bluuebunny/crossref_metadata_2025_split huggingface dataset:
Alternative Complement Pathway-Mediated Lysis of Measles Virus Infected Cells: Induction by IgG Antibody Bound to Individual Viral Glycoproteins and Comparative Efficacy of F(ab′)2 and Fab′ Fragments
Authors: Sissons J. G. Patrick , Cooper Neil R. , Oldstone Michael B. A. 
Published: December 2022

Abstract
Lysis of measles virus-infected cells by human serum is dependent on IgG antibody and an intact alternative complement pathway. The binding sites of IgG measles virus antibody, or its fragments, on the surface of measles virus-infected cells were determined by cell surface radioiodination and immunoprecipitation. It was found that IgG bound to either of the two measles virus surface glycoproteins was effective in inducing cell lysis by the alternative pathway. Thus, IgG bound only to the viral hemagglutinin (HA) or only to the fusion protein (F) could induced lysis in C4-depleted human serum; the same number of IgG molecules bound to either HA or F induced equivalent dose-related lysis. Divalent antibody bound preferentially to HA on the surface of intact cells; this enabled antibody to F to be obtained free of antibody to HA by adsorption. Quantitative studies of binding and lysis showed that lysis of measles virus-infected cells by whole or C4-depleted serum required binding of at least 10 times more Fab′ than F(ab′)2 or IgG molecules per cell. In these experiments, C4-depleted serum was quantitatively and kinetically equivalent to whole serum in mediating lysis of measles-infected cells. Hence, the binding of IgG to either of the two measles viral polypeptides (HA, F) expressed on the surface of infected cells induces lysis by the alternative pathway, and there is a clear requirement for divalency for effective induction of lysis.
The Beige Mutation in the Mouse
Authors: Roder John C. , Lohmann-Matthes Marie-Luise , Domzig Wolfgang , Wigzell Hans 
Published: December 2022

Abstract
In the preceding paper we showed that the beige mutation in the mouse leads to a complete impairment in NK cell function, which is predetermined at the level of progenitor cells in the bone marrow. The observations in this report suggest that the beige defect is selective for NK cells since killer function in promonocytes, macrophages, and alloimmune or lectin-generated T cells is relatively normal. In addition, spleen cell-mediated ADCC against chicken erythrocytes and promocyte ADCC against tumor cells was also identical in homozygous mutants or their heterozygous littermate controls. However, spleen cell-mediated ADCC against tumor cells was impaired in bg mice as well as antibody-independent NK cytolysis. When combined with the work of others these results suggest that NK and K cells are identical and one effector can kill by an antibody-dependent or independent mechanism. Since the beige defect lies within the lytic machinery of the cell rather than at the level of target recognition, these results also suggest that the NK lytic mechanism as well as the effector cell type is distinct from that in T cells, macrophages, promonocytes, or nonlymphocytic effectors participating in ADCC. These mice should provide a valuable model for studies of in vivo resistance to tumors, grafts, and virus-infected cells in the absence of functional NK cells.
Early Damage of Herpes-Infected Cells by Antibody-Dependent Cellular Cytotoxicity: Relative Roles of Virus-Specified Cell-Surface Antigens and Input Virus
Authors: Shore Steven L. , Cromeans Theresa L. , Norrild Bodil 
Published: December 2022

Abstract
Previous studies in our laboratory indicated that tissue culture cells acutely infected with type 1 herpes simplex virus (HSV) could be recognized at 2 hr post-infection (PI) by antibody-dependent cellular cytotoxicity (ADCC) and damaged by ADCC at 3 hr PI. Since the design of these studies did not preculde the possibility that damage occurs even earlier in the infectious cycle, we performed the present series of experiments to assess this possibility as well as to gauge the relative contributions of virus-specified cell-surface antigens and input virus in rendering target cells susceptible to cytotoxic attack.
When present during both the first 2 hr of infection as well as during the following 4-hr ADCC assay period, both actinomycin D (1 µg/ml), an inhibitor of RNA synthesis, and cycloheximide (5 µg/ml), an inhibitor of protein synthesis, strongly reduced the cytotoxic reaction (by 70 and 90%, respectively). The inhibitory effects of these agents indicate that the bulk, but not all, of the ADCC reaction was directed at target cells synthesizing virus-specified surface antigens. Between 10 and 30% of the injured cells were attacked because of input virus that had not penetrated the target cell. This was shown by a) the small portion of ADCC that resisted actinomycin D and cycloheximide, b) the biphasic ADCC kinetic reaction curve of target cells exposed to antibody and effector cells immediately after virus adsorption, and c) the small but reproducible cytotoxic reaction against target cells exposed to virus inactivated by ultraviolet light. A small portion of the infected cell population could be injured by ADCC within the first hour after viral adsorption due to input virus.
Mechanisms of Action of Hyperimmune Serum in Mediating Protective Immunity to Rodent Malaria (Plasmodium Berghei)
Authors: Quinn Thomas C. , Wyler David J. 
Published: December 2022

Abstract
Little is known of the mechanisms whereby antibody can transfer protective immunity to malaria. We therefore examined in vivo the effects of the administration of hyperimmune serum to nonimmune rats on 1) the course of parasitemia after inoculation of P. berghei-infected erythrocytes, 2) the clearance of 51Cr-labeled infected erythrocytes, 3) the intraerythrocytic maturation of parasites, and 4) the invasion of erythrocytes by merozoites. Parasitemia rose in rats treated with normal serum but decreased in rats treated with hyperimmune serum. This protective effect of hyperimmune serum was not associated with a significant increase in the rate of clearance or the total reticuloendothelial organ uptake of 51Cr-labeled infected erythrocytes. Hyperimmune serum had no effect on the early intraerythrocytic development of the parasites since parasite maturation from ring form to trophozoite stage progressed normally. However, ring forms failed to increase after schizont rupture in recipients of hyperimmune serum, whereas an increase was observed in recipients of normal serum. Our in vivo study therefore supports in vitro evidence that the major protective role of antibody is to interfere with the interaction of the extracellular form of the parasite (merozoite) and the erythrocyte during the invasion process. Specifically, our data exclude opsonization of infected erythrocytes or interference with early parasite maturation as major mechanisms of antibody-mediated protection. Finally, our study suggests that the spleen may play an additional role by removing opsonized merozoites, since hyperimmune serum administered to splenectomized rats was only partially effective in preventing merozoite invasion of erythrocytes.
Isolation of the Fourth Component (C4) of Rat Complement
Authors: Daha Mohamed R. , van Es Leendert A. 
Published: December 2022

Abstract
The fourth component of rat complement was purified to homogeneity by sequential chromatography of rat plasma in benzamidine on QAE-A50, SP-C50, hydroxyapatite, and gel filtration on Bio-Gel A 1.5. The final material was homogeneous on SDS-PAGE analysis and had a calculated m.w. of 198,000. A monospecific antibody against rat C4 was obtained from immunized rabbits. The concentration of rat C4 in the plasma of normal 4-month-old Wistar rats was 190 ± 34 µg/ml (mean ± 1 S.D.).
Preservation of C3, C5, and C5a Functional Activities by a New Radiolabeling Method; Demonstration of C5 Products in Complement-Activated Serum
Authors: Kreutzer Donald L. , Kunkel Steven , Ward Peter A. , Showell Henry , McLean Robert H. 
Published: December 2022

Abstract
A new method is described for the radiolabeling of C3, C5, and C5a. Using a methylation procedure we have labeled C3, C5, and C5a with 14CH2O, achieving high specific activity, without loss of hemolytic activity in the case of C3 and C5, and in the case of C5a, full retention of chemotactic and enzyme releasing activities. The distribution of radiolabel in the α and β chains of C5 was 75 and 25%, respectively. Using intact labeled C5 the fragmentation pattern in zymosan-activated human serum, assessed by gel filtration and electrophoresis, is complex, yielding multiple fragmentation products. The labeling method described in this paper yield materials that have many advantages over the currently used labeling procedures.
Characterization of the IgE Receptor Isolated from Human Basophils
Authors: Hempstead Barbara L. , Parker Charles W. , Kulczycki Anthony 
Published: December 2022

Abstract
The IgE receptor of human basophils was purified by using simple and repetitive affinity chromatography on human IgE-Sepharose. Basophils were partially purified from peripheral blood of patients with chronic myelogenous or basophilic leukemia. Cells were labeled with 125I by using the lactoperoxidase method and were solubilized with nonionic detergent. Elution of IgE-Sepharose with 0.5 N acetic acid, 1% NP-40 allowed recovery of active IgE receptor. Analysis of human IgE receptor by SDS polyacrylamide gel electrophoresis with 10% gels demonstrated one major radioactive peak with an apparent m.w. of 58,000 to 68,000, somewhat larger than rat IgE receptor. The purified human IgE receptor was active since ∼10 to 42% of labeled receptor could specifically rebind to insolubilized human IgE. Rebinding was blocked by nanomolar concentrations of soluble human IgE or rat IgE but not by human or rat IgG, heat-inactivated human IgE, or heat-aggregated human IgG; thus it appears that rat IgE as well as human IgE binds specifically to human IgE receptor. The relative abilities of active rat IgE and active human IgE to inhibit human IgE receptor rebinding could not be precisely determined because of the limitations in assessing the proportion of human IgE that retains receptor-binding activity.
Presence of Circulating Anti-H-2 Antibody and Cell-Mediated Precursors in Successful DBA/2J-Plus-(DBA/2J × C3H/HeJ)F1 Parabiosed Mice
Authors: Drell Daniel W. , Carlson George A. , Wegmann Thomas G. 
Published: December 2022

Abstract
Joining adult DBA/2J (H-2d) mice with adult (DBA/2J × C3H/HeJ)F1 (H-2d/H-2k) mice in parabiosis results in long-term survival of 44% of such pairs, the complete takeover of the F1 lymphoid and erythropoietic systems by parent type cells, and specific unresponsiveness toward C3H/HeJ skin grafts on the parental partner. In an adoptive transfer system measuring the killing of an 131I-iododeoxyuridine-labeled tumor that is H-2 compatible with C3H/HeJ, parabiont spleen cells show full reactivity upon sensitization to C3H/HeJ alloantigens and do not inhibit the sensitization of normal DBA/2J cells to C3H/HeJ. The adoptively transferred parabiont cells do not appear to be active unless reexposed to C3H/HeJ antigen. However, intact parabionts show accelerated tumor destruction in situ even when not deliberately restimulated with C3H/HeJ spleen cells. Parabiont serum gives accelerated tumor clearance on passive transfer. Parabiont serum also shows complement-dependent antibody titers in vitro of 1:10 to 1:20 against lymphoid target cell antigens coded for by genes in the interval from K to I-E of the H-2 complex. Most of this activity is sensitive to 2-mercaptoethanol. Although circulating antibody is present in the parabionts, F1 parabiont skin shows no delay in rejection by normal DBA/2J recipients.
Antibody Responses of Tumor-Bearing Mice to Their Own Tumors Captured and Perpetuated As Hybridomas
Authors: Simrell Charles R. , Klein Paul A. 
Published: December 2022

Abstract
Experiments were performed to determine whether antibody-producing hybridomas could be derived by fusion of BALB/c P3X63Ag8 myeloma cells and spleen cells from C57BL/6 mice either immune to or bearing a syngeneic methylcholanthrene (MCA)-induced fibrosarcoma (M3). Mice were immunized by tumor amputation followed by injections of tumor cells, purified tumor cell membranes, or murine leukemia virus isolated from M3 cells (M3-MuLV). Tumor-bearer spleens were obtained at weekly intervals over a 6-week course of tumor bearing. Hybridoma cultures showing the best growth in selective HAT medium were screened by radioimmunoassay for antibody production against a) viable and glutaraldehyde-fixed cultured M3 cells, b) glutaraldehyde-fixed in vivo-passaged M3 cells, c) purified M3 cell membranes, and d) M3-MuLV. Stable antibody-producing cloned hybridomas were obtained only from fusions involving tumor-bearer spleen cells. One clone (2C2-A1) secretes an IgM antibody that binds well to purified M3-MuLV and M3 membranes, but only weakly to intact M3 cells. Another cloned line (3A5-L1) secretes an IgG1 antibody that binds to M3 cells but not to M3-MuLV. Further analysis of specificity by binding and absorption studies involving more than 30 other normal and transformed murine cell types indicates that 3A5-L1 antibody binds to some, but not all, MCA-induced murine fibrosarcomas. 3A5-L1 antibody demonstrates little, if any, binding to murine tumors of other histologic types or to normal tissues. The results demonstrate that it is possible to capture and perpetuate part of the tumor-bearing host's antibody repertoire in the form of hybridomas secreting monoclonal antibodies specific for tumor-associated antigens.
Mechanism of Action of Human B Cell-Activating Factor
Authors: Wood David D. 
Published: December 2022

Abstract
Human monocytes have been reported to release factors that can elicit distinct responses from a number of different target cells. Three of these activities are compared in this report. Supernatants of endotoxin-stimulated monocytes have been shown to stimulate the direct plaque response of T cell deficient splenocytes in vitro (BAF assay), to stimulate the thymidine incorporation by thymocytes (mitogen assay), and to synergize with phytohemagglutinin in the stimulation of thymocytes (PHA-enhancing assay). By gel filtration, plaque stimulating activity was recovered as a single peak with a m.w. of approximately 15,000 daltons when tested on nude splenocytes. When assayed on normal C57BL/10CR splenocytes, a second peak of plaque-stimulating activity was observed near 35,000 daltons. Mitogenic activity was observed both within the albumin peak and coincident with the 15,000 dalton plaque stimulator. Finally, PHA-enhancing activity was found to be broadly distributed between 8000 and 66,000 daltons, and included molecules that co-eluted with the 35,000 and 15,000 dalton plaque stimulators. Upon further purification of the 15,000 dalton material by isoelectric focusing, the plaque-enhancing, mitogenic, and PHA-enhancing activities co-eluted. This observation raised the possibility that all three activities of the 15,000 dalton material were due to the effects of a single factor on T cells. Some investigators favor this possibility based upon the study of the supernatants of human mixed leukocyte cultures (XRF). Here it is shown that spleen cell populations can be prepared that responded to BAF but not XRF, and vice versa. Therefore, studies on XRF cannot be used as evidence for the mechanism of the action of BAF. These results raise the possibility that this single factor may effect more than one target cell and may be responsible for a number of the sequellae of the stimulation of the reticuloendothelial system by bacterial products.
Purification and Characterization of a Single Chain Precursor C3-Protein (PRO-C3) from Normal Human Plasma
Authors: Minta Joe O. , Ngan Bo-Yee , Pang Anthony S. D. 
Published: December 2022

Abstract
Fresh human plasma was treated with proteinase inhibitors and passed through an immunoadsorbent column of Sepharose anti-C3 globulin. The insolubilized C3 was eluted with 5 M guanidine and, after extensive dialysis, was reduced with 0.2 M 2-mercaptoethanol and electrophoresed on SDS-polyacrylamide gels. The bulk of the eluted C3 dissociated into two major protein bands, m.w. 125,000 and 75,000 corresponding to the α-and β-chains of C3. In addition, a nonreducible single polypeptide chain (SPC) with a m.w. value of 197,000 ± 2,000 similar to the apparent m.w. of unreduced C3 was consistently present. SPC has been purified by elution from SDS (SPC*) and found to remain a single polypeptide chain upon re-electrophoresis on SDS gels in the presence of 0.2 M 2-Mercaptoethanol. The purified SPC reacted with antisera to denatured C3, C3α, and C3β chains. Additionally, antisera to SPC*, also reacted with denatured C3, C3α-, and C3β-chains, revealed a reaction of identity between SPC and C3, and detected partial identity between SPC and C3α- as well as C3β-chains. This suggested that SPC and C3 are antigenically related. The amino acid compositions and tryptic peptide maps of SPC and C3 were similar. Based on these findings, it is suggested that SPC must represent a single-chain precursor C3 (pro-C3) in plasma that escaped post-synthetic proteolytic cleavage into a two-subunit chain C3 molecule.
Synthesis of a Lipopolysaccharide Designed to Conjugate Haptens and Proteins to Erythrocytes for Use as Target Cells in Passive Hemagglutination and Hemolytic Assays
Authors: Bankert Richard B. , Mayers George L. , Pressman David 
Published: December 2022

Abstract
The synthesis of a reagent that is capable of conjugating a wide variety of haptens and proteins to cell membranes is presented. A very large chemically reactive glucose polymer (i.e., oxidized dextran), to which fatty acid side chains of myristic acid have been added, covalently binds to amino groups of haptens or proteins forming a Schiff base. The protein or hapten myristoyl dextran compounds attach directly to cell membranes when incubated with erythrocytes. The attachment of the ligands to the cell presumably occurs via the intercalation of the lipid portion of the myristoyl dextran compounds into the hydrophobic lipid bilayer of the plasma membrane. The conjugation of haptens and proteins to plasma membranes via this synthetic lipopolysaccharide reagent has several significant advantages over other coupling procedures: 1) conjugation of ligands occurs rapidly at conditions that are physiologic with respect to the cell, 2) both haptens and proteins can be conjugated via the same procedure, 3) the conjugating reagent, i.e., hapten-myristoyl dextran or protein-myristoyl dextran, is chemically stable and can be stored for extended periods of time at 4°C, 4) target cells prepared via myristoyl dextran detect picogram levels of antibody in hemolytic assays without any detectable nonspecific lysis, and 5) a very small amount of the hapten or protein myristoyl dextran reagent is required to produce target cells (i.e., as little as 1 to 10 µg of hapten).
The following compounds have been successfully conjugated to cell membranes by using myristoyl dextrans as the linking reagent; N-ε-DNP-lysine, 3-aminopyridine, 4-aminopyridine, 4-aminobenzenearsonate, 4-aminobenzoate, 4-aminophthalate, 5-aminoisophthalate, Bence Jones protein, human γ-globulin, bovine γ-globulin, rabbit anti-human Fab antibody, and mouse anti-phthalate antibody. The effective use of myristoyl dextran-conjugated erythrocytes as target cells in the passive hemagglutination assay, the hemolytic spot test, the Jerne plaque assay, and an antigen-mediated hemolytic assay is established and the sensitivity of the target cells is quantitatively compared in these assays to target cells prepared by other commonly used procedures.
Induction of Lysosomal Enzyme Secretion by Alveolar Macrophages in Response to the Purified Complement Fragments C5a and C5a des-arg
Authors: McCarthy Kathleen , Henson Peter M. 
Published: December 2022

Abstract
Purified C5a and its “inactive” form, C5a des-arg, were shown to induce secretion of acid hydrolases from rabbit alveolar macrophages (AM) in a concentration-dependent manner. Secretion increased with time to 5 times above controls by 72 hr. Concentrations of these enzymes in the cell lysates did not decrease during the incubation, suggesting that synthesis of new enzyme was occurring.
The lysosomal enzyme secretion was accompanied by increased pinocytosis and release of proteolytic enzymes from the macrophages. At no time was significant lactic dehydrogenase liberated, indicating that secretion was selective and not due to cell death.
Data presented also suggest that C5a des-arg induced secretion from the macrophages of a chemotactic factor for neutrophils. It was concluded that C5a and C5a desarg may play a role in lung injury by interactions with AM, inducing the secretion of acid hydrolases and proteolytic enzymes that can cause tissue damage, and by regulating the influx of other inflammatory cells into the interstitium and air spaces.
Macrophages in Resistance to Rickettsial Infection: Macrophage Activation in Vitro for Killing of Rickettsia Tsutsugamushi
Authors: Nacy Carol A. , Meltzer Monte S. 
Published: December 2022

Abstract
Rickettsia tsutsugamushi, strain Gilliam, replicates in cultures of resident peritoneal macrophages from BALB/c mice. Macrophage cultures treated with culture supernatants of spleen cells from rickettsial-infected mice stimulated with heat-killed rickettsiae markedly suppressed macrophage infection by rickettsiae. Rickettsiacidal activity of activated macrophages was dependent upon both lymphokine concentration and time of incubation in lymphokines. Treatment of macrophage cultures with lymphokines before exposure to viable rickettsiae resulted in an immediate decrease in percent macrophages infected and numbers of viable intracellular rickettsiae. In these cultures, enhanced intracellular killing was also apparent with further incubation (24 hr). The immediate effect of lymphokine-pretreated macrophages was dissociated from intracellular killing by infecting macrophage cultures first and adding lymphokines after infection. In these cultures, both percent macrophages infected and titers of viable intracellular rickettsiae were dramatically reduced as well.
Sex-Related Immunocompetence of BALB/c Mice
Authors: Krzych Urszula , Thurman Gary B. , Goldstein Allan L. , Bressler Joseph P. , Strausser Helen R. 
Published: December 2022

Abstract
The responses of lymphoid cells from the thymus, lymph nodes, and spleen of male and female BALB/c mice were evaluated to determine if sex-related variations in immune expression could be found. Immunologic assays used included blastogenic responses to mitogens, mixed lymphocyte responses, and direct and indirect measurement of plaque-forming cells against soluble and particulate antigens. The results indicated that responses of spleen cells from young adult female mice were higher than those of males in all comparative tests. Little or no differences between the sexes were observed in the mitogenesis of lymph nodes and thymuses. Newborn mice did not demonstrate the sex-associated immune differences. Among the weanling mice slight differences between male and female spleen cells responsiveness to mitogenic agents were observed.