bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumor-Specific and Allospecific Immunogenicity of Soluble Extracts from Chemically Induced Murine Sarcomas
Authors: Pellis Neil R. , Tom Baldwin H. , Kahan Barry D.
Published: December 2022

Chemically induced tumors possess vastly polymorphic tumor-specific transplantation antigens (TSTA)2 on their cell surfaces (1–4). Elucidation of the nature of tumor immunity will require detailed analysis of these antigenic markers. One step in such an endeavor is the solubilization of the marker from the cell membrane. There are several techniques to release cell surface antigens, namely, treatment with detergents or organic solvents, enzymatic hydrolysis, exposure to sonic energy, and salt extraction (5). Tumor-specific activity of the solubilized materials has been demonstrated by two types of immunologic assays: 1) in vivo immunogenicity, which is the capacity to protect syngeneic hosts against subsequent tumor challenge, and 2) elicitation of specific immune performances by sensitized cells, such as delayed-type cutaneous reactions, antigen-specific lymphocyte proliferation in vitro, and inhibition of the spontaneous migration of macrophages from sensitized hosts. Materials obtained from chemically induced guinea pig hepatomas by 3 M KCl extraction displayed tumor-specific antigenicity by the latter criteria (6, 7).
The Production of Antibody by Bursal Lymphocytes
Authors: Waltenbaugh Carl R. , Van Alten Pierson J.
Published: December 2022

Lymphocytes of the bursa of Fabricius have been found to actively bind an enteric form of Escherichia coli resulting in the formation of bacterial adherence colonies (BAC). The absolute number of binding lymphocytes appears to increase as the chicken reaches sexual maturity. Treatment of bursal cells with antichicken immunoglobulin results in the blocking of BAC formation. Furthermore, a modified plaque assay technique reveals that a portion of the bursal lymphocyte population is actively secreting antibody against the flagella of E. coli. Introduction of antigen into the bursal or cloacal lumen does not appear to significantly affect circulating antibody levels. Therefore, these data suggest that an additional functional role of the bursa after hatching is localized immunologic defense of the organism.
Suppressor Cells in the Rabbit Appendix
Authors: Kamin Roberta M. , Henry Claudia , Fudenberg H. Hugh
Published: December 2022

Rabbit appendix cells added to cultures of spleen cells, primed in vivo to sheep red blood cells, horse red blood cells, or azo-phenyl β-lactoside-keyhole limpet hemocyanin suppressed the secondary in vitro response of the spleen cells to the test antigen. In the sheep red blood cell system, suppression of the IgM response was found to be less than that of the IgG response and was independent from it. Although suppression of both the IgG and IgM responses could be demonstrated when appendix cells were added at the onset of culture or 24 hr after culture initiation, suppression of the IgM response was less when appendix cells were added 48 hr after culture initiation. Two lines of evidence indicated that a T cell population was involved in the suppression observed: a) complement-dependent cytolysis by antithymocyte serum of appendix cells reduced the degree of suppression produced, and b) appendix cell suspensions, enriched for T cells by nylon wool column adsorption of B cells, had enhanced suppressor activity. Suppressor cell activity was also found in spleen and Peyer's patches. The observation that normal cell populations of appendix, spleen, and Peyer's patches were capable of suppressing the secondary in vitro response to azo-phenyl β-lactoside, indicated a general rather than an antigen-specific suppression. It is proposed that the suppressor activity of the appendix may be a key function of this organ.
Tolerance and Contact Sensitivity to DNFB in Mice
Authors: Phanuphak Praphan , Moorhead John W. , Claman Henry N.
Published: December 2022

Mice were made tolerant to DNFB contact sensitization by pretreatment with DNB-SO3. Lymph node (LN) or spleen cells from such tolerant mice could specifically suppress the development of contact sensitivity to DNFB when adoptively transferred into normal (lightly irradiated) recipients. Mice receiving LN or spleen cells from tolerant donors were skin-painted with DNFB but failed to develop significant skin reactions when challenged with DNFB on the ears. LN cells from these same recipients also did not show antigen-driven DNA synthesis in vitro when stimulated with DNB-SO3. Tolerance was shown to be T cell dependent as prior treatment of tolerant LN cells with anti-ϑ serum and complement inhibited the adoptive transfer of tolerance while passage of these cells over an anti-immunoglobulin column had no such effect. Adoptive transfer of tolerance could not be accomplished via transfer of serum from tolerant donors. While lymphocytes from tolerant donors could prevent the development of contact sensitivity by normal recipients, they could not inhibit the expression of contact sensitivity of previously sensitized cells. Thus, cells from tolerant animals appear to inhibit the afferent limb but not the efferent limb of contact sensitivity. In this context, they may be termed “suppressor T cells.”
Association of Receptors for Mouse IgE with the Plasma Membrane of Rat Mast Cells
Authors: König Wolfgang , Ishizaka Kimishige
Published: December 2022

Evidence was presented that mouse reaginic antibody can be absorbed with rat peritoneal mast cells. After disruption of whole peritoneal cells by ultrasonication in a hypotonic medium and subsequent differential centrifugation, the 20,000 × G supernatant blocked passive cutaneous anaphylaxis with mouse reaginic antibody. The same fraction from mast cell-depleted preparation failed to block the passive cutaneous anaphylaxis reaction. Further purification was achieved by centrifugation on 35% Ficoll and by gel filtration through Sepharose 6B column. The blocking activity and membrane-specific enzymes were eluted from Sepharose column in the void volume. Chromatography on DEAE cellulose separated the blocking fraction from the major bulk of the enzymatic membrane markers. Rechromatography of the active DEAE fraction on Sepharose 6B revealed a different elution pattern for the active component. Activity was recovered from the fractions between void volume and blue dextran (2000) elution thus indicating an apparent cleavage of cell membranes to a smaller size. Similar data were obtained with purified mast cells.
Graft-Versus-Host and Cytotoxic Activity of Rat Thoracic Duct Lymphocytes Fractionated by Zonal Centrifugation after Culture on Allogeneic Fibroblasts
Authors: Bowers William E.
Published: December 2022

Large cytotoxic lymphocytes were generated by culturing syngeneic rat thoracic duct lymphocytes on a monolayer of allogeneic fibroblasts. Lymphocytes removed from culture at 5 days were fractionated by zonal centrifugation, and as expected, cytotoxicity was found to be a property of the large cells. At the same time, graft-vs-host activity of the small lymphocytes purified by zonal centrifugation was assayed by a quantitative method. Activities similar to controls were found in both specific (Ag-B-identical to those of fibroblasts) and nonspecific (Ag-B-different from those of fibroblasts) semiallogeneic F1 hybrids. Cultures were also terminated after 3 and 4 days, and graft-vs-host assays were performed on small lymphocytes purified by zonal centrifugation. The activities also did not differ markedly from the controls. Hence, large cytotoxic lymphocytes are not generated at the expense of small lymphocytes capable of specific graft-vs-host activity; various possibilities for this finding are discussed.
Interaction of IgE with Rat Basophilic Leukemia Cells
Authors: Carson Dennis A. , Metzger Henry
Published: December 2022

Rat leukemic basophils with surface-bound IgE were reacted with fluoresceinated anti-IgE. The results were qualitatively similar to those obtained in analogous studies with lymphocytes, normal human basophils, and other cells: local aggregation (patch formation) was only moderately temperature-dependent and was uninhibited by NaN3. Polar cap formation was significantly more sensitive to temperature and was completely inhibited by 0.01 to 0.1 M NaN3 and cytochalasin B. The amount and rate of capping could be altered by varying either the number of bound IgE molecules or the anti-IgE concentration indicating that lattice formation is required for redistribution. Monovalent Fab′ fragments of the anti-IgE caused no redistribution. No endocytosis of the antibody-surface determinant complexes was observed. Under conditions adequate to inhibit cap formation on mouse lymphocytes, concanavalin A did not inhibit cap formation on the leukemic basophils.
Macrophage-Lymphocyte Synergy in the Production of Osteoclast Activating Factor
Authors: Horton John E. , Oppenheim Joost J. , Mergenhagen Stephan E. , Raisz Lawrence G.
Published: December 2022

Supernatants from cultures of activated normal human peripheral blood leukocytes contain a factor which stimulates osteoclastic bone resorption in vitro. The present study indicates that osteoclast activating factor (OAF) is produced by phytohemagglutinin-activated mononuclear cells within 6 hr of activation, and that peak production is associated with increased amino acid incorporation into protein but precedes increased thymidine incorporation into DNA. Granulocytes, platelets, and red cells were not necessary for the production of OAF, since mononuclear cells purified on Ficoll-Hypaque gradients and devoid of these elements produced as much OAF as unseparated leukocytes. Granulocyte extracts did not show bone-resorbing activity. The production of OAF by mononuclear cells was impaired by irradiation, suggesting that it is made by radiosensitive lymphocytes rather than radioresistant macrophages. However, OAF was not produced by lymphocytes purified on glass-bead columns although these cells could respond to phytohemagglutinin with increased thymidine and amino acid incorporation. Purified macrophages also failed to produce substantial bone-resorbing activity. However, when a small number of irradiated but viable macrophages (5 × 104 cells/ml) was added to glass bead-purified lymphocytes (2 × 106 cells/ml), OAF production was as great as with unseparated mononuclear cell populations. These results suggest that OAF is produced by lymphocytes, but that its production is dependent on macrophage-lymphocyte interaction. OAF is a unique bone-resorbing factor which may play an important role in bone lesions associated with chronic inflammation and neoplasia. The cellular mechanisms for its production participate in the pathogenesis of these disorders.
Human Lymphotoxin Obtained from Established Lymphoid Lines: Purification Characteristics and Inhibition by Anti-Immunoglobulin
Authors: Amino Nobuyuki , Linn Edward S. , Pysher Theodore J. , Mier Richard , Moore George E. , DeGroot Leslie J.
Published: December 2022

Human lymphotoxin obtained from established lymphoid cell lines (HLT-LCL) was studied in detail and purified using ammonium sulfate fractionation, gel filtration, ion-exchange column chromatography, and polyacrylamide gel electrophoresis. HLT-LCL is stable at 20°C for over 1 year, at 4°C for 3 weeks, and at 37°C for 1 week. HLT-LCL showed a decrease in activity at pH 2.0 and 3.0, but was stable at a pH range of 5.0–11.0 HLT-LCL, which contained fetal calf serum, was purified 50-fold. HLT-LCL obtained from serum-free culture medium yielded greater purity, although complete purification was not achieved. Antiserum to HLT-LCL neutralized the cytotoxic activity of HLT-LCL, HLT induced by PHA (HLT-PHA), and HLT induced by PPD (HLT-PPD). Antiserum to human IgG, λ-type L chain, and Fc fragment partially but significantly inhibited HLT-LCL activity. These data, together with previous reports, indicate that the HLT-LCL molecule is immunologically similar to HLT-PHA and HLT-PPD and suggest that there is immunologic cross-reactivity with IgG globulin.
Heterogeneity of Antinucleolar Antibody and IgE Antinuclear Antibody in Patients with Systemic Rheumatic Diseases
Authors: Miyawaki Shoji , Ritchie Robert F.
Published: December 2022

Antinucleolar antibodies (ANoA) were found most frequently in patients with systemic sclerosis and atypical illnesses manifesting Raynaud's phenomenon. These usually were found to be of more than two immunoglobulin classes. Over 70% of sera with ANoA fixed C3 and C4. IgE ANoA was found in 56% of our patients. IgE antinuclear antibodies were found in 48% of patients with systemic lupus erythematosus, 58% of patients with procainamide-induced systemic lupus erythematosus, and in only 5% of patients with rheumatoid arthritis.
Immunochemical Classification of Nonisolated Light Chains of Immunoglobulins
Authors: McLaughlin Carla L. , Solomon Alan
Published: December 2022

Antigenic sites of the light chain are often masked in the intact immunoglobulin (Ig) molecule; and because many of our anti-light chain antisera have stronger reactivity with the free light chain than with the intact Ig molecule, a microtechnique for reduction and alkylation of homogeneous immunoglobulins which does not require isolation of the light chain before immunochemical typing was used in preparation of type K myeloma proteins and M macroglobulins for immunochemical analyses without group-specific antisera. Antisera specific for κI, κII, κIII, and κIV light chains were used in immunodiffusion analyses of the reduced and alkylated Ig; antigenic identity between the treated Ig and homologous isolated light chain was readily demonstrable. The classification of the light chains of proteins of known antibody activity, e.g., cold agglutinins, as well as the study of proteins available in small quantity, was facilitated by the use of the microtechnique for reduction and alkylation.
Clonal Expansion of Lipopolysaccharide-Stimulated B Lymphocytes
Authors: Quintáns José , Lefkovits Ivan
Published: December 2022

The effect of lipopolysaccharide from Gram-negative bacteria (LPS) on the activation and clonal expansion of B cell precursors specific for SRC has been studies in a microculture system.

Results indicate that LPS can activate the precursor cells to antibody production in the absence of antigen and without significant cell proliferation. Clonal expansion will ensure only if LPS and SRC are present.
Characterization of Lymphocytes in Bronchial Lavage Fluid from Monkeys
Authors: Kazmierowski John A. , Fauci Anthony S. , Reynolds Herbert Y.
Published: December 2022

At least two populations of lymphocytes can be identified in primate lung washings, one forming rosettes with SRBC and another forming rosettes with erythrocyte-antibody-complement complexes. The response to mitogens by cells from lavage fluid is similar to spleen and blood lymphocytes. The distribution of lymphocytes in pulmonary lavage fluid is similar to that found in the peripheral blood. The identity of lymphocyte populations in respiratory fluids has relevance in understanding the immunologic capabilities of the primate respiratory tract.
Comparative Studies on Monotypic IgMλ and IgGκ from an Individual Patient
Authors: Hopper John E. , Noyes Claudia , Heinrikson Robert , Kessel James W.
Published: December 2022

Amino terminal sequence analyses were performed on the H and L chains of the idiotypically related IgMλ and IgGκ paraproteins isolated from the sera of a patient, Br. The N-terminal 41 residues of the Br κ-chain belonged to the VKIII subgroup, and L chains derived from BrIgMλ revealed a blocked N-terminus characteristic of λ-chains.

Comparative sequence analysis of the Br µ- and γ-chains indicated that, although both possessed an unblocked N-terminal glutamic acid, the respective VH regions belonged to separate subgroups. The N-terminal 27 residues of the Br µ-chain reflected a typical VHIII subgroup sequence. The Br γ-chain sequence demonstrated a VHI pattern with an unblocked N-terminus. These structural data stand ir contrast to previously reported serologic evidence, which indicated that the BrIgMλ and BrIgGκ proteins possessed highly similar VH-associated idiotypic determinants. Final interpretation of these findings of similar VH idiotypic determinants expressed by H chains belonging to separate VH subgroups must await complete sequence analysis of the hypervariable segments of the Br γ- and µ-chains.
The Role of Plasmin on the Double Antibody Radioimmunoassay of Carcinoembryonic Antigen in Human Blood Samples
Authors: Das Saroj , Das B. R.
Published: December 2022

Double antibody radioimmunoassay (RIA) of carcinoembryonic antigen (CEA) on a series of freshly drawn out concurrent plasma and serum samples of normal human blood donors showed that the serum CEA values were invariably higher than the corresponding plasma CEA values. Extraneous addition of fibrinogen brought down the serum-CEA level to a value comparable to or less than the corresponding plasma value. The effect of certain factors associated with blood clotting, particularly Ca++, fibrinogen, and the fibrinolytic enzyme plasmin, was investigated. Ca++ was shown to play no role whereas the effect of fibrinogen was shown to be indirect in that it served as a specific substrate for plasmin, thereby preventing the plasmin degradation of the primary antibody used in the RIA. The finding stresses the role of enzymes like plasmin in double antibody RIA in general and may explain some of the anomalous results obtained when testing biologic material containing plasmin-like substances.
Phytohemagglutinin-Induced Proliferation of Guinea Pig Thymus-Derived Lymphocytes
Authors: Lipsky Peter E. , Ellner Jerrold J. , Rosenthal Alan S.
Published: December 2022

The accessory cell requirement for mitogen-induced T lymphocyte proliferation has been investigated by using a population of guinea pig lymph node lymphocytes enriched in T cells and markedly depleted of macrophages and B lymphocytes. We have found that effective phytohemagglutinin-induced proliferation of T cells is dependent on the participation of accessory cells. Augmentation of PHA responsiveness was noted when cultural conditions were manipulated to increase cell density, suggesting that physical proximity between T cell and accessory cell is required for efficient triggering. Both syngeneic and allogeneic macrophages, as well as syngeneic fibroblasts, serve as accessory cells in this response whereas polymorphonuclear leukocytes or thymocytes do not. Thus, although PHA-induced T lymphocyte proliferation requires accessory cells, the specificity of these cells is strikingly less stringent than for antigen-mediated triggering of immune guinea pig T cells, a response which is dependent upon participation of syngeneic macrophages.
Idiotypic Specificities of Anti-Rh Antibodies
Authors: Natvig J. B. , Kunkel H. G. , Rosenfield R. E. , Dalton J. F. , Kochwa S.
Published: December 2022

Idiotypic antibodies were produced in rabbits to Rh antibodies isolated from the serum of an individual with high Rh antibody titer. These antisera after absorption with pooled IgG failed to react with immunoglobulins lacking anti-Rh activity. They agglutinated cells coated with the immunizing antibody to high titers. Two other anti-Rh coats out of 22 tested also were agglutinated but to considerably reduced titers indicating a degree of anti-Rh cross specificity. Four other antisera made to red cells coated with different anti-Rh antibodies showed definite but weaker idiotypic specificity.

Both the idiotypic and cross-idiotypic antigens of the Rh antibodies were completely blocked when univalent fragments of the Rh antibodies were bound to antigen on the red cell.
The Formation of Septate-Like Junctional Complexes between Lymphoid Cells in Vitro
Authors: McIntype John A. , Pierce Carl W. , Karnovsky Morris J.
Published: December 2022

During the initial 48 hr of incubation in immunized Mishell-Dutton spleen cell cultures, most lymphocytes exist as single cells or an occasional pair of cells (doublets). Ultrastructural examination of the area between those cells forming doublets revealed a septate-like junctional complex occurring over large portions of the plasmalemma. This junctional zone persisted in areas where extensive cytoplasmic interdigitation among cell processes was evident. Approximately 1% of the lymphocytes isolated from the top fraction of a discontinuous fetal calf serum gradient were involved in formation of doublets. The majority of these doublets showed evidence of junctional interaction. Moreover, this septate-like zone of adhesion was seen using several different fixatives and stains and remained intact after osmotic cell disruption. The junction was not demonstrable by using freeze-fracture techniques and therefore must be limited to interactions within the surface of the cells.
Identification of Heavy Chain Tyrosine 33 in the Binding Site of Myeloma Protein McPC 603 by Paired Label Iodination
Authors: Grossberg Allan L. , Roholt Oliver A. , Pressman David
Published: December 2022

The phosphorylcholine binding mouse myeloma protein McPC 603 has been shown to have tyrosyl residues in its binding sites by the fact that iodination of the protein causes extensive loss of binding activity which can be substantially retained when the protein is iodinated with sites occupied by ligand. Paired label iodination of McPC 603 protein allowed identification of the tyrosine involved and showed the tyrosine to be in the heavy chain. Gel filtration of heavy chain peptides enabled the tyrosyl-containing peptide of interest to be identified as the N-terminal 33 residue peptide in which the only tyrosine is Tyr 33. Thus H chain Tyr 33 was shown to be a contact amino acid residue in the site of McPC 603 protein. These results provide chemical evidence confirming previously reported x-ray crystallographic identification of H chain Tyr 33 in the site of McPC 603 protein.
Purification of Homogeneous Murine Immunoglobulins with Anti-Fructofuranan Specificity
Authors: Vrana Mark , Tomašić Jelka , Glaudemans Cornelis P. J.
Published: December 2022

Homogenous IgAκ immunoglobulins ABPC-47 new, EPC-109, UPC-61 (anti-2,1 fructofuranan specificity) and homogeneous IgAκ ABPC-48 and IgGκ UPC-10 (anti-2,6 fructofuranan specificity) were purified by affinity chromatography by using inulin-and grass levan-Sepharose columns, respectively.
Release of Slow Reacting Substance of Anaphylaxis (Srs-A) from Human Leukocytes by the Calcium Ionophore A23187
Authors: Conroy Mary Carol , Orange Robert P. , Lichtenstein Lawrence M.
Published: December 2022

The ability of the calcium ionophore A23187 to release slow reacting substance of anaphylaxis (SRS-A) from human leukocytes was studied. About 25 times more SRS-A activity was released from aliquots of leukocytes by ionophore stimulation than by antigen stimulation, although comparable amounts of histamine were released. Cell separation studies revealed that granulocytes other than basophils were also capable of releasing SRS-A. The contractile activity released after challenge with ionophore appeared physicochemically identical to the SRS-A of rat or human origin released by antigen challenge in terms of its stability to base hydrolysis, inactivation by arylsulfatase, and chromatographic behavior on silicic acid and Sephadex LH-20 columns. We suggest that some mediators of allergic reactions previously associated, in man, only with antigen-IgE antibody interaction on mast cells or basophils may be released by other stimuli and from other cell types.
The Role of Factor D for the Interaction of Zymosan with Components of the Properdin System: Studies with D-Depleted Guinea Pig Serum
Authors: Brade V. , Burger R. , Bitter-Suermann D. , Hadding U.
Published: December 2022

Serum was depleted of factor D by gel filtration and immunoabsorption with unsolubilized anti-D IgG. Substitution of this reagent (RD) with purified D̄ was found to fully reconstitute its function. In the following experiments the reaction of zymosan (Z) with RD was investigated; therefore these studies are concerned with the role of Factor D for the interaction of Z with components of the properdin system.

RD was incubated for 30 min at 0°C, or for 30 min at 37°C, with Z (10 mg/ml) in the presence of EGTA. After centrifugation both supernatants, as well as the corresponding Z particles, were analyzed. Both supernatants were found to contain identical amounts of B, whereas C3 was slightly reduced in the 37°C supernatant. After addition of Z and of purified D̄, both supernatants supported the formation of the C3-cleaving ZX-complex with the same efficiency as untreated RD. Thus, incubation of Z with RD was only followed by a slight loss of C3; no other factors of the properdin system appeared to be reduced in their activity.
Studies on the Activation of C1 by a Univalent Hapten-Antibody Complex
Authors: Goers J. , Ziccardi B. , Glovsky M. , Schumaker V.
Published: December 2022

The clasical serum complement system is efficiently activated by a complex formed between a monovalent DNP-polylysine hapten (nonadeca lysyl ε-DNP-lysine) and rabbit anti-DNP antibody. The hapten-antibody complex sediments as a single antibody molecule, but activates complement in the same concentration range as heat-aggregated human IgG, or immune aggregates (J. Biol. Chem. 250:4918, 1975). Activation by similar, but shorter, DNP-polylysine haptens occurs only when the polylysine chain is 9 to 10 lysyl residues in length. The activation of C1 by the hapten-antibody complex does not appear to be a result of an increased efficiency of the hapten-antibody complex to bind C1q. Ultracentrifugation studies show no difference in antibody-binding affinities to C1q with or without the DNP-polylysine hapten. However, activation of C1s to C11̄s by the hapten-antibody complex does occur at high antibody concentrations. Furthermore, the hapten-antibody complex and the hapten alone are both capable of activating a mixture of C1r and C1s.
On the Mechanism of Cytolysis by Complement: Further Studies on the One-Hit Characteristics of the C9 Reaction
Authors: Mayer Manfred M. , Vuozzo Eugene F. , Paznekas William A.
Published: December 2022

Since C9 is the terminal complement component it is important to determine whether its reaction displays one-hit or multi-hit characteristics. This question has been studied previously by kinetic methods. We now report studies by endpoint titration of C9. We have developed a two-step procedure in order to attain the endpoint of the C9 reaction with respect to time. In the first step EAC1-8 are allowed to react with C9 for 10 to 20 hr at 0°C. During this interval all of the C9 combines with the cells. In the second step the cells are incubated for 2.5 hr at 37°C. During this interval the events leading to release of hemoglobin proceed to completion. By this technique it has been found that the dose-response curve for the reaction of C9 with EAC1-8 is entirely concave to the abscissa, as would be expected for a one-hit process. Also, titrations of C9 performed with different quantities of EAC1-8 support the one-hit interpretation.
Thin Layer Isoelectric Focusing for Final Purification of Human C3
Authors: McLean Robert H.
Published: December 2022

Chromatographic isoelectric focusing of human complement (C) components by other investigators has shown the isoelectric point (pI) for human C3 to be pH 5.7. C3 activity (immune adherence and hemolytic function) was separable from the protein B1C when reacted with anti-B1C/B1A antisera and B1C had a pI of pH 6.1. We have used thin layer isoelectric focusing on Sephadex gel for the final purification step in the isolation of human C3. After the final chromatographic purification step of C3, protein (without detectable C5) was predialyzed in 0.0005 M phosphate buffer, pH 7.5. Several hundred micrograms in volumes of 0.3 to 0.5 ml were applied either at the anode or to the center of the isoelectric focusing slab (Sephadex G-75 superfine). Hemolytic C3 was measured by three methods: by specific C3 assay, using purified terminal human C components and EA1̄ 4̄ 2̄ intermediate cells; by potassium bromide-treated normal serum and EA4̄ intermediate cells; and by monospecific goat anti-B2C/B1A antiserum.
Ultrastructure and Chemistry of C1t Subcomponent of C1: Similarities to Amyloid P-Component
Authors: Painter Robert H. , Pinteric L. , Hofmann T. , Kells D. I. C. , Katz A.
Published: December 2022

We have recently isolated a 9.5S α1 glycoprotein from human serum and shown it to be a Ca++-dependent subcomponent of the first component of complement and which we have named C1t. It is believed that C1t serves as a bridge between C1q and C1s enhancing and amplifying the ability of C1q to bind C1s and present it to C1r for activation.

The molecular weight of C1t was determined by sedimentation equilibrium analysis by using the meniscus depletion method. In neutral solvents, 0.1 mM Na3HEDTA, a molecular weight of 233,000 was found. In 5.95 M guanidinium chloride, a molecular weight of 23,000 was obtained. This value was unaffected by prior reduction and alkylation with 0.1 M mercaptoethanol and 0.22 M iodoacetamide. The molecule appears to be composed of 10 similar subunits each composed of a single polypeptide chain.
Complement in the Induction of IgA and IgE Antibody Production
Authors: Pepys M. B. , Wansbrough-Jones M. H. , Mirjah D. D. , Dash A. C. , Brighton W. D. , Hewitt B. E. , Bryant D. E. W. , Pepys J. , Feldmann M.
Published: December 2022

The IgA-antibody forming cell response to sheep erythrocytes in BALB/c and CBA mice depleted of complement by cobra factor (CoF) was suppressed to 10 to 40% of the level in controls. The IgE response to alum-precipitated ovalbumin was measured in NIH mice by passive cutaneous anaphylaxis in rat skin. Reaginic antibody was scarcely detectable in CoF-treated animals whereas controls had titers of up to 1:2000. Fourteen days after boosting with ovalbumin, control mice had peripheral eosinophilia whereas animals which had received CoF before priming 6 weeks earlier had levels only minimally raised above normal. Studies of the generation of T cell blasts, “education” of thymus cells, and priming of splenic T cells indicated that activation of T cells by antigen proceeded normally under conditions in which thymus-dependent antibody production was suppressed by in vivo complement depletion.

Pure sheep IgG anti-mouse C3 and pure F(ab′)2 anti-mouse C3 antibody both inhibited the in vitro T-dependent antihapten response of mouse spleen cells to TNP-KLH.
A C5 Fragment with Chemotactic Activity for Tumor Cells
Authors: Romualdez A. , Ward P. A.
Published: December 2022

The chemotactic behavior of several animal tumor cells (Novikoff hepatoma cells, Walker carcinosarcoma cells, and murine mastocytoma cells) was studied in modified Boyden chambers. Leukotactic factors derived from complement (C3 and C5 fragments) or from bacteria (Escherichia coli) had no chemotactic activity for tumor cells. Incubation of normal rat or human serum with a crude extract from Walker tumor cells resulted in the generation of a factor chemotactic for tumor cells but not for leukocytes. Experiments using complement deficient (acquired or genetic) serum, complement-specific antiserums, and preparations of purified complement components indicated that the factor chemotactic for tumor cells was a fragment of the fifth component of complement (C5) produced by a C5-cleaving enzyme extractable from tumor cells. Sucrose density gradient analysis and Sephadex gel filtration chromatography indicated that the C5 fragment chemotactic for tumor cells had physicochemical features different from the C5 fragment chemotactically active for neutrophils.
The Role of Properdin As a Modulator of the Alternative Pathway
Authors: Spitzer R. E. , Stitzel A. E. , Urmson J. R.
Published: December 2022

When activated properdin convertase () is added to serum, C3–C9 consumption occurs without Factors B or D. Only properdin is required. When zymosan is added to serum, PC is activated but C3–C9 consumption requires not only properdin but both Factors B and D. An apparent discrepancy, therefore, exists. The present studies illustrate that for C3–C9 consumption, a critical concentration of exists in serum relative to the amount of zymosan added. If 2.5 µg/ml of are added to serum, C3–C9 hemolytic activity is rapidly consumed. If zymosan is added to that mixture, even in small amounts (0.4 mg/ml of serum), this consumption is 90% inhibited. As the amount of zymosan is increased (greater than 3.0 mg/ml), however, the inhibition can be completely overcome. In the first instance of C3–C9 consumption (in the absence of zymosan) Factor B is utilized to only a small extent.
Species Specificity in the Isoelectric Spectra of Immunoglobulin Light Chains
Authors: Gibson David M.
Published: December 2022

Light chains isolated from normal immunoglobulin give rise to a finite number of discrete bands when analyzed by isoelectric focusing in polyacrylamide gel. The positions and relative intensities of the bands were identical in light chains isolated from different individuals of a species, but clear differences were evident when light chains of several mammalian species were compared.
Purification of a Chlamydia Trachomatis-Specific Antigen by Immunoadsorption with Monospecific Antibody
Authors: Caldwell Harlan D. , Kuo Cho-Chou
Published: December 2022

This study describes the isolation and partial characterization of a Chlamydia trachomatis specific antigen. A species-specific antigen of C. trachomatis (antigen-0.65) was identified by two-dimensional immunoelectrophoresis. Antiserum specific for antigen-0.65 was prepared in rabbits by immunizing with agarose-gel precipitates excised from two-dimensional immunoelectrophorograms. Purified γ-globulins from antigen-0.65 specific serum were coupled to the N-hydroxysuccinimide ester derivative of agarose which was then used for the immunoadsorbent purification of antigen-0.65 from Triton X-100 solubilized lymphogranuloma venereum (L2/434/Bu) organisms. The isolated antigen was immunochemically pure when tested against rabbit antiserum prepared to LGV-434 organisms by using rocket and two-dimensional immunoelectrophoresis. Antigenicity was destroyed by protease treatment and heating at 56°C for 30 min, but the antigen was stable to ribonuclease, deoxyribonuclease, periodate oxidation and pH extremes of 2.2 and 10.6. Polyacrylamide gel electrophoresis of purified antigen showed a major protein band with an apparent m.w. of 155,000.
The Nature of the Local Immune System of the Bovine Mammary Gland
Authors: Newby Timothy J. , Bourne John
Published: December 2022

The origin of the immunoglobulins of bovine colostrum and milk were examined by using radiolabeled protein. Colostrum was found to consist mainly of immunoglobulin derived from serum, although there was some local synthesis. In lactation the immunoglobulins of milk were almost entirely serum derived. The local immune system was very inactive but could be stimulated to activity by local immunization. The immunoglobulin profiles of the secretion from the stimulated gland were still relatively deficient in IgA compared to other species, notably the pig.
Antibody-Mediated Suppression of Tumor Growth
Authors: Johnson Robert J. , Pasternack Gary R. , Shin Hyun S.
Published: December 2022

The tumor suppressive activity of murine IgG1 antibody was studied in vivo. IgG1 was isolated from hyperimmune alloantisera against a murine lymphoma, 6C3HED, by absorption with heat-killed, formalinized Staphylococcus aureus. Cowan strain I, followed by DEAE ion exchange chromatography and Sephadex G-200 gel filtration chromatography. The isolated IgG1, which had no detectable IgM, IgA, IgG2a, IgG2b or IgG3, could suppress the growth of the tumor. In addition, DEAE and Sephadex G-200 column profiles of the in vivo tumor suppressive activity showed good correlation with the profiles of total IgG1 immunoglobulin and anti-tumor antibody assayed for IgG1 heavy chains and kappa light chains. The IgG1 tumor suppressive activity was not diminished after heating at 56°C for 30 min.
Histaminase Release from Human Eosinophils
Authors: Zeiger Robert S. , Colten Harvey R.
Published: December 2022

Phagocytosis of opsonized zymosan by human eosinophils results in a dose-dependent noncytotoxic release of histaminase as well as arylsulfatase and β-glucuronidase. The calcium ionophore A23187 also stimulates release of eosinophil histaminase at concentrations of ionophore which barely release arylsulfatase and β-glucuronidase. Zymosan-induced histaminase release from eosinophils but not from neutrophils was abolished or markedly reduced in the presence of cytochalasin B, suggesting a difference in the mechanisms of histaminase release from the two granulocyte cell types.
Partial Characterization of Nervous System-Specific Cell Surface Antigen(s) NS-2
Authors: Yuan Dorothy , Vitetta Ellen S. , Schachner Melitta
Published: December 2022

Partial biochemical characterization of several neural tissue specific antigens isolated from a murine glioblastoma cell line was accomplished by means of radioiodination of intact cells followed by immunoprecipitation of the cell lysate with a rabbit serum specific for neural tissue antigens. Polyacrylamide gel electrophoresis of the immunoprecipitate in sodium dodecyl sulfate resolved the labeled antigens into several major components: two proteins (or glycoproteins) having apparent m.w.'s of 84,000 and 120,000 and lipid associated components which may be heterogeneous. The protein and lipid associated components apparently possess independent antigenicity because after chloroform-methanol extraction the protein components can be immunoprecipitated from the aqueous phase and the lipid associated component can be immunoprecipitated from the organic phase. Despite their independent antigenicity it is not known whether the components may be noncovalently associated on the cell surface.

Although some of these antigens can be isolated from brain or glioma cells (a related tumor), none can be demonstrated in lymphoid tissues or C1300 neuroblastoma cells using identical methods. Therefore, these studies confirm our previous findings concerning the specificity of the anti-NS-2 antiserum by using cytotoxicity tests.
Reduction in Basal Adenylate Cyclase Activity during the Immunologic Release of Histamine from Guinea Pig Lung
Authors: Hitchcock Margaret
Published: December 2022

Cyclic AMP has been implicated in the regulation of the immunologic release of histamine from lung and other tissues and cell types. The mechanism whereby intracellular levels of cAMP are altered during mediator release was investigated. Measurements of histamine, adenylate cyclase, and cAMP phosphodiesterase activities were made in actively and passively sensitized guinea pig lung after challenge with antigen. A transient decrease in basal adenylate cyclase activity occurred which returned to control levels after histamine release. There was no change in cAMP phosphodiesterase activity determined at substrate concentrations of 1 mM and 0.01 mM. The adenylate cyclase response did not occur under the following conditions: 1) incubation of nonsensitized lung with antigen, 2) incubation of sensitized lung with antigen in the absence of extracellular calcium, and 3) incubation of nonsensitized lung with compound 48/80. These observations indicate 1) the adenylate cyclase response and the immunologic release of histamine are intimately related, and 2) the reduction in intracellular levels of cAMP which have been reported to occur during immunologic histamine release are mediated via adenylate cyclase.
Experimental Cell-Mediated Interstitial Nephritis Induced with Exogenous Antigens
Authors: Van Zwieten Matthew J. , Leber Paul D. , Bhan Atul K. , McCluskey Robert T.
Published: December 2022

The capacity of intrarenal injections of bovine γ-globulin (BGG) to elicit hypersensitivity reactions was studied in guinea pigs immunized with DNP BGG or BGG immune complexes in CFA or in rats immunized with BGG in CFA. In guinea pigs it was found that heat-aggregated BGG elicited inflammatory reactions in the renal cortex, whereas soluble BGG did not. In rats only aggregated BGG was used, and this was found to be effective. The reactions were characterized by a predominantly mononuclear cell infiltrate. Transfer experiments were performed in rats and it was found that reactivity was transferrable with lymph node cells but not with serum.
Immunologic Reactivity of Purified Human Urinary Kallikrein (Urokallikrein) with Antiserum Directed against Human Pancreas
Authors: Ole-MoiYoi Onesmo , Spragg Jocelyn , Halbert Seymour P. , Austen K. Frank
Published: December 2022

Donkey antiserum to normal human pancreas absorbed with lyophilized human plasma recognizes human urokallikrein in concentrated crude urine or after an approximately 500-fold purification. The urokallikrein antigen was associated with kinin-generating and α-N-p-tosyl-L-arginine methyl ester (TAMe) cleaving activity on isoelectric focusing, with the isoelectric point being 3.8 to 4.4. Both kiningenerating and esterolytic activity were removed from the purified urokallikrein by an immunoadsorbent prepared by coupling the IgG fraction of the absorbed donkey antiserum to Sepharose 6B. The failure of anti-plasma kallikrein to react in immunodiffusion with purified urokallikrein indicates that urinary kallikrein is distinct from plasma kallikrein although antigenically related to glandular kallikreins.
Immunity to Lymphoid Tumors Induced in Syngeneic Mice by Immunization with Mitomycin C-Treated Cells
Authors: Benjamini Eli , Fong Sherman , Erickson Caroline , Leung Cherry Y. , Rennick Donna , Scibienski Robert J.
Published: December 2022

Immunization of mice with syngeneic mitomycin C-treated lymphoid tumor cells (EL-4 and S49A) conferred a high degree of immunity to transplantation with viable tumor cells in syngeneic animals. The development of this immunity was paralleled by the development of specific cell-mediated cytotoxicity; no anti-tumor antibodies could be detected in the immunized animals. In contrast to the high immunoprophylactic capacity of mitomycin C-treated cells, attempts to utilize these cells for immunotherapy were unsuccessful. Preliminary experiments did not reveal antigenic differences between mitomycin C-treated and untreated tumor cells.
Effects of Local Immunization with Glucosyltransferase Fractions from Streptococcus Mutans on Dental Caries in Rats and Hamsters
Authors: Taubman Martin A. , Smith Daniel J.
Published: December 2022

The effect of local immunization with glucosyltransferase enzymes of Streptococcus mutans on dental caries in conventional rats, hamsters, and gnotobiotic rats was studied. Injection of these animals with crude or defined glucosyltransferase enzyme preparations incorporated into complete Freund's adjuvant consistently resulted in the presence of antibody in saliva demonstrable by functional inhibition of enzymatic activity and binding of radioactive enzyme. Serum antibody was also present. Two experiments have been completed in conventional rats and one in gnotobiotic rats in which the animals were immunized with crude enzyme antigens. The immunized group of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. Hamsters immunized with a defined enzyme preparation, containing no more than three antigenic components (two of which were enzyme), also demonstrated significant reductions in mean caries scores. The numbers of lesions were also always lower in immunized animals. In some cases there were reductions in the numbers of S. mutans that could be recovered from the teeth of immunized, infected animals. The reductions in dental caries and lesions were greater on smooth dental surfaces than on occlusal surfaces, which might be explained as interference with adherence phenomena demonstrated by S. mutans. It is proposed that antibody interference affects dental caries caused by this organism.
On the Antigenic Substances of the Tubercle Bacillus
Authors: Dienes L. , Freund J.
Published: December 2022

It has been shown that beside the non-protein precipitable substance present in the tubercle bacillus extracts the protein substances of the extracts are potent antigens in the complement fixation test.

All three of the antigens demonstrated in the tubercle bacillus, the protein substances, the alcohol soluble specific substance, and the carbohydrate precipitable substance of Mueller and of Laidlow and Dudley, as they occur in our preparations, are distinct and separate antigens.

A short description is given of the properties of the known antigens of the tubercle bacillus and of the response of the animal organism to them. Some related questions are briefly discussed.
Structural Differences between the Mouse H-2D Products of the Mutant B10.D2.M504 (H-2da) and the Parental Nonmutant Strain B10.D2 (H-2d)
Authors: Brown J. Lynne , Nairn Roderick , Nathenson Stanley G.
Published: December 2022

The H-2K and H-2D glycoproteins from the mutant mouse strain B10.D2.M504 (H-2Kd,Dda) were compared with the H-2K and H-2D molecules from the parent nonmutant strain B10.D2 (H-2Kd,Dd). Whereas in every respect the K molecules of the mutant and nonmutant cells were identical, the Dda molecule from the mutant showed notable differences from the Dd molecule of the nonmutant parent. First, smaller amounts (ca. 25%) of the Dda glycoprotein appeared to be present in M504 cell extracts as compared to the amount of Dd glycoprotein present in nonmutant cell extracts. Second, a proportion of the Dda glycoprotein molecules showed an abnormal interaction with the lectin from lentil beans (LcH), suggesting an altered carbohydrate chain on some of the molecules. Third, comparative tryptic peptide analysis showed a significant degree of diversity between the mutant Dda molecule and the nonmutant Dd molecule. Thus, analysis of arginine-labeled peptides showed at least two peptides unique to H-2Dd and at least three peptides unique to H-2Dda. Analysis of lysine-labeled peptides showed at least three peptides unique to H-2Hd and greater than two peptides unique to the mutant H-2Dda. These data thus document that the Dda glycoprotein has an altered primary amino acid sequence as compared to the nonmutant Dd molecule. The finding of differences in greater than one-third of the total peptides recovered correlates with the strong serologic and histogenic reactivity between the cells of the M504 mutant and the cells of the B10.D2 parent. This peptide diversity is in contrast to the small degree of variation (ca. 10%) noted for the mutants of the H-2Kb locus (e.g., H-2Kba, H-2Kbd) which have only histogenic reactivity and essentially no serologic differences between mutant and nonmutant cells. The multiple structural differences noted between H-2Dd and H-2Dda suggest that a complex mutation has taken place.