bluuebunny/crossref_metadata_2025 huggingface dataset:
Experimental Allergic Uveitis
Authors: Wacker W. B. , Lipton M. M.
Published: December 2022

Guinea pigs and rabbits given injections of homologous retina emulsified in Freund's adjuvant developed ocular lesions typically affecting the uveal tract. Injections of both species with heterologous retina also resulted in uveitis in some instances, depending on the species of retina used. These results were obtained by both single and repeated injections of retinal tissue. In contrast, single injections of many other homologous or heterologous intraocular tissues, including uvea itself, were ineffective in production of ocular disease.

A series of repeated injections of the guinea pig with homologous uvea resulted in mild uveitis demonstrated histologically, but not clinically. Similar injections of the guinea pig with heterologous uvea were ineffective in production of uveitis, as were injections of both homologous or heterologous uvea in the rabbit.

Immunization of the rabbit with homologous optic nerve elicited uveitis and encephalomyelitis which occurred simultaneously in the same animal. Similar immunization of the guinea pig resulted in encephalomyelitis only. These results were correlated with demonstration of encephalitogenic antigen in rabbit retina, but not in guinea pig retina. The interrelation and differentiation of these models of EAU are discussed.
Modulation of TL (Thymus-Leukemia) Antigens by Fab-Fracments of TL Antibody
Authors: Lamm Michael E. , Boyse Edward A. , Old Lloyd J. , Lisowska-Bernstein Barbara , Stockert Elisabeth
Published: December 2022

Cytotoxic antibodies of H-2 and TL specificities were located in the same 7 S γ-globulin fraction of mouse antiserum. The anti-TL and anti-H-2 activities occurred together in antiserum fractions prepared by Sephadex G 200 gel filtration or by electrophoresis in a Geon block. The modulating activity of TL antiserum (i.e., its capacity to cause the TL + → TL - phenotypic change in cells exposed to the antiserum in the absence of lytic C′) also occurred in the 7 S γ-globulin fraction. When γ-globulin prepared from antiserum by Geon block electrophoresis was digested with papain, cytotoxic activity of both H-2 and TL specificities was lost, but TL modulating activity remained. Thus attachment of univalent TL antibody fragments (Fab) lacking the complement-fixing and other biologic properties of the Fc-fragment was sufficient to induce modulation. Papain digests of H-2 antibody prepared in the same way did not induce modulation of H-2 antigen or of TL antigen, indicating further that modulation of TL antigen is a specific response to TL antibody.
Immunodiffusion Reactions with Rubella Antigens
Authors: Schmidt Nathalie J. , Styk Bohumil
Published: December 2022

Immune precipitates were demonstrable in gel double diffusion tests with the viral and soluble antigens of rubella using agarose and a “chamber” technique employing a plastic matrix over a thin layer of gel. Soluble antigens gave heavier lines of precipitate than did viral antigens, possibly due to better diffusion of the smaller antigen. The immunologic relationship between viral (large-particle) and soluble (small-particle) antigens which was suggested by previous studies using complement fixation, hemagglutination inhibition and neutralization tests was confirmed by the results of immunodiffusion studies. Reactions of immunologic identity were obtained between viral and soluble antigens and between antibodies produced to either of these antigens.
Ontogeny of the Immune Response
Authors: Kim Yoon Berm , Bradley S. Gaylen , Watson Dennis W.
Published: December 2022

Germfree, immunologically virgin piglets were used to study the ontogenic development of immunoglobulins. In the true primary immune response, the first antibody synthesized was 19 S γG-immunoglobulin which was antigenically identical to the late immune 7 S γG-immunoglobulin but distinct from 19 S γM. In addition, there was a sequential synthesis of 7 S γG-immunoglobulin before the appearance of γM or γA. It was, therefore, possible to isolate 19 S γG free from 19 S γM-macroglobulin and 7 S γG free from γA.

The purified 19 S γG-immunoglobulin of the true primary immune piglet sera had a sedimentation constant of 18.5 S (s020,w) and a molecular weight of 900,000. The purified 7 S γG-immunoglobulin of piglet and sow had a sedimentation constant of 6.6 S (s020,w) and a molecular weight of 170,000.

The specific antibody activity was demonstrated only in 19 S γG- and 7 S γG-immunoglobulins but not in 19 S γM as determined by radio-immunoelectrophoretic technique using I131-labeled actinophage and keyhole limpet hemocyanin.

Evidence presented indicates that the 19 S γG is not an artificial aggregate of 7 S γG but is synthesized as a true immunoglobulin of the 19 S class.

Because of the physiochemical similarities of 19 S γG and 19 S γM, the difficulties involved in their separation and characterization when both are present in the sera are discussed.
The Production of Thyroid Autoantibodies in Mice
Authors: Twarog Frank J. , Rose Noel R.
Published: December 2022

Circulating thyroid autoantibodies have been produced in mice by the injection of either homologous or heterologous (rat and rabbit) thyroid extract incorporated in complete Freund adjuvant. Rat thyroid extract injected without adjuvant also was capable of eliciting antibodies directed against both rat and mouse thyroid extract. Even though normal animals did not respond to aqueous mouse thyroid extract (MTE), animals previously sensitized with the same antigen in complete Freund adjuvant, or syngeneic recipients of lymphoid cells from sensitized donors, readily responded to a similar injection. These results imply the production of a new population of responsive cells following initial stimulation. Histologic examination of thyroid glands from inbred mice sensitized under a wide variety of conditions revealed no pathologic changes despite the presence over a long period of time of circulating antibody demonstrated by tanned cell hemagglutination and passive cutaneous anaphylaxis.
The Transformation of Column-Purified Lymphocytes with Nonspecific and Specific Antigenic Stimuli
Authors: Oppenheim Joost J. , Leventhal Brigid G. , Hersh Evan M.
Published: December 2022

Phytohemagglutinin (PHA)-stimulated cultures of column-separated “purified” human peripheral blood lymphocytes incorporated tritiated thymidine to the same extent as cultures of unseparated leukocytes. However, they incorporated less tritiated thymidine when stimulated with the nonspecific staphylococcal filtrate (SF), specific antigens or with low doses of PHA. Furthermore, SF, antigens, and homologous leukocytes stimulated a median of 10, 5 and 5% respectively of the small lymphocytes in unseparated leukocyte cultures to transform, whereas this decreased to 2, 0.5 and 0.5% respectively in the cultures of “purified” lymphocytes.

Prior incubation of the leukocytes with antigens followed by column separation resulted in an intermediate proportion of transforming lymphocytes. The addition of autologous phagocyte-rich, lymphocyte-poor leukocytes to the purified lymphocytes also partially restored the response to antigens. Restoration was also achieved by growing the “purified” lymphocytes at a higher cell density. However, growing the purified lymphocytes in conjunction with WI-38 human embryonic lung monolayers failed to restore their antigen-stimulated transformation.

The data suggest that the phagocytic cells facilitate the in vitro transformation of lymphocytes with antigenic as well as “nonspecific” stimuli.
Studies on Heterologous Antilymphocyte and Antithymocyte Sera
Authors: Barth Rolf F. , Southworth Joanne , Burger Gerald M.
Published: December 2022

Rabbit antisera prepared against mouse lymphocytes, thymocytes or fibroblasts showed similar patterns of serologic reactivity with cell surface antigens as indicated by leukoagglutinin, hemagglutinin, cytotoxic and hemadsorption titers.

Mice treated with 0.25 ml of antilymphocyte or antithymocyte sera successively on days -1, 0, and +1 had a striking reduction in both the total number of splenic plaque-forming cells and serum hemagglutinin and hemolysin titers 4 days following immunization with sheep erythrocytes. The activity of these antisera was independent of the presence or absence of passively transferred anti-sheep cell antibody. Rabbit anti-mouse fibroblast serum also depressed the immune response but to a lesser degree than ALS or ATS.

A single injection of antithymocyte serum given on day 0, 1, 2 or 3 did not result in a significant reduction in PFC, but did if given on day -1, -2, or -3. The time of treatment, therefore, was a critical factor in determining immunosuppressive activity. These observations suggest that the anti-mouse sera may have exerted their primary effects on early events associated with the immune response.

Mice treated with heat-inactivated normal rabbit serum had fewer splenic plaque-forming cells than those immunized with sheep erythrocytes alone, and it is thought that this depression may have been due to antigenic competition between simultaneously administered immunogens. All serum-treated mice developed demonstrable precipitating antibody to a rabbit serum protein which presumably was neither albumin nor rabbit γG which were still detectable at 11 days.
Prolongation of Skin Homografts by Rabbit Anti-Mouse Lymphocyte Serum in Mice Rendered Tolerant to Rabbit γ-Globulin
Authors: Howard Richard J. , Dougherty Suanne F. , Mergenhagen Stephan E.
Published: December 2022

Balb/C mice readily form antibodies to the γ-globulin portion of rabbit anti-mouse lymphocyte serum (ALS) as indicated by immune elimination of 131I-rabbit γ-globulin. Mice were rendered tolerant to rabbit γ-globulin (RGG) by injection of “biologically filtered” RGG prior to grafting with tail skin from C3H mice. Animals were shown to be tolerant by their failure to demonstrate immune elimination of 131I-RGG and by the lack of precipitating antibodies to RGG in their sera. Tolerant and nontolerant mice were given either a single injection or a series of injections of ALS after grafting. In all cases, whether single or multiple injections of ALS were given, animals tolerant to RGG retained their grafts for a longer period than nontolerant animals. These results were attributed to a decreased rate of elimination of the active γ-globulin component of ALS in mice rendered tolerant to RGG. However, both tolerant and nontolerant mice showed the same suppression of white blood cell count and lymphocyte count after an injection of ALS and both groups showed the same rate of return toward normal.

Of interest was the finding that heat-treated (63°C) RGG, as well as “biologically filtered” RGG, induced immunologic tolerance to the γ-globulin fraction of ALS and prolonged skin grafts after administration of ALS. On the other hand, active immunization with RGG in complete Freund's adjuvant did not shorten graft survival in ALS-treated mice.
Antibody to Brandykinin: Effect of Carrier and Method of Coupling on Specificity and Affinity
Authors: Talamo Richard C. , Haber Edgar , Austen K. Frank
Published: December 2022

In order to establish a reliable method for production of antibody to bradykinin for use in radioimmunoassay, eight compounds were prepared in which bradykinin as a hapten was coupled to either a polypeptide or a protein carrier. Specific antibody was elicited by four of these compounds, as detected by binding of tritiated acetyl-N-bradykinin, and the antibody response was followed for 6 months.

The various antisera obtained were compared in competition studies between unlabled bradykinin and tritiated acetyl-N-bradykinin or [14C]-2,3-proline-bradykinin. Binding affinities and antibody titers of the sera were determined from binding curves with the [14C]-bradykinin.

Coupling of bradykinin to ovalbumin via toluene diisocyanate results in an immunogen which reliably elicits specific antibradykinin antibody of specificity and affinity appropriate for use in radioimmunoassay.
Immunofluorescent Studies of Immunoglobulins in Human Lymphoid Cells in Continuous Culture
Authors: Finegold Ira , Fahey John L. , Dutcher Thomas F.
Published: December 2022

Immunoglobulin formation was assessed in 19 human lymphoid lines in continuous culture using fluorescein- and rhodamine-labeled antiserums specific for IgG, IgA, IgM heavy chains and κ and λ light chains. Fifteen of these lines demonstrated cytoplasmic localization of one or more heavy and light chain immunoglobulin molecules. In the cell lines which gave positive fluorescent tests only part of the cell population contained detectable immunoglobulins.

Both γ- and µ-classes of immunoglobulin heavy chain were identified in the cytoplasm of individual cells of the cell line IM-1, in which both IgG and IgM molecules are synthesized. This finding indicates that in these cells the genetic information for synthesis of two classes of immunoglobulins (IgG and IgM) was expressed. This finding contrasts with the evidence that most normal and malignant immunoglobulin-synthesizing cells in vivo express genetic information for only a single immunoglobulin class. Whether this feature of the cells in culture corresponds to the normal in vivo function of the culture precursor cell(s) or whether it reflects special conditions of culture will have to be answered by further studies.
An Immuno-Cytopathogenic Interaction between Sensitized Leukocytes and Epithelial Cells Carrying a Persistent Noncytocidal Myxovirus Infection
Authors: Speel Lawrence F. , Osborn June E. , Walker Duard L.
Published: December 2022

Experiments in vitro have demonstrated that splenocytes from mice immunized 4 to 5 days previously with mumps virus destroy and/or inhibit the growth of epithelial cells persistently carrying a non-cytocidal mumps virus infection. Because this cytotoxic reaction is immunologically specific and appears to be induced by contact between immune leukocytes and virus antigen associated with target cell plasma membranes, it can be evaluated as a manifestation of the efferent arc of a homograft response to viral infection, and is therefore considered to be a component of cellular immunity.
Studies of Human C-Reactive Protein in an in Vitro Phagocytic System
Authors: Williams Ralph C. , Quie Paul G.
Published: December 2022

The interaction of human C-reactive protein and phagocytic mechanisms was studied using an in vitro phagocytosis assay with human polymorphonuclear leukocytes, isolated γG opsonins, or various opsonic serum fractions and living bacteria. No consistent phagocytosis potentiating or blocking effect was seen with isolated human C-reactive protein either added to the phagocytic mixtures or after preincubation with either bacteria or fresh serum fractions.
Studies on Human Peripheral Blood Lymphocytes in Vitro
Authors: Chessin L. N. , Glade P. R. , Asofsky R. , Baker P. D. , Reisfeld R. , Terry W.
Published: December 2022

Circulating human lymphocytes synthesize immunoglobulins in vitro. Immunoglobulin molecules with IgG (γ), IgA (α), IgM (µ), IgD (δ), type K (κ) and type L (λ) polypeptide chain specificity was demonstrated by isotopic labeling with 14C-amino acids and identified by radio-immunoelectrophoresis with class- and type-specific antisera.

Cell fractionation studies on glass wool columns revealed that there were two functionally distinct populations of immunoglobulin synthesizing cells in the peripheral blood: 1) non-phagocytic, small lymphocytes, and 2) a phagocytic cell fraction containing granulocytes, monocytes and up to 10% small lymphocytes. Donors displayed marked individuality with regard to the cell concentration necessary for detection of newly synthesized immunoglobulin molecules, the intensity, position, distribution of radiolabel in specific precipitin arcs, and the number of classes and types of molecules synthesized.

Circulating lymphocytes taken from the same individual on separate occasions displayed distinctive biosynthetic profiles with regard to the specific classes and types of immunoglobulin molecules synthesized. Time-course biosynthesis studies revealed that cells maintained in tissue culture retain the capacity to synthesize immunoglobulins in vitro and display distinctive biosynthetic patterns after various periods of time in culture medium.
Is Delayed Sensitivity a Preparation for Antibody Synthesis?
Authors: Leskowitz Sidney
Published: December 2022

Guinea pigs exhibiting delayed sensitivity to the azobenzenearsonate (ABA) hapten or the bovine serum albumin (BSA) carrier showed little evidence of an enhanced antibody response to the hapten when given a booster injection of ABA-BSA conjugate. Rabbits treated similarly did show some enhancement of antibody formation to the hapten. However, when delayed sensitivity to the hapten was present prior to the booster injection of ABA-BSA the ABA-specific antibody formed was mercaptoethanol-sensitive. Moreover, enhanced antibody formation to the carrier BSA was also found. These results do not lend weight to the suggestion that delayed sensitivity per se is preparation for an accelerated antibody response, but rather suggest that nonspecific enhancement of antibody production may be produced to all antigenic determinants present at the site of a delayed reaction.
Immunosuppression by Leukemia Viruses
Authors: Ceglowski Walter S. , Friedman Herman
Published: December 2022

Infection of adult BALB/c mice with Friend disease virus resulted in marked suppression of the ability of the animals to respond to a potential immunizing inoculum of sheep erythrocytes as detected both on the cellular and humoral antibody level. Whereas normal noninfected animals had a vigorous immune response to erythrocytes, with the appearance of relatively large numbers of antibody plaque-forming cells in their spleens, infected mice generally had a markedly depressed response. The timing of virus infection in relation to immunization was an important factor since the greatest immunosuppression occurred in mice infected 3, 8 or 21 days before immunization. Less suppression occurred in mice infected the same day as immunization. The peak response was depressed in all mice, regardless of the day of antigen injection, when the number of PFC was calculated per million leukocytes rather than per whole spleen. Appearance of 7 S IgG PFC late in the immune response was markedly inhibited in all infected mice, even those receiving FDV after immunization.

The “background” counts of 19 S PFC preexisting in spleens of both nonstimulated control and FDV-infected mice were similar. Serum hemagglutinin and hemolysin titers were only slightly, if at all, suppressed in mice infected the same day as immunization or 2 days later. The peak serum titer was markedly suppressed in animals infected 3 or 8 days prior to immunization. The amount of 2-mercaptoethanol (2ME) resistant 7 S antibody in serum specimens of infected mice was less than in control mice.

Immunosuppression appeared to be related to leukocytosis and splenomegaly. However, immunosuppression was usually detectable prior to overt splenomegaly in mice infected simultaneously or after immunization. Although the mechanisms involved in FDV induced immunosuppression are unknown, the effects of leukemogenic viruses such as FDV on antibody-forming cells per se, on progenator or stem cells, and/or antigen-processing cells, such as macrophages, were discussed in relation to both immunogenesis and virus tumorigenesis.
Ferritin-Antibody Localization of Erythrocyte Antigenic Sites in Immunohemolytic Anemias
Authors: Davis William C. , Douglas Steven D. , Petz Lawrence D. , Fundenberg H. Hugh
Published: December 2022

Ferritin-labeled rabbit anti-human γ globulin (FE-RAHγG) and anti-C′3 (FE-RAHC′3) were assayed by means of a chromic chloride hemagglutination technique. These reagents were used in an electron microscopic study of localization of various erythrocyte antigenic sites, e.g., Rh isoantigens and the antigens presumably associated with idiopathic and drug-induced antibody hemolytic anemias. Qualitative and quantitative differences in the periodicity and intensity of label have been observed. These findings suggest that discrete sites are involved in a non-random binding of exogenous antigens to erythrocytes; these may be related to specific but yet unknown biochemical determinants.
Nonspecific Stimulation of Antibody Production in Surgically Bursectomized Chickens by Bursa-Containing Diffusion Chambers
Authors: Dent Peter B. , Perey Daniel Y. E. , Cooper Max D. , Good Robert A.
Published: December 2022

Implantation of diffusion chambers containing bursa from 5-day-old chicks resulted in an enhanced antibody response in surgically bursectomized chickens immunized with human erythrocytes 3 days after implantation. A similar effect was obtained with bowel in the chambers. Bursas from 19-day embryos caused no enhancement. Bacterial cultures confirmed that chambers containing bursa from 5-day-old chicks were contaminated whereas those containing embryonic bursa were sterile. The existing evidence for a bursal humoral factor which is important in controlling immunoglobulin and antibody production must be re-evaluated with respect to the possibility that the observed enhancement of antibody production may be a function of bacterial products released from contaminated diffusion chambers.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Colten H. R. , Gordon J. M. , Borsos T. , Rapp H. J.
Published: December 2022

We have presented evidence that the first component of guinea pig complement (C′1gp) is synthesized by columnar epithelial cells of the small intestine. We now report that isolated segments of human colon and to a lesser extent ileum are capable of synthesizing hemolytically active C′1hu. This conclusion is based on the following evidence: After elimination of C′1hu from tissue with EDTA, we found that segments of the intestinal tract in short-term organ culture showed a 100- to 1000-fold increase in C′1hu activity. The rate of production of C′1hu in human intestine was highly temperature dependent; C′1hu production was reversibly inhibited by puromycin and actinomycin D. Furthermore, 14C-labeled amino acids were incorporated into molecules which behaved like C′1. No significant C′1hu synthesis was observed in isolated segments of jejunum, stomach, liver, kidney, lung, spleen or lymph node.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Carpenter C. B. , Ruddy S. , Austen K. F. , Müller-Eberhard H. J. , Brigham Peter Bent , Brigham Robert B.
Published: December 2022

Radioiodinated C′4 (β1E globulin) was given intravenously to 11 renal allograft recipients and one anephric patient prior to transplantation. The catabolic and synthetic rates were determined by analysis of plasma, urine and stool radioactivities over the course of 6 to 9 days. The purpose of this study was to define the metabolic behavior of normal (ranging from 27 to 100% of specific functional activity of native C′4) and inactivated C′4 and to determine the method of analysis appropriate for such investigations. In patients with normal renal function, there was good agreement between catabolic rates derived from a multiexponential analysis of the plasma protein-bound radioactivity curve and the urinary excretion rate of free iodide. Impairment of renal function per se did not influence catabolic rates. In patients without evidence of rejection at the time of study, the catabolic rates (% of plasma pool/hour) ranged from 1.2 to 2.6% (mean, 1.9 ± 0.6%).
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Gigli I. , Mason J. W. , Colman R. W. , Austen K. F.
Published: December 2022

The development of a hemolytic assay for the measurement of the inhibitor of the first component of complement (C′1aINH) and the isolation of physically homogeneous and functionally pure human plasma kallikrein have permitted studies evaluating the possible interrelation of the complement and kinin-forming systems. It was demonstrated that C′1aINH inhibited the action of kallikrein on the synthetic substrate, p-toluene sulfonyl l-arginine methyl ester (TAMe). It was further observed that consumption of C′laINH exhibited a concentration and time dependence similar to its interaction with C′1 esterase. Kinetic analysis suggested the formation of an inactive stoichiometric complex of kallikrein and C′1aINH. Similar observations were made utilizing a highly purified preparation of C′1aINH supplied by Dr. J. Pensky. In contrast, C′laINH failed to inhibit plasmin, trypsin, and thrombin; and inactivation of C′1aINH by plasmin did not occur except at very high enzyme concentrations. Sera from patients lacking C′laINH failed to inhibit the TAMe esterase activity of plasma kallikrein, while sera deficient in α1-antitrypsin but containing normal amounts of C′1aINH were equally inhibitory as normal sera.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Dalmasso A. P. , de Bracco M. M. E.
Published: December 2022

Sheep red cells can be of high or low potassium concentration, which is under genetic control. It was proposed that the difference in potassium concentration depends on the level of membrane Na-K dependent ATPase (Tosteson, D. C., Fed. Proc., 22: 19, 1963). Experiments were performed to ascertain whether high and low potassium sheep red cells could be employed as a model to study the participation in immune hemolysis of cell, or cell membrane factors. High and low potassium red cells were treated with constant amounts of rabbit antibody and guinea pig complement for 1 hr at 37°. When the concentration of these reagents was adjusted to obtain 45.0% (±7.0) lysis of low potassium erythrocytes, only 20.7% (±3.6) lysis was obtained with high potassium erythrocytes. By contrast, osmotic fragility was similar in both types of cells. Several methods, including immune adherence, showed that the high and low potassium erythrocytes reacted similarly with antibody and with the complement components C′1, C′2, C′3, and C′4.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Feinman L. , Cohen S. , Becker E. L.
Published: December 2022

Carbamylation of rabbit antibody results in modifications which render it incapable of participating in C′ fixation reactions but which allow for an undiminished capacity to combine with and precipitate antigen. This effect may be used to help evaluate the role of C′ in biologic reactions. In the present study, the PCA reaction, elicited in guinea pigs or rabbits by a hyperimmune rabbit anti-egg albumin antibody, was investigated. This antibody produces the classical short-latency PCA in guinea pigs which mimics that elicited by guinea pig γ1 antibody. Extensive carbamylation of the antibody had no effect on the PCA reaction. In rabbits the same antiserum produces a PCA reaction which requires a latent period of at least 48 hr. The antibody responsible for this reaction is heat stable and mercaptoethanol insensitive, and thus differs from the homocytotropic rabbit anaphylactic antibody which appears early in immunization.
The Heterogeneity of Combining Sites of Anti-Benzylpenicilloyl Antibodies Obtained from Individual Rabbits: Changes in Combining Sites of γG and γM Antibodies During the Immune Response
Authors: Atsumi Tsuyoshi , Adachi Masaaki , Kinoshita Yoshio , Kawasaki Morio , Horiuchi Yoshihiko
Published: December 2022

Nine rabbits were immunized with benzylpenicilloyl-human γG conjugate (BPO-HGG). Radioiodinated γG and γM were prepared from the anti-BPO-HGG sera of individual rabbits collected 10 days and 10 weeks after the first immunization. These antibodies were charged on a specific immunoadsorbent and eluted step by step with three haptens corresponding to constituents of the BPO molecule. Three antibody fractions were separated. The antibody fractions were adapted to 6-acetylaminopenicilloic acid (nuclear portion), phenylacetylglycine (acyl side chain portion) and benzylpenicilloyl-ε-aminocaproate (whole BPO molecule), respectively.

The binding properties of these antibodies for BPO antigens were examined by equilibrium dialysis. The binding affinity of antibodies directed to the whole BPO molecule was stronger than that of antibodies directed to smaller parts of the BPO molecule. By PCA reactions the responses of antibodies complementary to the smaller parts of the BPO molecule were shown to be weaker than those of antibodies complementary to the whole BPO molecule.

The combining sites of γG and γM antibodies changed with time after the immune response. At a late stage in the immune response a significant increase was observed in antibodies adapted to the whole BPO molecule. These changes of combining sites were more characteristic in γG than in γM. However, at an early stage of immune response no significant differences were observed between γG and γM. It is concluded that changes in the binding affinity and heterogeneity of antibodies during the immune response are caused by changes in the combining sites of the antibodies.
Studies on Organ Specificity
Authors: Shulman Sidney , Ghayasuddin Mohammad , Witebsky Ernest
Published: December 2022

Rabbits were immunized with rabbit thyroid extract, using several types of complete Freund's adjuvant, and the resulting antisera were evaluated by tanned cell hemagglutination and gel diffusion. For several rabbits that achieved high titers of activity, several late bleedings (about 53 to 75 days from first injection) were then taken and fractionated into the major molecular forms of antibody by means of Sephadex gel filtration. With the accompanying immunochemical and physicochemical tests to indicate that the 19 S and 7 S pools were essentially free of contamination by each other, it was possible to show that there were low but significant levels of 19 S autoantibody, in addition to the very high levels of the 7 S form.

In a second group of rabbits, it was feasible to study much earlier bleedings (7 to 14 days from first injection) by using a slightly denatured antigen, which gave higher antibody responses at an earlier time. Again it was shown that the 19 S globulin pool had significant, although low, antibody activity, in contrast to a higher level of activity in the 7 S form.

Although both 19 S and 7 S forms of autoantibody to thyroglobulin have thus been shown to occur, it cannot be stated from these data which form appears first. They may well be produced at the same time, but in changeable proportions. Comparisons have been drawn with the results reported in the literature on the molecular form of this kind of autoantibody in the sera of human cases of thyroiditis. Several observations were also made on the phenomenon of “double-line” (or “split-line”) precipitation in gel diffusion.
A Study of Variables in the Assay of Cytotoxic Antisera
Authors: Miller Gail Lorenz , Wilson Jean E.
Published: December 2022

A study of variables in the cytotoxicity assay showed the importance of the order of addition of reacting constituents, the temperature at which reactants are added to one another, the concentration and source of complement, the initial dilution of the antiserum, the composition of the medium, the time of incubation, and the concentration and viability of the target cells. A modified procedure was developed in which serial dilution at 4°C of antiserum in a mixture of complement and target cells, used as the diluent, was shown to be feasible.
Communications
Authors: Paterson Philip Y. , Drobish Diane G.
Published: December 2022

Discussion and Summary

The remarkable adjuvant properties of pertussis vaccine in augmenting and intensifying responses to nervous tissue-adjuvant and in accelerating the induction of EAE in mice (5) and rats (3, 6) are well known. During our studies, Levine and Wenk (7) described accelerated production of adrenalitis in rats and Hargio et al. (8) reported enhanced induction of aspermatogenesis in mice sensitized to adrenal gland and testes combined with pertussis vaccine, respectively. Our observations (Table I) indicate that pertussis vaccine also has a dramatic influence on the production of another experimental auto-immune disease, EAT, in the rat. Pertussis vaccine offers not only a specific and practical means for rapid induction of hyperacute experimental autoimmune diseases, but also a promising means for further probing the pathogenesis of auto-allergic sensitization and tissue injury.
Effect of Anti-Hamster Thymocyte Serum on Papovavirus SV40-Induced Transplantation Immunity
Authors: Tevethia Satvir S. , Dreesman Gordon R. , Lausch Robert N. , Rapp Fred
Published: December 2022

The effect of anti-hamster thymocyte serum (AHTS) on SV40-induced transplantation immunity was investigated. The AHTS, when administered to hamsters during the period of virus immunization, was effective in preventing the sensitization of animals to virus-induced transplantation antigens; the treated animals failed to reject the cells transformed by SV40. Animals immune to SV40-transformed cells also failed to reject a challenge of the transformed cells following treatment with AHTS. The animals treated with AHTS and bearing tumors induced by the SV40-transformed cells synthesized antibodies against SV40 T antigen in a manner comparable to the synthesis of such antibodies by untreated animals bearing tumors.
Delayed Hypersensitivity and Arthus Reactivity in Relation to Host Resistance in Salmonella-Infected Mice
Authors: Collins F. M. , Mackaness G. B.
Published: December 2022

Mice vaccinated with living S. enteritidis or S. gallinarum but not with S. pullorum showed a typical delayed-type hypersensitive reaction when injected in the footpad with a protein-containing fraction obtained from cultures of the three strains. Mice immunized with killed suspensions of the three strains developed Arthus but not delayed sensitivity. The active material in the cell-free culture medium was partially purified by chromatography and high speed centrifugation. Delayed reactivity was transferable to normal mice by spleen cells but not by the serum obtained from mice vaccinated with living S. gallinarum. A close temporal relationship was formed between the development of delayed hypersensitivity and the emergence of effective antibacterial immunity. Arthus sensitivity showed no such correlation. S. pullorum whether given as a single dose of living organisms or repeatedly as daily intravenous injections, failed to induce a state of delayed hypersensitivity or any demonstrable antibacterial immunity to an intravenously-injected challenge by S. enteritidis. However, this organism contained an antigen that elicited delayed reaction in mice immunized with S. enteritidis or S. gallinarum. The significance of this observation is discussed.
Quantitative Passive Hemagglutination: Adaptation of the Cell Migration Technique to Measurement of Antibodies to E. Coli
Authors: Johnson Howard M. , Peeler James T. , Hall Herbert E.
Published: December 2022

A simple and sensitive technique for quantitating red cell agglutinins has been adapted for the measurement of antibodies to the O antigen of E. coli 0111. The procedure involves centrifugation of 0111-sensitized red cells to packed pellets in sealed capillary tubes. The distance migrated by the packed cells down the inclined surface of the tubes was related to the agglutinin content of the serum. The technique is highly quantitative while possessing the specificity and sensitivity of the tube tests. It is adaptable to quantitative inhibitions. A statistical evaluation of the data indicated a linear relationship between cell migration and antiserum dilution.
Studies on the Immunogenicity of Antigen-Antibody Precipitates
Authors: Henney Christopher S. , Ishizaka Kimishige
Published: December 2022

Guinea pigs immunized with antibody excess immune precipitates of human γG globulin (HGG) and rabbit or guinea pig anti-Fab or anti-Fc antibody make little antibody to HGG. The small amount of anti-HGG antibody produced in each case was directed against antigenic determinants present in both the Fab and Fc portions of the HGG molecule, and thus bore no reflection on the specificity of the antibody in the complexes. This finding suggests that the suppression is to the whole antigen molecule rather than its individual antigenic determinants.

The mechanism of the suppression of antibody formation to complexes comprised of immunogenic components cannot be explained in terms of antigenic competition for antibody-producing cells, nor in terms of “blocking” of antigenic determinants. It was shown that the combination of HGG with specific anti-Fab antibody at the equivalence point hinders only 21% of the Fc antigenic determinant, while in 4-fold antibody excess precipitates 66% of the Fc antigenic determinants are still capable of reacting with homologous antibody. Similarly, equivalence precipitates of HGG with anti-Fc antibody “block” only 16% of Fab determinants, whereas in 4-fold antibody excess precipitates 43% of the Fab determinants are “hidden.”

Quantitatively, HGG bound in a 4-fold antibody excess precipitate with either anti-Fab or anti-Fc antibody has approximately 0.5% of the immunogenicity of the uncombined antigen. It thus appears that antigen in antibody excess immune precipitates is non-immunogenic, and that the antibody produced against HGG can be explained in terms of a 0.5% dissociation of the complex in vivo.
A Structural Study of Human Exocrine IgA Globulin
Authors: Newcomb Richard W. , Normansell David , Stanworth Dennis R.
Published: December 2022

Exocrine γA globulin was isolated from human colostrum in a pure state. Measurement of s020,w and d20,w indicated a MW of about 393,000. Dissociation of the subunits of this 11.7 S molecule was effected by reduction and alkylation, with or without treatment with acid or sodium dodecyl sulfate. The results suggested that the 11.7 S exocrine γA molecule comprises two molecules of monomer γA globulin and one of secretory piece (SP) and is stabilized by both disulfide and noncovalent bonds. A protein was identified in normal colostrum with the same antigenic and size characteristics as the SP dissociated from exocrine γA globulin. The MW of SP was estimated to be 76,000 on the basis of gel filtration data.
Modification of Antigenic Structure in Vivo: Quantitative Studies on the Processing of Streptococcal Cell Wall Antigens in Mice
Authors: Schwab John H. , Brown Roger R.
Published: December 2022

Radioantibodies were employed to measure specific antigenic groups on streptococcal cell wall antigens. The processing of these precise chemical structures was followed in the homogenates of tissues of A.Sw mice at intervals following injection. There was an initial absolute increase in the number of antigenic structures as the macromolecule was degraded, suggesting that functional immunogenic groups are those structures which escape digestion. The rate and extent of the increase and decay was independent and characteristic of each group. This model also demonstrated the modification of structure by the animal to reveal “new” antigenic groups which were not detectable on the original molecule. These quantitative and qualitative changes of structural groups varied between different tissues and also in a given tissue depending upon the route of injection.
Further Characterization of the Lymphoid Cell Transfer System for the Study of Antigen-Sensitive Progenitor Cells
Authors: Bosma M. J. , Perkins E. H. , Makinodan T.
Published: December 2022

Further characterization of a lymphoid cell transfer system, a system in which heavily x-rayed syngeneic mice served as passive recipients for the antibody expression of transferred spleen cells, revealed the following: Most of the serologically detectable hemagglutinins and hemolysins released into the blood during the 1st week after cell transfer were synthesized by the repopulated spleen.Only a small fraction of the transferred immunocompetent cells colonized initially in the spleen. This fraction was estimated to be about 10% on the basis of the double spleen cell transfer method. This serologic appraisal was in striking concordance with the estimate based on the number of hemolysin plaque-forming cells in the repopulated spleen.The accuracy of these estimates was dependent upon spleen cell dose and, in the case of nenprimed spleen cells, also on the time when the repopulated spleen was removed from the primary recipient.The effect of the colonizing density on the ratio of the maximum number of hemolysin plaque-forming cells in the spleen to the unit number of spleen cells transferred was as follows: the ratio was large and constant at optimal spleen cell doses, but relatively small and variable at limiting spleen cell doses.
Antigenic Heterogeneity of Reduced and Alkylated Subunits of Human Monoclonal Macroglobulins
Authors: Dominguez Carlos J. , Caggiano Vincent , Kochwa Shaul , Wasserman Louis R.
Published: December 2022

Twenty-two isolated human monoclonal macroglobulins were studied by several immuno- and physicochemical methods. The size of the subunits dissociated by thiols and alkylated by monoiodoacetamide depended on the concentration of the reducing agent and the time of reduction. Immunochemical analysis performed with an antiserum prepared in rabbits against one of the proteins (reagent 17a) indicated that the subunits of individual macroglobulins are antigenically heterogeneous. Two antigenic determinants detected by this antiserum can be found on different subunits of native macroglobulins following mild reduction and alkylation. These antigenic determinants vary in their relative concentration and are destroyed by high concentrations of thiol. Alkylation of the reduced protein leaves an additional, common, and separable antigenic determinant, which persists after heavy reduction and alkylation and is independent of the light chain type of the native macroglobulin.
Bat Immunoglobulins Formed in Response to Experimental Japanese B Encephalitis (JBE) Virus Infection
Authors: Leonard Larry L. , Allen Rae , Sulkin S. Edward
Published: December 2022

The immune response of the big brown bat (Eptesicus f. fuscus) to Japanese B encephalitis (JBE) virus was characterized in terms of the influence of the environmental temperature and the route of inoculation on the length of the antibody induction period and the types of immunoglobulins synthesized. Sucrose density gradient analyses, supported by data derived from DEAE-cellulose chromatography and immunoelectrophoresis, indicated that the bats produced at least two types of immunoglobulins. Hyperimmune bat plasma and hyperimmune guinea pig serum against JBE virus were employed in an examination of the virus-antibody interaction. The results are discussed in terms of the influence of the antibody response on the potential role of bats as reservoir hosts for arboviruses in nature.
Lack of Relationship between Virus-Specific Surface and Transplantation Antigens in Hamster Cells Transformed by Simian Papovavirus SV40
Authors: Tevethia Satvir S. , Diamandopoulos George Th. , Rapp Fred , Enders John F.
Published: December 2022

The relationship between SV40 surface (S) and transplantation antigens in hamster cells transformed by SV40 was investigated by carrying out transplantation rejection tests in weanling hamsters and by ability of x-irradiated cells to prevent viral tumorigenesis in newborn animals. Only the cells which contained detectable levels of virus-specific tumor (T) and S antigens were found to contain detectable levels of transplantation antigens. The cells which had only the S antigen and cells which synthesized neither S nor T antigens did not contain detectable levels of transplantation antigens. These results strongly suggest that the S antigen detected in vitro by the immunofluorescence test in cells not synthesizing the T antigen is not identical with the SV40 transplantation antigen involved in the rejection or protection phenomena.
Initiation of Primary-Type and Secondary-Type Antibody Responses to Bacteriophage φX 174 in Vitro
Authors: Tao Tien-Wen
Published: December 2022

Rabbit lymph nodes were maintained in organ culture for 2 to 3 weeks. Tissue integrity and organization were well preserved with survival and formation of all cell types, e.g., blast cells, lymphocytes, plasma cells and macrophages. A primary-type anti-φX 174 response was initiated in vitro by φX 174 stimulation in cultures of lymph nodes removed from normal rabbits. On the other hand, a secondary-type response was initiated and produced in cultures of lymphoid tissues from rabbits immunized with φX 174. The kinetics of these two types of antibody responses were presented and are summarized as follows: The average peak k representing antibody activity was 0.1 in the primary response, whereas it was 124 in the secondary response, i.e., a difference of approximately 1000.In the primary response, 19 S antibody formation preceded that of 7 S, whereas 7 S antibodies were produced throughout the secondary response.In the primary response, 19 S antibodies were the predominant antibody type synthesized and 7 S, if any, only in small quantities. In the secondary response, 7 S antibodies were the only antibody type detected.Peak antibody production in the primary response was reached more than 3 days earlier than that in the secondary response.In the primary response, antibody production usually decreased steadily after the peak, whereas in the secondary response, it continued at a high level.The primary antibody response did not appear to be accompanied by increased mitotic activity, whereas, in the secondary response, mitotic activity increased prior to peak antibody production.There was a direct relationship between the concentration of φX 174 and the initiation of primary and secondary anti-φX 174 responses. The higher the concentration of the antigen, the greater was the induced antibody response. Lower concentrations of φX 174 were required to initiate a secondary response, e.g., 105 PFU of φX 174/ml failed to initiate a primary response but did initiate a secondary response. Lower concentrations of φX 174 also elicited a maximal secondary response, e.g., 108 PFU of φX 174/ml elicited a maximal secondary response, while 109 PFU of φX 174/ml failed to elicit a maximal primary response.

No primary anti-φX 174 response could be detected in cultures of adult rabbit spleen and thymus under the present experimental con-

The antibody responses initiated and produced in vitro and in vivo were compared and discussed.
Studies on Heterologous Antilymphocyte and Antithymocyte Sera
Authors: Barth Rolf F. , Southworth Joanne
Published: December 2022

The effects of rabbit antilymphocyte (ALS) and antithymocyte (ATS) sera on the early primary and secondary response of mice to sheep RBC were studied using the hemolytic plaque technique as an assay system for enumerating antibody-producing cells.

Immunosuppressive effectiveness was related to the routes of immunization and serum treatment. Intraperitoneal administration of ALS or ATS was the most effective in reducing the number of splenic plaque-forming cells (PFC), and intravenous administration was the least effective when associated with i.p. immunization.

Mice treated successively on days -1, 0 and +1 with 0.25 ml of ALS or ATS had a peak response of 4000 PFC by 76 hr following primary immunization compared to 387,000 for nontreated controls at 96 hr. There was a prolongation of the lag phase and a truncation of the exponential phase which may have been due to a reduction in the number of stimulated antigen sensitive precursor cells.

Mice treated with ALS or ATS at the time of primary immunization had a reduction in both unfacilitated and facilitated splenic PFC 3 days following a second injection of SRBC. Serum hemagglutinin and hemolysin titers were identical to controls, and it was suggested that peritoneal lymph nodes may have compensated for the decreased splenic response.

Infusion of ALS-treated and immunized mice with 50 × 106 spleen cells from normal syngeneic donors resulted in some restoration of immunocompetence both in terms of splenic PFC and serum antibodies.