bluuebunny/crossref_metadata_2025 huggingface dataset:
Organ and Isotype Distribution of Plasma Cells Producing Specific Antibody after Oral Immunization: Evidence for A Generalized Secretory Immune System
Authors: Weisz-Carrington Paul , Roux Maria E. , McWilliams Michael , Phillips-Quagliata Julia M. , Lamm Michael E.
Published: December 2022

Mice were induced to produce IgA antibodies against ferritin after oral immunization. Such antibodies were detected by immunofluorescence in plasma cells in the intestinal mucosa as well as in secretory sites located elsewhere, such as the lactating mammary gland, salivary gland, and respiratory tract. The observation suggested that cells immunized locally via the gut could home to distant secretory sites. To confirm this hypothesis, lymphocyte transfer studies were done with mesenteric node (MN) versus peripheral node (PN) cells from orally immunized donors into nonimmunized recipients. IgA anti-ferritin cells from MN homed to exocrine targets, whereas IgM and IgG anti-ferritin cells homed to PN. The findings overall support the concept of a generalized and interrelated secretory immune system.
Histocompatibility-Linked Susceptibility for Hepatosplenomegaly in Human Schistosomiasis Mansoni
Authors: Salam Ekram Abdel , Ishaac Soad , Mahmoud Adel A. F.
Published: December 2022

In schistosomiasis mansoni, the pathogenesis of hepatosplenic disease has been shown to be due primarily to immune mechanisms. The present study was designed to examine the relationship between the development of schistosomal hepatosplenomegaly in Egyptian school children and the HLA antigens. Two groups of schistosome-infected children with similar fecal egg counts were examined: one group (23 children) had no clinically demonstrable hepatosplenomegaly whereas all the children (28) in the second group suffered from liver enlargement. Furthermore, 13 of the 28 individuals in the latter group had splenomegaly as well. Our results show that hepatosplenomegaly was related to the presence of two HLA antigens: HLA A1 and B5. The average relative risk of developing hepatomegaly is 29 for HLA A1 and 18.9 for B5 and in the presence of both antigens it increases to 55.6. Furthermore, the severity of hepatomegaly was correlated with the presence of these two HLA antigens. These findings represent a step toward elucidating the factors controlling the pathogenic mechanisms in human schistosomiasis mansoni.
Fcγ Receptor of the Murine Macrophage-Like Cell Line P388D1
Authors: Haeffner-Cavaillon Nicole , Klein Michel , Dorrington Keith J.
Published: December 2022

Studies of the binding of monomeric murine IgG2a and IgG2b to the macrophage-like cell line P388D1 confirmed the existence of at least two classes of Fc receptors. One receptor binds IgG2a preferentially, whereas the second interacts better with IgG2b. Approximately equal numbers of each class of receptor were present. The receptors showed a marked cross-specificity toward IgG2 subclass proteins but did not recognize murine IgG1. The P388D1 receptors were used to study the phylogenetic emergence, the conservation, and evolution of the molecular structures necessary for the expression of cytophilic activity by IgG. Heterocytophilic activity of plasma, sera, or purified immunoglobulins from modern representatives of the major phylogenetic families was assessed by competitive displacement of specifically bound radiolabeled IgG2a or IgG2b. The P388D1 Fc receptors showed marked species specificity. Fish IgM and non-IgM 7S immunoglobulins from amphibians, reptiles, and avians lacked cytophilic activity toward P388D1; only turtle plasma could displace murine IgG2 proteins from their receptors. In higher vertebrate species, rat, rabbit, and human IgG competed for the two classes of receptor equally well. In contrast, both guinea pig IgG1 and IgG2, and also IgG from bovine, sheep, goat, horse, and opossum lacked the ability to interact with P388D1. Canine IgG could displace murine IgG2a but not IgG2b from their receptors. These results, when compared to the data available on macrophage receptors for homologous and heterologous IgG in most of the above species, indicate that cytophilic structures have diverged in parallel with the cell-surface receptors and that mouse, rabbit, and human macrophage Fc receptors are structurally related.
Studies on the Fcγ Receptor of the Murine Macrophage-Like Cell Line P388D1
Authors: Haeffner-Cavaillon Nicole , Dorrington Keith J. , Klein Michel
Published: December 2022

Binding studies of human IgG proteins to murine P388D1 cells indicated that they bind to an apparently homogeneous Fc receptor population. The association constant was 0.89 × 106 M-1 at 22°C and was comparable to the binding affinities of homologous murine IgG2a and IgG2b. The number of receptor sites was found to be approximately 6 × 105/cell. Fcγ1 and Fcγ3 fragments bound with an affinity comparable to that of the parent proteins. The P388D1 receptors could discriminate between the human IgG subclasses; the relative cytophilic activity was IgG3 > IgG1 > IgG4 and IgG2 was devoid of binding activity. Fragments corresponding to the Cγ2 and Cγ3 domains of human IgG1 were both unable to bind to the P388D1 receptors either alone or in equimolar combination. This suggests that the cytophilic site may be formed cooperatively by interaction between the two domains. The integrity of the hinge region appeared to be essential for full expression of cytophilic activity since reduction of the hinge-region disulfides in both human IgG1 and its Fc fragment markedly decreased their binding affinity. In addition, a mutant IgG1 molecule lacking the hinge region was significantly less cytophilic than its normal counterpart.
Genetically Restricted Immune Responses in Guinea Pigs Primed in Vivo with Antigen-Bearing Macrophages
Authors: Yokomuro Kozo , Rosenthal Alan S.
Published: December 2022

Guinea pigs injected intradermally with antigen pulsed macrophages generate a population of immune T cells that proliferate in vitro on second exposure to antigen. T cells from F1 (2 × 13) guinea pigs immunized with DNP-OVA on one parental macrophage respond in vitro only to DNP-OVA on macrophages identical to those used for immunization and not to DNP-OVA associated with the other parental macrophages. These results demonstrate that the immunogenicity of antigen is dependent upon the macrophages used for priming in that, with this approach, strain 2 or 13 guinea pigs immunized with allogeneic macrophages pulsed with antigen do not respond to either allogeneic or syngeneic antigen-bearing macrophages. However, lysates of antigen-pulsed macrophages can still immunize either allogeneic or syngeneic recipient via their own macrophages. F1 (2 × 13) guinea pigs are immunized by insulin B chain pulsed strain 13 macrophages (responder) but not by strain 2 macrophages (nonresponder) suggesting that whether a F1 (nonresponder × responder) guinea pig recognizes antigen bound to a parental macrophage is genetically restricted before immunization to the same extent as the donor parental macrophages used for immunization.
Avian Antigen-Binding Cells
Authors: Higgins Gloria , McDermott Christine , Choi Yong Sung
Published: December 2022

Antigen-binding B and T cells from chicken spleens were selected on plates of antigen-derived gelatin. Hapten-specific B cells from DNP-immune normal chicken spleens were selected on DNP-gelatin. As much as 165-fold functional enrichment of precursors of anti-DNP antibody-producing cells, as measured in an adoptive transfer system, was achieved. However, the enrichment of DNP-binding cells assessed by rosette formation and autoradiography was no more than 25-fold. HGG-specific T cells from bursectomized agammaglobulinemic chickens immunized with deaggregated HGG were selected on HGG-gelatin. In the fraction adherent to HGG-gelatin, at least a 20-fold enrichment of suppressors of the antibody response to TNP-HGG, as measured by adoptive transfer, was accomplished. In contrast, no more than 6-fold enrichment of HGG-binding cells was detected by autoradiography. Antigen-specific depletion and enrichment of suppressor T cells and of HGG-binding cells occurred in parallel, suggesting that suppressor cells can bind soluble antigen and can be isolated on antigen coupled to a solid support.
The Beige Mutation in the Mouse
Authors: Roder John C.
Published: December 2022

A point mutation, called beige, on linkage group 14 in C57BL/6 mice leads to a marked impairment in natural killing and antibody-dependent, cell-mediated cytolysis (ADCC) of tumor cells. The defect in NK cytolysis was predetermined at the level of progenitor cells in the bone marrow as revealed in radiation chimeras. This impairment in NK function could not be accounted for by an altered organ distribution, target selectivity, or ontogenesis. Interferon did not fully restore the response, which suggests that the defect may not result solely from a lack of endogenous interferon stimulation in beige mice. The frequency of target-binding cells was normal in all lymphoid organs, which suggests that the defect is intrinsic to the NK cell and does not involve an altered population size or an inability to recognize and interact with the target. Rather, the defect may lie within the lytic pathway subsequent to target cell contact. These mice should provide a useful NK-deficient system for studies of T cell or macrophage immunity and in addition they provide a means for testing the in vivo significance of NK cells in resistance to tumors and virusinfected cells.
Activation of the Classical Complement Pathway in Human Serum by a Small Oligosaccharide
Authors: Dieminger Lothar , Schultz Duane R. , Arnold Patricia I.
Published: December 2022

Two small oligosaccharides were hydrolyzed from a native polysaccharide by polysaccharidases indigenous to the venom of the tropical ant Pseudomyrmex sp. after incubation for 5 hr at 37°C. They were purified by passage through an ultrafiltration membrane (m.w. cut-off, 500 daltons), cation exchange chromatography at pH 3 to separate the oligosaccharides from peptides, and gel filtration chromatography. The highly purified oligosaccharides contained no amino acids and no polar or nonpolar lipids.

The larger oligosaccharide had an estimated m.w. of 840 daltons, and the smaller 430 daltons. The larger species was a potent activator of C1 in normal human serum at 37°C, resulting in the consumption of its two natural substrates C4 and C2. It had no effect on the C1 INH, and its action was destroyed by treatment with sodium periodate.

Analysis of both oligosaccharides by gas chromatography revealed five sugars: glucose, mannose, galactose, N-acetyl-glucosamine, and N-acetyl-galactosamine. Chemical analyses revealed the presence of reducing sugars in the larger species, but neither contained hexuronic acids. The latter were found in the native polysaccharide. The smaller oligosaccharide may be a monomer of the larger species. After electrophoresis on paper at pH 1.9, the 840-dalton oligosaccharide separated into two species: the first, which remained at the origin, contained reducing carbon atoms. The second migrated toward the cathode and contained no reducing sugars. Both species caused the consumption of C2 in human serum. The oligosaccharide(s) with a m.w. of 840 daltons is the smallest carbohydrate described that causes activation of the classical complement pathway.
Cell-Mediated Immunity in Interstitial Nephritis
Authors: Neilson Eric G. , Phillips S. Michael
Published: December 2022

Guinea pigs immunized with renal tubular antigens develop a severe interstitial nephritis. The complete expression of disease depends on a complex interplay between cellular and humoral mechanisms. The present studies were undertaken to analyze the lymphocyte diversity engendered during the course of this nephropathy. The results demonstrated a strong cellular recognition of the sensitizing antigens, as well as the concomitant development of a potent nonspecific suppressor system. Sequential analyses indicated that reactive T cells from different anatomic compartments maintained these distinctive functions. Judged from isotope incorporation studies, lymphocytes harvested from peritoneal exudates were more likely to recognize sensitizing renal tubular antigens than either spleen, lymph node, or peripheral blood cells. Lymph node cells, however, were more efficient in mediating the intense polyclonal immunosuppression that was typically present in nephritic animals after 1 week of disease. Delayed hypersensitivity measurements confirmed the in vivo presence of this immunosuppression. All of these observed T cell functions were fully developed before the appearance of antitubular antibodies in the kidneys of diseased animals. Eventually, however, the cellular recognition of the sensitizing antigens started to decline in the face of continued suppression and pathologic damage along cortical tubules. These findings are discussed in terms of the complex role of T lymphocytes in the pathogenesis and regulatory modulation of interstitial kidney disease.
A Cell Surface Antigen, TER, Expressed by Embryos and Germ Cells
Authors: Michaelson James , Artzt Karen , Bennett Dorothea , Caldwell Jane , Heath John K.
Published: December 2022

An antiserum prepared in rabbits against the C3HeB/FeJ mouse ovarian teratocarcinoma E6496 was absorbed in vivo in C3HeB/FeJ mice. This absorbed antiserum identified an antigen, denoted TER, that is present on sperm, ova, embryonic germ cells, and cells of the early mouse embryo. TER was absent from all adult somatic cells tested, but found on several murine tumors.
Immunologic Tolerance to Allergenic Protein Determinants: Properties of Tolerance Induced in Mice Treated with Conjugates of Protein and a Synthetic Copolymer of D-Glutamic Acid and D-Lysine (D-GL)
Authors: Liu Fu-Tong , Bogowitz Cheryl A. , Bargatze Robert F. , Zinnecker Mark , Katz Lee R. , Katz David H.
Published: December 2022

Conjugates of proteins and the synthetic copolymer of D-glutamic acid and D-lysine (protein-D-GL) reproducibly induce significant unresponsiveness to the protein antigens in experimental mice. Proteins studied include ovalbumin and antigen E of ragweed extract, the major allergen of ragweed pollen. The unresponsive state 1) can be induced in both unsensitized and previously sensitized experimental animals, 2) is selectively confined to responses of the IgE antibody class, 3) is long lasting, and 4) is highly antigen specific. IgE antibody responses can also be suppressed by administering comparable doses of unconjugated protein alone; however, the unresponsive state induced in this manner is only transient and rebound production of IgE antibody is often observed.

Results from the studies of the cellular basis of the protein-D-GL induced unresponsiveness demonstrate that 1) protein-D-GL conjugates do not induce unresponsiveness at the level of protein-specific B cells, 2) tolerance is not induced by virture of a detectable antigen-specific suppressor T cell mechanism, 3) tolerance is most probably induced in the antigen-specific helper T cell populations.

The significant IgE-selective and antigen-specific tolerogenic activity of protein-D-GL conjugates make these compounds potential candidates for use as therapeutic agents in the treatment of IgE-mediated human allergic disorders induced by protein allergens.
Modulation of Platelet Activation by Native DNA
Authors: Fiedel Barry A. , Schoenberger John S. , Gewurz Henry
Published: December 2022

Native DNA (dsDNA) was found to induce the aggregation of isolated human platelets and the release of platelet 5HT; this activation was inhibited by both theophylline and TYA, suggesting a role for cAMP and metabolic products formed from arachidonate. By contrast, nonaggregating amounts of dsDNA inhibited platelet activation induced by collagen or thrombin. This inhibition, which could be overcome by use of greater amounts of the stimulatory agents, was not associated with the loss of platelet viability. Activation of platelets by dsDNA was not observed in plasma or in isolated platelet systems to which small amounts of cell-free plasma were added. However, dsDNA maintained in plasma its ability to inhibit platelet aggregation induced by collagen and thrombin. RNA and single-stranded DNA failed to induce platelet aggregation or release of 5HT and to block the platelet activation stimulated by dsDNA. Further, dsDNA did not significantly inhibit platelet aggregation in platelet-rich plasma stimulated by ADP or epinephrine. These data implicate dsDNA as a selective and potentially important activator and modulator of platelet responsiveness.
The Role of Complement in the Binding and Degradation of Immunoglobulin Aggregates by Macrophages
Authors: Kijlstra Aize , van Es Leendert A. , Daha Mohamed R.
Published: December 2022

The role of complement (C) in the processing of soluble immune complexes by guinea pig peritoneal macrophages was studied in a homologous system in vitro by using isolated stable IgG2 aggregates as a model for immune complexes.

Degradation of immunoglobulin aggregates by macrophages is markedly stimulated in the presence of fresh serum, an effect that was shown to be mediated via the classical pathway of C. After treatment with serum, aggregates were isolated from the serum proteins by sucrose gradient ultracentrifugation and subsequently presented to the macrophages; it appeared that the stimulating effect was mediated via the binding of C to the aggregates and is probably therefore not due to generation of free C split products in the medium.

Degradation by macrophages of serum-treated aggregates with comparable sedimentation rates increased as the concentration of the serum in which the aggregates had been incubated increased. Furthermore, a stimulated degradation of aggregates could be correlated with the ability of the aggregates to activate the C system as assessed by C4 consumption. Enhanced degradation of the aggregates was eliminated when the C receptors of the macrophages were removed by trypsin treatment before incubation with the aggregates. These results thus suggest that the effect observed is dependent on the ability of the aggregates to bind and activate C components and requires the presence of intact cellular C receptors. Experiments in which the rate of binding of the aggregates to the macrophage cell membrane was compared with the ingestion and digestion of membrane-bound aggregates showed that C increased the rate of binding but had no effect on the subsequent processing of the aggregates.

These studies thus demonstrate that C plays an important role in the clearance of soluble immunoglobulin aggregates by mononuclear phagocytes.
Biosynthesis and Post-Synthetic Modification of a Precursor (PRO-C5) of the Fifth Component of Mouse Complement (C5)
Authors: Ooi Yuet M. , Colten Harvey R.
Published: December 2022

Mouse peritoneal macrophages synthesized and secreted a precursor (pro-C5) of the fifth component of serum complement (C5) in short-term tissue culture. Approximately 0.2% of the newly synthesized intracellular protein and 0.8% of secreted protein were precipitable with antiserum to mouse C5. The precursor is similar in size to the native serum protein (210,000 daltons), but consists of a single polypeptide chain. In contrast, serum C5 consists of two polypeptide chains (m.w. 125,000 and 83,000) linked by disulfide bridges. An electrophoretic variant of pro-C5 distinct from intracellular C5 was detected in medium from macrophage cultures.
Propagation of Antigen-Specific T Cell Helper Function in vitro
Authors: Schrier Rachel D. , Skidmore Barry J. , Kurnick James T. , Goldstine Steven N. , Chiller Jacques M.
Published: December 2022

Antigen-induced proliferation of primed lymph node cells was abrogated by treatment with anti-Ly 1 serum and complement (C) but not with anti-Ly 2 serum and C. Lymph node cells from animals primed to ovalbumin were activated with antigen in vitro, followed by propagation in an antigen-free supernatant fluid obtained from lectin-induced normal spleen cells. T cells processed in this manner displayed a stepwise enrichment of helper activity for antibody production as measured in a secondary hapten-carrier response. The sequential increase in antigen-specific help was paralleled by a rise in the antigen-induced proliferative response, a phenomenon whose expression was dependent on the presence of syngeneic or semi-syngeneic irradiated filler cells.
Cell-Mediated Lympholysis by Human Maternal and Neonatal Lymphocytes: Mother's Reactivity against Neonatal Cells and Vice Versa
Authors: Granberg Christer , Hirvonen Toivo , Toivanen Paavo
Published: December 2022

Maternal reactivity in cell-mediated lympholysis (CML) against cells from her own child is, in average, half of the maternal reactivity against unrelated adult cells. This finding remains the same when cells from a newborn or from an older child are used, suggesting that the reduced maternal reactivity is based rather on the one haplotype identity between the mother and child than on the occurrence of specific maternal tolerance. Consistently, CML-capacity of the child, directed against cells of own mother, is half of the control values, again independently of the child's age.
Differences between Syrian Hamster Strains in Natural Killer Cell Activity Induced by Infection with Pichinde Virus
Authors: Gee Sydney R. , Clark David A. , Rawls William E.
Published: December 2022

Adult MHA hamsters are susceptible to a fatal Pichinde virus infection and are unable to limit viremia. Adult hamsters of other strains, such as LVG and LSH, limit viremia and do not develop fatal disease. Both survival and the ability to limit viremia are each controlled by a dominant gene. During studies to delineate the role of cytotoxic cells in Pichinde virus infection, it was observed that MHA hamsters exhibited appreciable levels of cytotoxic activity against syngeneic and allogeneic tumor cells and that this activity increased after Pichinde virus infection. In contrast, LSH hamsters showed lower endogenous cytotoxicity and less enhancement after virus infection. The cytotoxicity was primarily mediated by a cell with characteristics of natural killer (NK) cells. The separation of MHA hamster spleen cells by velocity sedimentation revealed that fractions of the gradient containing maximum NK activity also contained numerous infectious centers, whereas the corresponding fractions from gradients of LSH hamster spleen cells showed less cytotoxicity and contained one-tenth the number of infected cells. These observations suggest that a population of cells, defined by NK activity, may be a target for Pichinde virus replication, and that the activity of these cells is only minimally expressed in the resistant LSH strains.
Suppression of Antigen and Mitogen Induced Human T Lymphocyte DNA Synthesis by Bacterial Lipopolysaccharide: Mediation by Monocyte Activation and Production of Prostaglandins
Authors: Ellner Jerrold J. , Spagnuolo Philip J.
Published: December 2022

These studies further characterize the effects of bacterial lipopolysaccharide (LPS) on circulating human mononuclear cells. Escherichia coli LPS did not induce 3H-thymidine incorporation in unseparated mononuclear cells from healthy subjects during 5 days of culture. However, in co-culture experiments, LPS 2 to 20 µg/ml suppressed the response to suboptimal concentrations of phytohemagglutinin and streptokinase-streptodornase by 36 to 84%. Depletion of adherent cells or phagocytic cells resulted in a T lymphocyte-enriched population highly responsive to stimulation by antigens and mitogens but refractory to suppression by LPS. Add-back experiments confirmed adherent cell dependence of the suppressive activity of endotoxin. Moreover, 5 × 104 adherent cells cultured with LPS for 24 hr and then washed inhibited the response of 1.5 × 105 T lymphocytes to antigen by 44 to 67%. Larger numbers of untreated adherent cells had a similar effect.

Endotoxin was shown to activate monocytes under these cultural conditions by the criteria of increased attachment to plastic surfaces (by 2.0-fold) and increased release of immunoreactive prostaglandin E2 (by 6.6-fold). The prostaglandin release of cells cultured with LPS was reversed by indomethacin. Indomethacin and RO-20-5720 also blocked inhibition of lymphocyte function by large numbers of adherent cells and endotoxin-activated adherent cells. Moreover, supernatants of macrophages cultured with or without LPS suppressed T lymphocyte function; this activity was diminished when indomethacin was present during supernatant generation. These findings suggest that bacterial LPS activates human monocytes to produce prostaglandins that suppress T lymphocyte function.
B Cell Function in Human Marrow Transplant Recipients Assessed by Direct and Indirect Hemolysis-in-Gel Assays
Authors: Ringden Olle , Witherspoon Robert , Storb Rainer , Ekelund Elsie , Thomas E. Donnall
Published: December 2022

B cell function was studied in 35 healthy controls and 42 individuals who were recipients of bone marrow transplants for aplastic anemia or hematologic malignancy. Lymphocytes were stimulated in vitro by killed Staphylococcus aureus bacteria (Staph. aureus). Antibody secretion was measured as plaque forming cells (PFC) in a direct hemolysis-in-gel assay by using fluorescein isothiocyanate (FITC)-coupled sheep red blood cells (SRBC) and in an indirect assay with protein A coupled SRBC and antisera against human IgG, IgA, and IgM. A pH of 7.5 in culture was crucial for optimal antibody secretion to occur. Lymphocytes from patients studied during the first 3 months after transplantation (short-term patients) had lower cell survival after 6 days in culture than that of lymphocytes from healthy controls, whereas cell survival of lymphocytes from patients studied more than 1 year after transplantation (long-term patients) was equal to that of controls. Short-term patients tested in the direct assay showed a mean of 12 ± 4 [S.E.] PFC/106 cells compared to a mean of 300 ± 91 PFC/106 in the controls (p < 0.01). The mean number of PFC/106 in long-term patients was 310 ± 222.

In healthy controls, the direct assay detected about 4% of the IgM-producing B cells found in the indirect assay. Twelve short-term patients were studied with both assays. None of the patients had PFC with the direct assay, but 11 showed some PFC in the indirect assay. The indirect PFC responses after stimulation by Staph. aureus were significantly lower for patients than for controls (IgG p < 0.05, IgA p < 0.01, IgM p < 0.01).

In summary, short-term marrow graft recipients had deficient B cell responses in vitro, but in long-term patients this function was normal.
Cross-Reactivity of Rat, Mouse, and Human IgD
Authors: Golding Hana , Cuchens Marvin A. , Leslie Gerrie A. , Rittenberg Marvin B.
Published: December 2022

Highly purified sheep anti-rat lymphocyte membrane IgD (mIgD) was used to detect cross-reactivity with the putative murine-δ chain on mouse lymphocytes. Cross-reactivity is demonstrated by indirect immunofluorescent staining and by immunoprecipitation of 125I-labeled lymphocyte membrane extracts followed by electrophoresis on 10% polyacrylamide gels. In addition, cross-reactivity of anti-rat-δ with human IgD is shown by gel diffusion analysis.

The anti-rat-δ reagent stained both Ig5a+ and Ig5b+ lymphocytes. Preincubation of Ig5b+ (but not Ig5a+) cells with monoclonal allotype-specific antibodies (anti-Ig5b) under capping conditions caused inhibition of staining by the sheep anti-rat-δ reagent, indicating that it is the δ-chain that is recognized on mouse lymphocytes and that the anti-rat-δ reagent does not distinguish between mouse-δ allotypes. Furthermore, absorption of the sheep anti-rat-δ serum with purified human IgD reduced subsequent staining of mouse lymphocytes by approximately 50%; staining was not affected by absorption with human IgM. This xenogeneic anti-δ antiserum appears to detect determinants on the δ-heavy chain, which are shared by at least three species of mammals, suggesting that these determinants represent important molecular features conserved during evolution.
Humoral Immunostimulation
Authors: Shearer William T. , Mettes Hal J.
Published: December 2022

Anti-L antibody and cytosine arabinoside acting synergistically produced an inhibition of growth of L cells in vitro. This effect was dependent upon both antibody and cytosine arabinoside concentrations with a 1:200 dilution of antiserum and a 0.10 µg/ml (4 × 10-7 M) concentration of drug being optimal. There was a 50% reduction in the growth rate of such 24-hr cell cultures, and the effect was due primarily to inhibition of cell growth rather than cell lysis. Replicate long-term cell cultures showed that antiserum (IgG fraction) stimulated uptake of 3H-cytosine arabinoside into nuclear DNA of cells. In acute 3H-cytosine arabinoside uptake studies, antibody had no discernable effect upon the transport of drug into the cell but, rather, stimulated an event subsequent to transport, presumably phosphorylation of drug. Indeed, the antibody-enhanced uptake of 3H-cytosine arabinoside appeared first in the total isotope pool of the cells, followed by appearance in the nondiffusible pool, and, finally, in the acid-insoluble pool of cells. Moreover, antibody stimulation of labeled drug into the nondiffusible pool was exquisitely dependent upon energy sources. In total, these observations suggest that antibody stimulated cytosine arabinoside uptake by activation of phosphorylation pathways with subsequent incorporation of the phosphorylated drug into DNA. Enhanced incorporation of drug into nuclear DNA of antibody-treated cells was thus correlated with inhibition of cell replication. Therefore, it is possible to modulate tumor cell replication in vitro by the judicious use of antibody and cytosine arabinoside.
Production of Human Lymphocyte Mediators after Activation with Periodate or Neuraminidase and Galactose Oxidase
Authors: Greineder Dirk K. , Rocklin Ross E. , David John R.
Published: December 2022

Treatment of human mononuclear cells with sodium metaperiodate (NaIO4) or neuraminidase and galactose oxidase (NGO) results in lymphocyte activation and subsequent generation of supernatants rich in migration inhibitory factor (MIF) and leukocyte inhibitory factor (LIF). Preliminary characterization of these mediators by Sephadex G-100 gel filtration suggests that they are similar to antigen- and concanavalin A-induced MIF and LIF, eluting in the 25000 m.w. and 68000 m.w. regions, respectively. The possibility of galactose oxidase carryover into the supernatants has been studied and conditions are described that minimize this eventuality. A method is presented for producing control and lymphokine-rich supernatants both of which have been exposed to identical concentrations of NGO although in the control activation is blocked by the addition of 0.1 M galactose to the incubation. These findings establish NaIO4 and NGO as useful mitogens for generating human lymphokine-rich supernatants that can be used without further purification.
Platelet Activating Factor (PAF) Induced Release of Platelet Factor 4 (PF4) in vitro and during IgE Anaphylaxis in the Rabbit
Authors: McManus Linda M. , Morley Cindy A. , Levine Shirley P. , Pinckard R. Neal
Published: December 2022

Basophil-derived platelet activating factor (PAF) was shown to induce the release of platelet factor 4 (PF4), a specific platelet protein, from isolated, washed rabbit platelets. The kinetics and dose response of this release were similar to that of PAF-induced 3H-serotonin secretion, and indicated that PF4 release was a reliable and sensitive estimate of platelet stimulation by PAF. Plasma PF4 concentrations were therefore used as a marker for in vivo platelet release during systemic anaphylaxis induced by the i.v. injection of antigen (horseradish peroxidase, HRP) into rabbits conditioned to produce only IgE antibody against HRP. Antigen administration was accompanied by the loss of circulating stainable basophils, acute thrombocytopenia and neutropenia, and a significant increase in PF4 levels. The increase in plasma PF4 began approximately 90 sec after antigen challenge, corresponding to 60 sec after the intravascular aggregation of platelets. However, maximum levels of plasma PF4 were not reached until 3 to 5 min after antigen challenge. Concomitant measurement of circulating PAF during anaphylaxis indicated that PAF release into the circulation preceded the decrease in circulating platelet numbers by about 30 sec and preceded the increase in plasma PF4 levels by about 60 sec. Thus, this study documents that platelets undergo a release reaction during anaphylaxis at a time when they are sequestered in the microvasculature. The temporal relationship of this release suggests that the following series of events occur during IgE anaphylaxis: antigen stimulates circulating IgE-sensitized basophils to release PAF, PAF causes intravascular platelet aggregation and subsequent sequestration, and finally, sequestered platelets release their intracellular constituents.
Evidence That Thymocytes Require at Least Two Distinct Signals to Proliferate
Authors: Beller David I. , Unanue Emil R.
Published: December 2022

Optimal receptivity of thymocytes to the mitogenic stimulus in macrophage culture fluid (MCF) is limited to the time in culture when these cells are spontaneously proliferating. By providing a brief, nonmitogenic lectin pulse, nonproliferating cells can be “activated” and their ability to respond to MCF totally restored. This procedure provides a model system for evaluating thymocyte proliferation and, in particular, the mechanism of action of MCF.
Inhibition of Human Lymphocyte E-Rosette Formation by Oxygenated Sterols
Authors: Streuli Rolf A. , Chung Jiwhey , Scanu Angelo M. , Yachnin Stanley
Published: December 2022

25-Hydroxycholesterol, 20α-hydroxycholesterol, 7α-hydroxycholesterol, and 5α-hydroxy-6-ketocholestanol, when added to cultures of human lymphocytes in lipoprotein-depleted medium (LPDM) at a concentration of 2.5 × 10-5 M, inhibit E-rosette formation with sheep red blood cells. 20α-Hydroxycholesterol, 7α-hydroxycholesterol, and 5α-hydroxy-6-ketocholestanol are more potent inhibitors than 25-hydroxycholesterol. The inhibitory effect of 5αhydroxy-6-ketocholestanol on E-rosette formation appears after 15 min of exposure; with the other three compounds, an exposure time of 18 hr is necessary. The inhibitory effect of E-rosette formation can be abolished by addition of free cholesterol, low-density lipoprotein, or high-density lipoprotein to the LPDM or by incubation of the cells in normal AB serum, but not by the addition of mevalonic acid to the LPDM. These observations suggest that the capacity of oxygenated sterol compounds (OSC) to inhibit E-rosette formation is independent of their inhibitory effect on sterol synthesis. It is possible that OSC inhibit E-rosette formation as a consequence of their insertion into the lymphocyte membrane as cholesterol analogues.
Cytotoxic Properties of the Eosinophil Major Basic Protein
Authors: Gleich Gerald J. , Frigas Evangelo , Loegering David A. , Wassom Donald L. , Steinmuller David
Published: December 2022

The eosinophil major basic protein (MBP) damages cells including those from organs whose dysfunction is associated with eosinophil infiltration. Concentrations of MBP in the toxic range were found in sputa from asthmatics, in pleural fluid and in sera. These findings suggest the hypothesis that the eosinophil produces damage to cells in disease.
Identification of ganglio-N-tetraosylceramide as a new cell surface marker for murine natural killer (NK) cells.
Authors: Young W W , Hakomori S I , Durdik J M , Henney C S
Published: December 2022

The BCG-induced NK cell activity of murine peritoneal exudate cells was abolished by preincubation of effector cells with anti-ganglio-N-tetraosylceramide (anti-asialo GM1) and C but not with other anti-glycolipid antibodies, anti-ganglioside GM1, anti-globoside, and anti-ganglio-N-triosylceramide (anti-asialo GM2). In contrast, the cytotoxic activity of alloimmune T cells was not affected by treatment with anti-asialo GM1 antisera. These findings suggest that asialo GM1 display may be characteristic of NK cell populations and aid in the isolation of this population of cytotoxic cells.
Distribution of IgG subclasses in human B lymphocytes: evidence for dual expression of subclasses in surface and cytoplasmic IgG in minor B lymphocyte subpopulations.
Authors: Yount W J , Fuller C R , Simmons J G
Published: December 2022

The distribution of B lymphocytes expressing surface IgG subclasses has been defined by using immunochemically purified, subclass-specific, F(ab')2 fragments. Studies in normal adults showed 1.16% of peripheral lymphocytes to be positive for IgG. Percentages of IgG subclasses were IgG2, 0.50%; IgG4, 0.40%; IgG1, 0.33%; and IgG3, 0.14%. Dual label experiments indicated that minor subpopulations of normal lymphocytes may express more than one IgG subclass, both on surface IgG as well as in the cytoplasm. Cord blood showed significantly less IgG4-positive cells than were present in adults. Preliminary studies of patients with common variable immunodeficiency showed shifts in cell populations paralleling those previously noted in serum. Some patients with multiple myeloma were noted to exhibit striking shifts in the distribution of B lymphocytes toward a population of cells showing a similar IgG subclass to that of the M-component.
Delineation of tolerance-sensitive and tolerance-insensitive B cells in normal and immune defective mice.
Authors: McKearn J P , Quintáns J
Published: December 2022

B cells responsive to TNP on various thymus-independent carriers acquire immunocompetence at different times during ontogeny and elicit varying levels of responsiveness in immune-defective CBA/N mice. Such B cells have now been found to display differential tolerance susceptibilities in a hapten-specific in vitro tolerance assay. B lymphocytes from normal neonatal or immune-defective adult CBA/N mice responsive to TNP-lipopolysaccharide were shown to be hypersusceptible to tolerogenesis whereas B cell responses to a second TI antigen, TNP-Brucella abortus, were not rendered tolerant until high concentrations of tolerogen had been presented. Therefore, tolerance hypsersusceptibility is characteristic of a fraction of neonatal and immune-defective adult CBA/N antigen-reactive B lymphocytes.
A restricted component of T cell help in pokeweed mitogen-stimulated humans peripheral blood cell cultures.
Authors: Macy E , Stevens R H
Published: December 2022

Limiting dilution analyses using human T and B cells from the peripheral blood of individuals recently (2 to 5 weeks) boosted with tetanus toxoid or a combination of diphtheria and tetanus toxoids were performed in order to investigate the specificity of T cell help during in vitro pokeweed mitogen (PWM) stimulation. Limiting numbers of irradiated T cells were added to an excess of B cells and their ability to help 1) IgG anti-tetanus toxoid antibody or 2) IgG anti-tetanus toxoid and IgG antidiphtheria toxoid antibody production were compared. The helper activity for anti-tetanus toxoid antibody production did not dilute out at the same point as help for total IgG production. Cultures were observed at limiting dilutions of added helper T cells that produced anti-tetanus toxoid IgG or anti-diphtheria toxoid IgG but not both even though a sufficient number of potential specific IgG-producing B cells were present. A distribution analysis of the IgG anti-tetanus and IgG anti-diphtheria helper T cells indicated that the two subpopulations did not segregate together. Thus in the presence of PWM, individual helper T cells appeared capable of supplying help for a restricted number of B cells.
Tegument-associated antigen modulation by adult male Schistosoma mansoni.
Authors: Kemp W M , Brown P R , Merritt S C , Miller R E
Published: December 2022

Adult Schistosoma mansoni from mice were incubated in rabbit anti-mouse immunoglobulin class or subclass-specific sera. The rabbit antibody binding was then visualized by incubation of the labeled worms in fluoresceinated Staphylococcus aureus, which associated with the Fc portion of the rabbit antibody molecule by means of bacterial cell wall Protein A. The assay system was found to be simple, inexpensive, specific, repeatable, and rapidly accomplished. By using this technique, previous work documenting the presence of mouse IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM on the tegumental surface of adult worms of mouse origin was confirmed. It was further confirmed that the complexing of rabbit antibody to the mouse antigen led to the shedding of that antigen from the parasite's surface membrane within 20 min at 37 degrees C. This shedding phenomenon was specific for only those antigens interacting with the ligand and was inhibited by incubation with sodium fluoride, 2-deoxy-D-glucose, cytochalasin B, cytochalasin D, caffeine, or at 4 degrees C. These data indicate a sophisticated control mechanism exercised by the parasite over its interface with the host and may indicate a complexity of the host-parasite relationship not heretofore recognized.
T cell-mediated immunity to oncornavirus-induced tumors. II. Ability of different T cell sets to prevent tumor growth in vivo.
Authors: Leclerc J C , Cantor H
Published: December 2022

These studies demonstrate that two different types of protective immunologic reactions are generated in mice after inoculation with M.MSV. Type I protection, mediated by Ly1 cells, prevents MSV-induced sarcomas but has no effect on the growth of MLV+ lymphomas. This anti-MSV protection reflects induction of antivirion antibody and delayed type hypersensitivity reaction. Type II protection is mediated by Ly23 cells and prevents the growth of an MLV+ lymphoma, but has little or no protective effects against MSV-induced sarcomas. This "type II" reaction is directly related to CTL activity. The implications of these two protective reaction are discussed.
Human T lymphcytes are enriched for transfer factor.
Authors: Tomar R H , John P A
Published: December 2022

Using a canine local skin assay, we tested transfer factor prepared from three subpopulations of human peripheral blood mononuclear cells. T lymphocytes are enriched for transfer factor whereas adherent cells and nonadherent/nonrosetting cells are not. Thus, it appears as if transfer factor is a T cell lymphokine, the immunobiology of which is still unclear.
Lymphotoxin production by subsets of T cells.
Authors: Eardley D D , Shen F W , Gershon R K , Ruddle N H
Published: December 2022

Lymphotoxin is produced by T cells sensitized to antigen upon re-exposure in vitro. It is also elicited by mitogen treatment. Its production has been correlated with delayed-type hypersensitivity and it may be a mediator of that phenomenon. We have examined the Ly phenotype of the subset(s) of T cells that produce lymphotoxin in order to investigate the relationship of lymphotokin killing to allo-killing mediated by Ly2+ T cells. We have found that Ly1 T cells sensitized to ovalbumin secrete more lymphotoxin than Ly2 cells. The ovalbumin-sensitized T cells do not lyse their target in a short term 51Cr-release assay even when "glued" to the target with Con A. Thus, lymphotoxin-producing cells differ phenotypically from the previously defined cytotoxic T cells that bear the Ly2 differentiation marker.
IA antigens and antigen-presenting function of thymic macrophages.
Authors: Beller D I , Unanue E R
Published: December 2022

Thymic adherent cells were isolated after their enrichment on density gradients. The predominant cell type found was the macrophage, as determined by morphology, surface receptors for Fc and C3, phagocytosis and esterase activity. There was, in addition, a minor fraction of cells with a distinctive dendritic morphology. These dendritic cells had surface properties similar to macrophages but limited phagocytic capacity. Approximately 50% of thymic adherent cells bear Ia antigens detected by immunofluorescence by using either A.TH anti-A.TL or monoclonal anti-I-A antibodies. These cells were also found to be an extremely effective antigen-presenting source for macrophage-depleted immune T cells, supporting the idea that the Ia antigens detected are of functional significance. Our data indicate that the macrophage is the predominant adherent cell type in general, and the principal Ia-bearing cell in particular, isolated by either physical disruption of the thymus or by collagenase dissociation of thymis stroma.
Human T lymphocyte surface antigens: partial purification and characterization utilizing a high-titer heteroantiserum.
Authors: Pratt D M , Schlossman S F , Strominger J L
Published: December 2022

A high-titer human T lymphocyte-specific antiserum (A99) has been prepared by immunization of rabbits with a preparation partially purified from membranes of the T cell line, CEM. The specificities recognized by this antiserum included those recognized by TH1 and HTL antisera. Several experiments indicated that these antisera recognized several different proteins. A partial purification of material recognized by A99 serum was obtained by 1) isolation of plasma membranes, 2) detergent solubilization and chromatography on DEAE-cellulose, and 3) affinity chromatography on a wheat germ lectin-Sepharose 4B column. These three steps yielded material about 100-fold purified, and an estimate was made in several ways that on overall purification of 3500-fold would be required to achieve homogeneity. Immunoprecipitation of 125I-labeled 100-fold purified material showed that the major bands recognized by A99 serum had m.w. of 52,000, 96,000 (seen only after reduction), 120,000, 152,000, and 195,000. These data are compared to other published reports of human T lymphocyte-specific proteins.
Characterization of guinea pig lymphocyte-derived chemotactic factor for fibroblasts.
Authors: Postlethwaite A E , Kang A H
Published: December 2022

An assay that measures guinea pig fibroblast chemotaxis in vitro is described. Application of this assay has revealed that stimulation of guinea pig blood or lymph node lymphocytes in vitro by T cell antigen (horseradish peroxidase) or mitogen (phytohemagglutinin) is associated with production of a chemoattractant for guinea pig fibroblasts. This lymphocyte-derived chemotactic factor for fibroblasts (LDCF-F) has a m.w. or 80,000 and is distinct from guinea pig lymphocyte-derived chemotactic factor for monocytes (LDCF-M). LDCF-F is heat labile and trypsin sensitive. The production of LDCF-F in vitro correlates with lymphocyte transformation to antigen or mitogen and with the presence of in vivo-delayed hypersensitivity to antigen. LDCF-F might function in vivo to attract connective tissue fibroblasts to sites of cell-mediated immune reactions.
Structural studies on the murine Ia alloantigens. VII. Further evidence for two-gene control of the E/C alloantigens.
Authors: Cook R G , Vitetta E S , Uhr J W , Capra J D
Published: December 2022

By utilizing comparative tryptic peptide analysis of the E/C alloantigens from additional H-2 recombinant strains, the gene encoding the E/C beta subunit was localized to the I-A or B subregions. By peptide mapping techniques, the E/C alpha- and beta-chains showed little structural homology with the A alloantigen alpha and beta subunits. In addition, comparisons of the E/C beta subunits from F1 strains (b x d) with those from both parental strains revealed that in a heterozygote the beta subunit from either allele (b or d) can associate with the alpha-chain from one allele (d). Thus, the formation of hybrid E/C antigens by cis-trans gene complementation increases the E/C alloantigen repertoire in heterozygotes.