bluuebunny/crossref_metadata_2025 huggingface dataset:
Recognition of Hepatitis B Antigen Determinants by a Double Label Radioimmunoassay: A Sensitive Means of Subtyping Hepatitis B Antigen
Authors: Aach Richard D. , Hacker Edwin J. , Parker Charles W.
Published: December 2022

A sensitive radioimmunoassay is described for subtyping hepatitis B antigen (HB Ag) in human serum. The method utilizes antisera (HB Ab), prepared in immunized guinea pigs, which contain predominantly anti-d or anti-y antibodies, and radioiodinated HB Ag of either D or Y subtype. Sera with HB Ag of subtype D (D) were strongly positive by RIA, employing anti-d HB Ab and 125I-HB Ág (D), whereas specimens with HB Ag of Y subtype were negative or only weakly positive. Converse results were observed when antigenemic sera were evaluated by RIA with anti-y HB Ab and 125I Y. By combining guinea pig HB Ab possessing high-titered anti-d and anti-y with 131I D and 125I Y, it was possible to screen for and subtype HB Ag simultaneously. One hundred and five antigenemic sera were evaluated; 93 or 88% could be readily subtyped by the combined RIA, including 31 of 43 samples with HB Ag in amounts too low to be detected by agar gel diffusion (AGD) or counter immunoelectrophoresis (CEP). Subtype D-HB Ag was found primarily in the serum of blood donors and patients with posttransfusion hepatitis, whereas subtype Y antigen predominated in individuals whose hepatitis was associated with the illicit use of drugs. Samples showing only “a” specificity by AGD also possessed “d” or “y” determinants by RIA. The relative antigenic reactivity of “a” and “d,” or “a” and “y” determinants appeared to vary in different specimens. This could not be explained simply by the HB Ag subtype or the number of particles in the specimens evaluated.
Preferential Radioassay of Influenza-Specific 7S (IgG) Over 19S (IgM) Class of Antibodies
Authors: Daugharty Harry
Published: December 2022

Binding affinities of immunoglobulins for A2 Hong Kong influenza viral antigen have been studied as another aspect of antibody heterogeneity. Immunoglobulin G (IgG) binding exceeds immunoglobulin M (IgM) binding in antigen-antibody reactions of precipitation, dissociation, and absorption. Evidence now indicates that IgG is more active than IgM in solid phase radioimmunoassay (RIA) inhibition as well. The preferential binding capacity of either 7S (IgG) or 19S (IgM) radioiodine-labeled globulin was quantitated with the other globulin serving as inhibitor. The range of ratios of concentrations of 7S and 19S globulins indicated that at a 7S/19S weight ratio of approximately five, the inhibition of 19S by 7S globulins appeared to be almost complete. Serologically negative 7S globulins had no such effect upon binding of 19S globulins. Conversely, the effect of serologically positive 19S globulins upon binding of 7S globulins was not apparent even at a 5-fold ratio of 19S globulin concentration. Alternatively, binding of the 7S globulins could be completely inhibited by preincubating the 7S globulins with anti-IgG before solid-phase RIA.
Human Skin Responses to Products of Concanavalin A-Activated Lymphocytes
Authors: Valdimarsson Helgi , Gross Nicholas J.
Published: December 2022

A partially purified “lymphokine” preparation derived from concanavalin A-stimulated human lymphoid cells inhibited macrophage migration in vitro. Intradermal injection of this material into 55 human volunteers produced erythema and induration typical of delayed hypersensitivity except that the response was accelerated. It began after 4 to 6 hr, was maximally indurated after 16 to 20 hr, and subsided within 36 hr. Skin window tests revealed that the glass adherent cells of this reaction were predominantly macrophages as early as 5 to 6 hr after application of the preparation under the coverslips. By contrast, 10 to 24 hr were needed for this degree of macrophage accumulation when antigens were used. Thus, when compared with antigens, the mononuclear phase of the lymphokine reaction was accelerated to a similar extent as was the inflammatory response. A control preparation derived from unstimulated lymphocytes had no migration inhibitory activity and it produced a slight inflammatory reaction, notably erythema.

This observation is in keeping with the idea that lymphocytes are capable of producing factors which preferentially attract monocytes to the site of antigenic stimulation. It also suggests that production of lymphokines, responsible for the accumulation of macrophages, may be a rate-limiting step early in the delayed hypersensitivity reaction.

The migration inhibitory factor active material was used for analyzing skin anergy. Two patients with granulomatous candidiasis and skin anergy failed to develop an inflammatory response to the preparation. Similarly, of eight tuberculin negative patients with sarcoidosis, six failed to respond. Most of these patients responded, however, when the material was injected together with cortisone.
Allograft Immunity in the Mouse
Authors: MacDonald H. R. , Phillips R. A. , Miller R. G.
Published: December 2022

An in vitro procedure for studying cell-mediated immunity in the mouse is described in which one can obtain cells (cytotoxic lymphocytes or CL) capable of lysing foreign (target) cells. The lysis is specific to antigens present during culture and is directed primarily against H-2 antigen differences although it is shown that non-H-2 antigens can also contribute. The relative number of CL produced in the cultures is also measured in vitro by a 51Cr release technique. By measuring the cytotoxicity produced by cells from each culture at several multiplicities of sensitized cells to target cells, it is shown that appropriate analysis can yield the actual ratio of CL in different cultures rather than the more qualitative measure obtained by merely measuring percentage of chromium release at fixed multiplicity. The procedure is extended to enable one also to quantitate relative numbers of progenitors of CL.
Allograft Immunity in the Mouse
Authors: MacDonald H. R. , Phillips R. A. , Miller R. G.
Published: December 2022

The cellular requirements for the initiation of an allograft response in mice have been investigated with in vitro procedures for both the initiation of the response and for the measurement of the response obtained. The progenitor of the cytotoxic lymphocyte (CL) was found to be a lymphocyte of thymic origin. Various analytical cell separation and cell-mixing experiments failed to show any evidence for cooperation of this cell with another lymphocyte of either the bone marrow or thumus-dependent lineage. However, evidence was found for cooperation with a nonspecific “macrophage-like” cell. Radiation survival and analytical cell separation experiments performed to follow the development of CL after initiation indicate that they develop through proliferation and “blast transformation,” and suggest that they revert back to small lymphocytes, physically distinguishable from the original progenitors, at late times (2 to 4 weeks). Attempts to demonstrate immunologic memory in this latter population of cells were unsuccessful.
Heterologous Adoptive Cutaneous Anaphylaxis: a Method for Detecting Reaginic Antibody Formation by Cells of the Mouse
Authors: Kind L. S. , Macedo-Sobrinho B.
Published: December 2022

One approach to a rational therapy of allergic reactions is the acquisition of a greater knowledge of the factors which control reaginic antibody synthesis. We now report a method for detecting cells producing reaginic antibody in the mouse.

Waltzer and Glazer first demonstrated local passive cutaneous sensitization of normal individuals by the intracutaneous injection of viable leukocytes obtained from atopic patients (1). Our procedure is based on the subsequent findings of Chase (2), Rosenberg et al. (3), and Battisto (4) that spleen, thymus, lymph node, and peritoneal exudate cells from immunized guinea pigs could be transferred to the skin of isologous or homologous guinea pigs with the subsequent detection of antibody at the site of the deposited cells. Presumably antibody from the transferred cells bound to skin mast cells and a typical passive cutaneous anaphylaxis (PCA)3 (5) reaction resulted upon injection of antigen and Evans blue dye. The technique was termed adoptive cutaneous anaphylaxis (ACA) by Battisto (4).
A Third Non-Adherent Cell Required for the in Vitro Immune Response to Sheep Erythrocytes
Authors: Talmage David W. , Hemmingsen Helen
Published: December 2022

The requirement of nonspecific cells for the induction of antibody-forming cells (PFC)1 during in vitro culture has been reported by several investigators. Mosier first demonstrated the requirement for adherent cells (1) and Roseman demonstrated the radioresistance of the adherent cell (2). Hoffman and Dutton reported that peritoneal macrophages or supernatants from macrophage cultures could substitute for the adherent cells in this system (3). More recently, Chen and Hirsch reported that the inclusion of mercaptoethanol in the medium led to the restoration of the PFC response of non-adherent cells (4).

In 1969, we reported that the spleen cells from mice x-rayed and reconstituted with bone marrow cells (BMSC) effectively reconstituted the response of non-adherent cells (5). Later we demonstrated the x-ray resistance of the reconstituting cells (6). Because the activity of BMSC resided at least partly in the adherent cell and because of a similar x-ray resistance it was concluded that the activity of BMSC could be explained by its content of adherent cells.
Retention of Graft-vs-Host Activity in Non-Adherent Spleen Cells After Depletion of Cytotoxic Activity by Incubation on Allogeneic Target Cells
Authors: Mage Michael G. , McHugh Louise L.
Published: December 2022

Antibody-forming cells (1) and their precursors (2) readily bind to antigen, presumably through immunoglobulin receptors on their surfaces (3). Absorption occurs at 4°C, within minutes (4). On the other hand, thymusderived lymphocytes (T-cells),1 which have been implicated in cellular immune phenomena such as cell-mediated killing of allogeneic target cells (5), and in the graft-vs-host (GVH) reaction (6), do not have readily demonstrable immunoglobulin on their surfaces (7), and specific binding to cell surface antigen in vitro has generally required incubation at 37°C for cytotoxic effector cells (5, 8), or in vivo absorption for GVH antigen-reactive cells (GVH-ARC) (9).

On immunization across a strong histocompatibility barrier in vivo (10, 11) or in vitro (12), there is a large increase in the level of cytotoxic activity (10) but little or no change in GVH activity (11, 12), implying that different populations of T-cells mediate these two reactions.
Further Evidence for the Involvement of Thymus-Processed Lymphocytes in Syngeneic Tumor Cell Cytolysis
Authors: Plata Fernando , Gomard Elisabeth , Leclerc Jean-Claude , Levy Jean-Paul
Published: December 2022

The chromium-51 release test was used to study syngeneic tumor cell lysis by spleen lymphocytes from MSV tumor-bearing animals throughout the various stages of tumor growth and regression. Thymus dependent (T) cells were found to be necessary for tumor cell immune lysis to occur, and no requirement for non-T cells was detected.
Effects of Sublethal Irradiation on the Plaque-Forming Cell Response in Mice
Authors: Schmidtke Jon R. , Dixon Frank J.
Published: December 2022

X-ray (440 R) administered 4 days after a primary injection of 1 × 108 sheep red blood cells (SRBC) into normal BALB/c mice resulted in increased numbers of indirect plaque-forming cells per million spleen cells (PFC/106) 3 days after a secondary challenge of 1 × 108 SRBC. The results were compared to control, non-x-rayed mice. The time of x-ray in relation to SRBC injection was critical as x-ray 3, 5, or 6 days after primary SRBC injection did not increase the number of PFC/106 cells. The state of increased responsiveness induced by x-ray was dependent on the dose of SRBC used for primary injection and could not be transferred by spleen cells to syngeneic recipient mice. The number of primary direct PFC/106 cells was increased when the response was measured 8 days after injection of SRBC and mice x-rayed 48 hr after antigen injection when compared to control non-x-rayed mice; this increase was probably due to a delay in the response induced by x-ray.
The Role of Complement in Human Immediate Hypersensitivity: Evidence against Involvement of the Alternate Pathway of Complement Activation
Authors: Grant J. Andrew , Lichtenstein Lawrence M.
Published: December 2022

A role for the alternate pathway of complement activation in immediate hypersensitivity reactions was investigated following a number of recent observations which suggested this possibility. The ability of normal human serum to enhance allergen-mediated histamine release from the leukocytes of atopic human donors was not reduced by heating serum to 100°C, nor did the addition of purified C3 to allergen-challenged leukocytes change the percentage of histamine released. To ascertain if cell-bound C3 was involved, we incubated leukocytes with anti-C3: no histamine was released by a preparation of anti-C3 which had been made with a highly purified C3 antigen, nor did preincubation of leukocytes in this anti-C3 inhibit the subsequent allergen-mediated release of histamine. Moreover, most of the anti-C3 antibodies could be recovered in the supernatant at the end of the incubation period. We have been able to confirm previous reports that some anti-C3 preparations can cause the release of histamine and/or can inhibit allergen-mediated release of histamine from human leukocytes. However, further investigation suggested that these latter results were obtained with preparations of anti-C3 contaminated with antibodies against human light chains. Anti-light chain antibodies would be capable of releasing histamine directly from leukocytes or of “desensitizing” leukocytes to subsequent allergen-challenge.

It is concluded that the release of histamine from human leukocytes challenged by allergens is independent of free or cell-bound C3, and that there is little evidence that the alternate pathway of complement activation is involved in this type of response.
The Cellular Basis of Cross-Tolerance
Authors: Ruben Thomas J. , Chiller Jacques M. , Weigle William O.
Published: December 2022

Mice made unresponsive to human γ-globulin demonstrate a marked cross-tolerance to porcine γ-globulin and equine γ-globulin but react normally to the more phylogenetically distinct chicken γ-globulin. This cross-tolerance exists in spite of an antigenic cross-reactivity at the indirect plaque-forming cell level of only 1 to 3%. This cross-tolerance status can be transferred to thymectomized, irradiated, bone marrow-reconstituted recipients by tolerogen-treated thymus cells. It is suggested that antigen recognition by the thymus-derived cell manifests a wider range of cross-reactivity than that evidenced by the bone marrow-derived cell.
Detection of Idiotypic Determinants on Human Monoclonal Proteins with Specific Rabbit Antisera and Hemagglutination-Inhibition
Authors: Wells J. Vivian , Wang An-Chuan , Thornsvard Charles T. , Schanfield Melvin S. , Bleumers Jan F. , Fudenberg H. Hugh
Published: December 2022

Idiotypic antigenic determinants on human monoclonal immunoglobulins were studied by hemagglutination inhibition (HAI) by using human O Rh-positive erythrocytes coated with the proteins by the chromic chloride method. The rabbit antisera to the purified proteins required initial absorption with washed normal human erythrocytes and the hemagglutinating titer of anti-idiotypic antibodies after absorption varied from 1:1,300 to 1:328,000 for different antisera. Specificities against non-idiotypic V-region determinants, such as V-region subgroups, could not be demonstrated in the antisera.

The HAI method is both sensitive, with inhibition detected at inhibitor concentrations of 0.001 mg/ml, and specific, since heterologous proteins did not produce significant inhibition. In studies with a single myeloma protein, inhibition was only obtained with the whole homologous protein and fragments thereof which contained the combining site, e.g., F (ab′)2 and Fab. HAI is a satisfactory method for studying idiotypic specificities in human monoclonal immunoglobulins and should permit analysis of sharing of specificities and should prove suitable for the detection of residual myeloma protein in the serum of patients with multiple myeloma during their response to therapy.
The Skeletal Muscle Binding Site of Antistriated Muscle Antibody in Myasthenia Gravis: An Electron Microscopic Immunohistochemical Study Using Peroxidase Conjugated Antibody Fragments
Authors: Mendell J. R. , Whitaker J. N. , Engel W. K.
Published: December 2022

The skeletal muscle-binding site of antistriated muscle antibody (ASMAb) from individuals with myasthenia gravis (MG) was established ultrastructurally by means of an indirect immunohistochemical technique with purified sheep antihuman IgG (PSAHG) conjugated to horseradish peroxidase (HRP). Penetration of the HRP-antibody conjugate was greatly enhanced by preparing fragments, the Fab or F(ab′)2 of the PSAHG before HRP conjugation. ASMAb from nine patients with MG localized to the sarcoplasmic reticulum surrounding the I-band. No binding with the myofilaments could be demonstrated.
Cross-Reactions of Additional Extracellular Polysaccharides of Klebsiella
Authors: Heidelberger Michael , Dutton Guy G. S.
Published: December 2022

Cross-reactions are recorded of the K polysaccharides of Klebsiella types 5, 7, 18, 20, 21, 24, and 62 and of deacetylated K5 in a number of antipneumococcal and anti-Salmonella sera. In several instances these confirm structures already established. Findings likely to emerge from further chemical study are also pointed out on the basis of previously established relations between chemical constitution and immunological specificity.
Studies of Mopc 104E Plasmacytoma Cells Forming Rosettes with Autologous and Syngeneic Erythrocytes
Authors: Hiramoto Raymond N. , Ghanta Vithal K.
Published: December 2022

MOPC 104E plasmacytoma cells secreting an IgM antibody to dextran 1355 S1,3 were shown to make mixed rosettes with both dextran-coated chicken erythrocytes (D-CRBC) and syngeneic mouse erythrocytes from tumor-bearing mice (T-MRBC). D-CRBC rosettes were readily blocked by soluble dextran, monospecific goat anti-mouse IgM, anti-mouse IgG2 but not anti-mouse IgG1. T-MRBC rosettes were blocked by monospecific anti-mouse IgG2, but not with anti-mouse IgM or soluble dextran. These results indicate that the plasmacytoma carries an IgG2-like receptor to T-MRBC.
Selective Cytotoxicity of Hapten-Substituted Cells with an Antibody-Enzyme Conjugate
Authors: Philpott Gordon W. , Shearer William T. , Bower Richard J. , Parker Charles W.
Published: December 2022

Affinity iodination and cytotoxicity of hapten-substituted tumor cells were accomplished with an anti-hapten antibody conjugated to glucose oxidase (which generates H2O2) and lactoperoxidase (which iodinates cells in the presence of H2O2 and I-). Polynitrophenylated HeLa and Hep-2 cells, obtained by brief incubation with 2,4,6, trinitrophenyl (TNP)-sulfonic acid, were selectively iodinated and killed when treated briefly with an anti-TNP antibody-glucose oxidase conjugate (Ab-GO) and cultured in media containing lactoperoxidase (LP) and iodide. Cell killing was determined by three methods: 1) a microcytotoxicity assay, 2) the loss of cells prelabeled with 125IUdR and 3) the degree of incorporation of 125IUdR after treatment (postlabeling). Results with the three independent methods showed cytotoxicity, ranging from 19% to 98% in TNP-cells incubated with Ab-GO (0.6 to 57 µg/ml), LP (50 µg/ml), and iodide (20 µM). ε-DNP-lysine (100 µM) significantly reduced the amount of killing and minimal cytotoxicity (0 to 35%) was seen with unsubstituted cells or if Ab-GO, LP, or I- were omitted.
Extraction of Complement Inhibitory Factors from the Erythrocytes of Non-Human Species
Authors: Hoffmann E. M. , Etlinger H. M.
Published: December 2022

Extracts from the erythrocyte stromata of seven species were examined for the presence of a factor capable of accelerating the rate of decay of sheep erythrocytes in the intermediate state .3 Only the extracts from rabbit and guinea pig erythrocytes possessed decay accelerating activity. Gel filtration (Sephadex G-200) of the active crude extracts gave partial purification with the active fraction passing through the column near the void volume. The materials from rabbit and guinea pig red cells were capable of decreasing the lytic susceptibility of sheep E, EA, EAC1̄G P, EAC4G P and EAC1̄G P4 as well as sheep . Diluted rabbit and guinea pig erythrocyte stromata extracts, which were incapable of affecting any component of the sheep EAC1̄G P4G P complex, were still capable of accelerating the normal rate of decay of sheep .
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Tada Tomio , Okumura Ko , Taniguchi Masaru
Published: December 2022

An antigen-specific subcellular component, possibly from thymus derived (T) cells that regulate homocytotropic antibody (HTA) formation against dinitrophenylated Ascaris suum extract (DNP-As), was extracted from mechanically disrupted thymocytes and spleen cells of rats hyperimmunized with the homologous antigen. This cell free extract, when injected into HTA-producing animals that had been 400R x-irradiated and immunized with DNP-As and Bordetella pertusis vaccine so as to produce high and persistant HTA formation, exerted a strong inhibitory effect on the ongoing HTA production of the recipients. A similar thymocyte extract obtained from rats hyperimmunized with the carrier (Ascaris extract, As) alone also showed the inhibitory activity, whereas that from rats hyperimmunized with the hapten coupled to a different carrier (DNP-bovine serum albumin) or with Freund's complete adjuvant alone did not. The inhibitory activity of the thymocyte extract was completely abolished by absorption with immunoadsorbents composed of DNP-As or As, but not with that of DNP-BSA, indicating that the active component has an affinity to carrier molecules. However, the inhibitory activity was not absorbed with antisera directed to rat Fab, µ chain and polyvalent immunoglobulins, suggesting that the active component is not likely to be an immunoglobulin. The activity was significantly reduced by absorption with an immunoadsorbent composed of anti-rat thymocyte serum. These results suggest that the inhibitory cell interaction in the rat HTA formation, which has previously been reported by us, is mediated by this subcellular component of primed T cell which is not immunoglobulin in nature but has an affinity and specificity to the carrier molecule.
Immunologic Responses in the Gastrointestinal Tract of the Guinea Pig I. Characterization of Peyer's Patch Cells
Authors: Levin Douglas M. , Rosenstreich David L. , Reynolds Herbert Y.
Published: December 2022

Peyer's patches are prominent gut-associated lymphoid aggregates the function of which is as yet unknown. Recent articles have reviewed data suggesting either a central lymphoid (bursal equivalent) function or second-level, peripheral lymphoid function (1, 2). The existence of both thymus-dependent lymphocytes (T cells)1 and bone marrow-dependent lymphocytes (B cells) in the Peyer's patches of some species, including mice, rats, rabbits, and hamsters, is suggested by morphologic (3–7) and some functional (8–13) evidence. However, although the presence of B cells (or B cell precursors) (9, 11, 13) seems clear, the presence of T cells has been questioned by recent data showing an absence of graft-vs-host response with donor cells from Peyer's patches; these findings imply that, at least in mice and rats, few or no functional T cells reside in Peyer's patches (14, 15). In order to characterize further the Peyer's patch, and particularly to detect any T cell population, single cell preparations were made from this tissue in adult guinea pigs.
The Pathogenesis of Corneal Inflammation Due to Herpes Simplex Virus
Authors: Meyers Roberta L. , Pettit Thomas H.
Published: December 2022

Corneal immune rings were produced with purified herpes simplex virus (HSV) antigen in both systemically and locally (intracorneal) HSV-sensitized rabbits. The ringlike corneal opacity consisted of a local accumulation of inflammatory cells, HSV antigen, host-antibody, and complement. A similar linear opacity was produced by injection of purified HSV antigen and anti-HSV antibody simultaneously at opposite sides of a normal rabbit cornea in a nonsensitized rabbit.

Immunohistopathologic studies of experimental stromal herpes keratitis in rabbits induced by corneal infection with HSV revealed antigen, host-antibody, complement (C3), and inflammatory cells at the site of the stromal infiltrate. The tissue injury occurring in stromal herpes keratitis appears to result from HSV antigen-HSV antibody interaction; this attracts serum complement and results in the leukotaxis of polymorphonuclear leukocytes which constitute the major corneal infiltrate seen in stromal keratitis.
The Proliferative and Anamnestic Antibody Response of Rabbit Lymphoid Cells in Vitro
Authors: Thorbecke G. Jeanette , Bell Melvin K.
Published: December 2022

The anamnestic responses of the right and left popliteal lymph nodes of rabbits after a single injection of diphtheria toxoid (DT) into the left footpad were studied in vitro. The draining node always produced a higher response, even at intervals up to 11 months after priming.

Weekly injections of rabbit anti-DT, either into the right footpad, or in larger amounts, intravenously, significantly increased the difference in responsiveness between the two nodes by lowering the response on the uninjected side.

It is suggested that, aside from memory cells reaching the right node, priming of this lymph node is partly due to antigen carried over from the injected side.
Association of Tumor and Histocompatibility Antigens in Sera of Lymphoma-Bearing Mice
Authors: Fujimoto Shigeyoshi , Chen Ching Hong , Sabbadini Edris , Sehon Alec H.
Published: December 2022

Soluble tumor-associated antigens and histocompatibility antigens were detected in sera of A/J mice bearing a spontaneous lymphoma (L1117) by the inhibition of the cytotoxicity of rabbit anti-L1117 serum and of alloantisera to normal A/J antigens, respectively; the rabbit antiserum was rendered specific for lymphoma antigens by extensive absorption with normal A/J cells and alloantisera were produced in C57BL/6J and C3H mice. Both types of antigen were isolated together from serum of tumor-bearing animals, with reverse immunosorbents prepared by insolubilization of either rabbit anti-L1117 antibodies or of the anti-A/J alloantibodies, and were also present in the supernatant of the medium used for culturing L1117 cells. It is suggested that the determinants responsible for these two antigenic specificities were associated with the same molecular moiety.
Anti-Alloantibodies: A de Novo Product
Authors: Binz Hans , Ramseier Hansruedy , Lindenmann Jean
Published: December 2022

(Lewis × DA)F1 hybrid rats were injected once or twice with a small, subsaturating dose of DA anti-Lewis posttransplantation alloantiserum. The time course of antialloantibody response was followed over a period of 30 days by using the PAR assay. After a single injection, antialloantibody titers became first detectable on the 2nd day, they increased until the 11th day and reached a plateau between days 11 and 14; thereafter titers slowly declined. A second injection given 2 weeks after the first was followed, without measurable latency, by rapid exponential increase of antialloantibody titers lasting 6 days. The plateau reached 6 to 8 days after the second injection was 500 times higher than that attained after a single injection. Between the 8th and the 16th day after the booster injection titers slowly declined. We conclude that the response measured represents the de novo synthesis of antialloantibody and not a complex of alloantibody and antigen.
Carrier Independent T Cell Helper Effects in Antigenic Stimulation
Authors: Doughty Robert A. , Klinman Norman R.
Published: December 2022

Anti-ϑ serum treatment of cells from immunized mice before their transfer to irradiated non-immune recipients decreased the ability of such cells to be stimulated by hapten on both homologous and heterologous carriers over a wide range of antigen doses. The anti-ϑ treatment was shown to affect only thymus-dependent (T) cells, since the number of bone marrow-derived (B) cell precursors was unaltered. Thus, specific T cells contribute significantly, not only to stimulation by the hapten on the homologous carrier, but also when hapten is presented on heterologous and even poorly immunogenic carriers. These results are interpreted as indicating a role for collaborative interactions of hapten-specific T cells with B cells specific for the same determinant. Such interactions were found at low antigen concentration. Results are discussed with relation to the mechanism of B cell stimulation.
Purification of Mouse Interferon by Affinity Chromatography on Anti-Interferon Globulin-Sepharose
Authors: Ogburn Clifton A. , Berg Kurt , Paucker Kurt
Published: December 2022

Mouse interferon produced in L cells was subjected to affinity chromatography on Sepharose-bound anti-interferon globulin. The hyperimmune anti-interferon serum was specifically adsorbed to remove antibodies against proteins derived from normal L cells, medium, and inducer virus preparation. The method permitted selective elimination of identified contaminant antigens from crude interferon preparations. Recovery of interferon was quantitative, and purification during this step was from 20- to 50-fold. Specific activities in peak fractions ranged from 1 to 2.7 × 108 National Institutes of Health reference units per mg protein. Interferons induced by irradiated Newcastle disease virus and poly I:C were purified to the same degree.
Regulation of the Immune Response
Authors: Kappler John W. , van der Hoven André , Dharmarajan Umarani , Hoffmann Michael
Published: December 2022

The immune response of mouse spleen cell suspension to heterologous erythrocyte antigens and the hapten, trinitrophenol, can be inhibited by the presence of antibody specific for these antigens. The properties of the suppression are: 1) It is antigen specific in that the suppressive antibody must recognize at least one determinant on the antigen. 2) It is not determinant specific in that antibody directed against as few as one determinant on the antigen can suppress the response to the rest of the determinants. 3) The participation of the Fc fragment of the suppressive antibody is indicated by the ineffectiveness of (Fab)′2 and chicken antibody in suppression. 4) The mechanism of the suppression does not appear to be central in that cells preincubated with antigen-antibody complexes for as long as 24 hr are not impaired significantly in their ability to respond to a subsequent challenge with antigen. These findings are discussed in view of the various models which have been proposed for the mechanism of antibody-mediated suppression of the immune response.
The Mechanism of a Hapten-Specific Helper Effect in Mice
Authors: Janeway Charles A.
Published: December 2022

Mice immunized with 2,4-dinitrophenyl-bovine serum albumin (DNP-BSA) show a greater anti-ovalbumin (OVA) antibody response to subsequent immunization with DNP-OVA than do mice initially immunized with BSA alone. In order to determine the mechanism of this DNP-specific augmentation of anti-OVA antibody response to DNP-OVA, we performed cell and serum transfer experiments. Irradiated recipient mice received spleen cells from donor mice which had been primed with OVA and subsequently treated with heterologous anti-mouse lymphocyte serum; these cells served as precursors of anti-OVA antibody-forming cells. Such recipients made greatly augmented anti-OVA responses to DNP-OVA if they also received serum from mice primed with DNP-keyhole limpet hemocyanin (KLH). Spleen cells from DNP-KLH primed donors transferred little or no augmentation of this response, and such activity as they possessed appears to be due to the anti-DNP antibody they secrete. Anti-DNP antibody purified by elution from a solid immunoadsorbent possessed all the augmenting activity of anti-DNP-KLH serum. These experiments provide no evidence for functionally active, thymus-derived, hapten-specific “helper” cells. The implications of this result are discussed.
Immunoglobulins in the Developing Amphibian, Rana Catesbeiana
Authors: Geczy Carolyn L. , Green Patrick C. , Steiner Lisa A.
Published: December 2022

Rana catesbeiana tadpoles contain high molecular weight and low molecular weight immunoglobulins that closely resemble, in structural and antigenic features, the corresponding immunoglobulins in adult bullfrogs. Although the low molecular weight immunoglobulins are antigenically heterogeneous, all the antigenic species found in frogs also occur in tadpoles. Thus, in these amphibia, the immunoglobulin classes develop before metamorphosis is completed.
Local Transfer of Delayed Hypersensitivity by T Lymphocytes
Authors: Jaffer Adrian M. , Jones Gerald , Kasdon Earl J. , Schlossman Stuart F.
Published: December 2022

It is now well established that lymphocytes consist of two major subpopulations, thymus dependent (T)2 cells and thymus independent (B) cells (1). These populations may be distinguished by the presence of readily demonstrable surface immunoglobulin on B cells compared with T cells (2). Although cellular immune reactions, such as skin graft rejection, are known to be thymus dependent, there is no direct evidence that T cells mediate delayed skin reactions. This report describes the production of delayed skin reactions in guinea pigs by lymphocytes depleted of Ig-bearing (B) cells. An insoluble anti-Ig column was used to remove B cells (3) and the ability of the B-depleted cell population to mediate delayed hypersensitivity was assessed by a local transfer technique (4).

Antiserum to guinea pig Ig was raised in rabbits and purified antibody was prepared with the aid of an insoluble immunoabsorbent of guinea pig globulin covalently bound to Sepharose 4B (5).
Human Immunoglobulin A: Production of an Fc Fragment by an Enteric Microbial Proteolytic Enzyme
Authors: Mehta Satish K. , Plaut Andrew G. , Calvanico Nickolas J. , Tomasi Thomas B.
Published: December 2022

An Fc fragment has been prepared in good yield by proteolytic cleavage of human secretory and serum myeloma IgA with an enzyme recovered from the human gastrointestinal tract. The naturally occurring enzyme is of microbial origin and its activity is metal dependent.
Comparative Chemical Study of α- and µ-Chains from a Single Patient (SC)
Authors: Seon B.-K. , Yagi Y. , Pressman D.
Published: December 2022

In a previous report (1), we have shown that an IgA(κ) and an IgM(κ) myeloma protein derived from the same patient (SC) shared individually specific determinants which were found in the Fab portion of the molecules. We have also shown by various criteria that the light chains of these two immunoglobulins are identical (2). In the present work, we have compared the N-terminal amino acid sequence of the variable portion of the heavy chain of the IgA molecule with that of the IgM molecule and have found them both to be Glp-Met-Gln-Leu, a sequence not previously reported, but close to the VH1 heavy chain subgroup sequence, Glp-Val-Gln-Leu. In addition, peptide maps of chymotryptic digests showed appreciable identity between substantial portions of the variable portions of the α and µ heavy chains.

Materials and Methods. IgA and IgM and the Fab fragments of IgA and IgM were prepared as described previously (1).
Independent Movement of IgD and IgM Molecules on the Surface of Individual Lymphocytes
Authors: Knapp W. , Bolhuis R. L. H. , Rádl J. , Hijmans W.
Published: December 2022

It is now generally accepted that immunoglobulins (Ig) on the membrane of lymphocytes are mobile in the plane of the membrane itself. The reaction of divalent antibodies with the Ig molecules, which were originally diffusely distributed, induces cross-linkage and aggregation of the Ig molecules. These aggregates are then actively transported to one pole of the cell where they form the so-called caps. This capping process is markedly inhibited in the cold and can be blocked by different drugs (1, 2).

We used this phenomenon for the investigation of Ig molecules of two different classes (IgM and IgD) on individual lymphocytes.

Materials and Methods. Cord blood lymphocytes from 20 healthy babies and peripheral lymphocytes from 20 adults were investigated for the presence of membrane-bound Ig in immunofluorescence. Lymphocytes were obtained by gradient centrifugation with the Isopaque Ficoll technique (3). Cells were tested for viability and stained in suspension.
Antinucleolar Antibodies in Human Sera
Authors: Pinnas Jacob L. , Northway James D. , Tan Eng M.
Published: December 2022

Antibodies to mammalian cell nucleoli occur spontaneously in certain autoimmune human diseases and have been demonstrated by immunofluorescence. In this study, nucleoli were isolated from fresh calf thymus and antinucleolar antibodies were used as reagents to identify a reactive nucleolar antigen. The antigen was a low molecular weight 4–6S RNA, and the precipitating activity of this antigen was destroyed by treatment with spleen phosphodiesterase. Sera were selected for this study because of predominance of nucleolar staining by immunofluorescence and were found to be from patients with scleroderma, Sjogren's syndrome, and undiagnosed illnesses manifesting Raynaud's phenomenon.
The Role of Humoral Factors in the Initiation of in Vitro Primary Immune Responses
Authors: Watson James
Published: December 2022

Immune responses in thymus derived (T) cell-depleted mouse spleen cultures were stimulated by the addition of either syngeneic T cells immunized with antigen, or allogeneic T cells reactive against precursor antibody-forming cells. Both require the presence of specific antigen in cultures. At low T cell concentrations, (< 106), allogeneic T cells are more effective in stimulating immune responses than the syngeneic T cells, however, at higher cell numbers (> 106), allogeneic T cells suppress the response. The stimulation of in vitro immune responses to heterologous erythrocyte antigens is mediated by nonspecific activity released from antigen-stimulated T cells in culture supernatants. Fractionation of culture supernatants prepared in serum-free media, by Sephadex G-100 chromatography, revealed the presence of three thymus cell-replacing factors (TRF). The fractionation profiles of supernatant activities prepared from syngeneic and allogeneic T cells were identical. TRF-I from both T cell sources has a molecular weight around 150,000, is labile to heat, periodate, and trypsin, and requires the presence of θ-positive cells for production in culture. TRF-I does not appear to be immunoglobulin in nature. TRF-II (m.w. 15,000 to 40,000), and TRF-III (m.w. < 7000) do not require the presence of θ-positive cells for production in culture. Syngeneic and allogeneic T cells also released a factor (m.w. 10,000 to 20,000) that suppressed the induction of normal in vitro immune responses.
Mechanisms of Endotoxin Tolerance
Authors: Greisman Sheldon E. , Young Edward J. , DuBuy Bernard
Published: December 2022

Acclimatized rabbits were passively immunized with graded volumes of antisera against endotoxins from either smooth wild type Gram-negative bacteria possessing intact O-antigenic side chains, or rough mutants deficient in these O-antigens. The recipients were challenged with test doses of endotoxins that evoked fever within the sensitive dose-response range. Pyrogenic tolerance conferred with antisera to smooth endotoxins was consistently directed toward the O-specific antigen; minimal protection extended to endotoxins from heterologous Gram-negative bacterial species, or from homologous O-deficient rough mutant strains. In contrast, antisera to O-deficient rough mutant strains transferred significantly more effective protection against heterologous endotoxins. The level of such heterologous protection, however, was markedly less than the O-specific protection conveyed by the smooth anti-endotoxin sera.

The protective factors in antiserum to smooth endotoxin were separable by either pyrogen-free Sephadex G-200 columns or ammonium sulfate precipitation into 2-mercaptoethanol (2-ME) resistant and sensitive O-antibody-containing fractions. In the latter, 2-ME virtually abolished O-agglutinating, but not O-combining or protective activities. Both O-combining and protective activities could be reduced by absorption of antiserum with spleen cells but not with erythrocytes.

It is concluded that: 1) the humoral components of endotoxin tolerance are antibodies to both O-specific and common core antigens. Such antibodies are both IgM and IgG, protect by primary (binding) rather than secondary (agglutinating) activity, and are in part cytophilic; 2) the relative importance of O-specific and common core antibodies is dependent upon both the immunizing and test endotoxin preparations. Antisera to smooth endotoxins afford the highest levels of passive tolerance, but such protection is restricted primarily to the O-specific endotoxin. Antisera to rough O-deficient endotoxins provide less tolerance but afford the better approach to broad passive endotoxin protection; 3) the findings support the hypothesis that common core antigens are partially masked by the O-specific antigens in the endotoxin molecule.
The Reaction of Zymosan with the Properdin System in Normal and C4-Deficient Guinea Pig Serum
Authors: Brade Volker , Lee Gerald D. , Nicholson Anne , Shin Hyun S. , Mayer Manfred M.
Published: December 2022

On incubation with serum, zymosan (Z) combines with several serum components, collectively designated X, to form an intermediate, ZX, which is capable of activating the complement system by cleavage of components C3 and C5. We now report kinetic studies on the formation of ZX, as well as the reaction of this intermediate with purified C3 or C5, which indicate that the splitting of these complement components is mediated by two multi-unit enzyme complexes on the zymosan particles, one which acts on C3, and another which splits C5. These two enzymes share in common factor B of the properdin system, but differ in other respects. On extended incubation at 37°C they decay to a form designated ZXd by release of the factor B subunit. ZX can be regenerated from ZXd by treatment with factor B but this process is efficient only with respect to C5 cleavage, presumably because a subunit of the C3-cleaving enzyme, other than factor B, is also lost during decay at 37°C. The reaction of factor B with ZXd to yield ZX is believed to be enzymatic on the basis of its temperature-dependence and its kinetic behavior. Presumably, an enzyme on ZXd activates factor B, probably by cleavage, thus enabling it to combine with ZXd to form ZX. This process of activation and fixation of factor B is believed to be the final step in the complex series of reactions leading to formation of the C3- and C5- cleaving enzymes on zymosan particles. Evidence is also presented that the formation of these enzymes on zymosan proceeds much more rapidly in normal guinea pig serum than in C4-deficient serum, which indicates a marked accelerating effect of the early-acting complement components.
Evidence for a Macrophage-Mediated Effect of Poliovirus on the Lymphocyte Response to Phytohemagglutinin
Authors: Soontiëns F. J. C. J. , Van Der Veen J.
Published: December 2022

Poliovirus inhibits the response of human peripheral leukocytes to phytohemagglutinin (PHA). The inhibitory activity requires the presence of macrophages. There is no evidence that lymphocytes are capable of supporting viral replication. On the other hand, the virus multiplies in macrophages derived from monocytes. Further, macrophages enhance the response of lymphocytes to PHA, but the cooperation between these cell types is absent in virus-infected cultures. The findings support the suggestion that poliovirus inhibits the response of leukocytes not by direct action on lymphocytes but by suppressing the enhancing effect of the macrophage.
Lymphocyte-Derived Eosinophil Chemotactic Factor
Authors: Torisu Motomichi , Yoshida Takeshi , Ward Peter A. , Cohen Stanley
Published: December 2022

When cultured in the presence of specific antigen, lymphocytes from delayed hypersensitive guinea pigs release a precursor substance (ECFp) which can be activated by specific immune complexes to yield an eosinophil chemotactic factor (ECF). The generation of ECFp like that of migration inhibition factor (MIF) is carrier specific. The immune complexes which activate ECFp must contain IgG2 antibody with specificity directed against the antigen which was used to stimulate the lymphocytes to produce ECFp. Previous studies have shown that this reaction does not involve complement.

It was not possible to dissociate MIF activity from ECFp activity by a combination of sephadex chromotography and agarose block electrophoresis. However, columns containing sepharose beads conjugated with specific antibody removed ECFp but not MIF from the supernatant fluids. Antigen-conjugated columns did not affect either activity. If the supernatants were activated with immune complexes before passage through the columns, then eosinophil activity could not be removed by either column. These results all suggest that ECFp co-precipitates with immune complexes by virtue of an antigenic fragment it contains. The activation of ECFp to ECF involves loss of this fragment.
A Mathematical Model for the Quantitation of Tumor and Antibody-Forming Cell Populations within an Animal's Body
Authors: Hiramoto Raymond N. , Ghanta Vithal K. , McGhee Jerry R. , Hamlin N. Marlene
Published: December 2022

The application of a mathematical model described by Hege and Cole was tested by a murine myeloma model. The plasmacytoma MOPC 104E secretes an IgM antibody to bacterial dextran B-1355. By using this system the various parameters required in the equation could be quantitated. The model relates the number of tumor cells in the population to antibody produced under conditions of logarithmic growth. Experimental values of antibody measured and estimation of starting population (An) of tumor cells showed that 2.24 × 106 cells grew when 3 × 106 cells were injected. This indicates that approximately 75% of the cells grew when 3.0 × 106 tumor cells were given i.p. into each mouse. The experimentally determined antibody levels were in agreement with the computer calculated values.