bluuebunny/crossref_metadata_2025 huggingface dataset:
Homotropic Interactions in the Binding of C1̄ to Antigen-Antibody Complexes
Authors: Thompson J. J. , Hoffmann L. G.
Published: December 2022

The mechanism of guinea pig C1̄ binding to IgM and IgG antigen-antibody complexes on sheep erythrocytes has been examined by quantitative studies with independent assays of bound and free C1̄ in the reaction mixtures. Simple and heterogeneous binding play no important role; cooperative binding occurs. The cooperative binding mechanism apparently involves no interactions between domains on the red cell surface or interactions between C1̄ molecules in solution. The validity of an allosteric model for the binding mechanism is currently being tested by both curve-fitting and statistical methods.
Studies of Immunologic Function in C4 Deficient Guinea Pigs
Authors: Frank M. M. , Ellman L. , Green I.
Published: December 2022

Several in vivo parameters of immunologic function were examined in a strain of guinea pigs with a total deficiency of hemolytically active C4. Development of delayed hypersensitivity to three antigens (ovalbumin, DNP-BGG and Mycobacterium tuberculosis) was normal. The animals had a normal exudative response to a foreign body implanted in a subcutaneous pocket. Normal passive cutaneous anaphylaxis reactions were elicited with both guinea pig and rabbit antibodies. Unexpectedly, C4 deficient guinea pigs immunized with ovalbumin and DNP-BGG displayed normal direct Arthus reactions to these antigens. They demonstrated normal reverse passive Arthus reactions to intravenous injection of human serum albumin (HSA) following intradermal injection of guinea pig or rabbit anti-HSA. These findings suggest that these animals possess a mechanism to by-pass C4 and directly activate late acting complement components. Such a mechanism was clearly demonstrated in experiments performed in vitro.

In a limited number of studies, findings with regard to antibody response were complex.
Purification and Reaction Mechanism of Guinea Pig C9
Authors: Rommel F. A. , Mayer M. M.
Published: December 2022

C9 was isolated in a highly purified state by chromatography of guinea pig serum pseudoglobulin on DEAE-cellulose, hydroxyapatite and Sephadex G-200, as well as by disc electrophoresis on polyacrylamide gel. The final product, obtained in a yield of about 2%, gave a single band on analytic disc electrophoresis on polyacrylamide gel and a single arc on immunoelectrophoresis with an antiserum against crude C9. About 1 to 2 molecules of C9 were required for lysis of one cell in the state EAC ∼ 8.

The initial reaction velocity between C9 and EAC ∼ 8 at 0°C was found to be directly proportional to the concentration of either of the reactants. It was essentially unaffected by change of pH within the range 6.2 to 8.2. The initial reaction rate increased approximately twofold between 0°C and 30°C. The optimal ionic strength was 0.074. C9 is consumed in the reaction with EAC ∼ 8, presumably by fixation.
Complement and Immunologic Reactions in Vivo
Authors: Cochrane C. G. , Henson P. M.
Published: December 2022

The role of complement in immunologic inquiry of the glomerulus has been studied. Sheep antibody to rabbit renal basement membrane (ARBM), when injected into rabbits, induced glomerulitis and proteinuria dependent in part upon the participation of host neutrophils. Neutrophil depletion diminished the level of proteinuria but the degree varied from donor to donor and with the time of immunization of the sheep. Fractionation of the ARBM on DEAE-Sephadex revealed different zones of antibody activity: in the fast γ region, antibody was obtained that was greatly dependent upon neutrophils for its action, while in the slow γ regions, antibody was found that produced glomerular injury not dependent upon neutrophils. The injury produced by the neutrophil independent antibody was not affected by C3 depletion with cobra venom factor (CoF) and occurred equally well in C6 deficient rabbits.
Blocking Serum Lysis of Brucella Abortus by Hyperimmune Rabbit Immunoglobulin A
Authors: Hall Wendell H. , Manion Robert E. , Zinneman Horace H.
Published: December 2022

Immunization of rabbits with concentrated Brucella abortus antigens for 10 weeks led to nearly total loss of serum bactericidal activity for this organism. β and γ globulin fractions of hyperimmune serum obtained by starch block electrophoresis had bactericidal activity. Sephadex G-200 gel filtration of the β globulin fraction obtained by starch block electrophoresis yielded a fraction consisting of IgM and another containing IgG and IgA; both had bacteriolytic activity. The latter subfraction was separated by column chromatography on DEAE-Sephadex into IgG and IgA (probably contaminated by 15% to 20% IgG). Bacteriolytic tests indicated that IgA had a prozone of inhibition of bacteriolysis, whereas IgM and IgG did not. Addition of excess IgA inhibited bacteriolysis produced by IgG and IgM. Bactericidal kinetics disclosed little kill by whole hyperimmune serum. Hyperimmune IgM and IgG killed Br, abortus rapidly but IgA fractions gave a lag of more than 3 min before any kill was measured. Addition of excess IgA markedly retarded the bactericidal kinetics of both IgG and IgM.
Biopolymer Membrane: A Model System for the Study of the Neutrophilic Leukocyte Response to Immune Complexes
Authors: Hawkins David
Published: December 2022

A biosynthetic collagen polymer membrane on which specific antibody was linearly deposited provided an in vitro model of anti-glomerular basement membrane antibody nephritis. Exposure of neutrophilic leukocytes to this membrane induced adherence and marked spreading of the cells over the polymer surface. This was associated with release of bioactive substances from the neutrophils and indicated that immunologically induced release of inflammatory factors could occur in the absence of phagocytosis of immune complexes.
The Immunosuppressive Effect of Rabbit Anti-Mouse Macrophage Serum
Authors: Isa Abdallah M.
Published: December 2022

The suppressive effect of anti-macrophage serum (AMS) on the serum antibody response to trachoma agent antigens was studied in white Swiss mice. When given on days -1, 0 and +1, the antigen being given on day 0, AMS caused clumping of peritoneal macrophages but not peritoneal lymphocytes or spleen cells and interfered with the phagocytic activity of peritoneal macrophages. AMS also inhibited the primary immune response to trachoma agent antigens and specifically inhibited the production of IgM, IgA and IgG2 (7S γ2) antibodies.
Presidential Address
Authors: Waksman Byron H.
Published: December 2022

Fellow members of the Society, friends, and guests: The addresses of retiring presidents, which I have had the pleasure of hearing over the last 25 years, have fallen into several predictable categories. Three quarters have been reviews of a field or have dealt with the individual speaker's personal research and its consequences. The remainder have dealt with broader topics or with the state of our society. I have chosen to prepare an essay of the latter type.

Science at present is confronted by a crisis of public confidence. Basic research appears to many thoughtful citizens increasingly remote from applications in the sphere of daily life and the applications, when they are visible, often seem to harm more than they help. In the face of the numerous problems created by increasing population, increasing urbanization of our society, and increasing individual consumption and waste, it is not surprising that the priority of basic investigations in the competition for public funds has declined (1).
Increased Complement Efficiency through Control of Ionic Strength and Temperature
Authors: Linscott William D.
Published: December 2022

When complement (C) was in relative excess and IgG antibody (Ab) was limited, a great increase in hemolytic efficiency was obtained by incubating sensitized red cells with C for 1 hr at 0°C, µ = 0.037, followed by 1 hr at 37°C, µ = 0.094. When IgG Ab was in relative excess and C was limited, this effect was much smaller. C1 binding affinity seemed to be the most important factor in determining the hemolytic efficiency. With IgM Ab, a smaller but still significant (65%) increase in hemolytic efficiency was obtained by a one-step incubation at 37°C, µ = 0.094. When direct lysis by whole C was compared with C1 fixation and transfer, up to 20% of the IgG sites capable of fixing C1 could be converted to S* under optimal conditions of temperature and ionic strength.
Cell Separation on Affinity Columns: The Preparation of Pure Populations of Anti-Hapten Specific Lymphocytes
Authors: Wofsy Leon , Kimura John , Truffa-Bachi Paolo
Published: December 2022

Procedures were developed for purifying anti-hapten specific lymphocytes on an immunoadsorbent. Mouse spleen cells were filtered through large polyacrylamide beads with covalently attached β-lactoside hapten groups. Specifically binding cells were eluted with hapten from washed affinity beads. Hapten-eluted spleen cell populations derived from non-immune mice were much smaller in number than those from mice immunized against hemocyanin azophenyl-β-lactoside. Pure populations of anti-hapten specific cells from immune spleens were greatly enriched in indirect plaque-forming cells against β-lactoside-coupled sheep red blood cells.
Arthritis Associated with Induced Rubella Infection
Authors: Ogra Pearay L. , Herd J. Kenneth
Published: December 2022

Rubella antibody activity in serum, nasopharynx and joint aspirates, and recovery of rubella virus from joint aspirates was studied in three previously well children, and one child with pre-existing rheumatoid arthritis, who developed repeated episodes of arthritis with effusion for several months following parenteral immunization with live attenuated rubella vaccine (HPV-77 DK/12). Three months after immunization, rubella antibody activity in serum was present in the γG and to a smaller extent in γM class of immunoglobulin. No rubella antibody response was demonstrable in the nasopharynx. The response in joint aspirates was limited to the γG class of immunoglobulin in all children studied. Rubella virus was recovered from the joint aspirates of all previously well children for as long as 3 to 4 months after immunization with rubella virus vaccine. It is suggested that repeated episodes of arthritis which follow rubella immunization may be related to the continued presence of rubella virus in the affected joints, and to a selective distribution of rubella virus antigen in synovial and other body fluids.
Degradation of Human IgM by Pepsin: Characterization of a High Molecular Weight Fragment
Authors: Schrohenloher Ralph E. , Bennett J. Claude
Published: December 2022

Digestion of a Waldenström's IgM by pepsin for 3 hr at 37°C in pH 4.1 buffer resulted in the production of a fragment having an so20 w value of 15S. The molecular weight of the 15S fragment, as determined from its sedimentation and diffusion coefficients, was approximately 700,000, while that for the undigested IgM was 840,000. Smaller quantities of F(ab′)2µ and Fab′µ fragments were also found. Reductive cleavage of the high molecular weight fragment demonstrated the pressence of 7S subunits and a small quantity of lower molecular weight materials. The similarity of the subunits derived from the fragment to those from the undigested IgM was established by sedimentation, diffusion and molecular weight studies. The data indicated the presence of four subunits in the 15S fragment. Immunoelectrophoresis and Ouchterlony agar diffusion analysis failed to reveal differences in the subunits from the fragment and the undigested IgM. Tyrptic peptide mapping studies of the chains isolated from the undigested IgM and the 7S subunit of the fragment after extensive reduction and alkylation demonstrated only minor differences. The results indicate that the initial phases of the digestion of IgM by pepsin involve an attack at a single subunit of the molecule.
Characteristics of the Lambda Chain Class in Mice
Authors: Weigert Martin , Cesari Italo , Geckeler William R. , Blomberg Bonnie , Cohn Melvin
Published: December 2022

Mouse λ chains exhibit a simple pattern of variability. This is shown by a comparison of the amino acid sequences of the variable region of 12 λ chains produced by plasmacytomas of BALB/c mice. Eight of the 12 λ chains are identical, two differ from these by one residue, one by two residues, and one by three residues. Our interpretation of this finding is that λ chains are coded for by a single germ-line gene for both vλ and cλ that determines the structure represented by the eight identical sequences, and the variant sequences are the result of single point mutations. We postulate that these somatic mutants in vλ are selected for in a sequential fashion by antigen because all of the amino acid substitutions fall into that region of the L-chain that contributes to the antibody combining site (Wu, T. T. and Kabat, E. A., J. Exp. Med., 132: 211, 1970).
Amino Acid Sequence Studies of the Hypervariable Regions of Human Immunoglobulin Heavy Chains
Authors: Capra J. Donald , Kehoe J. Michael
Published: December 2022

Hypervariable regions of immunoglobulins are thought to be the areas of the immunoglobulin molecule involved in antibody complementarity. Three hypervariable regions have been described in immunoglobulin light chains (24 to 34, 50 to 56, 89 to 97). Since immunoglobulin heavy chains are presumed to share a common evolutionary origin with light chains it has been presumed that similar hypervariable areas would be found in the heavy chains as well. Eight human immunoglobulin heavy chains of the HVIII subgroup were sequenced to step 40 utilizing a Beckman model 890 Protein Sequencer. From the N terminus to residue 30, variation was minimal. Residues 31 through 35 showed extreme variability. A gap was inserted in position 36 and 37 to maximize homology with HVII proteins. Positions 38 to 40 were invariant. By CNBr cleavage a fragment was obtained from several HVIII proteins starting at position 85.
Physicochemical, Immunological and Structural Studies on Two M-Components from a Single Individual
Authors: Freedman M. , Goldrosen M. , Pruzanski W.
Published: December 2022

Serum from a patient with multiple myeloma contained two elevated monoclonal IgG M-components which differed with respect to electrophoretic mobility. Prolonged melphalan therapy reduced the serum concentration of the slow component (γ-electrophoretic mobility, pI 7.1) to normal levels but had no effect on the fast component (β-electrophoretic mobility, pI 6.8). The two M-components were isolated by column chromatography, on DEAE-cellulose and Sephadex G-200, and by isoelectric focusing. Both isolated heavy chains belonged to γG1 subgroup (a-b-f+ allotypes), possessed glutamic acid as the N-terminal amino acid, and were indistinguishable by acid and alkali urea starch gel electrophoresis, amino acid analysis, peptide mapping, and amide content. The isolated light chains of both M-components were λ(III) [Oz (-), St (-) II], possessed tyrosine as the N-terminal amino acid and were indistinguishable by their initial N-terminal sequence [tyr, valleu, leu, thr], circular dichroism spectra (240 to 310 nm), amino acid analysis, and mobility in acid urea starch gel electrophoresis.
The Immediate Allergic Response: in Vitro Separation of Antigen Activation, Decay and Histamine Release
Authors: Lichtenstein Lawrence M.
Published: December 2022

Antigen-induced, IgE-mediated histamine release can be separated into two stages: an antigen (or anti-IgE)-dependent, calcium-independent first stage, and an antigenindependent, calcium-dependent second stage. In the first stage at 37°C the cells are rapidly activated and, if the reaction proceeds longer than the optimal time (2 to 4 min), the activated state decays and leaves the cells unresponsive to further challenge by the homologous antigen or by a non-cross-reacting antigen. A 4°C first stage can also be generated which includes the antigen-antibody interaction but lacks a temperature sensitive chlorphenesin inhibitable step or steps. If separated from antigen after 2 to 4 min of the first stage, the activated cells are “stable” at 4°C for at least 30 min. The second or histamine release stage appears to begin immediately upon providing the activated cells with calcium. These experiments demonstrate the generation, by antigen, of an activity, presumably an enzyme or enzyme product, which is coupled to subsequent release stages by calcium; if uncoupled by calcium lack the enzyme/product is wasted and the cell becomes “desensitized.” They also provide a number of cell intermediates for experimental manipulation.
Antigen-Binding Cells
Authors: Unanue E. R.
Published: December 2022

Spleen, thymus and peritoneal cells of immunized mice were incubated with radioactive antigens and studied by autoradiography. A small number of lymphocytes in the spleen (0.1% to 0.5%) but very few in the thymus (0.01%) bind radioactive hemocyanin. The binding of antigen to lymphocytes apparently takes place by antibody receptors on their membranes. Exposure of most lymphocytes to antibodies to Ig blocked the binding. It was also found that the number of cells binding weak or strong immunogens was similar. Lymphocytes have been found that bind an autologous antigen, murine growth hormone. No lymphocytes bind mouse albumin. Incubation of spleen cells from immune or non-immune mice with highly radioactive KLH in vitro suppressed the capacity of the cells to adaptively transfer an immune response in x-irradiated mice. The inactivation of the adoptive immunity was specific since the treated cells made antibody to unrelated antigens. Macrophages took up radioactive antigen and this uptake was not blocked by exposure to anti-Ig.
Univalent Arsanilic Acid Derivatives as Elicitors of Passive Cutaneous Anaphylaxis
Authors: Rosenberg Leon T. , Amkraut Alfred A. , Corp Richard B. , Raffel Sidney
Published: December 2022

A number of monovalent p-azobenzenearsonate derivatives of tyrosine elicit passive cutaneous anaphylaxis (PCA) reactions in guinea pigs sensitized with some, but not all, rabbit or guinea pig antisera raised by immunization with haptenated protein. Our data suggest that certain populations of cell-binding antibodies mediate these reactions. Reversibility of antibody-hapten interaction is indicated by failure of an eliciting injection of hapten to desensitize a site. In two hapten systems so far studied, this one and a series of dinitrophenylated compounds, certain monovalent antigens were found to be capable of eliciting cutaneous anaphylactic reactions.
Mechanisms of Passive Sensitization
Authors: Tomioka Hisao , Ishizaka Kimishige
Published: December 2022

E myeloma protein was injected intracutaneously into monkey skin to produce passive sensitization. The sensitized and nonsensitized sites were challenged with 125I-labeled purified anti-IgE. The results of autoradiography of the tissues indicated that IgE was present on mast cells. When the E myeloma protein was injected intravenously into monkeys, the antigen was detected on mast cells in small intestine, bronchi and omentum but not on other tissue cells. The E myeloma protein was detected on basophil granulocytes in the blood, but not on other leukocytes. Evidence was obtained that both mast cells and basophil granulocytes contain monkey IgE, and that E myeloma protein combines with the same cells upon passive sensitization. IgG was detected on neutrophil granulocytes, monocytes and macrophages but not on mast cells.
Transformation of Lymphocytes from Animals Sensitized to Trichinella Spiralis
Authors: Kim C. W. , Jamuar M. P. , Hamilton L. D.
Published: December 2022

Guinea pigs sensitized once or hypersensitized four times at bi-weekly intervals in the footpads with Trichinella extract, emulsified in complete adjuvant, were examined for serum antibody content and tissue cellular changes. The following modifications were found in the two groups. Those sensitized, when homologously skin-tested at 1 week, showed a typical delayed hypersensitivity reaction, whereas the hypersensitized group, when similarly skin-tested 2 weeks after the fourth injection, showed an accelerated reaction which appeared somewhat like the Arthus type. The difference in skin reactivity of the two groups was not associated with the kind of cells present at the site but rather with a marked difference in antibody content of the serum and in the absolute and relative number of mononuclear cells present at the site and in the lymph nodes. The cellular content of the subcutaneous tissue at the site of the skin test was characteristic of delayed hypersensitivity in both groups as evidenced by a predominant perivascular infiltration of mononuclear cells. The hypersensitized group, as compared with the sensitized group, showed 1) more circulating antibody, as measured by passive cutaneous anaphylaxis and the precipitin ring test, 2) a greater proportion, as compared to small lymphocytes, of mediumand large-sized lymphocytes, macrophages, plasma cells and eosinophils, 3) a greater antibody content per mononuclear cell in the lymph nodes, as measured by fluorescent staining, and 4) more deaths (50% vs 0%) when tested by active systemic anaphylaxis. The four differences between the sensitized and hypersensitized groups were evident whether the groups were homologously skin-tested or not. Primed lymphocytes when incubated for 5 days in vitro with the Trichinella antigen, however, showed more transformation into “blast-like” cells in the skin-tested as compared with the non-skin-tested groups of both sensitized and hypersensitized guinea pigs; but there was no difference between the sensitized and hypersensitized groups.
The Reactions of Pepsin and Pepsin-Trypsin Digestion Products of Bovine Serum Albumin with Antisera from Rabbits Ingesting This Protein
Authors: Wright Ruel N. , Rothberg Richard M.
Published: December 2022

The effect of digestion by alimentary enzymes on the antigenicity of bovine serum albumin (BSA) was studied by the sequential exposure of this food antigen to pepsin and trypsin under conditions simulating mammalian digestion. The physical degradation was assessed by G-100 chromatography which demonstrated three different molecular weight groups. The first peak of the pepsin digest eluted in the same area as undigested BSA and the last peak was composed mainly of dialyzable peptides. The major alteration caused by trypsin digestion was a decrease in the amount of peak 1. A comparison of the antigenicity of these digests with that of BSA demonstrated that they contained all the antigenic sites of BSA, but in concentrations approximately 1/100 that in the intact molecule. Peak 1 also appeared to have all these sites in an unaltered form whereas the sites in peak 2 appeared to be altered by the digestion. New antigenic sites were demonstrated in peaks 1 and 2 by using antisera from rabbits parenterally immunized with the pepsin digest. However, antisera from animals orally immunized to BSA reacted with the intact protein and the digestion products in a manner identical to antisera from rabbits subcutaneously immunized with BSA. These data suggest that enough surface antigenic sites survive alimentary digestion to sensitize the lamina propria of rabbits. In addition, the recognition of only surface antigenic sites by antisera from orally immunized rabbits implies that these animals are not immunized to the internal determinants uncovered by alimentary digestion.
Effect of Selected Lipids and Surfactants on Immunogenicity of Several Bacterial Antigens
Authors: Agarwal Manjul K. , Neter E.
Published: December 2022

Several biogenic and non-biogenic lipids and two surfactants were tested to determine their effect on the immune response to two common bacterial antigens. After incubation in vitro, antigen and test substances were injected intravenously into groups of rabbits, and the antibody response was quantitated by means of the passive hemagglutination test. The effect on priming was ascertained by the documentation of an enhanced antibody response 3 or 4 days following the secondary injection of a minimally effective dose of immunogenic antigen. The following results were obtained: (1) Triton X-100, in amounts of 4 to 500 µg/ml, when injected together with the common enterobacterial antigen, inhibited the immune response as well as immunologic priming. Immunosuppression also occurred when antigen and Triton X-100 were administered separately, although concurrently. However, the compound was ineffective, or only minimally effective, when given 24 hr before or after the antigen. (2) Methyl palmitate also inhibited the primary immune response, when administered together with the antigen. In contrast to Triton X-100, an undiminished immune response followed separate, albeit concurrent, injection of drug and common enterobacterial antigen. (3) Cholesterol reversed the immunosuppressive effect of both Triton X-100 and methyl palmitate. (4) Triton X-100 suppressed the antibody response also to the common antigen of Gram-positive bacteria, but reduced only slightly the antibody response to O antigen of Escherichia coli O111. (5) Both Tween-20 and gangliosides also interfered with the hemagglutinin response of the rabbit to common enterobacterial antigen, but neither of these two agents, nor Triton X-100 and methyl palmitate, inhibited attachment of the common enterobacterial antigen to red blood cells. Furthermore, these compounds did not alter the antigenic determinant, as indicated in hemagglutination inhibition tests by undiminished antibody-neutralizing capacity of the antigen in the presence of the drugs. (6) Glyceryl trioleate, sphingomyelin, B-carotene, cerebrosides, and cephalin (brain extract) did not significantly inhibit the hemagglutinin response to common enterobacterial antigen. (7) It is concluded that gangliosides, Triton X-100, methyl palmitate, and Tween-20 belong to the group of antigen-associated immunosuppressants, and the former two compounds, in addition, inhibit immunologic priming by certain bacterial antigens.
Noncovalent Interaction Between Immunoglobulin (Ig) Polypeptide Chains
Authors: Zimmerman Barry , Grey Howard M.
Published: December 2022

A number of studies have demonstrated the feasibility of recombining light and heavy chains from a variety of different sources to yield four chain molecules having a conformation indistinguishable from native molecules by several independent criteria. Thus, although competitive recombination experiments have demonstrated preferential reassociation of autologous chains (1–4), hybridization with heterologous light chains, including light chains derived from Igs of different mammalian species, was also readily accomplished (5).

Other experiments have shown that the reconstitution of rabbit half molecules results in random hybridization of γ heavy chains creating heteroligating 7S antibody molecules (6) and hybrid H chain allotype pairings (7).

The present investigation grew out of published data supporting the concept that during its antibody-secreting lifetime a single cell may be capable of “switching” the class of its antibody product from IgM to IgG (8, 9). This premise raised the possibility that during the period of “switching over” immunoglobulin “monsters” might be synthesized containing both µ and γ heavy chains.
Response of Peripheral Blood Lymphocytes from Normal, Hypogammaglobulinemic and Chronic Lymphocytic Leukemic Patients
Authors: Rodey G. E. , Davis T. , Quie P. G.
Published: December 2022

Staphylococcal protein A (SPA) derived from Cowan I strain of Staphylococcus aureus was cultured in vitro with peripheral blood leucocytes of normal donors and of patients with various diseases. Cells from the majority of normal donors proliferate in the presence of SPA. Maximum tritiated thymidine incorporation occurs with 25 to 50 µg SPA per culture and when cultures are incubated 7 to 9 days. Removal of glass adherent cells prior to culture markedly reduces the proliferative response to SPA. Cells of three patients with hypogammaglobulinemia responded vigorously to SPA. These findings suggest that the response to SPA is antigen-specific, rather than nonselective, and that responding lymphocytes are thymus-derived, T-cells. The in vitro model described may provide a tool to study the role of cellular immunity in defense against staphylococci
Anti-Hapten Antibodies of Restricted Heterogeneity: Studies on Binding Properties and Component Chains
Authors: Hoffman Donald R. , Grossberg Allan L. , Pressman David
Published: December 2022

Antibodies against p-azobenzoate (Xp) from two rabbits that exhibited restricted heterogeneity were studied by isoelectric focusing of whole antibody and of light and heavy chains.3 One antibody (2709) showed two light chain bands and five heavy chain bands. Binding studies on 2709 showed two homogeneously binding components with a 100-fold difference in binding constant for p-iodobenzoate. The other antibody (2883) had about 15 components detectable by isoelectric focusing with three major light and five heavy chain bands. A sample of antibody 2883 was fractionated by sucrose gradient isoelectric focusing and seven of the fractions were further characterized. Binding studies with four haptens indicated that fractions A, B and C contained antibodies with similar binding properties and fractions F and G contained other antibodies with similar binding properties. Isoelectric focusing of mildly reduced and alkylated samples of the fractions showed that the antibodies in fractions A, B and C gave a light chain band not found in the other fractions and that the antibodies in F and G gave a large amount of the third light chain band. Although some of the fractions gave a single significant band on gel isoelectric focusing, each of them gave at least three heavy chain bands. This indicated that post-synthetic charge modifications are not necessarily symmetrical. Isoelectric focusing of mildly reduced and alkylated antibodies has much higher resolution than disc electrophoresis of either light or heavy chains.
The Immunosuppressive Effects of Rauscher Leukemia Virus (RLV) upon Spleen Cells Cultured in Cell-Impermeable Diffusion Chambers
Authors: Borella Luis
Published: December 2022

In mouse spleen cell cultures, keyhole limpet hemocyanin (KLH) inhibited the leukemogenic process induced by Rauscher leukemia virus (RLV). Spleen cells obtained from normal or RLV-infected BALB/C mice were cultured with or without KLH in cell-impermeable diffusion chambers that were implanted in isogeneic hosts. The effects of RLV and KLH upon these cells were determined by quantitative analysis of cell morphology and by measurement of in vitro immunoglobulin synthesis. Increases in blast, myeloid, erythroid and monocytoid cells were found in RLV-infected cultures that were not restimulated with KLH. In RLV-infected cultures that were restimulated with KLH, numbers of blasts and hematopoietic cells were within normal range, but there was a marked increase of plasma cells and of in vitro immunoglobulin synthesis. It is postulated that antigenic stimulation by KLH of RLV-infected cultures shifts the differentiation pathways of a pluripotent stem cell from a blast-hematopoietic cell line into a lymphoid-plasma cell line.

The finding that antigenic stimulation of infected cell cultures alters the cellular pathway induced by a leukemogenic virus may provide a new approach to the investigation of regulatory mechanisms and control of cell proliferation and differentiation during leukemogenesis.
Differential Stimulation of Precursor Cells and Carrier-Specific Thymus-Derived Cell Activity in the in vivo Response to Heterologous Erythrocytes in Mice
Authors: Falkoff Reuben , Kettman J.
Published: December 2022

We have found that sub-immunogenic doses of sheep red blood cells (SRBC) in vivo increase the plaque-forming cell (PFC) responses to trinitrophenyl and to SRBC when spleen cells are challenged in vitro using TNP-SRBC as antigen.

By mixing irradiated spleen cells from low dose-primed mice with normal spleen cells, the enhancing activity is shown to be radiation resistant. Treatment with anti-BAϑ abolishes the activity. We therefore infer that it resides in a thymus-derived helper-cell population.

While the enhancing activity is maximal for the carrier to which the mice were low dose primed, priming with SRBC gives a substantial enhancement of the response when burro RBC are used as carrier, but little enhancement when chicken RBC are used as carrier. There is little cross-reactivity between SRBC and burro RBC at the direct PFC level in this system, demonstrating that cross-reactivity at the T cell level need not parallel that at the B cell level.
Cell Interactions in Antibody Formation in Vitro
Authors: Cosenza Humberto , Leserman Lee D.
Published: December 2022

Spleen cells from DBA/2J and CBA/J mice were separated into adherent and nonadherent cell populations; adherent cells obtained from either strain may be recombined with DBA nonadherent cells and the kinetics and magnitude of the plaque-forming cell response to SRBC are equivalent. An alloantiserum directed against CBA spleen cells was used to study the length of time that the adherent population was required for a maximum in vitro response. The alloantiserum abolished the response when added during the first 2 days of culture, indicating that the adherent cell was required at least during the inductive phase of the response. This was true whether adherent cells were prepared from either spleen cells or peritoneal exudate cells of CBA mice.

Nonadherent cells from mice immunized 1 week earlier required the cooperative participation of adherent cells for a shorter period than nonadherent cells from normal or remotely immunized mice. This result is consistent with the idea that most of the adherent cell dependent events have occurred in the nonadherent population following exposure to antigen in vivo. The present study demonstrates that a cell type in the adherent population is an active participant in the cell interactions leading to antibody formation in both primary and secondary in vitro responses. It is proposed that the adherent cell is an “antigen-focusing” cell which functions by providing foci of increased cell density thereby promoting interactions between thymus and bone marrow-derived cells.
The Use of Killed Vaccines in Immunization against an Intracellular Parasite: Toxoplasma Gondii
Authors: Krahenbuhl James L. , Ruskin Joel , Remington Jack S.
Published: December 2022

The pretreatment of mice with antigens prepared from killed Toxoplasma gondii induced long term protection against challenge with this organism. Resistance was measured in terms of length of survival and number of deaths following challenge with graded doses of the relatively avirulent C56 strain of Toxoplasma, and was observed when antigen was administered alone or incorporated into Freund's incomplete adjuvant. Freund's complete adjuvant also afforded significant protection. A protective role for humoral antibody was observed as significant resistance following the passive transfer of high titer anti-Toxoplasma antisera.
Immunization with R Mutants of S. Minnesota
Authors: McCabe William R. , Greely Anna
Published: December 2022

Recent studies have demonstrated that the basal core structure of the lipopolysaccharides of most Gram-negative bacilli are of similar or identical chemical structure. The current investigations evaluate the protective effect of immunization with these shared antigens against challenge with two heterologous Gram-negative bacilli, K. pneumoniae and E. coli 107. Active and passive immunization of mice with the Ra, Rb, Rc and Rd1 mutants of S. minnesota failed to protect against challenge with K. pneumoniae. In contrast, active and passive immunization with the Rd2 and Re mutants of S. minnesota afforded significant protection against challenge with K. pneumoniae. Similarly, active immunization with Re and passive immunization with Ra, Rc, Rd2 and Re provided significant protection against challenge with E. coli 107. In all instances, immunization with Re afforded greater protection than any of the other rough mutants of S. minnesota. Active immunization with the Re mutant was considerably less effective than type specific immunization. Immunization with Re increased the LD50 of E. coli 107 by 10-fold and K. pneumoniae by 100-fold over that of controls, while type specific immunization increased the LD50 to these organisms by greater than 100-fold and 10,000-fold respectively. The specificity of the protection induced was demonstrated by the loss of the protective activity of rabbit antiserum to Re after adsorption of the serum with dead Re mutants.
Cell Surface Immunoglobulin
Authors: Baur Sonia , Schenkein Isaac , Uhr Jonathan W.
Published: December 2022

Enzymatic radioiodination has been used to investigate the cell surface of three murine myeloma tumors that secrete Ig. In each cell type studied (IgG1, IgG2a and IgA), radiolabeled Ig can be obtained from the cell surface by specific precipitation. In one cell line, surface IgG was also identified by digestion with trypsin followed by fingerprinting as well as by digestion with papain into characteristic Fab and Fc fragments. The Fc fragment contained a significant amount of radioactivity consistent with the concept that the IgG molecule is fully exteriorized. In each plasma cell line studied, Ig was associated with a molecular weight equal to or greater than 200,000 daltons which may be the complex between Ig and the molecule(s) on the plasma membrane to which it is attached.
Effect of C3 Inactivator on Bound C3 Antigen
Authors: Leonard Edward J. , Borsos Tibor
Published: December 2022

The effect of C3 inactivator (C3ina) in human EDTA plasma on C3 antigen bound to various immune complexes was determined. C3ina greatly decreased the amount of C3 bound to human erythrocytes sensitized with IgM anti-I cold agglutinin, and also decreased erythrocyte-bound C3 of a patient with deficiency of C3 inactivator, but had little effect on C3 bound to human red cells sensitized with Donath-Landsteiner IgG cold agglutinin. C3ina did not alter the amount of human C3 antigen bound to sheep E, to pneumococcal polysaccharide-antibody precipitate and to Sepharose-2B beads. Although C3ina did not alter the amount of C3 antigen on sheep E, a change in the molecule must have occurred since immune adherence was abolished. These data along with evidence from the literature show that several stages of bound C3 inactivation can occur.
Phylogeny of Immunocompetent Cells
Authors: Goldshein Stuart J. , Cohen Nicholas
Published: December 2022

Toad splenic lymphocytes undergo blastogenic conversion in vitro in response to both phytohemagglutinin (PHA) and allogeneic splenocytes. At 26°C, maximum incorporation of tritiated thymidine occurs on day 3 in response to PHA, and on day 7 in a mixed lymphocyte culture (MLC). The response to PHA is concentration-dependent. The maximum response to PHA and to alloantigens varies from toad to toad, and is influenced by the type of serum used to supplement the culture medium. More lymphocytes cultured with xenogeneic serum and PHA undergo mitosis than those cultured with PHA and allogeneic serum. In contrast, allogeneic serum suppresses transformation in the MLC.
Receptors for Immunoglobulin on B Lymphocytes and Cells of a Cultured Plasma Cell Tumor
Authors: Cline Martin J. , Sprent Jonathan , Warner Noel L. , Harris Alan W.
Published: December 2022

Mouse B lymphocytes and cells of a plasma cell tumor bound immune complexes and IgG but not IgA or IgM immunoglobulins or light chains. Binding required an intact Fc piece but was independent of complement. T lymphocytes did not bind immune complexes.

The presence of surface receptors for IgG immunoglobulin on B lymphocytes and a plasma cell variant are consistent with the concept of an ancestor-progeny relationship between these cell lines.
A Receptor for Fc on Mouse B-Lymphocytes
Authors: Paraskevas F. , Lee S-T. , Orr K. B. , Israels L. G.
Published: December 2022

Antigen-antibody complexes derived from rabbit antibodies bind to mouse spleen lymphocytes. This binding was detected by the inhibition of reverse immune cytoadherence (RICA), a technique detecting surface γ globulin and suggests that these complexes bind to γ globulin carrying B-cells. Complement is not a requirement for binding. Complexes made from 5S bivalent antibodies do not bind nor does 7S antibody or normal γ globulin. Chemically produced microaggregates of ferritin show no binding. Mouse complexes made with mouse γG (7Sγ2a) antibodies also bind to mouse spleen cells.

Fc fragments isolated by papain digestion of rabbit antibodies also inhibit RICA and Fc crystals can form around the mouse spleen lymphocytes. Fc from normal rabbit γ globulin is less inhibitory. These results are presented as evidence of the existence of a receptor, provisionally called the Fc receptor, on the surface of mouse spleen lymphocytes which are probably B-cells.
Association of Synthetic Polypeptides with Ribonucleic Acid of Immunocompetent Tissues
Authors: Smyth Robert D. , Maurer Paul H.
Published: December 2022

The association of 125I-labeled synthetic polypeptides with ribonucleic acid (RNA) of rabbit peritoneal exudate (p.e.) cells in in vitro systems has been shown to be a cell-mediated reaction, related to the charge, state of aggregation and optical configuration of the polymer. Although slightly higher levels of several D-amino acid polymers were associated with p.e. cells in in vitro systems, the concentrations of p.e. cell RNA-associated 125I-polypeptides of the L-configuration were significantly greater than the D-form. Intraperitoneal administration of the L or D configuration of the polymer—(Glu60Ala30Tyr10)n—(GAT) to oil-stimulated rabbits resulted in p.e. cell uptake and RNA-association of both isomeric polypeptide forms. Whereas the levels of cellular and RNA-associated L-polypeptide declined rapidly over a 24-hr period, the respective concentrations of the D-polymer significantly increased during this period. Similar observations were noted following i.v. administration of D- or L-GAT to normal or hyperimmune animals. Levels of 125I L-GAT declined rapidly over a 24-hr period in kidney, bone marrow, liver, spleen and lymph node. Tissue and RNA-associated levels of 125I D-GAT remained fairly constant for 7 days after administration. There was essentially no correlation between in vitro and in vivo studies with respect to formation of RNA-antigen complexes. The significance of tissue distribution and polypeptide metabolism in the intact animal, its role in the formation of RNA-antigen complexes and induction of the immune response appear to preclude any oversimplification in attempting to correlate data from a unicom-partmental in vitro cell system with the dynamic multicompartmental in vivo systems.
Synthesis of a Photo-Activated Menadione (VIT K1) Affinity Label and Its Reaction with a Menadione-Binding Myeloma Protein
Authors: Rosenstein Robert W. , Richards Frank F.
Published: December 2022

Photolytically derived carbenes were first used by Westheimer's group (1) as labeling agents for proteins. The carbenes and the related nitrenes (2) are useful agents for labeling receptor sites on proteins where a primary, non-covalent complex can be formed between protein and label. Such complexes can subsequently be triggered by near-UV light to give a carbene or nitrene capable of reacting covalently with amino acid side chains. Diazoketone reagents have been used with myeloma proteins and antibodies (3, 4) and supplement the information given by other types of affinity reagents labeling residues present at or near the combining site (5–7).

A γA myeloma protein (protein 460) derived from the mouse tumor line MOPC 460 binds menadione with a K° in the order of 104 l/mol. This myeloma protein also binds other haptens with higher binding constants elsewhere in the combining region (8). We have used an affinity label derived from menadione to study the primary and three-dimensional structure of immunoglobulin-combining sites.
Complement-Mediated Tissue Damage: Contribution of the Classical and Alternate Complement Pathways in the Forssman Reaction
Authors: May Joseph E. , Frank Michael M.
Published: December 2022

The contribution of the classical and alternate complement pathways to complement-mediated tissue damage in the Forssman reaction was studied in a strain of C4D guinea pigs with a nonoperative classical pathway, but an intact alternate pathway. It was found that the C4D guinea pigs were resistant to the Forssman reaction, but that reconstitution of the classical pathway by the administration of normal guinea pig serum as a source of C4 restored the deficient animals' susceptibility to the toxic effects of injected Forssman antiserum. C4D animals with a restored C4 level only developed the Forssman reaction if the Forssman antiserum was injected many hours after the injection of C4. Thus, a period of time was required for C4 to distribute within the extravascular compartment. These results indicate that interaction of anti-Forssman antibody with extravascular Forssman antigen and complement is necessary for lethality, and that vascular endothelial damage cannot alone account for the pathogenesis of the Forssman reaction, as previously believed. The results also indicate that intravascular C4 is not freely available to tissue sites.
The Effect of Immunoregulatory α Globulin (IRA) upon Lymphocytes in Vitro
Authors: Cooperband Sidney R. , Badger Alison M. , Davis Robert C. , Schmid Karl , Mannick John A.
Published: December 2022

An immunosuppressive factor has been extracted from α globulin containing fractions of human plasma proteins, which we have called “immunoregulatory α globubin” (IRA). Cohn fraction IV has been found to contain this material. The IRA has been assayed by its ability to inhibit human blood lymphocyte proliferation induced in vitro by phytohemagglutinin (PHA). The IRA has been effective in reducing PHA-induced proliferation by 75% at doses as low as 0.2 mg/106 cells in 1 ml of culture. Most fractions, however, were active at 2 to 3 mg/ml. We have also found the IRA to suppress lymphocyte proliferation induced by anti-human thymocyte serum and specific antigens. The concentration of IRA necessary to inhibit these weaker mitogenic stimuli was considerably less than that necessary to inhibit PHA response.

The IRA does not act by cytotoxicity, contains no cortisol and does not appear to interact sterically with the stimulating agents. The IRA is not firmly bound to lymphocytes. In inhibiting the PHA response, IRA behaves as a noncompetitive antagonist of the early metabolic events which lead to “lymphocyte activation.”
Rapid Micro-Radioimmunoassay for the Measurement of Antiviral Antibody
Authors: Rosenthal Joel D. , Hayashi Kozaburo , Notkins Abner Louis
Published: December 2022

The binding of radiolabeled antiviral antibody to the surface of virus-infected cells can be used to detect viral antigens and measure antiviral antibody (1, 2). The binding of radiolabeled anti-γ globulin to unlabeled antiviral immunoglobulins on the surface of infected cells also can be used to quantitate antiviral antibody (3). This indirect technique is highly sensitive and has the advantage of employing a single reagent (125I-labeled anti-γ globulin) to detect antibody against a variety of viruses. Both the direct and indirect techniques, however, require fairly large quantities of materials and a monolayer of cells. We now describe an indirect radioimmunoassay which requires only small amounts of material and employs antigens prepared from virus-infected cells.

Materials and Methods. Viral antigens. Monolayers of primary rabbit kidney cells, grown in Eagle's minimal essential medium with 10% heat-inactivated calf serum, were infected with type 1 herpes simplex virus (HSV) or vaccinia virus (CVI-79) (4–6).