bluuebunny/crossref_metadata_2025 huggingface dataset:
Bone marrow transplantation across major histocompatability barriers in mice. III. Treatment of donor grafts with monoclonal antibodies directed against Lyt determinants.
Authors: Vallera D A , Soderling C C , Kersey J H
Published: December 2022

We studied the effect of eliminating T cells from donor grafts of mice in a system in which bone marrow was transplanted across major histocompatibility barriers. BALB/c bone marrow (added as a source of hematopoietic stem cells) combined with equal volumes of spleen cells (added as a source of GVHD-promoting cells) was pretreated in vitro with monoclonal anti-Lyt-1.2 or Lyt-2.2 plus absorbed rabbit complement before injection into C57BL/6 total-body-irradiated recipients. Functional activity of anti-Lyt monoclonal antibodies was determined in CML assay. Treatment with anti Lyt-1.2 plus C did not have any anti-stem cell activity, as measured by CFU-S assay, and protected recipients from the onset of lethal GVHD. Treatment with Lyt-2.2 plus C also did not reduce CFU-S; however, mice receiving treated marrow did develop GVHD and were all dead by 2 mo, as were untreated control mice. Surviving "anti-Lyt-1.2 + C chimeras" demonstrated a high percentage of donor mononuclear cells in their peripheral blood. Similar results were obtained when C3H/HeN donor BMS was treated with monoclonal anti-Lyt-1.1 plus C and injected into C57BL/6 recipients. These findings show that monoclonal antibodies directed against determinants unrelated to Thy-1 can eliminate T cells in the presence of C and successfully protect transplanted mice from lethal GVHD. They also suggest that these anti-Lyt antibodies may be useful tools in determining subpopulations of T cells that contribute to the development of GVHD.
Monoclonal anti-deoxyribonucleic antibodies. I. Isotype and specificity studies.
Authors: Jacob L , Tron F
Published: December 2022

Ten monoclonal anti-DNA antibodies generated in three separate fusion experiments performed using nonimmunized B/W spleen cells were studied. Their antigenic specificities were demonstrated to be identical and directed against a conformational determinant of the B helical form of double-stranded DNA (dsDNA). These data suggest that autoantibodies to dsDNA in B/W mice could constitute a homogeneous population.
T cells produce TRF in response to Con A and factors in T cell hybridoma supernatants.
Authors: Raulet D H , Hünig T , Parker D C
Published: December 2022

In recent experiments, a soluble factor (TRF) that mediates the differentiation of anti-immunoglobulin (Ig)-activated B cells to Ig-secreting cells has been identified. TRF works in concert with a growth factor, probably IL 2, in the induction of activated B cells. In previous studies, TRF was identified in culture supernatants of activated T cells and accessory cells, and thus the cellular source (T cell or accessory cell) of the factor was not determined. In the present studies, we succeeded in inducing the production of TRF by T cell populations from which accessory cells had been vigorously depleted. Lymph node cells were depleted of accessory cells by nylon wool adherence and anti-Ia and complement treatment; these cells were activated with Con A and a T cell hybridoma supernatant that contains IL 2. Supernatants from these activated T cell cultures supported the differentiation of anti-Ig-activated B cells to Ig secreting cells. These results show that T cells produce the differentiation factor, and further that they do so in response to ligand (Con A) plus a T cell-derived factor.
Preferential induction of autoantibody secretion in polyclonal activation by peptidoglycan and lipopolysaccharide. I. In vitro studies.
Authors: Dziarski R
Published: December 2022

Peptidoglycan (PG) and lipopolysaccharide (LPS) are bacterial cell wall components that are B cell mitogens and activators of polyclonal antibodies. Using the hemolytic plaque assay and its modifications with protein A-, IgG-, or DNA-coated erythrocytes, we determined the proportions of cells that secrete antibodies specific to various autologous and heterologous antigens in PG- and LPS-stimulated cultures of mouse splenic lymphocytes. Of all PG- and LPS-activated IgM-secreting cells, 47 to 75% produced antibodies specific to mouse IgG; 2.7 to 21% produced IgM antibodies that reacted with bovine IgG; 4.4 to 17%, with single-stranded (ss) DNA; 0.18 to 5.9% with double-stranded (ds) DNA; 0.16 top 3.3%, with bromelin-treated mouse red cells; 0.006 to 0.02%, with intact mouse red cells; and 0.35 to 1.9%, with sheep red cells. A similar distribution of cells secreting these antibodies was observed in unstimulated cultures. However, the absolute numbers of cells secreting these antibodies were substantially higher in the PG- and LPS-stimulated cultures. Similar results were obtained in BALB/c, CBA/H, and C57BL/6 mice. Since these strains of mice have different H-2 types, there was no association of a single H-2 type with the ability to form hgh numbers of autoantibody-secreting cells in vitro. The production of autoantibodies closely resembled polyclonal activation of all immunoglobulin-secreting cells in terms of the kinetics and dose-response. It did not involve any substantial specific anti-PG or anti-LPS response. Further studies on the specificities of polyclonally induced antibodies confirmed the specificity of anti-mouse IgG and anti-ssDNA antibodies. However, the formation of plaques with bovine IgG- or dsDNA-sensitized red cells was due to the cross-reacting anti-mouse IgG or anti-ssDNA antibodies, respectively. These results do not support the popular hypothesis of an equal polyclonal activation of lymphocytes secreting antibodies of all different specificities. Also, if preferential activation of cells secreting rheumatoid factor and anti-ssDNA antibodies occurs in vivo, it may have important pathologic consequences.
Analysis of cerebrospinal fluid and serum of patients with multiple sclerosis by means of anti-idiotypic antisera.
Authors: Nagelkerken L M , van Zoonen-van Exel M , van Walbeek H K , Aalberse R C , Out T A
Published: December 2022

Twelve antisera were prepared, each specific for idiotype(s) of IgG from the cerebrospinal fluid (CSF) of an individual patient with multiple sclerosis (MS). Idiotype-positive (Id(+)) IgG represented 24 to 33% of the CSF-IgG. Radioautographic analysis showed that Id(+)IgG in CSF and in serum had a restricted electrophoretic mobility. From serum of three patients, at least two different noncross-reacting Id(+)IgG fractions were isolated. The anti-Id antisera, which detect IgG considered to be of importance in MS, were used to measure Id(+)IgG in CSF and in serum. Relative to the total IgG, Id(+)IgG was always enriched in CSF, which suggests intrathecal synthesis of such IgG. However, in all patients the absolute concentration of Id(+)IgG was higher in serum than in CSF. Studies attempting to demonstrate cross-reactivity between the IgG of different patients were negative.
Genetic control of experimental autoimmune myasthenia gravis in mice. III. Ia molecules mediate cellular immune responsiveness to acetylcholine receptors.
Authors: Christadoss P , Lennon V A , Krco C J , David C S
Published: December 2022

When MHC congenic and recombinant mice are inoculated with Torpedo acetylcholine receptors (AChR) with adjuvants, the magnitude of autoantibody responses to muscle AChR and the defect of neuromuscular transmission closely parallel in vitro lymphocyte proliferative responses to Torpedo AChR. All of these responses are controlled by gene(s) at the I-A subregion of the H-2 complex. Data presented in this report confirm in back-cross mice that T lymphocyte proliferative responses to AChR are controlled by a Mendelian dominant gene linked to H-2, at the I-A subregion. Lymphocyte responses were eliminated by blocking Ia antigens on lymph node cell surfaces with appropriate anti-I-A alloantisera and by removal of adherent cells. A spontaneous mutation at the I-A subregion in the B6 strain, which resulted in structural alteration of the A beta chain of Ia, converted high responsiveness to AChR to a state of low responsiveness. These data implicate a macrophage-associated Ia molecule in induction of autoimmune responses to AchR, probably in the presentation of AChR to helper T lymphocytes that thereby help B lymphocytes to differentiate into anti-AChR antibody-forming cells.
Mac-2, a novel 32,000 Mr mouse macrophage subpopulation-specific antigen defined by monoclonal antibodies.
Authors: Ho M K , Springer T A
Published: December 2022

Two monoclonal antibodies, M3/31 and M3/38, were obtained by fusion of mouse myeloma cells with rat spleen cells immunized to immunoadsorbent-purified macrophage glycoproteins. Co-precipitation experiments show that antigenic determinants recognized by these two antibodies reside on the same molecular species, termed Mac-2, Mac-2, an antigen of 32,000 Mr, is synthesized by and expressed on the surface of thioglycollate-elicited macrophages as shown by [35S]-methionine and 125I labeling. Saturation binding experiments show that thioglycollate-elicited macrophages express 1.7 X 10(5) Mac-2 sites/cell. Thioglycollate-elicited macrophages are strongly absorptive for 125I-labeled M3/38 MAb. Kidneys are also absorptive; however, evidence is presented pointing to the nonspecificity of this absorption. Lymph node and thymus are negative, whereas spleen and bone marrow are weakly absorptive, probably due to stromal cells. Nonlymphoid tissues, such as lung, liver, heart, and brain, exhibit slight or no absorbing capacity. Cell suspensions from spleen, bone marrow, thymus, and peripheral lymph node are greater than 99% Mac-2- by immunofluorescent flow cytometry. In contrast, thioglycollate-elicited macrophages are greater than 96% strongly positive for Mac-2. Only 20% of peptone-elicited cells are weakly positive, whereas resident peritoneal macrophages and other macrophage elicited by Listeria monocytogenes, Con A, or LPS are greater than 98% negative. SDS-PAGE of [35S]-methionine-labeled Mac-2 shows that thioglycollate-elicited macrophages synthesize 10- to 30-fold more Mac-2 than other peritoneal macrophage subpopulations, whereas all types of peritoneal macrophages synthesize and express on their surfaces similar amounts of the Mac-1 antigen. Mac-2 antigen is therefore induced in macrophages only in response to specific differentiative signals.
Fine specificity of a T cell line reactive to bovine insulin.
Authors: Reske-Kunz A B , Rüde E
Published: December 2022

A continuous T cell line was established from lymph node T cells of (high responder X low responder) hybrid mice ((B10 X B10.BR)F1) immune to bovine insulin (BI). T cells were expanded by restimulation with BI presented on irradiated syngeneic spleen cells and by culture in interleukin 2-containing medium. The proliferative response, measured in a 3H-thymidine incorporation assay, was specific for the selecting antigen BI; unrelated proteins did not induce DNA synthesis. However, in addition to BI the T blasts were efficiently restimulated by pig and sheep insulin, which are identical to BI except for the amino acid sequence in the A chain loop. This suggests that the T blasts recognized an antigenic determinant(s) different from the A chain loop epitope of BI, the essential antigenic moiety for cells of high responder parental H-2b mice. Induction of the blasts to DNA synthesis by the three insulins required the presence of F1 spleen cells. Parental strain B10 or B10.BR splenocytes failed to support a proliferative response, even when present as a mixture. The antigen-presenting determinants unique to F1 cells were mapped to the I-Ab/Kb and I-Ak loci. These data demonstrate that, although at least the majority of responding T cells in H-2b high responder mice recognize the A chain loop determinant of BI in the context of restriction elements of the b haplotype, T cells can be isolated from (high responder X low responder)F1 mice, which require recognition of other determinants on the insulin molecule in the context of F1 unique restriction structures for stimulation to occur.
Hapten-specific T cell responses to 4-hydroxy-3-nitrophenyl acetyl. XIII. Characterization of a third T cell population involved in suppression of in vitro PFC responses.
Authors: Sherr D H , Dorf M E
Published: December 2022

The involvement of a third-order suppressor T cell population (Ts3) in the suppression of in vitro PFC responses was analyzed. It was shown that Ts2 effector-phase suppressor cells, induced by the i.v. injection of NP-coupled syngeneic spleen cells, require a third suppressor T cell population to effect NPb idiotype-specific suppression of an in vitro B cell response. This Ts3 population was shown to be present in NP-primed but not unprimed donors. The Ts3 population specifically binds NP and is Lyt-1-, Lyt-2+, I-J+ and bears NPb idiotypic determinants. The involvement of the Ts3 population in a suppressor pathway that requires recognition of idiotypic determinants is discussed.
Biosynthesis and processing of a human T lymphocyte antigen.
Authors: Bergman Y , Levy R
Published: December 2022

The biosynthesis and processing of Leu-1, a human T lymphocyte antigen, has been studied with the use of a monoclonal antibody. This molecule exists on the cell surface as a 67,000 m.w. glycoprotein. Through a series of pulse-labeling studies, in conjunction with the use of the antibiotic tunicamycin and the enzyme Endo-H, the details of glycosylation, processing, and deposition at the cell membrane were examined. The protein backbone of the molecule is 58,000 m.w. High-mannose sugars are added to asparagine residues during synthesis. Within 20 min, these high mannose sugars are converted to complex type carbohydrates, including fucose. The fully processed glycoprotein appears at the cell surface within 30 min after synthesis. This sequence of events is similar to that for other cell surface glycoproteins, including HLA and vesicular stomatitus virus glycoprotein.
Kinetics of extravasation and redistribution of IgA-specific antibody-containing cells in the intestine.
Authors: Husband A J
Published: December 2022

Double Thiry-Vella loops were prepared in rats, each locally immunized with a different antigen, and the kinetics of appearance in the loops of antibody-containing cells (ACC) specific for each antigen was recorded. The results confirm, under more rigorously controlled conditions, the conclusions of previous work that the initial appearance of IgA lymphoblasts in the intestinal lamina propria occurs independently of antigen. In addition, these findings are extended by demonstrating 1) that there is no specialized site within the intestinal lamina propria for ACC to enter the tissue; 2) that in immunized intestine, specific ACC progressively accumulate around the crypt regions; and 3) that a proportion of this specific accumulation can be accounted for by ACC proliferation within the lamina propria.
Chemotactic factor inactivation by stimulated human neutrophils mediated by myeloperoxidase-catalyzed methionine oxidation.
Authors: Clark R A , Szot S
Published: December 2022

Leukocyte chemoattractants were inactivated when exposed to human neutrophils and either ingestible particles or phorbol esters. Loss of biologic activity was time- and temperature-dependent, required physiologic concentrations of viable neutrophils and a halide, and was inhibited by azide or catalase. Neutrophils from patients with either hereditary myeloperoxidase deficiency or chronic granulomatous disease failed to inactivate the chemoattractants unless purified myeloperoxidase or H2O2, respectively, was added. Susceptibility to inactivation by neutrophils correlated with the presence of methionine in the attractant. Loss of chemotactic activity was blocked by low concentrations of methionine and by higher concentrations of other reducing agents, but was unaffected by oxidized methionine. Paper chromatography demonstrated that exposure of a formyl-methionyl peptide chemotactic factor to either the cellfree myeloperoxidase system or stimulated neutrophils resulted in its conversion to a molecular species whose location in the chromatographs was identical to that of the peptide containing oxidized methionine. Thus, stimulated human neutrophils inactivate peptide chemoattractants by secretion of myeloperoxidase and H2O2, which combine with halides to form oxidants that react with a critical methionine residue. We suggest that myeloperoxidase-catalyzed oxidation of thioethers may constitute an inflammatory control mechanism as well as a general means of modifying the functional properties of biologic mediators.
Macrophage surface component gp160: sensitivity to plasmin and other proteases.
Authors: Remold-O'Donnell E , Lewandrowski K
Published: December 2022

The correlation between activation of macrophages and increased secretion of plasminogen activator suggests that macrophages are exposed to the protease plasmin. Incubation of 125I-labeled, caseinate-elicited guinea pig peritoneal macrophages with plasmin cleaves a surface protein, gp160, characterized previously by its sensitivity to trypsin. The gp160 fragments produced by plasmin (fr85 and fr71), which remain disulfide-bonded in the membrane, comigrate with the fragments produced by trypsin, indicating close or identical cleavage sites. No other detectable 125I-labeled surface component is cleaved by plasmin. Neither gp160 nor any other detectable 125I-labeled surface component was cleaved by a series of other proteases associated with inflammation including thrombin, collagenase, pancreatic elastase, leukocyte elastase, cathepsin G, and urokinase. Analysis with the use of homogeneous plasmin from guinea pig plasma shows that concentrations as low as 50 micrograms/ml cause measurable cleavage of gp160 in 30 min.
Factors that modulate clearance and ultimate fate of a specific schistosome antigen (GASP) in schistosome infections.
Authors: Nash T E
Published: December 2022

To understand factors that modulate the amount and removal of antigen found in the circulation of schistosome-infected mice, a specific schistosome antigen, GASP, found in the circulation of infected animals, was radiolabeled with Bolton-Hunter reagent and injected into infected and control mice. The rates of clearance, levels of antigen binding, and the fate of the injected antigen were determined. Within 1 to 3 wk after injection, infected animals developed enhanced antigen clearance that paralleled the increase in antibody to GASP. Lightly infected animals showed faster clearance rates and increased antibody levels compared with heavily infected mice. The level of antibody in the sera, irrespective of the duration or intensity of infection, correlated with enhanced clearance. Heavily infected animals removed preformed immune complexes as rapidly as lightly infected mice. This indicates that the relative inability to clear antigen was not due to blockade of Kupffer cells. Circulating GASP antigen, determined by countercurrent electrophoresis, was detected in heavily infected animals in the presence of increased antibody, suggesting that circulating GASP was in immune complexes. Both free and complexed antigen were primarily cleared by the liver and then excreted in the urine as low m.w. nonantigenic products. With sensitive radioimmunoassays, GASP could not be detected in infected human or mouse urine.
Isolation, characterization, and mechanism of action of rat beta 1H.
Authors: Daha M R , van Es L A
Published: December 2022

beta 1Hrat was purified to homogeneity from fresh rat plasma by precipitation with 28.6% ammonium sulfate followed by sequential chromatography on DEAE-Sephacel, Biorex-70, and gel filtration on Sephacryl-S300. The final material was homogeneous on SDS-PAGE analysis and had an apparent m.w. of 150,000. Reduction with dithiothreitol did not affect its m.w., suggesting that the molecule is composed of one polypeptide chain. The recovery of beta 1H was approximately 10%. A monospecific antibody against beta 1Hrat was obtained from immunized rabbits, which recognized beta 1Hrat as a protein with beta-electrophoretic mobility upon immunoelectrophoresis of fresh rat plasma. The concentration of beta 1H in plasma of normal 4-mo-old Wistar rats was 243.5 +/- 36.3 micrograms/ml (mean +/- S.D.). beta 1Hrat in this study was detected by its capacity to inhibit formation of the P-stabilized cell-bound amplification C3/C5 convertase composed of cell-bound C3bhu and Bbhu. Purified beta 1Hrat produced a dose-related, first-order loss of convertase function and release of 126I-Bbhu from the P-stabilized C3bhuBbhu convertase, indicating a mechanism of action by decay-dissociation of Bbhu from the complex C3bhuBbhuP. beta 1Hrat was at least four times less effective than beta 1Hhu in release of 125I-Bbhu from the homologous convertase composed of C3bhu and Bbhu. On the other hand beta 1H was twice as effective in releasing 125I-Bbrat from the convertase composed of C3bratBbratP when compared to beta 1Hhu. These differences are presumably dependent upon the species-specific affinity of beta 1H from humans or rats for C3bhu or C3brat, respectively.
Properties of a surface antigen expressed on activated human thymus-derived lymphocytes.
Authors: Udey M C , Jendrisak M D , Parker C W
Published: December 2022

We have identified previously a quantitatively minor membrane protein (p28) with an apparent reduced m.w. of 28,000, which is biosynthetically labeled in activated human lymphocytes. Rabbit antisera with activity directed against p28 (alpha-ATC) were prepared and p28 was identified by immunoprecipitation in NP-40 extracts of activated, extrinsically labeled lymphocytes. p28 was not expressed in appreciable amounts by unstimulated T cells, stimulated or unstimulated B cells, null cells, or adherent cells. Protein p28 was only minimally represented on resting thymocytes but was easily detected on 4-hr activated thymocytes and the T lymphoblastoid cell lines HSB2 and MOLT-4. Absorption and immunoprecipitation studies with alpha-ATC indicated that p28 was not present on erythrocytes, platelets, neutrophils, six B cell lines, six null cell lines, and seven other T lymphoblastoid cell lines. Protein p28 from HSB2 cells was absorbed by lentil lectin, concanavalin A, and wheat germ agglutinin affinity columns and was eluted with the appropriate sugars. Gel filtration column chromatography of unreduced p28 in the presence of 0.5% NP-40 or 0.1% deoxycholate gave elution characteristics consistent with a m.w. of approximately 60,000 to 100,000. In preparative isoelectric focusing (IEF) studies the isoelectric point (pI (p28) = 5.2 to 6.1) was similar or identical to that described for the reduced and denatured protein in two-dimensional polyacrylamide gels (pI = 5.5 to 6.2). Protein p28 was eluted from DEAE-cellulose (Whatman DE-52) ion exchange columns at 0.05 to 0.15 M NaCl. Experiments with monoclonal antibodies or heteroantisera specific for other T cell and B cell antigens and various lymphoblastoid cell lines and normal peripheral blood cells indicated that p28 is distinct from the human Ia-like antigens, from T3, T4, T5, T8, and from several other reported human T cell antigens that appear to correspond to Thy-1, the sheep erythrocyte receptor, and a human thymus-leukemia antigen.
Human helper T cell factor(s) (ThF). II. Induction of IgG production in B lymphoblastoid cell lines and identification of T cell-replacing factor- (TRF) like factor(s).
Authors: Teranishi T , Hirano T , Arima N , Onoue K
Published: December 2022

IgG-PFC was induced in Epstein-Barr virus-transformed B lymphoblastoid cell lines (LCL) by the addition of allogeneic T cells. T cells involved in the induction of IgG-PFC were shown to belong to the Leu 3a+/2a- T cell subset. Furthermore, partially purified soluble factors obtained from the culture supernatant of PPD-stimulated pleural T cells or PWM-stimulated tonsillar mononuclear cells was shown to induce IgG-PFC in LCL across the major histocompatibility complex barrier. The induction of IgG-PFC was observed only in surface IgG-positive LCL cell populations and was not accompanied by the increase in the number of LCL cells. The factors with such a TRF-like activity were found in two fractions corresponding to the m.w. range of 18,000 to 25,000 (22K fraction) and 28,000 to 38,000 (36K fraction) by gel filtration. Isoelectric focusing of these fractions revealed that TRF-like activity of both 22K and 36K fractions distributed in the pI range of 5.0 to 6.0, and both fractions were found to be devoid of TCGF activity. These results appear to indicate that the factors act on the B cells in terminal stages to trigger final differentiation to Ig-producing cells.
Differences in expression of MHC products between several H-2-restricted CTL clones.
Authors: Hauptfeld V , Braciale T J , Shreffler D C
Published: December 2022

Five cytotoxic T lymphocyte (CTL) clones, all specific for influenza virus A/JAP/57 and all H-2Kd-restricted, were typed for expression of MHC products. All express H-2K and H-2D encoded antigens, but they differ in their expression of I region products. One clone, 14-2, is positive with antisera directed against serologically detectable products apparently related to three different I regions, namely I-A, I-J, and I-E. Two clones, 11-1 and 14-1, lack I-E-associated antigens but express I-A and I-J products, and two other clones, 14-7 and 14-13, lack detectable I region products. These observations show that although discrete CTL clones may be homogeneous in their function, they differ in their expression of I region products.
Immunosuppression of experimental allergic encephalomyelitis. III. In vitro evidence for induction of suppressor T lymphocytes in draining lymph node cells of animals immunized with myelin basic protein complexed to lipopolysaccharides.
Authors: Raziuddin S , Kibler R F , Morrison D C
Published: December 2022

We recently demonstrated that Lewis rats immunized with bacterial lipopolysaccharides (LPS) precomplexed to guinea pig myelin basic protein (BP) in complete Freund's adjuvant were less effective in inducing experimental allergic encephalomyelitis (EAE) than BP-immunized controls. When tested in vitro both lymph node cells (LNC) and spleen cells (SpC) of animals immunized with BP-LPS were less effective in proliferative responses to various mitogens, which included phytohemagglutinin, concanavalin A, purified protein derivative of tuberculin, LPS, and BP. Of importance immunization of rats with BP complexed to LPS results in the generation of cells in lymph nodes of these animals that suppress the mitogenic response of BP-immunized LNC and also SpC in mixed lymphocyte cultures. The suppressive effect of these cells in mixed lymphocyte culture reaction was found specifically in response to BP and to a lesser extent to LPS in LNC. SpC of BP-LPS immunized animals did not suppress the proliferative response to SpC of BP-immunized animals. Treatment of these LNC with antithymocyte serum and complement abolished this suppressive effect of LNC, suggesting that the immunoregulatory cells in LNC of BP-LPS immunized animals are suppressor T lymphocytes. The parallel between the in vitro induction of suppressor T lymphocytes in the draining LNC and the function of LPS in the development of EAE in Lewis rats suggests a possible immunologic significance of the effect.
Induction in mice of cell-mediated immunity to Pseudomonas aeruginosa by high molecular weight polysaccharide and vinblastine.
Authors: Pier G B , Markham R B
Published: December 2022

The effect of the cytotoxic drug vinblastine on the development of immunity to high m.w. polysaccharide (PS) isolated from culture supernates of Pseudomonas aeruginosa was investigated. One microgram of PS, a normally nonimmunogenic, nonprotective dose, plus 75 micrograms of vinblastine were administered to BALB/c mice, and afforded protection to live organism challenge with the homologous strain. The kinetics and serotype specificity of the immune response indicated an active immunization had occurred. Analyses of serum antibody levels of mice given the PS-drug regimen in a sensitive, radioactive antigen-binding assay (RABA) failed to show development of antibody to the immunizing PS. Immunity could be passively transferred with spleen cells but not by serum from PS-drug-immunized animals, and the effector cell was removed by antisera to the Thy-1.2 antigen. Nu/nu mice were also protected against challenge after immunization with PS and vinblastine, but this protection was observed in association with the development of serum antibody to PS in these mice, as measured in the RABA. Protective immunity could not be elicited in the BALB/c mice by PS plus cyclophosphamide. These data suggest that under certain conditions, PS antigens can elicit T cell-dependent immune phenomena, and this T cell-dependent immunity can protect mice from live organism challenge against an extracellular bacterial pathogen.
Murine epidermal cell-derived thymocyte-activating factor resembles murine interleukin 1.
Authors: Luger T A , Stadler B M , Luger B M , Mathieson B J , Mage M , Schmidt J A , Oppenheim J J
Published: December 2022

We have previously described an epidermal cell-derived thymocyte-activating factor (ETAF), which is produced by the murine PAM 212 keratinocyte cell line. ETAF appeared to be similar to macrophage-derived interleukin 1 (IL 1) in its biologic activities and biochemical characteristics. Both IL 1 and ETAF augment thymocyte proliferation, enhance lymphocyte production of interleukin 2 (IL 2), and are 15,000 m.w. polypeptides that are stable at pH 4 to 11 and from -70 degrees C to 60 degrees C. In this study we describe a quantitative microassay to obtain standardized assessment of ETAF activity, which enabled us to further define the characteristics of ETAF and its relationship to IL 1. Just as stimulated macrophages produce more IL 1 activity, Pam 212 keratinocyte production of ETAF activity was increased by stimulation with lipopolysaccharide (LPS) or silica. Increased levels were also obtained by mechanical disruption of confluent monolayers of keratinocytes and by blocking proliferation of the Pam 212 cells with hydroxyurea at the G1/S interphase. These observations in conjunction with a concomitant decrease in keratinocyte viability suggest that "injurious" stimuli that prolong the G1 phase of the cell cycle factor ETAF production. ETAF, like murine IL 1, has an isoelectric point of 5.2. The same subpopulations of PNA-thymocytes that respond to PHA and IL 1 are responsible for the enhanced proliferative response to ETAF. Furthermore, as in the case of IL 1, PNA- Lyt-2- thymocytes were most responsive to ETAF, but not PNA+ LYt-2+ thymocytes. Finally, ETAF activity, like IL 1, appears to be a mitogenic signal for fibroblasts. Although produced by different cell types, these observations continue to support the view that ETAF may be identical or closely related to IL 1.
Hepatobiliary transport of IgA immune complexes: molecular and cellular aspects.
Authors: Brown T A , Russell M W , Mestecky J
Published: December 2022

The molecular and cellular parameters of the transport of IgA immune complexes (IgA-IC) from the circulation to the bile in vivo were studied in mice. Polymeric MOPC 315 IgA was effective in the transport of 125I-DNP-HSA into the bile, whereas complexes containing monomeric IgA were not transported. The transport to IgA-IC could be blocked by excess from IgA of different antigen specificity, but not by IgG or IgM. Hepatobiliary transport did not appear to involve the galactose- or mannose-specific glycoprotein receptors of the liver. No differences in IgA-IC transport, relative to controls, could be seen in mice that had been treated with cobra venom factor or whose mononuclear phagocytic system had been blockaded with colloidal carbon. These data suggested that the hepatobiliary transport of IgA-IC proceeds by a mechanism similar to the selective transport of free polymeric IgA through the hepatocyte.
Extracts of pregnancy urine contain a mitogen for human peripheral blood lymphocytes (PBL).
Authors: Morse J H , Ehrlich P H , Canfield R E
Published: December 2022

A mitogen for human peripheral blood lymphocytes (PBL) has been found in commercial batches of crude human chorionic gonadotropin (hCG), which is obtained from pregnancy urine. These crude hCG preparations, as well as column eluates free of hCG and a previously noted immunosuppressive factor, are mitogen for normal human PBL cultured in fetal calf serum. This mitogen is designated pregnancy-associated growth factor (PAGF) because of its source. PAGF induces 3H-thymidine incorporation of mixed Ia+ non-T cells and T cells, probably confined to the helper subclass; proliferation is abolished by complement-mediated lysis using monoclonal antisera to Ia, mature T cells (OKT3), and the Th (OKT4) subset, but not by antisera to the Ts (OKT8) subset. The proliferating cell is a T cell because irradiation of these cells, but not the non-T cells, abolished PAGF mitogenicity. In addition, PAGF augments the autologus mixed lymphocyte reaction. Preliminary experiments indicate that its m.w., in the range of 18,000 to 22,000, is different from epidermal growth factor, the only previously described growth factor found in crude hCG preparations. In addition epidermal growth factor failed to stimulate PBL under similar conditions.
IgG subclass distribution of anti-sheep red blood cell plaque-forming cells in mice with the CBA/N defect.
Authors: Phillips N E , Campbell P A
Published: December 2022

The IgG subclass distribution of anti-sheep red blood cell (SRBC) plaque-forming cells (PFC) in CBA/N, CBA/Ca, and (CBA/N X CBA/Ca)F1 male and female mice was examined. The results show that the X-linked immunodeficiency of CBA/N mice affects the development of anti-SRBC PFC of all IgG subclasses tested.
Suppression of human B lymphocyte function by cyclophosphamide.
Authors: Cupps T R , Edgar L C , Fauci A S
Published: December 2022

The immune responses of 16 patients with nonneoplastic immune mediated diseases including Wegener's granulomatosis, systemic necrotizing vasculitis, cutaneous vasculitis, and relapsing nodular panniculitis were evaluated before and during therapy with chronic low-dose (2 mg/kg/day) cyclophosphamide. A striking selective suppression of B cell function was noted as measured by PWM-induced immunoglobulin secretion. This suppression was a direct effect on the B cells themselves because T cell function, measured by blastogenic responses to the mitogens PHA, Con A, and PWM, was not significantly suppressed. Furthermore, the ability of T cells from cyclophosphamide-treated patients to provide helper function in T cell-dependent B cell assays remained intact. Treated patients manifested a total lymphocytopenia without a selective depletion of relative proportions of B cells or T cell subsets. However, the spontaneous secretion of immunoglobulin by peripheral blood B cells that is elevated in untreated patients was suppressed back to normal levels during cyclophosphamide therapy. This selective effect on spontaneous and induced secretion of immunoglobulin by human B cells may help explain the efficacy of cyclophosphamide therapy in certain antibody and immune complex-mediated diseases.
Mucosal mast cells. I. Isolation and functional characteristics of rat intestinal mast cells.
Authors: Befus A D , Pearce F L , Gauldie J , Horsewood P , Bienenstock J
Published: December 2022

We have developed a procedure for the dispersion of mast cells from the intestinal lamina propria (LP) and epithelium of rats infected with the intestinal nematode, Nippostrongylus brasiliensis. The dispersed cells are morphologically and histochemically similar to intestinal mucosal mast cells (MMC) in situ and are distinguishable from peritoneal mast cells (PMC). MMC derived from the LP or epithelium of parasitized animals secrete histamine in response to the specific parasite antigens as well as anti-IgE. Unlike PMC, these cells are unresponsive to the basic secretagogues 48/80 and bee venom peptide 401. Similarly, bee venom peptide 401 conjugated with dansyl chloride binds to PMC and mast cells in the thymus and intestinal serosa, but not to mast cells in or derived from the intestinal LP and epithelium. Studies on PMC treated by the intestinal cell isolation procedure show that the functional characteristics of the MMC cannot be solely attributed to the isolation procedure. Thus, MMC have been isolated and shown to be morphologically, histochemically, and functionally different from PMC, as suggested by previous in vivo studies of the normal intestine.
Mucosal mast cells. II. Effects of anti-allergic compounds on histamine secretion by isolated intestinal mast cells.
Authors: Pearce F L , Befus A D , Gauldie J , Bienenstock J
Published: December 2022

Functional mast cells have been isolated from the lamina propria of the small intestine of rats infected with the nematode Nippostrongylus brasiliensis. The cells released histamine on challenge with specific antigen, anti-rat IgE, concanavalin A, and calcium ionophores but were less responsive than peritoneal mast cells (MMC) from the same animals. Intestinal mucosa mast cells (PMC) were refractory to the action of the basic secretagogues peptide 401 from bee venom and compound 48/80. The anti-allergic compounds disodium cromoglycate (less than or equal to 10(-3) M), AH 9679 (less than or equal to 10(-4) M), and theophylline (less than or equal to 10(-2)) did not inhibit antigen-induced histamine secretion by MMC, although these compounds were effective against PMC. In contrast, doxantrazole (10(-5) to 10(-3) M) inhibited the secretion of histamine from both MMC and PMC in a comparable dose-dependent fashion. Thus, we have established that mast cells from different sites are functionally heterogeneous not only in their response to various stimuli for histamine secretion, but also in their responses to different pharmacologic modulators of secretion. It cannot be assumed that anti-allergic compounds effective against mast cells in one tissue site or organ will be equally efficacious against mast cells in other sites. The extent of this functional heterogeneity must be established, and its investigation may provide new insights into the biochemical events involved in mast cell secretion.
Mixing two monoclonal antibodies yields enhanced affinity for antigen.
Authors: Ehrlich P H , Moyle W R , Moustafa Z A , Canfield R E
Published: December 2022

We observed that mixing monoclonal antibodies directed against various epitopes of human chorionic gonadotropin (hCG) can increase the sensitivity of antigen-binding assays. Depending on the antibody pair chosen, the affinity of the mixture was as much as 10-fold higher than either of the monoclonal antibodies assayed separately. Because hCG does not have any repeating sequences, these results do not reflect a change in avidity of an individual antibody. The increased avidity of the mixture can be detected in both solid-phase assays and liquid-phase double antibody radioimmunoassays. This was not a general property of all monoclonal antibodies raised against hCG. Certain antibodies, apparently reacting with the same region of the antigen molecule, do not bind simultaneously and do not result in enhanced affinity when mixed. In addition, certain other pairs of antibodies do not bind cooperatively even though they can bind simultaneously. The mechanism for the increase in affinity depends on the formation of a multicomponent complex. If one of the antibodies of a pair that results in enhanced affinity upon mixing is replaced by its F(ab) fragment, the enhancement is no longer detectable, indicating it is unlikely the enhancement is due to an allosteric effect. Although the F(ab')2 fragment shows some enhancement when mixed with another antibody, it is not as effective as the intact antibody.
Loss of surface-bound antibody accompanying the anti-complementary modulation of leukemic B cell immunoglobulin: contrasting effects of antibodies directed against idiotypic and constant regions.
Authors: Gordon J , Anderson V A , Stevenson G T
Published: December 2022

Guinea pig L2C leukemic lymphocytes display at their surfaces monoclonal IgM, which when compared with antibody undergoes rapid redistribution and variable endocytosis. One consequence of this is that the cells can prove resistant to lysis by complement subsequently added to the system, a process termed here anti-complementary modulation. We studied quantitatively the extent of antibody loss accompanying the modulation by radioimmunolabeling the cell surfaces with 125I-Fab' gamma fragments from an anti-antibody. Antibody directed against the constant region of the IgG light chain (anti-lambda) gave modulation effective against syngeneic (guinea pig strain 2) complement that closely paralleled the disappearance of anti-lambda from the cell surfaces. Antibody directed against the idiotypic region of the light chain (anti-Id) was as effective as anti-lambda in modulating against syngeneic complement. However, the bulk of the anti-Id was seen by radioimmunolabeling to persist on the surfaces of the resistant cells, even after prolonged exposure at 37 degrees C, and was shown by immunofluorescence to be in a patched configuration. In contrast to the results with syngeneic complement, modulation effective against rabbit complement appeared to have an absolute requirement for clearing of the antibody: thus anti-lambda could modulate, anti-Id could not. The differences observed between anti-lambda and anti-Id could not be accounted for by differences in their isotypic (Ig subclass) composition nor by the numbers of antibody molecules bound. Studies with directly fluoresceinated and 125I-labeled anti-lambda revealed endocytosis rather than shedding was the major route of antibody loss from the cell surfaces over the period of anti-complementary modulation. The findings are discussed in relation to mechanisms that enable leukemic B lymphocytes to escape destruction when confronted by antibody and complement.
Immune (y) interferon production by murine T cell lymphomas.
Authors: Landolfo S , Arnold B , Suzan M
Published: December 2022

Various cloned murine T cell hybridomas and T cell lymphomas were evaluated for their ability to produce interferon (IFN). Two T cell tumor clones, L12-R1 and L12-R4, derived from the spontaneously in vitro transformed cell lines L12 originally established from fetal calf serum-primed C57BL/6 spleen cells were found to produce high IFN amounts upon mitogen stimulation. Phorbol myristate acetate led to maximal IFN production (2187 IU) by L12-R4 cells at concentrations of 2 x 10(-7) M, whereas concanavalin A and phytohemagglutinin induced lower levels of IFN synthesis (160 to 243 IU). None of the cell lines tested produced IFN constitutively or upon lipopolysaccharides stimulation. The IFN was characterized as immune (y) by being labile at pH 2 and neutralized by two rabbit anti-murine IFN-y antisera but not by antiserum to murine leukocyte (alpha) and fibroblast (beta) IFN. Phenotypic characterization of IFN-y-producing cells showed the L12 clones to be Thy-1.2+, Lyt-1+, 23-, and Ig-. The L12-R4 tumor cell therefore provide a unique source of IFN-y for purification, and may represent a useful model for studying the molecular mechanisms involved in T cell differentiation leading to IFN-y production.
Inhibition of in vitro granulopoiesis by autologous allogeneic human NK cells.
Authors: Hansson M , Beran M , Andersson B , Kiessling R
Published: December 2022

This study demonstrates the ability of human NK cells to inhibit in vitro granulopoiesis of autologous and allogeneic BM cells. NK lytic activity and GM-CFC inhibition was present among nonstimulated lymphocytes from healthy donors and could be increased by treatment of PBL with IFN. Both the cytotoxic NK cells and the GM-CFC inhibitory cells could be enriched for among nonadherent, low-density cells. High-density cells were not cytotoxic, only inhibitory to a small extent, and could become neither cytotoxic nor more inhibitory after IFN treatment. In contrast, low-density cells showed an increased cytotoxic and GM-CFC inhibitory capacity after IFN treatment. The NK mediated GM-CFC inhibition was dependent on cell contact with BM cells, increased with longer preincubation times, and was most efficient against 7-day GM-CFC as compared with 14 day GM-CFC progenitors. In conclusion, these data provide new information about the human NK cell as a potent inhibitor of in vitro granulopoiesis and also as a possible regulator of hematopoiesis in vivo.
The modification of human immunoglobulin binding to staphylococcal protein A using diethylpyrocarbonate.
Authors: Haake D A , Franklin E C , Frangione B
Published: December 2022

Human IgG subclasses 1, 2, and 4, as well as proteins of the IgG3 subclass that are allotype G3m (s+t+), bind avidly to staphylococcal protein A by means of their Fc portion. Proteins of the IgG3 subclass that are allotype G3m (s-t-) do not bind. The importance of a histidine residue at position 435 has been implicated from comparison of amino acid sequences of immunoglobulins that bind with those that do not bind to staphylococcal protein A, as well as from crystallographic data. Modification of histidines at a low concentration of diethylpyrocarbonate successfully and reversibly alters the binding of immunoglobulins to staphylococcal protein A with only minimal change in the antigenic properties. This method provides strong evidence for the critical importance of histidine in the binding of immunoglobulins to staphylococcal protein A.
Local passive transfer of delayed-type hypersensitivity in the mouse.
Authors: Scovern H , Kantor F S
Published: December 2022

A means for local passive transfer of DTH in the mouse has been discovered and partially characterized. Briefly, peritoneal exudates (PE) are induced with incomplete Freund's adjuvant 5 to 7 days after immunization with antigen (usually picGPA) emulsified with complete Freund's adjuvant. Five days later, PE cells are harvested and transferred with soluble antigen into a hind footpad of a naive mouse. Antigen-specific swelling is defined as the change in diameter of footpads over the 24 hr after injection of PE cells plus immunizing antigen minus change in diameter of footpads injected with PE cells plus control antigen. Antigen-specific footpad swelling of 20 to 80% was found in contrast to meager swelling when lymph node or spleen cells were used. The magnitude was dependent on dose of both cells and antigen over a broad range (1 to 12 X 10(6) cells, 1 to 100 micrograms of antigen per recipient). Specific swelling was reduced to nil after treatment of PE cells with anti-theta serum, indicating T cell dependence. Carrier specificity of the response was similar to DTH in the guinea pig. Time course and histology are compatible with a DTH response as well. The superiority of PE cells in local transfer of DTH and the modest requirements of this murine local transfer system provide potent tools for identification of the cell(s) that mediate DTH and their interactions. This system, in addition, should provide a sensitive indicator of immunomodulating activity in cellfree supernatants.
Mouse mast cell activation and desensitization for immune aggregate-induced histamine release.
Authors: Fox P C , Basciano L K , Siraganian R P
Published: December 2022

Immune aggregate-induced histamine release and desensitization were studied in mouse mast cells. Maximal histamine release was rapid, occurred at 37 degrees C, and required the addition of alpha-L-phosphatidyl-L-serine and Ca2+. The amount of histamine released varied with the composition of the immune aggregates and was dependent on the antibody concentration. Saturation of mast cell Fc epsilon receptors with rat or mouse IgE had no effect on subsequent immune aggregate-induced release. The incubation of mouse mast cells with immune aggregates in the absence of cations of alpha-L-phosphatidyl-L-serine did not stimulate the release of histamine but resulted in desensitization of the cells for release with the addition of the same or unrelated immune aggregates. Such cells are capable, however, of IgE-mediated histamine release. Mast cells desensitized for IgE-mediated histamine release by incubation with anti-IgE were capable of immune aggregate-induced release. These data suggest that IgE-mediated and immune aggregate-induced triggering of mouse mast cells occurs through separate receptors.