antigenic recognition unit on the lymphoid cell surface, little is known of the mechanisms by which the cytolytic event is effected. One suggestion (3) has been that the interaction between the lymphoid and target cells leads to the release of a soluble factor, termed lymphotoxin, which is the mediator of cell lysis.
Exposure of Hidden Antigenic Determinants in Human IgG by Digestion with Spleen Proteases
Authors: Fehr Kurt , LoSpalluto Joseph
Published: December 2022

Fragments of human IgG produced by digestion with spleen proteases were tested for capacity to react with “agglutionators” present in human sera. Antigenic sites, apparently buried in the intact IgG, were exposed by these enzymes. Hemagglutination-inhibition tests indicated that the fragments produced were similar to but not identical with those produced by digestion with pepsin or papain.
Precipitating Antibody to Group a Streptococcal Polysaccharide in Humans
Authors: Zimmerman Robert A. , Auernheimer Arthur H. , Taranta Angelo
Published: December 2022

A simple method for detecting and quantitating antibodies against streptococal group A carbohydrate (A-CHO) is described. Its limits of sensitivity were determined as 0.84 µg Ab N/ml and 0.6 µg/ml A-CHO antigen (Ag). The precipitating activity was specifically inhibited by N-acetyl-glucosamine and was associated with the serum fraction precipitated by half-saturated ammonium sulfate and by the 7S pool obtained by gel filtration on Sephadex G-200. The method was highly reproducible using blind duplicates on the same and subsequent days. Subjects with detectable A-CHO antibody varied in population groups from 30% to 84%. The incidence of positive sera in patients with a history of rheumatic fever with or without rheumatic heart disease (RHD) was intermediate between that observed in populations studied under endemic and epidemic conditions. The antibody content was lower in patients with RHD than in those with history of rheumatic fever but no RHD. The age-specific distribution of A-CHO antibody differed from that of an antiprotein streptococcal antibody, antistreptolysin O (ASO), reaching a maximum titer during the teenage years rather than in the elementary school age group when challenge is greatest. There was evidence that the rise in A-CHO antibody following group A infections is lower than the concurrent rise in ASO.
The IgE-Mediated Release of an Eosinophil Leukocyte Chemotactic Factor from Human Lung
Authors: Kay A. B. , Austen K. Frank
Published: December 2022

Following antigenic challenge of human lung passively sensitized with serum from a ragweed-sensitive donor, the anaphylactic diffusate contained, in addition to histamine and slow reacting substance of anaphylaxis (SRS-A), a factor selectively chemotactic for human eosinophils (ECF-A). ECF-A evoked the migration of eosinophils obtained from the blood of a number of donors with eosinophilia. ECF-A was distinguished from histamine, bradykinin, serotonin, the prostaglandins PGE1, PGE2 and PGF2α and from SRS-A. The immunoglobulin mediating the release of ECF-A was IgE. Heating or absorption of ragweed-sensitive serum with a rabbit antibody specific for IgE removed its ability to passively sensitize lung for mediator release, and anti-IgE released ECF-A from normal lung. ECF-A, a unique mediator of the anaphylactic reaction, has been identified in human lung tissue.
Studies on Binding Sites of Myeloma Proteins
Authors: Grossberg Allan L.
Published: December 2022

Mouse IgA myeloma protein S-183 detected as binding the 3-nitro-4-hydroxyl-5-iodophenylacetyl (NIP) groups by precipitation of NIP-protein conjugate, was specifically purified on a NIP-immunoadsorbent (NIP-poly-RSA). The purified protein binds 2 moles NIP-ε-aminocaproate/150,000 g protein with a binding constant (K0) of ∼5.5 × 103 M-1. Fab fragments of specifically purified S-183 have one specific NIP-binding site/50,000 g protein, K0 ≈ 3.1 × 103 M-1. The binding sites on S-183 Fab contain arginyl residues, as do those of conventionally induced rabbit anti-NIP antibodies, as shown by loss of binding sites when 63% of S-183 Fab arginine is glyoxalated. Some protection of the sites from glyoxalation is afforded by 0.1 M NIP. Evidence for the degree of specificity of the S-183 sites for NIP obtains from comparison with NIP binding by MOPC-315 mouse IgA myeloma protein (K0 ∼2 × 103 M-1) which does not precipitate with NIP-protein and by several human IgG myeloma proteins (K0 ∼0.3–0.7 × 103 M-1).
Studies on a Biclonal Gammopathy: Evidence for a Unique Genetic Mechanism Controlling Immunoglobulin Synthesis
Authors: Fudenberg H. H. , Wang A.-C. , Pink J. R. L. , Levin A. S.
Published: December 2022

The partial amino acid sequences of the variable regions (V-regions) of 40 human immunoglobulin heavy chains have been examined by automatic sequencing methods. At least three distinct classes of the heavy chain sequences exist; members of any VH class may be found in association with different heavy chain constant regions (e.g., with constant regions of µ, α and γ chains. In addition, the proteins of a patient (Til) who was producing two serum paraproteins was studied: one protein was an IgG2-K and the other an IgM-K immunoglobulin. The light chains of the two proteins through the hypervariable region in positions 30 to 35 were identical. A hypervariable region (HYP-R) was noted in the sequence (30 to 35) of the heavy chains of eight VHIII proteins homologous to the HYP-R in κ chains. The Til IgG and IgM heavy chains were identical in amino acid sequence as far as we have gone to date including this hypervariable region.
Microheterogeneity of Light Chains Derived from a Biclonal Gammopathy
Authors: Grey H. M. , Penn G. , Kunkel H. G.
Published: December 2022

A patient, Mi., with a biclonal gammopathy of the λ IgM-λ IgG type was studied. Antisera made against the λ light chains showed idiotypic specificity that was common to both light chains so that by this criterion the light chains derived from the IgG and IgM proteins were identical. However, acid urea starch gel electrophoresis indicated a slight but definite difference in mobility, the light chain derived from the IgG having a slightly more cathodic migration. Amino acid composition analysis of the light chains revealed differences between the two light chains in that the IgM λ chain had one mol less Arg and 0.7 mol less Glu and 1 mol more alanine and tyrosine. All these differences were significant at the 0.05 percentile level. Peptide mapping revealed a single peptide difference in these two preparations. The IgG λ chain had a single spot, which was missing from the IgM and on amino acid analysis was shown to be free Arg.
Common Individually Specific Determinants in Sera with Multiple Monoclonal Proteins
Authors: Penn G. M. , Kunkel H. G.
Published: December 2022

Six sera with multiple homogeneous immunoglobulins—two γA-γG, three γM-γG and one γM-γA—were found to have shared individually specific determinants. Two of these sera showed certain points of interest. Patient Mi. had a γMλ and a γG3λ monoclonal protein. The two proteins had common individually specific determinants localized to the heavy chains as determined by hemagglutination inhibition and precipitin analysis using antisera prepared against the γG F(ab′)2 fragment, a mixture of the γM and γG, and isolated γM heavy chains. Patient Wei had γMλ, a γA2λ and a γGK monoclonal protein. The γA2λ and γMλ were shown to have common individually specific determinants, using antiserum prepared against the γA protein, and identical light chains by acid-urea starch gel electrophoresis. These two patients initially had elevated γM concentrations followed by decreasing γM and increasing γG or γA levels.
Production of 19S Antibodies with Restricted Heterogeneity from Sharks
Authors: Clem L. William
Published: December 2022

Four of twelve small nurse sharks responded with 7 to 10 mg/ml serum of antibody to the group specific carbohydrate between 1 to 3 months after initial immunization with streptococcal A-variant cells. Three animals responded less well (3 to 5 mg Ab/ml) and five failed to respond (<0.25 mg/ml). Specific antibodies were batch-purified from A-variant carbohydrate-Sepharose immunoadsorbent columns. These antibodies (75% to 95% precipitable with antigen) were shown to represent nurse shark 19S Ig by ultracentrifugal and antigenic analysis; the one exception was the presence of a small amount of 7S material after 181 days. Cellulose acetate electrophoresis revealed that some of these antibodies migrated in a considerably more restricted manner than did pooled immunoglobulin. L chains isolated from extensively reduced and alkylated material exhibited considerably restricted heterogeneity by disc electrophoresis in urea; one preparation showed predominantly a single band.
Cloning of Antibody-Producing Cells in Vivo
Authors: Williamson A. R. , Askonas B. A. , Wright B. E. G.
Published: December 2022

A method is described for analyzing serum by isoelectric focusing in thin layer polyacrylamide gels and detecting specific antibody bands by 131I-hapten binding and autoradiography. The isoelectric spectrum of a biosynthetically homogeneous immunoglobulin produced by a single clone of plasma cells is a characteristic pattern of a few bands; such patterns provide phenotypic markers for antibody-forming cell clones. Anti-DNP-forming cell clones were distributed from primed mice by spleen cell transfer at limiting dilution into irradiated syngeneic mice. Preliminary results did not indicate a limit to the number of different anti-DNP antibody molecules. Selected single clones could be propagated by serial spleen cell transfer. The history and properties of one clone (E9) producing IgG anti-DNP are described. Transfer was mediated by memory-precursor cells; antibody production was strictly dependent on antigen (DNP-BGG) at each generation.
The Characterization of Monofocal Antibodies
Authors: Klinman N.
Published: December 2022

Numerous monofocal anti-DNP antibodies produced by fragment cultures of spleens from irradiated mice reconstituted by the transfer of 106 cells from immunized syngeneic mice have been purified. These presumed monoclonal antibodies exhibit homogeneous hapten binding properties and show restricted patterns on cellulose acetate electrophoresis (Immunochemistry, 6:757, 1969; J. Immun., 106: 1345, 1971). In addition, equilibrium dialysis studies indicate complete regain of binding properties after separation and recombination of polypeptide chains derived from such antibodies. Isoelectrofocusing has been adapted to permit the further characterization and purification of such antibodies. Preparations containing monofocal antibodies are electrophoresed in 5 M urea in a pH range of 5–8. Two-millimeter slices of the gels are eluted in water. Antibody is detected in the eluates by a radioimmunoassay. All monofocal antibodies analyzed by this method showed activity restricted to a narrow region.
Cyclic Amp and Immune Responses
Authors: Ishizuka Masaaki , Braun Werner , Matsumoto Takao
Published: December 2022

In tests with sheep red blood cells as antigen, both poly A and poly A·poly U (= polyA:U) stimulated the appearance of antibody-forming mouse spleen cells under in vitro conditions, poly A:U being more potent than poly A. High concentrations of the single- or double-stranded polynucleotides were less effective than low concentrations, and very high concentrations of poly A:U inhibited the responses. Pretreatment of cells with poly A:U prior to cultivation produced as good or better responses than did the incorporation of polynucleotides into the medium, but such pretreatment was ineffective when poly A:U-treated cells were subsequently exposed to trypsin. Lymphocytes, rather than macrophages, seem to be the principal targets of stimulation. The modification of antibody formation in vitro can be attributed to an alteration of cyclic AMP-mediated events, particularly since cAMP itself as well as the supplementation of poly A:U with theophylline (a known stabilizer of cAMP levels) can increase the number of antibody-forming cells that can be activated in vitro in the presence of a given amount of antigen.
Cyclic AMP and Immune Responses
Authors: Braun Werner , Ishizuka Masaaki
Published: December 2022

The stimulatory effects of poly A:U and poly I:C on antibody formation can be potentiated by the concurrent administration of theophylline or caffeine. This potentiation is dose-dependent and is absent, or may even lead to inhibited responses, when excessively high levels of theophylline or caffeine are employed. Potentiation is most effective by the subcutaneous route and stimulation expresses itself both in terms of rate of activation of antibody-forming cells as well as in circulating antibody titers. Theophylline, in the absence of poly A:U, stimulates antibody formation only when given subsequent to the time of immunization. These effects of theophylline and caffeine, both known stabilizers of cAMP levels, further support the conclusion that cAMP-mediated events are a critical component of the activation and regulation of immunocompetent cells.
Assay of Australia Antigen and Antibody Employing Double-Antibody and Solid-Phase Radioimmunoassay Techniques and Comparison with the Passive Hemagglutination Methods
Authors: Hollinger F. Blaine , Vorndam Vance , Dreesman Gordon R.
Published: December 2022

Australia antigen (Au) and antibody were assayed by double-antibody (RIA-DA) and solid-phase radioimmunoassay methods and the results were compared with those of other serodiagnostic tests. Purified Au (72 µg protein/ml and approximately 1013 particles/ml) was prepared and labeled with 125I. The use of a 2-day RIA-DA procedure (anti-Au + unknown, 6 hr; labeled Au, 18 hr; immunoprecipitating antibody, 24 hr) resulted in Au titers that were usually 5- to 35-fold greater than those obtained by the complement fixation (CF) method, and 5- to 10-fold greater than those resulting from the passive hemagglutination (PHA) inhibition procedure. Shortening the RIA-DA procedure to 1 day or using antibody-coated tubes in a solidphase radioimmunoassay system resulted in Au titers that were two- to threefold less than those obtained by the longer RIA-DA method, although both immunoassays were more sensitive than the CF test. RIA-DA and PHA anti-Au titers were comparable.
Alterations in the Antibody Response to Bovine Serum Albumin by Capsular Polysaccharide of Klebsiella Pneumoniae
Authors: Nakashima Izumi , Kobayashi Takashi , Kato Nobuo
Published: December 2022

Immunologic paralysis was induced in mice by administration of large amounts of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K). When a paralyzing dose of CPS-K was injected into mice 3 to 30 days before immunization with bovine serum albumin (BSA)-Freund's adjuvant complex, the immune response of mice to BSA was markedly depressed. Conversely, when a paralyzing dose of CPS-K was injected simultaneously with or 3 days after immunization with BSA-Freund's adjuvant complex, the immune response to BSA was enhanced. Paralyzing doses of CPS-K had a marked adjuvant effect on antibody formation to BSA, which is a weak immunogen when administered without any adjuvant. The adjuvant effect of CPS-K was most marked when it was administered simultaneously with BSA. In mice paralyzed with CPS-K serum γ globulin, splenic weight and phagocytic activity of the reticuloendothelial system increased markedly.
Immunologic Investigations of Naturally Occurring Canine Thyroiditis
Authors: Mizejewski Gerald J. , Baron Jennifer , Poissant Gerald
Published: December 2022

Naturally occurring canine thyroiditis was characterized in the present study by immunodiffusion, passive hemagglutination, complement fixation, immunofluorescence and skin tests. Canine thyroglobulin and thyroid gland extract were used as sources of antigen in all tests. Thyroid histology was examined in all instances. The study mainly concerned 11 beagles, which were classified as thyroiditis colony dogs. Precipitating antibodies were not detectable, while low titers of PHA and varying titers of CF antibodies were present in the sera of the thyroiditis colony dogs. Among the serologic procedures employed, only the fluorescent antibody (FA) tests were consistently positive. The skin test was generally positive in the less progressive stages of the disease. The dog thyroid extract rather than purified thyroglobulin was required for PHA antibody detection. This latter observation, together with the FA staining patterns obtained, implied involvement of the second colloid antigen. Overall evidence from present and past studies suggested that the development of the canine disorder is similar in many respects to human thyroiditis.
Effects of Intraperitoneal Thymus Graft on Hemolytic Plaque Formation
Authors: Takada Akikazu , Takada Yumiko , Ambrus Julian L.
Published: December 2022

CBA/H mice with adult CBA/HT6T6 thymuses implanted intraperitoneally after irradiation and injection with marrow cells had more plaques in their spleens than did non-grafted controls. Chromosome analyses 2 or 7 days following injection of sheep red blood cells indicated that all dividing cells were of host (marrow) origin in the spleens and lymph nodes.
Antiserum to a Soluble Lymphoid Membrane Preparation Stimulates Human Leukocyte DNA Synthesis
Authors: Einstein Albert B. , Mann Dean L. , Ficker Susan , Terry William D.
Published: December 2022

Rabbit antiserum against soluble human lymphoid cell membrane glycoproteins containing histocompatibility (HL-A) antigens stimulated DNA synthesis in human peripheral blood leukocytes. Absorption of the antiserum with human lymphoid cells totally removed the stimulating activity. Stimulation was independent of the HL-A type of the leukocyte donor. The glycoproteins isolated from papain solubilized lymphoid membrane preparations, therefore, retain sufficient structure to serve as immunogens in the production of heterologous antisera against membrane components. Interaction of these components with antibody leads to increased DNA synthesis.
Reconstitution from Subunits of the Hapten Binding Sites and Idiotypic Determinants of Mouse Anti-Phosphorylcholine Myeloma Proteins
Authors: Sher Alan , Lord Edith , Cohn Melvin
Published: December 2022

Reconstitution experiments were employed to investigate the structure of the hapten binding sites and idiotypic determinants of mouse myeloma proteins with antibody specificity for phosphorylcholine. Quantitative reconstitution of both the phosphorylcholine binding affinity and idiotypic reactivity of these immunoglobulins was shown to require specific heavy and light chain combinations. Interaction of myeloma protein with hapten (phosphorylcholine or choline) had no effect on the expression of the idiotype. The subunits of the immunoglobulins possessing in common an idiotype and affinity for phosphorylcholine reconstituted indistinguishable antibodies. In addition, it was shown that the light chains of the proteins belong to the same κ subgroup. Since these indistinguishable immunoglobulins are found repeatedly, they may represent the expression of the same heavy and light chain variable region genes out of a small number present in the BALB/c germ-line.
Dual Pathways of Complement Interaction with Guinea Pig Immunoglobulins
Authors: Sandberg Ann L. , Osler Abraham G.
Published: December 2022

Further evidence is provided to document the availability of dual pathways for the interaction of C with guinea pig immunoglobulins. Preformed aggregates containing the γ1 molecules, bypass C1, C4 and C2 in their attack on C3. This reaction is rapid, and is inhibited by salicylaldoxime or dilution of the serum used as a source of C. Magnesium provides the essential divalent cation for this fixation process, which is more efficient at 37°C than at 4°C. Preformed aggregates containing γ2 antibodies fix C more slowly, a process which is not diminished as the serum C is diluted and is not inhibited by salicylaldoxime. Preformed complexes of these immunoglobulins also require magnesium for the C3 shunt pathway.
Fate and Immunogenicity of Antigens Endocytosed by Macrophages: A Study Using Foreign Red Cells and Immunoglobulin G
Authors: Cruchaud Andre , Unanue Emil R.
Published: December 2022

Peritoneal macrophages which had ingested 51Cr SRBC were transferred into syngeneic mice. The recipient mice made anti-SRBC antibodies indicating that immunogen of SRBC had become available for immunologic recognition. The immune response was directed both to antigen(s) bound to macrophages and to antigen released into the extracellular fluid. The antigen bound to macrophages induced a much stronger immune response. Treatment of the macrophages with antibody or trypsin prior to transfer abrogated to a great extent the immune response to macrophage-bound SRBC. Tissue culture studies with SRBC labeled with 125I indicated that subsequent to their ingestion by macrophages there was extensive release of non-protein bound I, a likely indication of catabolism in intracellular vesicles. However, small amounts of protein-bound I were found on the plasma membrane of macrophages (i.e., removed by trypsin) and in culture supernatants. Further experiments with macrophages that had ingested SRBC opsonized with 125I rabbit anti-SRBC IgG showed the release of a few IgG molecules, some of which were smaller in size than monomer IgG. It was apparent that some molecules of antigen ingested by macrophages could escape extensive catabolism and could become available for immunologic recognition either at the surface of the macrophage or at a distant site.
Induction of Thyroiditis in the Rabbit by Intravenous Injection of Papain-Treated Rabbit Thyroglobulin
Authors: Anderson Charles L. , Rose Noel R.
Published: December 2022

Rabbit thyroglobulin was partially digested with papain. Low molecular size fragments were separated by gel filtration and injected intravenously into rabbits. Circulating thyroid-specific antibodies appeared, as demonstrated by tanned cell hemagglutination, precipitation and passive cutaneous anaphylaxis. Lymphocytic and monocytic infiltrates were demonstrated in the rabbits' thyroid glands. Antisera to papain-treated thyroglobulin reacted more strongly with the papain-treated antigen, in contrast to rabbits immunized with untreated thyroid antigen. On analysis by precipitation in gel, the papain-treated preparation showed at least two additional precipitating components that gave reactions of partial identity in intact thyroglobulin. Papain-generated rabbit thyroglobulin fragments serve as effective immunogens in the same species following intravenous injection without adjuvant.
Isolation of 7S IgM and Kappa Chains from the Surface Membrane of Tissue Culture Cells Derived from a Burkitt Lymphoma
Authors: Eskeland Trond , Klein Eva
Published: December 2022

By homogenization or by freezing and thawing, mu and kappa structures were liberated from the surface of Daudi cells, a Burkitt lymphoma-derived culture line. After removal of the membranes and particles by ultracentrifugation, approximately one-third of the immunoglobulin structures remained in the supernatant. By density gradient ultracentrifugation and gel filtration on Sephadex G-200, it was established that the supernatant contained 7S IgM molecules and free kappa chains. Also, kappa structures were in excess on the sedimented membranes as compared with mu structures. The immunoglobulins were detected and quantitated by inhibition of passive hemagglutination.
Inheritance and Strain Distribution of a Rat Immunoglobulin Allotype
Authors: Gutman George A. , Weissman Irving L.
Published: December 2022

The inheritance and strain distribution of RI-1, a rat immunoglobulin allotype, was studied by passive hemagglutination. Two alleles, a and b, were shown to segregate as Mendelian co-dominants, and were unlinked to the major rat histocompatibility locus (Ag-B) and three coat color genes (A, C and H).
Aleutian Disease of Mink: Detection of Large Quantities of Complement-Fixing Antibody to Viral Antigen
Authors: McGuire T. C. , Crawford T. B. , Henson J. B. , Gorham J. R.
Published: December 2022

A number of proposals have been suggested to explain the mechanism of plasmacytosis and hypergammaglobulinemia in mink infected with Aleutian disease (AD) virus. That the virus induces a myeloma-like proliferation of plasma cells and the production of immunoglobulins not directed toward specific antigens was suggested by several workers (1–5). The transition from polyclonal to monoclonal gammopathies in 10% to 20% of the infected mink with genotypes Aa and AA (non-Aleutian mink) supports this suggestion (2). However, detection of anti-virus antibody in the form of infectious virus-antibody complexes (6) and by direct and indirect immunofluorescence on infected tissue (7; J. B. Henson and T. B. Crawford, manuscript in preparation) demonstrates that some of the hypergammaglobulinemia represents antibody to AD viral antigens. A previous attempt by other investigators to demonstrate complement-fixation (CF) between AD-affected sera and infectious organ homogenates was unsuccessful (1).
Neoantigenic Expressions Engendered by Plasmin Cleavage of Fibrinogen
Authors: Plow Edward F. , Hougie Cecil , Edgington Thomas S.
Published: December 2022

Cleavage of fibrinogen to its pathophysiologic fragments by plasmin is associated with the appearance of unique neoantigens recognizable by Ouchterlony and radioimmunochemical techniques. Recognition of these cleavage-specific neoantigens by antisera to fibrinogen D-fragment introduces additional molecular markers for the study of fibrinogen conformation and also may permit direct assay of plasma for fibrinogen cleavage.
The Immunologic Release of Constituents from Neutrophil Leukocytes
Authors: Henson Peter M.
Published: December 2022

The interaction of rabbit neutrophils with immune complexes induced release of constitutents to the outside of the cell. Two situations were studied. In one, the neutrophils adhered to immune complexes dispersed along nonphagocytosable surfaces; in the other, the cells were allowed to engulf phagocytosable particles coated with antibody or complement. In each case, release of constituents from the neutrophil granules occurred. IgG antibody was as effective in stimulating release of enzymes as was C3, although they react with different receptors on the neutrophil membrane. Smaller quantities of immunologobulins were required to induce release if bound to the nonphagocytosable surface than if phagocytosed by the neutrophils in suspension.
Quantitation of the Cell-Mediated Immune Response
Authors: Henney Christopher S.
Published: December 2022

Quantitative aspects of the cell-mediated immune response of C57BL mice to a mouse DBA/2 mastocytoma cell have been studied by an in vitro assay involving the cytolytic activity of splenic lymphocytes towards 51Cr labeled target cells. Cytolysis of the DBA/2 mastocytoma cells by “sensitized” C57BL lymphocytes was shown to be a “one-hit” phenomenon, i.e., that lysis of a target cell resulted from single interaction with specifically sensitized “effector” cells. Kinetic studies showed that the rate of target cell destruction fell markedly with time following mixing with lymphocytes, and thus suggested that there is reciprocal death of the interacting cells at least from a functional standpoint, if not morphologically. These findings suggested a means of calculating the number of effector lymphocytes in the splenic pool of sensitized animals, by determining the maximal number of target cells that a given number of lymphocytes could lyse under specified conditions in gross target cell excess. Such determinations were used to compare the effect of mode and dose of antigenic stimulation on the cell-mediated immune response. Optimal antigenic stimulation of C57BL mice with alloantigenic mastocytoma cells was found to be by the intraperitoneal route. Maximal responses were elicited with 107 cells and occurred 11 days following antigenic stimulation. Under these conditions it was estimated that approximately 4% (4 × 104 out of 106) of the lymphocytes were effector cells. Incorporation of the mastocytoma cells into adjuvants (either complete or incomplete Freund's) resulted in a marked diminution of both humoral and cell mediated immune responses. Using complete Freund's adjuvant the maximal number of effector cells produced following antigenic stimulation with 3 × 107 mastocytoma cells was approximately 5 × 103/106 lymphocytes. The number of effector lymphocytes resulting from stimulation with 3 × 107 mastocytoma cells rose exponentially from day 4 (when the cytolytic activity was first demonstrable) and when there were calculated to be 50 effectors/106 lymphocytes to day 11 (4 × 104/106 lymphocytes) and fell therafter. The generation time of the cell population involved in cytolysis was estimated to be approximately 17 hr.
The Specificity and Cross-Reactivity of Antisynaptosome Antibodies as Determined by Sequential Adsorption Analysis
Authors: Mickey Don D. , McMillan Paul N. , Appel Stanley H. , Day Eugene D.
Published: December 2022

An improved iso-osmotic, Ficoll-sucrose, density-gradient technique for isolating synaptosomes, synaptic membranes, brain mitochondria and three forms of myelin from adult rat brain is described. Antibodies raised in rabbits to isolated synaptosomes, synaptic membranes, brain mitochondria and liver mitochondria, when assayed by means of sequential adsorption analysis, showed that a major class of determinants is shared by brain mitochondria and synaptic membranes, that another major class is shared by myelin and synaptic membranes and that a third class is shared by liver mitochondria and the various brain fractions. Besides these crossreactivities synaptosomes also displayed a class of distinctive determinants separate from those of myelin; brain mitochondria displayed a class of distinctive determinants separate from those of synaptic membranes and from liver mitochondria, and liver mitochondria displayed a class of distinctive determinants separate from those of brain mitochondria. Synaptosomes also combined with a small but definite amount of antibody in two different antisera that did not cross-react either with isolated synaptic membranes or brain mitochondria at the given concentrations, thereby revealing a distinctive class of synaptosome determinants, possibly conformational in nature.
Complement-Requiring Neutralizing Antibodies in Hyperimmune Sera to Human Cytomegaloviruses
Authors: Graham Bettie J. , Minamishima Yoichi , Dreesman Gordon R. , Haines Harold G. , Benyesh-Melnick Matilda
Published: December 2022

High-titered antisera to two strains of human (C87 and AD169) and two strains of monkey (GR2598 and GR2757) cytomegaloviruses (CMV) were prepared in primates hand-reared in captivity and free of pre-existing antibodies. Unlike monkey CMV, which elicited the production of complement-independent neutralizing (CIN) antibodies, hyperimmune sera to the two human strains were found to contain chiefly complement-requiring neutralizing (CRN) antibodies. The CRN antibodies elicited by human CMV were specific for the immunizing virus and were not directed to host-cell components. Treatment of whole serum with 2-mercaptoethanol indicated that 7S antibodies comprised the major population of CRN antibody activity in both early and late anti-human CMV sera. Preparations of highly purified early and late IgG (6.4S) and IgM (18.6S) anti-human CMV immunoglobulins all required complement for neutralization. The complement dependency of late IgG appears to be a phenomenon unique for antisera against human CMV. Cross-neutralization tests indicated a two-way cross between the two human viruses and a two-way cross between the two monkey viruses with negligible interspecies reactivity.