bluuebunny/crossref_metadata_2025 huggingface dataset:
Lysis of Neuraminidase-Treated Paroxysmal Nocturnal Hemoglobinuria (PNH) Erythrocytes in Normal Human Serum
Authors: Koethe S. , Hause L. , Rothwell D.
Published: January 2023

PNH cells are known to lyse in normal human serum (NHS) at acid pH (Ham's test) but not (or very little) in serum at physiologic pH. When PNH cells are treated with neuraminidase (PNH-NA) their lysis in acid serum is increased (Yachin, S. et al., Blood 17 [1961] 83); in addition they lyse (10 to 58%) in ABO compatible NHS at physiologic pH. There are three possible explanations for the lysis of PNH-NA in NHS to consider: a) An antibody to PNH-NA is present in NHS; b) PNH-NA are inherently more susceptible to complement lysis than PNH; or c) PNH-NA actively participate in the lytic process by serving as an activator of the alternative complement pathway. Against a), absorption of the serum at 0°C three times with PNH-NA did not appreciably remove its lytic activity. And against b), PNH and PNH-NA lysed equally well in ABO incompatible serum (a source of antibody and complement), and they lysed equally well when used as indicator cells in a CoVF-reactive lysis assay.
IgG Behaves Like IgM Immunoglobulin in the Presence of Protein A
Authors: Langone John J. , Boyle Michael D. P. , Borsos Tibor
Published: January 2023

Protein A (PA) is a cell wall component isolated from Staphylococcus aureus that binds specifically to the Fc region of IgG from several species including IgG subclasses 1, 2, and 4 from man. We investigated the effects of PA on the ability of rabbit IgG antibody to Forssman antigen to sensitize sheep erythrocytes (E) to lysis by excess guinea pig complement (GPC). The slope of the hemolytic dose response curve and the hemolytic titer of IgG were measured in buffer alone or in the presence of differing amounts of PA under conditions where PA had no significant effect on C activity. In buffer alone and at high ratios of PA:IgG the slope of the dose response curve was the same (4.4); but at high ratios of PA:IgG the titer of the IgG was markedly reduced. At lower ratios of PA:IgG the slope of the dose response curve was close to 1, a characteristic of IgM-like antibody.
Translation in Vitro of Mouse Liver mRNA Into C3, C4 and Albumin in a Protein-Synthesizing System from Rabbit Reticulocyte Lysate
Authors: Minta J. O. , Patel Feisal , Wax Mark
Published: January 2023

In an attempt to define the molecular basis of the genetic lesions responsible for some of the inherited complement deficiency states we have initiated studies on the ability of isolated mRNA to direct the synthesis of complement components in vitro. Poly (A)-mRNA was isolated from the post-mitochondrial supernatant fraction of mouse liver homogenate by phenol-chloroform extraction and subsequent oligo (dT)-cellulose chromatography. Translation of the isolated mRNA in a reticulocyte lysate cell-free system, in the presence of 3H leucine and an energy-synthesizing system, greatly stimulated the incorporation of the labeled amino acid into protein in a time and dose-dependent fashion. Specific immunoprecipitation of the translated products with the double antibody technique showed that proteins antigenically related to C3, C4, and albumin had been synthesized. The immunoprecipitates were analyzed by SDS/polyacrylamide gel electrophoresis. The immunoprecipitated C4 radioactivity migrated predominantly in a single peak, m.w. 230,000.
Biosynthesis of the Subcomponents of C1 by Twelve Human Established Cell Lines
Authors: Morris K. M. , Bing D. H. , Andrews J. M. , Silverstein L. J. , Shohet R. V. , Attisano C. M. , Goran P. S.
Published: January 2023

Synthesis of the subcomponents of C1 was detected in human cell lines derived from fetal intestine, intestine of an agammaglobulinemic fetus, colorectal adenocarcinomas, normal bladder, and transitional cell carcinomas of the bladder. Criteria for synthesis were detection of hemolytic activity in the culure medium; immunoprecipitation of specifically labeled protein after incubation of the cells with medium containing 3H- or 14C-labeled amino acids; isolation from the immunoprecipitates of labeled proteins with m.w. approximating those of the subcomponents of C1; and identification on autoradiographs of labeled precipitin bands after immunoelectrophoresis of labeled extracellular or intracellular medium and reaction with monospecific antisera.

Fetal intestine and intestine derived from an agammaglobulinemic fetus synthesized similar amounts of C1q, C1r, and C1s. Both synthesized twice as much C1s as C1r and one-half as much C1q as C1r. Hemolytically active C1̄, isolated from IgG-Sepharose, gave m.w. analogous to published reports on SDS-polyacrylamide gels.
Studies of the Alternative Complement Pathway (AP) in Normal Children
Authors: Norman Michael E. , Taylor Arlene , Green Paul , Laster Larry , Nilsson Ulf R.
Published: January 2023

Increasing evidence suggests that activation of complement by AP contributes an important protective mechanism in early defense against bacterial invasion, before effective antibody levels are established. Since this is particularly important in children, we undertook a survey of AP in 83 normal children, ages 3 days to 15 years (x̄ = 54 months). There were approximately equal numbers of males and females, blacks and whites, and all children were free of fever and intercurrent infection. C3, Factor B, properdin,1 and β1H were quantitated by radial immunodiffusion with monospecific antisera; hemolytic function of AP was monitored by lysis of glutathione-treated human erythrocytes with inulin activated serum (N. Engl. J. Med. 286:180, 1973). Data were compared to normal adults. There was a wide range of values when compared to adults except for C3.
Biosynthesis of the First Component of Complement by Human Fibroblasts
Authors: Reid K. B. M. , Solomon E.
Published: January 2023

C1-hemolytic activity was synthesized and secreted by all nine human fibroblast cell lines examined. The magnitude of the hemolytic titer appeared to be partially dependent on the batch of fetal calf serum used to culture the cells. The fibroblast C1 hemolytic activity was destroyed by heating at 56°C and, after activation, by treatment with 2 mM-DFP. Rabbit antihuman C1q inhibited the fibroblast C1-hemolytic activity. Biosynthesis of fibroblast hemolytic activity was inhibited by the presence of cycloheximide and regained on its removal. Fibroblasts were grown in the presence of radioactive amino acids and the culture media examined for the incorporation of radioactivity into C1q, C1r, and C1s as judged by the formation of specific immunoprecipitates. Polyacrylamide gel electrophoresis in sodium dodecyl sulphate of the immunoprecipitates formed using antisera against C1r and C1s provided convincing physiochemical evidence for the de novo biosynthesis of these subcomponents.
Interaction of the Third Component of Human Complement (C3) with Erythrocytes and Leukocytes
Authors: Tack B. F. , Segal D. M. , Schechter A. N.
Published: January 2023

A direct binding assay has been used to assess the kinetic and equilibrium parameters of the interaction between human C3 and its cell surface receptors on human blood cells. The C3 protein was radiolabeled using Na 125I and chloramine-T by standard methods to a specific activity of approximately 1 × 106 cpm/µg. The method of assay in present use is performed in the following manner: a) the labeled protein and test cells are incubated at 37°C in isotonic Tris:HCl buffer (pH 7.4) until equilibrium has been reached, b) an aliquot of the cell suspension is then layered on a column of synthetic oil (ρ = 1.033 gm/cc) contained in a microfuge tube, and (c) a rapid separation of free and bound labeled protein is obtained by pelleting the cells. The free ligand concentration is determined by counting an aliquot of the supernatant and the specific and nonspecific cell associated counts by counting the tip of the microfuge tube.
Direct Demonstration and Quantitation of Native and Activated C1 in Human Serum
Authors: Ziccardi R. J. , Cooper N. R.
Published: January 2023

The first component of complement, C1, can be directly demonstrated in normal human serum by double immunodiffusion analyses since native macromolecular C1 gives a continuous line of precipitation with antisera to C1q, C1r, and C1s. All of the C1q, C1r, and C1s in normal serum were found to be in the C1 complex. These observations allow the quantitation of C1 in human serum by single radial immunodiffusion techniques.

Activation of C1 in human serum can be readily detected by double immunodiffusion studies with anti-C1q, anti-C1r, and anti-C1s because C1̄ exhibits a pattern quite different from that of native C1. In order to investigate this phenomenon, increasing doses of aggregated IgG were added to serum. Three major changes were apparent in the Ouchterlony patterns. First, spurring of the C1s precipitin line over that of macromolecular C1, indicating release of C1s from C1̄, was observed with the lowest doses of activator examined.
Selective Suppression by Adherent Cells, Prostaglandin, and Cyclic AMP Analogues of Blastogenesis Induced by Different Mitogens
Authors: Novogrodsky Abraham , Rubin Albert L. , Stenzel Kurt H.
Published: January 2023

Depletion of adherent cells markedly enhances blastogenesis induced by the galactosyl-directed lectins, soybean agglutinin, peanut agglutinin, and hepatic binding protein, but has little effect on lymphocyte blastogenesis induced by phytohemagglutinin or concanavalin A. Treatment of unfractionated lymphocytes with indomethacin and other agents that inhibit prostaglandin synthesis mimics the selective effect of removing adherent cells. After depletion of adherent cells, these agents no longer affect mitogen-induced blastogenesis. Treatment of lymphocytes that had been depleted of adherent cells with prostaglandin E1 selectively suppresses responses to hepatic binding protein, peanut agglutinin, and soybean agglutinin, whereas responses to phytohemagglutinin and concanavalin A are less affected. Cyclic AMP analogues and an inhibitor of cyclic AMP phosphodiesterase, methyl isobutyl xanthine, also selectively suppress responses to the galactosyl-directed lectins. Prostaglandin E1, cyclic AMP analogues, and methyl isobutyl xanthine are most effective when added early in the course of lymphocyte activation. Prostaglandin E1 levels are similar in the culture media of cells treated with the different mitogens, and are suppressed by both indomethacin and the removal of adherent cells. The data indicate that cells stimulated by different mitogens differ markedly in their sensitivity to suppression by similar levels of prostaglandin. The gradation of sensitivity to prostaglandin and to cyclic AMP of blastogenesis induced by the various mitogens studied, from most to least sensitive, is: hepatic binding protein ≧ peanut agglutinin > soybean agglutinin > concanavalin A > phytohemagglutinin.
Synthesis of Specific Lipids Associated with the Hormone-Induced Resistance of Tumor Cells to Humoral Immune Killing
Authors: Schlager Seymour I. , Ohanian Sarkis H. , Borsos Tibor
Published: January 2023

Selected polypeptide (insulin), catecholamine (epinephrine), and glucocorticosteroid hormones (hydrocortisone and prednisolone) that increase the resistance of line-1 guinea pig hepatoma cells to humoral immune killing were studied for their effects on the ability of the cells to synthesize lipids. Line-1 tumor cells incubated for 1 hr at 37°C with 10-7 to 10-9 M concentrations of any of these hormones became more resistant to killing by antibody plus GPC and exhibited increased levels of fatty acid incorporation into lipids compared to untreated cells. Cells incubated with hormone for 4 hr at 37°C became as susceptible to antibody-GPC killing as untreated cells; lipid synthesis in these cells likewise reverted to control levels. When the reverted cells were reincubated with a different, but not the same, hormone, they were again rendered more resistant to antibody-GPC killing and their lipid synthesis was restimulated. Thin layer chromatography of lipid extracts from hormone-treated as well as from restimulated cells indicated that the increased resistance of the cells to killing correlated with increased synthesis of phosphatidyl choline, phosphatidyl serine, and, in some instances, triglycerides. The cells' reversion to the susceptible state during prolonged incubation with hormone or restimulation with the same hormone was accompanied by a reduction in the synthesis of these lipids along with an increase in cholesteryl ester and cardiolipin synthesis. The synthesis of specific phospholipids and neutral lipids, therefore, rather than lipid synthesis in general, appears to be associated with the mechanism whereby these tumor cells resist humoral immune killing.
Serum Decay and Placental Transport of a Mutant Mouse Myeloma Immunoglobulin with Defective Polypeptide and Oligosaccharide Structure
Authors: Weitzman Stephen , Palmer Lucy , Grennon Margery
Published: January 2023

A mutant mouse myeloma cell line, M3.11, has been previously isolated from an IgG2b-producing wild type. The immunoglobulin (Ig) produced by M3.11 has been shown in other studies to have the following structural defects: 1) deletion of the Cγ3 domain; 2) failure to glycosylate 30 to 40% of its secreted heavy (H) chains; and 3) a block in assembly with approximately equal amounts of whole molecule (H2L2) and half-molecule (HL) being secreted. In addition, the sialic acid content of the wild type and the mutant is similar and minimal. Experiments comparing two biologic functions of these Ig demonstrated that M3.11 had a markedly increased serum decay and a more rapid placental transport than the wild type. The three distinct M3.11 species, H2L2, nonglycosylated HL, and glycosylated HL, all had indistinguishable serum half-lives. In addition, M3.11 in vitro exhibited an increased sensitivity to trypsin and a more acidic pI than the wild type. These results support several conclusions regarding the structural features influencing Ig serum decay. Firstly, the serum decay and placental transport of Ig seem to be determined by the H chain polypeptide and not by the presence or absence of carbohydrate in general or of sialic acid in particular. Secondly, those aspects of protein structure that do influence serum decay are not restricted to a single domain but probably represent a complex interaction of multiple sites on the Ig molecule.
Early Complement Components in NZB/NZW Mice
Authors: Olsen Carol T. , Gabrielsen Ann E.
Published: January 2023

Sera from periodic bleedings of B/W female mice were assayed for C1 hemolytic activity. It peaked at 3 to 4 months of age and declined to very low levels by 6 to 7 months. Activity remained low even in long-lived animals.
Isolation of a Type-Specific Antigen from Chlamydia Trachomatis by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis
Authors: Sacks David L. , MacDonald A. Bruce
Published: January 2023

This report describes the isolation of a type-specific antigen of a serotype A strain of Chlamydia trachomatis. The antigen could be identified in sodium dodecyl sulfate-polyacrylamide electrophoretic (SDS-PAGE) analysis of immunoprecipitates of homologously reacted lysates from Bolton-Hunter 125I-labeled elementary bodies, solubilized by sonication and treatment with Nonidet P40. The electrophoretic pattern of this precipitate revealed a peak of unique mobility that was not reproduced by heterologous or control precipitates. Immunoadsorbtion of test antigen with purified IgG fractions from homologous antisera completely removed this peak, whereas similar adsorbtion with heterologous IgG had minimal effect. Comparison of this antigen in SDS-PAGE with protein standards revealed an approximate m.w. of 27,000.
The Appearance of IgM and IgG Cold Agglutinins in Rabbits Hyperimmunized with Group C Streptococcal Vaccine
Authors: Colling Richard G. , Brown John Clifford
Published: January 2023

Rabbits hyperimmunized with Group C streptococcal vaccine developed both IgM and IgG erythrocyte autoantibodies reactive with human type O and rabbit red blood cells at reduced temperatures. Although antibody preparations exhibited maximum agglutinating activity toward human type O and rabbit erythrocytes when assayed at 4°C, significant agglutinating activity toward human type A and sheep erythrocytes was observed when the preparations were assayed at 37°C. The cold agglutinins isolated by thermal elution from rabbit erythrocytes were also effective hemolysins; less than 1.0 μg of IgM was required to sensitize rabbit erythrocytes for 50% lysis in the presence of guinea pig complement (C) in a biphasic-temperature lysis assay. When saccharide haptens, group-specific carbohydrate, and soluble blood group substances were examined for their ability to effect inhibition of lysis, the following pattern was observed. The most effective overall inhibitor was Group C streptococcal carbohydrate, whereas the most effective hapten inhibitor was N-acetylgalactosamine. Of the soluble blood group substances tested, hog mucin H-substance (first Smith degradation product) was slightly more inhibitory than hog mucin A+H-substance (first Smith degradation product) but substantially more active than human ovarian cyst material. Although the soluble blood group substances were capable of competing for rabbit erythrocyte determinants, none gave a visible precipitate in the presence of cold agglutinin when assayed by Ouchterlony double diffusion. In contrast, significant precipitation occurred when cold agglutinin was examined in the presence of Group C carbohydrate in precipitin assays and by Ouchterlony analyses. Results of these studies provide evidence to suggest that the cold agglutinins produced in rabbits hyperimmunized with Group C streptococcal vaccine are a population of group carbohydrate-specific antibodies that are cross-reactive with I antigen-like determinants present on rabbit and human erythrocytes.
The Distribution of Adenosine Deaminase Among Lymphocyte Populations in the Rat
Authors: Barton Randall , Martiniuk Frank , Hirschhorn Rochelle , Goldschneider Irving
Published: January 2023

Adenosine deaminase (ADA) activity was determined in young adult rat lymphocyte populations. The ADA-specific activity (per 108 cells and per milligram protein) was 3- to 10-fold higher in thymocytes than in lymphocytes from thoracic duct, lymph node, spleen, and bone marrow. The high ADA activity in thymocytes appeared to be preferentially associated with cortical thymocytes. Enrichment or depletion of cortical thymocytes by density gradient centrifugation, cortisone treatment, or selective lysis with anti-Thy-1 plus complement resulted in parallel increases or decreases in ADA levels. These results also suggested that medullary thymocytes have ADA levels similar to those of peripheral lymphocytes. “Immature” cortical thymocytes and thymocyte progenitors appeared to have low ADA activity; low enzyme levels were found in fetal thymus at 16 days of embryonic life, in the early phases of thymus regeneration, and in a “null” cell population isolated from bone marrow. This study demonstrates that ADA activity varies markedly during T lymphocyte differentiation and suggests that fundamental differences in nucleotide metabolism may exist in T cells at different stages of development.
Presence of Alpha-1-Antitrypsin on Mitogen-Stimulated Human Lymphocytes
Authors: Lipsky James J. , Berninger Ronald W. , Hyman Lawrence R. , Talamo Richard C.
Published: January 2023

Human lymphocytes that have undergone concanavalin A-induced blastogenetic transformation demonstrate surface alpha-1-antitrypsin by immunofluorescence when examined after 72 hr of culture, whereas unstimulated cells do not. These results may indicate a role for alpha-1-antitrypsin in lymphocyte blastogenesis.
Cell-Mediated Suppression of the Fifth Component of Complement in Mice
Authors: Goldman Margaret B. , French Deborah L. , Goldman John N.
Published: January 2023

Suppression of levels of circulating C5 in (C5-C5+)F1 hybrids by administration of (C5-C5-) parental lymphoid cells in the neonatal period has been accomplished with the three strain combinations tested ((SWR × RIII)F1, (A/He × RIII)F1, and (SWR × DBA/1)F1). Suppression was shown to be specific for C5 and not accompanied by reductions of C4, C2, C6, or other major groups of blood proteins. This demonstrated that the C5 reduction was not due to activation of complement (C) with resultant hypercatabolism of C components. When there was a concurrent chronic GVH reaction induced by lymphoid cells administered to offspring of H-2 incompatible parents, there was usually a resultant hypergammaglobulinemia that was also unrelated to the presence or absence of C5 suppression. Effective suppression required preimmunization of either the cell donor, the mother of the F1 hybrids, or both. This suggests that either two cell types or a single cell plus a humoral factor are required for suppression in this system.
Functional Studies on Lymphocytes in Adult Human Bone Marrow
Authors: de Gast Gÿsbert C. , Platts-Mills Thomas A. E.
Published: January 2023

In seven preparations from adult human bone marrow (BM), surface IgM-positive (SmIgM+ve) cells were detected with anti-IgM coated ox red blood cells (anti-IgM-OxRBC). On the average, SmIgM+ve cells in the lymphocyte rich fractions from BM bound fewer anti-IgM-OxRBC than peripheral blood (PB) SmIgM+ve cells. After isolation on a Ficoll/metrizoate gradient, BM sIgM+ve cells were cultured with mitomycin C-treated PB T cells and pokeweed mitogen (PWM). These cells produced 0.44 ± 0.52 μg IgM/ml and only 0.10 ± 0.11 IgG/ml. By contrast, PB sIgM+ve cells under similar conditions produced 11.5 ± 10.4 μg IgM/ml and 6.5 ± 7.0 μg IgG/ml. In addition, BM sIgM+ve cells showed very poor PWM-induced thymidine incorporation in cultures with PB Tm (mean stimulation index 2.5 ± 2.0) compared to sIgM+ve cells from PB (mean 29.9 ± 25.3). On the other hand, the stimulator capacity of BM sIgM+ve cells in the mixed lymphocyte reaction was similar to that of PB sIgM+ve cells (mean stimulation ratio, BM: 11.1 ± 7.2; PB: 18.9 ± 14.6). The results suggest that B lymphocytes in adult BM develop IgM surface markers and the ability to stimulate in the MLR without becoming fully responsive to PWM for Ig production or proliferation. In addition, these “immature” B cells appear to have less IgM on their surface than PB B cells.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Howard M. C. , Fidler J. M. , Baker J. , Shortman K.
Published: January 2023

The primary “virgin” B lymphocyte population of adult CBA mouse spleen was separated on the basis of physical properties and AFC response kinetics into two distinct functional subsets. The biphasic nature of the primary splenic AFC response to NIP-POL reflected the separate expression of these two subpopulations. The least mature subset consisted on atypical, medium to large sized B cells, which took 8 days after stimulation to produce a peak IgM AFC response, and which were preferentially responsive in adoptive transfer assays. The more mature subset consisted mainly of typical, small B cells, which took only 3 to 4 days after stimulation to produce a peak IgM AFC response, and which were preferentially responsive in cell culture assays. There was no evidence that these subsets represented T-independent versus T-dependent B cell subgroups. It is proposed that these subsets are sequentially related, and that they be termed “preprogenitor” and “direct AFC-progenitor” B cells. Their distinction from earlier, “pre-B” cell forms is discussed.

Secondary, “memory” B lymphocytes from long-term hapten primed adult CBA mouse spleen, initiating IgM or IgG responses to NIP-POL, could not be physically separated into two distinct subgroups, although some aspects of the kinetics of AFC-production suggested such a division might exist. Subsets of B cells initiating kinetically distinct “primary” responses to SRC were separable by physical criteria, and were compared with the B cells initiating primary and secondary responses to NIP-POL.

An important conclusion is that different assays, all designed to measure the ability of B cells to develop into AFC, can reflect the expression of quite different B cell subsets. The kinetics of the AFC responses are crucial to the interpretation of the results of such assays. Many differences in results between different laboratories may have their origin in this problem.
Immunochemical Characterization of the Ly-8.2 Murine Lymphocyte Alloantigen: Possible Relationship to Actin
Authors: Delovitch Terry L. , Fegelman Alan , Barber Brian H. , Frelinger Jeffrey A.
Published: January 2023

An immunochemical analysis of the Ly-8.2 antigen was performed. Reactivity of B10-radiolabeled T and B cell lysates with a C3H anti-AKR serum immunoprecipitates a major 43,000 m.w. protein detectable by one-dimensional gel electrophoresis. This protein can be radioiodinated, however, it does not appear to bind to lentil lectin. Two-dimensional gel electrophoresis and peptide-mapping studies indicate that this protein is structurally very similar to lymphocyte actin. The possible molecular relationship between Ly-8.2, other lymphocyte membrane antigens, and actin is discussed.
In Vivo Activity of a Nonspecific T Cell-Replacing Factor
Authors: Kindred Berenice , Bösing-Schneider Rita , Corley Ronald B.
Published: January 2023

Nonspecific T cell-replacing factors prepared as supernatants from mixed lymphocyte cultures or concanavalin A-stimulated spleen cells are active in vivo if injected into nude mice at least 3 days before antigen. The supernatants appear to act by enhancing the weak IgM responses that occur in untreated nudes. Secondary responses and IgG antibody were not found.
Structural Studies of Fc Receptors
Authors: Yagawa Katsuro , Onoue Kaoru , Aida Yoshitomi
Published: January 2023

The binding properties, solubilization, and sensitivity to enzymatic treatment of the Fc receptors of guinea pig peritoneal macrophages were studied. The receptors on the cell surface were saturable with soluble immune complexes, and the binding showed linearity in the Scatchard plot. The observed linearity probably reflected preferential binding of the immune complexes of relatively large sizes. When receptors on the cells were operationally saturated, the number of antibody molecules in the cell-bound complexes was calculated to be 6.5 × 105/cell. By inhibition test on the binding of 125I-labeled soluble immune complexes to macrophages, receptor activity was found in the 20,000 × G supernatant fraction of sonically disrupted macrophages, which was precipitable at 100,000 × G. Treatment of the 20,000 × G supernatant or intact cells with Nonidet P-40 solubilized Fc receptors. In the presence of detergent, these receptors existed as molecules with a Stokes radius somewhat larger than that of IgG but they reaggregated when the detergent was removed.

Treatment of the sonic lysate of macrophages with phospholipase C revealed that Fc receptor activity on 100,000 × G-percipitable cell fragments was lost as indicated by the binding inhibition test. The loss of activity was shown to be not due to release of the receptors from the cell fragment. By using surface-labeled macrophages it was demonstrated that the activity of the receptors was restored by treating the phospholipase C-inactivated cell fragments with Nonidet P-40 as indicated by direct binding of solubilized receptors to insoluble immune complexes. This restoration of activity was probably brought about by binding of detergent to the receptors. These results suggest the possibility that Fc receptors on cell membranes require the structural support of phospholipids for their activity.
Isolation of a Polysaccharide Antigen from Schistosoma Mansoni Eggs
Authors: Boctor Fouad N. , Nash Theodore E. , Cheever Allen W.
Published: January 2023

A polysaccharide antigen was isolated from Schistosoma mansoni egg homogenates by the phenol procedure. The crude preparation (CPEA) contained at least two antigens. The more purified antigen (PEA) was isolated by sequential enzymatic treatment of CPEA with DNase, RNase, Pronase, and α-amylase. PEA was resistant to boiling, freezing and thawing, mild acid and alkali, and chloroform, but was destroyed with periodate. It gave a positive reaction with anthrone reagent. PEA was eluted in the wash fraction from a DEAE cellulose column and in the void volume of a Sephacryl 200 column. After immunoelectrophoresis and polyacrylamide electrophoresis there was little or no migration. Amino acid analysis failed to reveal ninhydrin-positive material in the a hydrolyzate of PEA. These results suggested that PEA is a neutral polysaccharide with a m.w. of more than 200,000 and contains no amino acids or hexosamine. Antibodies against PEA were detected in sera obtained from mice infected with S. mansoni. PEA is different from previously described antigens derived from schistosome eggs.
Suppressor Activity of Splenocytes from Mice Treated with Ecteinascidia Turbinata Extract
Authors: Lichter Wolf , Wellham Larry L. , Sigel M. Michael , Ghaffar Abdul
Published: January 2023

Extracts of a marine tunicate, Ecteinascidia turbinata (Ete) were previously shown to be capable of suppressing humoral and cellular immune responses in vivo and in vitro. In the present work we have examined the mechanisms of suppression in Ete-treated DBA/2 and BALB/c mice. Treatment with Ete resulted in a significant splenomegaly accompanied by a diminished response to mitogenic stimulation, reaching coincident maxima of effect at days 4 to 6. Spleen size and blastogenic reaction returned to normal at 12 to 21 days. Cells from enlarged spleens inhibited blastogenic responses of normal splenocytes to Con A. Marker studies indicated that the suppressor activity was exerted by cells possessing T lymphocyte characteristics.
Suppression of Antibody-Sensitized Tumor Cells by Macrophages: Insufficient Supply or Activation of Macrophages within Large Tumors
Authors: Johnson Robert J. , Siliciano Robert F. , Shin Hyun S.
Published: January 2023

Systemic administration of antibody against the C3H lymphosarcoma 6C3HED caused complete suppression of the growth of 105 tumor cells. When the size of the tumor inoculum was increased to 106 tumor cells, excess antibody was unable to cause complete suppression unless exogenous macrophages were added to the tumor inocula. The decreased effectiveness of antibody was shown to be caused by a local deficiency in the supply or activation of macrophages within the large tumor grafts and not by a systemic deficiency.
Mouse Antibody-Dependent Eosinophil and Macrophage Adherence and Damage to Schistosomula of Schistosoma Mansoni
Authors: Kassis Amin I. , Aikawa Masamichi , Mahmoud Adel A. F.
Published: January 2023

The effects of sera from S. mansoni-infected mice together with eosinophil- and macrophage-rich peritoneal exudate cells (PEC) on the schistosomula were examined in vitro. Adherence to and killing of the parasite by PEC is mediated through a complement-independent opsonizing IgG. Destruction of schistosomula was assessed morphologically and by evaluating their infectivity to normal mice. By using ferritin-labeled anti-mouse immunoglobulin, the antibody was localized on the surface of the schistosomula and at the cell-organism areas of contact. The serum antibody activity appears with the onset of egg production, does not correlate well with the duration of infection, and is relatively specific among schistosome species. Eosinophils and macrophages were demonstrated by electron microscopy to adhere firmly to the surface of the organisms; the latter cells were shown invading the schistosomula. Furthermore, depletion of eosinophils by prior incubation of PEC with monospecific anti-eosinophil serum blocked the early adherence phase, but a substantial part of PEC ability to kill the schistosomula was retained.
Expression of an Unidentified Immunoglobulin Isotype on Rabbit Ig-Bearing Lymphocytes
Authors: Eskinazi Daniel P. , Bessinger Beverley A. , McNicholas Janet M. , Leary Anne L. , Knight Katherine L.
Published: January 2023

A non-IgM immunoglobulin molecule was found on most rabbit Ig-bearing lymphocytes isolated from mesenteric lymph nodes. Membrane bound immunoglobulin light chains and heavy chains were detected by immunofluorescence and by rosetting with antibody-coated erythrocytes on mesenteric lymph node cells stripped of IgM by anti-IgM allotype antibodies. The percentage of cells bearing these residual immunoglobulin molecules was similar to the percentage of cells bearing immunoglobulin before “stripping” with anti-IgM antibody. These residual immunoglobulin molecules were not IgA nor IgG and are believed to be the rabbit analogue of human IgD.
Early Steps in Specific Tumor Cell Lysis by Sensitized Mouse T Lymphocytes
Authors: Gately Maurice K. , Martz Eric
Published: January 2023

It has previously been shown by several investigators that Mg++ in the absence of appreciable Ca++ is sufficient to support the formation of specific adhesions between cytolytic T lymphocytes (CTL) and tumor target cells but that programming of the targets for lysis is strongly Ca++ dependent. However, we recently found that Ca++, in the presence of suboptimal concentrations of Mg++, is capable of synergizing with Mg++ in supporting adhesion formation. The latter observation raised the possibility that the structure of the adhesion in the absence of Ca++ might be unphysiologic and incompatible with programming for lysis. Thus it was conceivable that the role of Ca++ in programming for lysis might be limited to its demonstrated role in adhesion.

In the present report we have examined the abilities of other alkaline earth metals to substitute for calcium in supporting adhesion formation and programming for lysis. In the presence of Mg++ at a concentration optimal to support adhesion, Sr++ could replace Ca++ in programming for lysis. The concentration of Sr++ required for maximum cytolysis was 5- to 20-fold greater than the optimum Ca++ concentration; nevertheless when Sr++ was present at an optimum concentration, the amount of lysis was identical to that observed in the presence of an optimum concentration of Ca++. In contrast to its ability to replace Ca++ in programming for lysis, Sr++ could not substitute for Ca++ in potentiating adhesion formation in the presence of suboptimal Mg++. Ba++ could not substitute for Ca++ in either adhesion formation or programming for lysis but inhibited both in the presence of Ca++. Likewise, high concentrations of Mg++ in the absence of Ca++ did not enhance programming for lysis.

The ability of Sr++ to replace Ca++ in cytolysis but not in adhesion formation indicates that the role of Ca++ in programming for lysis is distinct from its demonstrated role in adhesion and the calcium is required only at the former site. It is possible that the role of Ca++ in programming for lysis is to act as a co-factor essential to the activity of a CTL surface protein involved in the lytic mechanism. Alternatively, Ca++ influxes into CTL may trigger activation of the lytic mechanism in a manner analogous to the stimulus-secretion coupling seen in many secretory phenomena. The observation that the order of efficacy of alkaline earth metals is supporting programming for lysis is Ca++ τ Sr++ τ Ba++ or Mg++ is compatible with either possibility.
Adoptive Transfer of Experimental Allergic Encephalomyelitis: Incubation of Rat Spleen Cells with Specific Antigen
Authors: Richert John R. , Driscoll Bernard F. , Kies Marian W. , Alvord Ellsworth C.
Published: January 2023

Spleen cells from myelin basic protein (BP)-sensitized donor rats appear to be incapable of adoptively transferring experimental allergic encephalomyelitis (EAE) directly to normal recipients. It has been reported that in vitro incubation with concanavalin A (Con A) activates rat spleen cells so that they are capable of transferring EAE. We report here that incubation with specific antigen, BP, also permits transfer of disease with spleen cells. Data are presented in which activation of EAE spleen cells by Con A is compared with activation by BP. Cellular proliferation does not appear to be necessary for in vitro activation with specific antigen.
Primary in Vitro Cytotoxic Response of F1 T Lymphocytes against Parental Antigens
Authors: Ishikawa Hiromichi , Dutton Richard W.
Published: January 2023

Spleen cells from adult female (AKR/J × BALB/c)F1 mice can respond to mitomycin C-treated spleen from AKR/J mice and can generate effector CTL in a 5-day primary in vitro culture. The response is comparable in magnitude to the response to allogeneic H-2K or H-2D antigens. The response is T cell mediated and is directed to antigen(s) present only on the parental cells. The target cell must be homozygous at H-2Kk to be lysed and H-2Dk antigens do not serve as a target in this response. Spleen cells from (B10.BR × B10.D2) hybrids that have been stimulated with AKR/J lyse B10.Br as well as AKR/J target cells. Similar H-2k/d hybrid F1 anti-H-2k parent responses are seen in certain other strain combinations. A number of possible interpretations of these responses are discussed.
An Interpretation of the Apparent Dual Specificity of Some Murine Myeloma Immunoglobulins with Inulinbinding Activity
Authors: Streefkerk Dirk G. , Manjula Belur N. , Glaudemans Cornelis P. J.
Published: January 2023

Five murine myeloma immunoglobulins have been studied for their binding with fructose-containing homopolysaccharides. The results indicate that the anti-(2→1)-D-fructofuranans are capable of binding (2→6)-D-fructofuranans with reduced affinity. An anti-(2→6)-D-fructofuranan (U10) was incapable of binding a (2→1)-D-fructofuranan. By using molecular models of fructofuranans of both linkage types, and a hypothetical molecular model of the combining region of E109, an anti-(2→1)-D-fructofuranan, a rationalization of these data can be proposed.
Hyporesponsiveness of Canine Bronchoalveolar Lymphocytes to Mitogens: Inhibition of Lymphocyte Proliferation by Alveolar Macrophages
Authors: Ansfield Michael J. , Kaltreider H. Benfer , Caldwell Joseph L. , Herskowitz Fredric N.
Published: January 2023

Unfractionated canine bronchoalveolar cells (65% macrophages, 26% lymphocytes, and 9% granulocytes) obtained by lavage were markedly hyporesponsive to stimulation with concanavalin A, phytohemagglutinin, or pokeweed mitogen. The mechanisms responsible for the lack of responsiveness of bronchoalveolar lymphocytes were investigated. Unfractionated bronchoalveolar cells were depleted of adherent cells by sequential adsorption on plastic (nonadherent cells) and by nylon wool filtration (column-purified cells). Nonadherent and column-purified cells were substantially depleted of macrophages, enriched for lymphocytes, and displayed marked increases in their responses to mitogens (up to 7-fold).

Preparations of unfractionated bronchoalveolar cells markedly suppressed the responses of blood lymphocytes to mitogens; nonadherent and column-purified cells displayed minimal suppressive activity. Plastic-adherent cells, prepared from unfractionated bronchoalveolar cells, contained up to 93% macrophages and these adherent cells were potent suppressors of mitogen-responses when co-cultured with blood lymphocytes. Suppressor activity of adherent cells was radioresistant and was reversed by the addition to cultures of low concentrations of indomethacin. The data indicate that alveolar macrophages are the adherent cells responsible for this suppressor activity.

The responses of nonadherent and column-purified bronchoalveolar lymphocytes, although substantially greater than those of equal numbers of unfractionated bronchoalveolar lymphocytes, were consistently less than 25% of the responses of similarly prepared blood lymphocytes. Differences in cell viability between blood and lung lymphocytes were not found. Neither the addition of indomethacin nor the addition of mitomycintreated blood mononuclear cells increased the responses of column-purified bronchoalveolar lymphocytes.

Our studies demonstrate two mechanisms that limit the responses of canine lung lymphocytes to mitogenic stimulation: a suppression by alveolar macrophages of lymphocyte proliferation and an inherent hyporesponsiveness of bronchoalveolar lymphocytes. These may be of importance in regulating the response of lung to inhaled antigens.
Idiopathic Paraproteinemia
Authors: Radl Jiri , De Glopper Elisabeth , Schuit Henrica R. E. , Zurcher Chris
Published: January 2023

Transplantation experiments in the C57BL/KaLwRij mouse model of idiopathic paraproteinemia (IP) showed that an IP-producing clone can be further propagated in young, lethally irradiated mice and also equally as well in nonirradiated recipients by a bone marrow and/or spleen cell transfer. The latency period before the original paraprotein was detected in the sera of recipients varied in different experiments between 1 and 9 months after transplantation. With subsequent transplantations, the “take” frequency gradually decreased. Propagation of IP for three to four generations seems to be the final limit. In comparison to age-matched control groups, no substantial influence of the transplanted IP on the survival of the recipients was observed. In contrast, transplantation of cells from mice with a B cell lymphoma or a myeloma led to continuous propagation of the malignancy, with a high “take” frequency, progressive development of the paraproteinemia, and a shortened survival time of the recipients. These findings indicate that IP represents in its final stage in the aging C57BL mice an intrinsic cellular defect within the affected B cell clone, which is, however, different from that found in B cell malignancies.
Characterization of a Congenitally LPS-Resistant, Athymic Mouse Strain
Authors: Vogel Stefanie N. , Hansen Carl T. , Rosenstreich David L.
Published: January 2023

C57BL/10ScN (nu/nu) mice have B cells and macrophages unresponsive to a phenol-water extracted preparation of Escherichia coli K235 LPS. This unresponsiveness was demonstrated in vitro by the inability of spleen cells to incorporate 3H-thymidine after a 48 hr incubation with LPS (Ph) and by the inability of LPS (Ph) to inhibit macrophage phagocytosis of 51Cr-labeled, opsonized sheep erythrocytes. Furthermore, macrophage cultures stimulated with LPS (Ph) produced low levels of LAF and PGE2 when compared with macrophages from the LPS-sensitive C3H/HeN and C3H/HeN (nu/nu) strains. Therefore, the C57BL/10ScN (nu/nu) strain is similar in its LPS unresponsiveness to the well-characterized C3H/HeJ and C57BL/10ScCR strains. The combination of endotoxin unresponsiveness and the athymic nature of this mouse strain may provide a powerful new tool for studying the cellular events mediating endotoxicity.
Transplantation in Miniature Swine
Authors: Lunney Joan K. , Sachs David H.
Published: January 2023

The complexity of the I region analog associated with the MHC of miniature swine has been probed by sequential antibody precipitation studies of Ia antigens. Treatment of solubilized lymphocyte preparations from MSLA homozygotes of the DD haplotype with excess AA anti-DD alloantiserum led to precipitation of only a portion of the total Ia antigens, as determined by secondary precipitation of the remaining material with CC anti-DD serum. The presumed I region of miniature swine must therefore code for more than one Ia antigen-bearing polypeptide chain. In addition, certain mouse alloantisera that had previously been shown to react with rat Ia antigens were tested for reactivity with swine Ia antigens. Anti-Iak mouse alloantisera precipitated Ia molecules from every swine extract tested, regardless of MHC type, precluding genetic mapping studies. However, sequential precipitation studies demonstrated that the cross-reactive mouse alloantisera reacted only with a subclass of swine Ia antigens, again suggesting genetic complexity of the pig Ia locus.
A Quantitative Study at the Single Cell Level of Immunoglobulin Antigenic Determinants Present on the Surface of Murine B and T Lymphocytes
Authors: Avrameas Stratis , Hösli Peter , Stanislawski Marc , Rodrigot Maryvonne , Vogt Esther
Published: January 2023

An enzyme-immunoassay for the quantitation at the single cell level of immunoglobulin antigenic determinants present on the membrane of murine lymphocytes (MIg) has been developed. This immunoassay makes use of specific Fab:anti-Ig, anti-κ, anti-µ, and anti-γ conjugated to Escherichia coli β-galactosidase of high specific activity in conjunction with the micromanipulation of lymphocytes and the measurement of enzyme activity with the fluorogenic substrate 4-methylumbelliferyl-β-D-galactopyranoside with a microassay system. With this technique MIg was detected and could be measured in substantial amounts on the surface of almost all splenic and lymph node lymphocytes examined. Depending on the specific Fab employed, 0 to more than 79,000 MIg per lymphocyte were measured, but a larger proportion of cells with higher amounts of MIg were found in spleen than in lymph nodes. However, when the same Fab′s were used to localize MIg by a conventional immunohistochemical staining, only a fraction of cells, corresponding to published values, could be scored as positive.

The fact that MIg was associated with the vast majority of lymphoid cells implied that T cells carried such antigens. In order to investigate this further, experiments were performed on splenic B and T cell fractions obtained either with an anti-Ig immunoadsorbent or after treatment with anti-θ serum plus complement. The B cell fraction possessed greater amounts of MIg (1500 to 79,000) than the T cell fraction (0 to 39,000). However, on the average, B cells carried only 2 to 2.5 times more µ, κ, and γ antigenic determinants than T cells. Thus, B cells carried a mean of 27,300 to 47,800 Ig, 17,400 to 30,400 κ, 13,000 to 23,000 µ, and 4200 to 7300 γ antigenic determinants on their surface. T cells carried 4600 to 8000 Ig, 7200 to 12,600 κ, 5600 to 9800 µ, and 2400 to 4200 γ determinants. Immunoabsorbent column-fractionated T cells from which MIg was removed by anti-mouse Ig stripping were able to regenerate MIg after 24 hr of culture. The quantities reexpressed were equal to those found on T cells before anti-Ig treatment. These results strongly suggest that T cells synthesize and express MIg.

Two nonmutually exclusive hypotheses are advanced that may account for the observations made: 1) Irrespective of the molecular nature of the T cell structure detected in this study, it is proposed that the difficulty in detecting MIg by standard immunohistochemical staining methods may reflect differences in the membrane composition of B and T cells. Local concentrations of MIg on B cells could be higher than on T cells, or a microredistribution of MIg may be achieved more easily on B than on T cells. 2) The T cell MIg may be conceived of as a molecule structurally different from the MIg on B cells.