bluuebunny/crossref_metadata_2025 huggingface dataset:
Suppression of lymphocyte proliferative responses: characterization of the suppressor and kinetics of suppression.
Authors: Stout R D , Fisher M
Published: December 2022

Culturing spleen cells for 2 or more days in the absence of mitogenic stimulation results in the generation of suppressor cells that can effectively inhibit the proliferative responses of freshly prepared spleen cells to mitogen or alloantigen stimulation. Suppression does not appear to be mediated by prostaglandins nor other soluble factors produced during the preculture period. The suppressor cell is described as a plastic-adherent Thy-1.2-, IgM-, FcR+ macrophage-like cell. Significant suppression of Con A responses can be detected at suppressor to target ratios as low as 1:100. The plastic-adherent suppressor is capable of terminating Con A-induced proliferation of spleen cells whether added at the onset of the Con A response or added as late as 48 hr after mitogenic stimulation. The suppressed spleen cell population displays an absence of large blast cells and a decrease in surface density of Thy-1.2 determinants.
Presence of circulating anti-idiotype-bearing cells after booster immunization with tetanus toxoid (TT) and inhibition of anti-TT antibody synthesis by auto-anti-idiotypic antibody.
Authors: Geha R S
Published: December 2022

We have previously documented the appearance of auto-anti-idiotypic (anti-Id) antibody in the serum of normal subjects after booster immunization with tetanus toxoid (TT) antigen. In the present study circulating auto-anti-Id-bearing cells and the regulatory role of auto-anti-Id antibody on anti-TT antibody synthesis were examined. Two adult volunteers were boosted with TT antigen, and IgG (Fab')2 anti-TT was prepared from plasma obtained 7 and 10 days after boosting. Auto-anti-Id antibody directed against autologous IgG (Fab')2 anti-TT became detectable in the serum starting 2 wk and up to 4 mo after booster immunization with TT. Circulating cells capable of specifically binding fluorescein-conjugated autologous IgG (Fab')2 anti-TT were identified starting 3 wk after immunization and throughout the 3- to 4-mo study period. Auto-anti-Id-bearing cells were predominantly B cells and secreted auto-anti-Id antibody after stimulation with pokeweed mitogen (PWM) in vitro. When added to cultures of peripheral blood lymphocytes (PBL) obtained 10 days after immunization, serum auto-anti-Id antibody inhibited the PWM-induced synthesis of IgG anti-TT. This inhibition was idiotype specific and affected predominantly the synthesis of anti-TT idiotypes recognized by the auto-anti-Id antibody. Auto-anti-Id antibody did not affect the synthesis of IgG anti-TT by PBL that co-circulated with the auto-anti-Id antibody. These results suggest that auto-anti-Id antibodies play a role in the regulation of the normal human immune response.
Cellular basis of defective cell-mediated lympholysis in atopic dermatitis.
Authors: Leung D Y , Wood N , Dubey D , Rhodes A R , Geha R S
Published: December 2022

Peripheral blood lymphocytes (PBL) from patients with atopic dermatitis (A.D.) were examined for their capacity to generate cytotoxic T lymphocytes (CTL) during mixed lymphocyte cultures (MLC). Cell-mediated lympholysis (CML) activity, but not proliferative response during MLC, was significantly decreased (p less than 0.01) in patients with A.D. (n = 19) compared with normal controls (n = 19) or patients with other skin diseases (n = 14). Regression analysis revealed a significant correlation (p less than 0.001) between CML activity and the percentage of circulating T8+ suppressor/cytotoxic T cells. Deficient CML activity in A.D. patients, however, was not corrected when isolated autologous T4+ and T8+ cells were recombined at a normal ratio (2:1) of T4+ to T8+ cells. Examination of CML activity in co-cultures of isolated T4+ and T8+ cells obtained from two A.D. patients and their respective HLA-identical healthy siblings indicated that the defect in CML resided in both the helper/inducer T4+ cells and the effector T8+ cells. The defective cytotoxic T cell function reported in the present study may account for the increased susceptibility of patients with A.D. to develop severe viral infections.
C1Q receptors on cultured human gingival fibroblasts: analysis of binding properties.
Authors: Bordin S , Kolb W P , Page R C
Published: December 2022

The interaction of the purified C1q component of human C with synchronized, quiescent human gingival fibroblasts was investigated, and the presence of a specific binding site was demonstrated. Quantitative binding studies with radioiodinated C1q showed that binding was specific, saturable, and reversible upon addition of unlabeled C1q or by increasing the salt concentration. Scatchard plot analysis of the data yielded an affinity constant of 2 X 10(7) M-1 for all cell strains examined. The capacity for C1q binding varied among the eight cell strains examined. The number of binding sites per cell ranged from 2.6 to 17.7 X 10(6) with an average of 8.4 X 10(6). The receptor was insensitive to trypsin digestion, and it bound the collagen-like portion of the C1q molecule. Specific immunofluorescence staining showed that virtually all the viable cultured fibroblasts were able to bind added C1q. Flow cytometric analysis demonstrated a spectrum of fluorescence intensity among the cell strains, and there was a positive correlation between fluorescence intensity and the number of binding sites detected by using radiolabeled C1q.
Immunologic and ionophore-induced generation of leukotriene B4 from mouse bone marrow-derived mast cells.
Authors: Mencia-Huerta J M , Razin E , Ringel E W , Corey E J , Hoover D , Austen K F , Lewis R A
Published: December 2022

Mouse bone marrow-derived mast cells differentiated in vitro and sensitized with monoclonal IgE respond to antigen-initiated activation-secretion with the generation of leukotriene B4 (LTB4), a highly potent chemotactic factor. The antigen-initiated net percent release of the secretory granule marker beta-hexosaminidase and the generation of immunoreactive LTB4 and of immunoreactive leukotriene C4 (LTC4) were not diminished by washing the cells before challenge, indicating that interaction of the antigen occurred with the IgE fixed on cell membranes and was not due to phagocytosis of immune complexes formed in the fluid phase. The parallel dose-response relationship for the antigen-induced release of the performed mediator and the generation of both leukotrienes along with the superimposable time courses of their extracellular appearance indicate the origin of these mediators from a common cell type with IgE receptors. Resolution by reverse phase-high performance liquid chromatography (RP-HPLC) of the leukotrienes released from unsensitized cells by ionophore A23187 and from sensitized cells by the specific antigen revealed that the generation of LTB4 was accompanied by the production of the two diastereoisomers of 6-trans-LTB4, which were not immunoreactive. The immunoreactive LTB4 eluted from RP-HPLC at the same retention time as synthetic LTB4 was present in similar nanogram quantities when measured by either radioimmunoassay or integrated absorbance at 269 nm and exhibited a chemotactic activity for human neutrophils on a weight basis comparable to that of synthetic LTB4. The overall recoveries after RP-HPLC of immunoreactive LTB4 released from ionophore A23187-activated, bone marrow-derived mast cells and of added tritiated synthetic LTB4 in separate experiments with ionophore-activated cells were comparable and greater than 78%. The maximum generation of LTB4 by ionophore A23187-activated cells of 7.9 +/- 2.5 ng/10(6) cells (mean +/- SD), occurring at 40 min, was greater than the maximum generation of 4.5 +/- 0.8 ng/10(6) cells, with immunologic stimulation occurring 5 min after antigen challenge of sensitized cells. The initial rate of LTB4 generation after perturbation of the IgE-Fc receptors, however, exceeded that following ionophore A23187 stimulation and may represent one important variable in establishing a significant chemotactic gradient.
Generation of C3a anaphylatoxin from human C3 by human mast cell tryptase.
Authors: Schwartz L B , Kawahara M S , Hugli T E , Vik D , Fearon D T , Austen K F
Published: December 2022

Tryptase, the dominant neutral protease of human pulmonary mast cell secretory granules, has the capacity in vitro to generate C3a anaphylatoxin from purified human C3. Only the alpha-chain of C3 is cleaved, and major fragments with apparent m.w. of 105,000, 39,500, 34,000, 29,000, and 9000 are detected by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis under reducing conditions. Fragments of 34,000 and 9000 m.w. are detected without reduction. A portion of the 9000 m.w. protein corresponds to C3a by virtue of its co-migration in SDS polyacrylamide gels with purified C3a and with trypsin-generated C3a, by its detection in a radioimmunoassay for C3a, and by its contractile activity on the guinea pig ileum bioassay. In the presence of heparin, another component of the mast cell secretory granule, the rate of appearance and the distribution of C3 cleavage fragments as assessed in SDS polyacrylamide gels are not appreciably changed with the exception that no C3a material can be detected in the SDS polyacrylamide gels or by radioimmunoassay and bioassay of the unresolved reaction mixture. Enhanced catabolism of authentic C3a by tryptase occurs in the presence of heparin and by analogy when C3a is generated from C3 by tryptase in the presence of heparin. Whereas tryptase secreted by activated human mast cells may generate C3a, a potentially important additional mediator of immediate hypersensitivity events, the concomitant release of heparin may serve to down-regulate C3a irrespective of its mechanism of generation.
Histamine modulation of human neutrophil oxidative metabolism, locomotion, degranulation, and membrane potential changes.
Authors: Seligmann B E , Fletcher M P , Gallin J I
Published: December 2022

Previous reports have suggested that histamine modulates neutrophil chemotaxis, but this has not been observed by all laboratories. We have re-addressed this controversial point and demonstrate that histamine and H1- and H2-receptor-specific agonists cause limited inhibition of chemotaxis while stimulating chemokinesis. Furthermore, using the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (f-met-leu-phe) as a stimulus, we demonstrate that histamine and H1/H2 agonists inhibit f-met-leu-phe-stimulated changes in membrane potential (monitored with the cyanine dye dipentyloxacarbocyanine), superoxide anion production (cytochrome c reduction), hydrogen peroxide formation (scopoletin fluorescence), and degranulation of granule contents (lysozyme and beta-glucuronidase) in a dose-dependent manner but have no effect on neutrophil functions stimulated by the secretagogues phorbol myristate acetate or A23187. All inhibitory effects of histamine and the H1/H2 agonists are reversed in a competitive manner by the H2 antagonist cimetidine. In addition, structure-activity studies using H1 and H2 receptor agonists and antagonists indicate that a single site with specificity for both H1 and H2 analogue structures modulates the various f-met-leu-phe-stimulated functions studied. Kinetic studies demonstrate that the inhibitory effects of histamine on neutrophil function are only observed when histamine is added before f-met-leu-phe and that inhibition occurs within 10 to 20 sec of histamine addition, does not persist after its removal, and is reversed by addition of cimetidine 10 to 20 sec before stimulation with f-met-leu-phe. Although the inhibitory effects of histamine are exerted early in the sequence of PMN activation by f-met-leu-phe, histamine does not affect the binding or internalization of f-met-leu-[3H]phe. The ability of histamine to modify the variety of neutrophil responses demonstrated in this report suggests an important and direct role for histamine in the regulation of inflammatory reactions in acute allergic settings or other disease states in which histamine release may occur.
Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation.
Authors: Bass D A , Parce J W , Dechatelet L R , Szejda P , Seeds M C , Thomas M
Published: December 2022

We have developed a quantitative assay to monitor the oxidative burst (H2O2 production) of polymorphonuclear leukocytes (PMNL) using single cell analysis by flow cytometry, and have examined whether PMNL respond to membrane stimulation with an all-or-none oxidative burst. During incubation with normal neutrophils, dichlorofluorescin diacetate diffused into the cells, was hydrolyzed to 2',7'-dichlorofluorescin (DCFH) and was thereby trapped within the cells. The intracellular DCFH, a nonfluorescent fluorescein analogue, was oxidized to highly fluorescent 2',7'-dichlorofluorescein (DCF) by PMNL stimulated by phorbol myristate acetate (PMA). That the oxidative product was DCF was shown by excitation/emission spectra and by mass spectrometry of the product from PMA-stimulated PMNL. Normal resting and PMA-stimulated PMNL oxidized 6.9 +/- 0.7 and 160 +/- 13 attomoles DCF per cell, respectively, in 15 min. Absence of calcium and magnesium ions and/or addition of 2 mM EDTA did not inhibit DCF formation by PMNL stimulated by 100 ng/ml PMA. Since EDTA prevented aggregation of PMNL (even when stimulated by 100 ng/ml PMA), which would prevent accurate flow cytometric analysis, further experiments were performed with EDTA in the medium. A close correlation between average DCFH oxidation and hexose monophosphate shunt stimulation was demonstrated using cells from patients whose PMNL had oxidative metabolic defects of varying severity. Intracellular DCFH was also oxidized by reagent H2O2 or oxygen derivatives generated by glucose oxidase + glucose or by xanthine oxidase + acetaldehyde; DCFH oxidation by these systems was inhibited by catalase but unchanged by superoxide dismutase. The data indicate that the DCFH oxidation assay is quantitatively related to the oxidative metabolic burst of PMNL, and they strongly suggest that the reaction is mediated by H2O2 generated by the PMNL. Incubation of PMNL with varying concentrations of PMA caused graded responses by all PMNL present; i.e., 1 ng/ml PMA caused a mean response of 34% maximal with a single population of responding PMNL (rather than 66% resting and 34% fully stimulated as predicted by the all-or-none hypothesis). Thus, with these assay conditions, oxidative product formation by PMNL occurs as a graded response to membrane stimulation by PMA.
Characteristics of a shared idiotype by two IgM anti-herpes simplex virus monoclonal antibodies that recognize different determinants.
Authors: Kennedy R C , Adler-Storthz K , Henkel R D , Dreesman G R
Published: December 2022

Two IgM monoclonal antibodies (McAb) that recognized distinct antigenic determinants of herpes simplex virus type 2 (HSV2) were shown to share idiotypes. Seven other IgM McAb generated to either HSV2 or the major envelope glycoprotein of HSV2, VP119, did not express this shared idiotype. The idiotypic determinants were associated with the antibody-combining site, because HSV2 inhibited the idiotype-anti-idiotype reactions. Anti-HSV2 sera raised in BALB/c mice also expressed the idiotype shared by the two anti-HSV2 McAb.
Non-antibody-mediated control of parasitemia in acute experimental Chagas' disease.
Authors: Trischmann T M
Published: December 2022

There appear to be two phases in the control of parasitemia in acute Chagas' disease in the mouse. The first phase occurs during the first few weeks after infection and control is achieved through a thymus-dependent, antibody-independent mechanism. Challenge of B cell-suppressed C3H and F1 (C57BL/6 X C3H) mice with the Brazil strain of Trypanosoma cruzi led to a course of parasitemia for the first 3 wk after infection similar to that seen in normal C3H or F1 mice and markedly lower than the parasite levels observed in the blood of nu/nu mice. Challenge of BXH-2 recombinant inbred mice resulted in a course of parasitemia similar to that seen in nu/nu mice up to day 16 despite the production of normal levels of antibody. The BXH-2 mice lack the ability to effect the early control of parasitemia. The second phase begins several weeks after infection with the rise in antibody titer, and the control is exerted through an antibody-mediated mechanism. In all B cell-suppressed mice, an inexorable rise in parasitemia occurred up to the time of death, which suggests that antibody is important for the eventual clearance of parasites from the blood. A comparison of the IgM and IgG antibody titers to T. cruzi in a series of resistant and susceptible strains showed that there was no correlation between the appearance of specific antibody or antibody titers and the levels of parasitemia observed. The level of parasitemia attained in the late acute phase may be primarily determined by the extent of parasite proliferation in the early acute phase.
The T cell response to allelic determinants on Igh-1-encoded IgG2a antibodies: dual recognition examined by using antigenic antibodies with binding affinity for Ia molecules.
Authors: Hedrick S M , Schwartz R H
Published: December 2022

In this report we examine the effects of binding antigen directly to Ia molecules during immunization and stimulation of T cells in culture. To accomplish this we characterize the T cell response to Igh-1j-encoded IgG2a antibodies in C57BL/10 H-2 congenic mice. In contrast to the response to the closely related Igh-1a-encoded IgG2a antibodies, high responder mice were (C57BL/10 X B10.A)F1 hybrids, and the important alleles appeared to map to I-Ab and I-Ak. C57BL/10 and B10.A(5R) strains were low responders (I-Ab), and B10.A and B10.A(4R) strains were nonresponders (I-Ak). The interest in using monoclonal IgG2a antibodies as T cell antigens was that different V-region-encoded binding specificities could be associated with the same IgG2a antigenic determinant. In our interpretation of the Langman and Cohn dual binding site model of the T cell receptor, the response to anti-Ia antibody should have some of the same characteristics as that to an allogeneic Ia molecule; in particular, it should be non-H-2-restricted. Although the response to anti-Ia antibody had several interesting properties, no difference in the restriction specificity was found for the T cell response to anti-Ia antibodies relative to unreactive IgG2a antibodies. Our conclusion is that T cell receptors must bind both Ia and antigen molecules for activation to occur. In a separate set of experiments, the T cell response resulting from immunization with anti-Ia antibody was examined. Because the antigen was shown to be associated with the Ia molecule during immunization, experiments were designed to detect an alteration or lack of H-2 restriction in the resulting T cell population. A long-term T cell line induced after such immunization showed no qualitative differences from a T cell line produced with unreactive IgG2a antibody. These results demonstrate that simply attaching a foreign antigen to a restriction element is not sufficient to lead to unrestricted T cell activation. This in turn suggests the T cell receptor must recognize the restriction element for cell-cell interactions to occur.
Suppression of in vitro maintenance and interferon-mediated augmentation of natural killer cell activity by adherent peritoneal cells from normal mice.
Authors: Brunda M J , Taramelli D , Holden H T , Varesio L
Published: December 2022

The ability of adherent peritoneal cells (APC) to inhibit murine natural killer (NK) cell activity was examined. Nylon wool-nonadherent splenic effector cells were incubated overnight with or without different numbers of APC. NK activity was then measured against YAC-1 in a 4-hr 51Cr-release cytotoxicity assay. Proteose peptone-elicited or unstimulated resident APC from normal mice markedly suppressed NK activity of splenic effector cells in the presence or absence of exogenously added interferon. The suppression was dependent on the number of APC added with 10% APC, relative to the number of effector cells, resulting in a greater than 65% inhibition of cytotoxicity. The effector phase of cytotoxicity was not the target of the suppressor cells, because APC did not suppress NK activity when they were present only during the cytotoxicity assay. The addition of APC to alloimmune cytotoxic T cells under similar conditions resulted in no inhibition of cytotoxicity. Both syngeneic and allogeneic APC suppressed NK activity, but several murine macrophage-like cell lines lacked this property. In contrast to APC, incubation of effector cells with adherent spleen cells from normal mice resulted in no inhibition of NK activity. APC from mice injected with C. parvum were less inhibitory for NK activity than normal resident APC. In contrast, C. parvum APC suppressed concanavalin A-induced lymphoproliferation and were directly cytotoxic to tumor target cells in vitro, whereas normal APC lacked these properties. The results indicate that the peritoneum of untreated mice contains suppressor cells that can inhibit the in vitro maintenance and IFN-mediated augmentation of NK activity. In addition, these results indicate a broader spectrum of immune reactivities regulated by APC and suggest that, depending on their level of activation, APC can preferentially inhibit different immune functions.
Recombinant mouse gamma interferon induces the priming step in macrophage activation for tumor cell killing.
Authors: Pace J L , Russell S W , Torres B A , Johnson H M , Gray P W
Published: December 2022

Mouse macrophages become activated to kill tumor cells by traversing a series of steps (1-3). The first of these does not cause the expression of cytolytic activity; instead, it primes macrophages to respond to a second signal(s) that then triggers the onset of killing (4-7). The mediator that is responsible for priming is contained, along with other lymphokines, in the culture supernatants of concanavalin A-stimulated spleen cells (5-7) cloned T lymphocytes (8), or some T cell hybridomas (9). Close association has been noted between macrophage priming activity and antiviral activity that is attributable to gamma interferon (10-12). However, unequivocal evidence that the two activities are products of the same molecule has not been available. We now how conclusively by using mouse gamma interferon (MulFN-gamma)3 produced by recombinant DNA technology that, in addition to antiviral activity, this lymphokine has the capacity to induce the priming step in the process of macrophage activation for tumor cell killing.
Functional heterogeneity of the Lyb-5- B cell subpopulation: mutant xid B cells and normal Lyb-5- B cells differ in their responsiveness to phenol-extracted lipopolysaccharide.
Authors: Ono S , Yaffe L J , Ryan J L , Singer A
Published: December 2022

In the present study, responses stimulated by phenol-extracted lipopolysaccharide (LPS(phenol)) and butanol-extracted LPS (LPS(butanol)) were used to assess the possibility that xid B cells might not be identical to the Lyb-5- B cells present in normal mice. It was found that xid B cells responded well only to LPS(butanol) whereas normal B cells responded well to both LPS(butanol) and LPS(phenol). Thus, LPS(butanol) appeared to be a TI-1 antigen and LPS(phenol) appeared to be a TI-2 antigen. In contrast to classical TI-2 responses, however, responses stimulated by LPS(phenol) did not exhibit a stringent requirement for accessory cells. Furthermore, if LPS(phenol) were a classical TI-2 antigen, it should only activate Lyb-5+ B cells. To determine if the responsiveness of normal B cells to LPS(phenol) were due, at least in part, to the stimulation of normal Lyb-5- B cells, the responsiveness of normal neonatal B cells and normal adult B cells that had been pretreated with anti-Lyb-5.1 + C was assessed. It was found that both normal neonatal B cells and normal adult Lyb-5- B cells did respond well to LPS(phenol). Thus, even though LPS(phenol) does not stimulate xid B cells, these data demonstrate that LPS(phenol) is different from other TI-2 antigens. More importantly, these data also demonstrate that xid B cells and normal Lyb-5- B cells are not identical. It is hypothesized that the normal Lyb-5- B cell subpopulation is heterogeneous, consisting of an Lyb-5(1)- and an Lyb-5(2)-B cell subset with the xid mutation blocking the differentiation of Lyb-5(1)-B cells into Lyb-5(2)-B cells.
Induction of suppressor T cells by monoclonal anti-idiotope antibody in strains of mice not expressing the idiotope in hyperimmune serum.
Authors: Thomas W R , Morahan G , Miller J F
Published: December 2022

A monoclonal antibody, reactive with an idiotope present on some but not all A/J anti-azobenzenearsonate antibodies bearing the cross-reactive idiotype, was used to induce suppressor T cells. These cells suppressed the effector phase of the delayed hypersensitivity response to ABA but not to oxazolone. They could be enriched on dishes coated with ABA but not with trinitrophenylated human gamma-globulin, and the enrichment could be blocked by the anti-idiotope. Suppressor activity was also enhanced by first incubating T cells with the anti-idiotope and then collecting immunoglobulin-coated cells on dishes coated with anti-mouse immunoglobulin. The suppressor cells could be induced in A/J and CBA mice, even though hyperimmune CBA mice did not produce antibodies bearing this idiotope.
Activation of cytotoxic T lymphocytes requires at least two spleen cell-derived helper factors besides interleukin 2.
Authors: Falk W , Männel D N , Dröge W
Published: December 2022

The dependency of induction of T cell cytotoxicity on lymphokines was studied. 1 X 10(5) nylon wool-purified thymic lymphocytes or 10(4) spleen cells were cultured with TNP-haptenated syngeneic UV-irradiated spleen cells in the presence of a variety of lymphokine preparations. Concanavalin A-induced spleen cell supernatants mediated strong cytotoxic responses in this system. Three other preparations, namely, a partially purified IL 2 preparation from PMA-stimulated EL-4 thymoma cells, a Con A-induced spleen cell supernatant that was absorbed with an IL 2-dependent cell line, and a Con A-induced supernatant that was dialyzed at pH 2 were all ineffective in mediating a cytotoxic response. In reconstitution experiments, cytotoxic responses were only obtained when either the absorbed preparation or the pH 2-treated preparation was mixed with the IL 2 preparation from EL-4 cells. No reconstitution occurred after mixing of the absorbed with the pH 2-treated preparation. pH 2 treatment of the absorbed preparation did not abolish its synergistic effect when added to the IL 2 preparation from EL-4 cells. These results led to the conclusion that activation of cytotoxic lymphocyte precursors requires at least two other lymphokines in addition to IL 2. One T cell cytotoxicity-inducing factor (TCF1) remained in Con A-induced supernatants after absorption with IL 2 receptor-bearing T cell line cells. It was pH 2-resistant and was not found in EL-4 supernatants. A second T cell cytotoxicity-inducing factor (TCF2) was pH 2-sensitive and was found in Con A-induced spleen cell supernatants as well as in interferon-free supernatants of PMA-stimulated EL-4 cells. This activity co-purified with IL 2. It was absorbed by the IL 2-dependent T cell line together with IL 2. IL 2 differs from TCF2 since it is pH 2-resistant.
Phorbol myristate acetate and in vitro T lymphocyte function. II. Influence of PMA and supernatants from PMA-treated P388D1 cells on the proliferation of cloned T cells.
Authors: Orosz C G , Roopenian D C , Bach F H
Published: December 2022

Neither the culture supernatants from P388D1 cells pulsed with phorbol myristate acetate (PMA), nor PMA itself in concentrations ranging from 10(-6) M to 10(-9) M, are directly mitogenic for murine T lymphocyte clones, yet both markedly augment the antigen-driven proliferation of many, but not all, cloned T lymphocytes. The component of PMA-induced P388D1 supernatant responsible for its co-mitogenic activity is probably PMA, rather than interleukin 1. For responsive clones, the co-mitogenic effect of PMA requires stimulator cells that display the specific allogeneic determinants recognized by the clones. This response persists after T cells are removed from the stimulating population, ruling out induction of mitogenic lymphokines from stimulator T cells by PMA as a primary mechanism for augmentation of clonal proliferation. Both splenocytes and thymocytes cooperate with PMA for enhanced clonal expansion, but heat treatment (45 degrees C, 45 min) of thymocytes destroys their cooperative capacity. PMA can also potentiate the lymphokine-driven proliferation of cloned T cells, indicating that PMA can, under certain conditions, influence T cell clonal expansion by a direct action on T lymphocytes.
Characterization of a suppressor cell-activating factor (SCAF) released by adherent cells treated with M. tuberculosis.
Authors: Wadee A A , Mendelsohn D , Rabson A R
Published: December 2022

Peripheral blood adherent cells ingesting killed Mycobacterium tuberculosis release a suppressor cell-activating factor (SCAF) into their culture supernatants. When adherent cells ingested 125I-labeled M. tuberculosis, radioactivity could be detected in the supernatant within 2 hr. When this supernatant was fractionated on a Sepharose 2B column, the fraction with suppressor cell-activating activity was also found to contain the majority of the radiolabel, which suggests that the macrophage processed bacteria (or bacterial product) constituted the major portion of the SCAF. This fraction also contained a high proportion of lipid, and the fraction with suppressor activity resided purely within the phospholipid fraction. By employing thin layer chromatography, the phospholipids responsible were identified as phosphatidylethanolamine and phosphatidylinositol. These results indicate that when macrophages ingest mycobacteria, they release phosphatidylethanolamine and phosphatidylinositol of bacterial origin into their culture supernatants, which are responsible for activating suppressor T cells.
Binding of phosphatidylethanolamine and phosphatidylinositol to OKT8+ lymphocytes activates suppressor cell activity.
Authors: Wadee A A , Rabson A R
Published: December 2022

The phospholipids phosphatidylethanolamine (PE) and phosphatidylinositol (PI) have been shown to activate a population of OKT8-enriched lymphocytes to become activated suppressor cells that result in the suppression of lymphocyte blastogenesis to a variety of mitogens and antigens. This suppression is dose dependent, and maximal suppressor activity is obtained at concentrations of 125 micrograms/ml PE and 25 micrograms/ml PI. Activation of the suppressor cell population is not associated with an actual increase in the number of cells expressing the OKT8 antigen, but the number of these cells expressing Dr antigens on their surface was increased. Both PE and PI bound to lymphocytes in a specific manner. Binding of radiolabeled PE could be inhibited by unlabeled PE but not by PI or phosphatidylserine (PS). Similarly, the binding of PI to lymphocytes was also found to be specific. Although radiolabeled PE bound to lymphocytes other than OKT8+ cells, and to other peripheral leukocytes, it bound to OKT8+ cells with a significantly greater affinity than it did to the other cell types. The Kd for PE was 1 x 10(2) nM and for PI was 1 x 10(3) nM, and receptor cell densities for these two phospholipids were estimated at 1 x 10(-8) nM and 3 x 10(-9) nM, respectively. The receptors for these two phospholipids were trypsin and heat sensitive, and the receptor sites could be regenerated after a 24-hr incubation after trypsinization.
Studies of antigen binding on human basophils. II. Continued expression of antigen-specific IgE during antigen-induced desensitization.
Authors: MacGlashan D , Mogowski M , Lichtenstein L M
Published: December 2022

We previously postulated that desensitization of human basophils was due to endocytosis or shedding of IgE and/or its Fc receptor. We report here, however, that during a 3-hr desensitization of purified basophils, there was no loss of cell surface antigen-specific IgE. The specific desensitization induced by antigen persists for at least 24 hr whereas simultaneously occurring nonspecific desensitization can recover during the same time interval. Therefore, we suggested that specific desensitization may result from a permanent modification of the Fc receptor or some closely associated protein. On the basis of elution studies of 125I-labeled BPO-HSA, we also suggest the antigen antibody reaction matures to aggregates of increasing size, and we propose that these large aggregates, unable to initiate a release signal, cause an additional component of desensitization.
Inactivation of human high molecular weight kininogen by human mast cell tryptase.
Authors: Maier M , Spragg J , Schwartz L B
Published: December 2022

Tryptase, the major neutral protease of human pulmonary mast cell secretory granules, rapidly inactivates human high m.w. kininogen (HMWK) in vitro. HMWK (5600 nM) lost 50% of its capacity to release kinin in response to kallikrein after a 5-min incubation with tryptase (31 nM), even though kinin activity was neither generated nor, when bradykinin was incubated with tryptase, destroyed by tryptase. The procoagulant activity of HMWK (51 nM) and the purified procoagulant chain (40 nM) that is derived from HMWK were each 72% inactivated after 7 min of incubation with tryptase (0.04 nM and 0.02 nM, respectively). Human urinary and pancreatic kallikrein did not inactivate this procoagulant activity under conditions in which kinin generation occurs. Complete cleavage of native single-chain HMWK by tryptase occurred in less than 10 min as analyzed by electrophoresis in sodium dodecyl sulfate polyacrylamide slab gels. The major products formed during the initial 2 min were proteins of 100,000 and 95,000 apparent m.w., and by 10 to 30 min were fragments of 74,000 and 67,000 apparent m.w. Reduction of these cleavage products yielded two major fragments of 67,000 and 66,000 apparent m.w. that were both present by 0.17 min. The presence of lower m.w. products, thought to be primarily from the carboxy-terminal procoagulant region of HMWK, were also detected with and without reduction. The capacity of tryptase to inactivate HMWK is consistent with the ability of other mast cell-derived mediators, such as heparin proteoglycan and prostaglandin D2, to suppress blood coagulation and thrombosis, and may play an important role in the biology of mast cell-dependent events in vivo.
Induction of immune responses by schistosome granuloma macrophages.
Authors: Sunday M E , Stadecker M J , Wright J A , Aoki I , Dorf M E
Published: December 2022

In mice, granuloma formation after Schistosomiasis mansoni infection is known to be a T cell-dependent response to schistosome eggs that peaks at 6 to 8 wk after infection (early) then regresses to a minimum by 20 to 32 wk (late). This decline in host responsiveness, termed modulation, has been attributed to T cell-mediated suppression. We now have investigated the macrophages from either Early or Late schistosome granulomas phenotypically and found no difference in expression of I-A subregion encoded antigens. We also tested granuloma macrophage (Gm) populations in their capacity to induce positive and negative antigen-specific immune responses to the hapten 4-hydroxy-3-nitrophenyl acetyl (NP). As few as 10(3) to 10(4) NP-coupled Early or Late Gm given s.c. were equally capable of inducing MHC-restricted NP-specific delayed-type hypersensitivity (DTH) responses and this DTH-inducing capability was equivalent to that of splenic adherent cell (SAC) controls. Further, both Early and Late Gm were able to generate NP-specific induction-phase suppressor cells (Ts1) when as few as 10(2) NP-coupled Gm were given i.v., the same as NP-SAC controls. Finally, NP-specific effector-phase suppressor cells (Ts3) were equally induced by Early Gm, Late Gm, or SAC controls. Therefore, macrophages derived from Early or Late schistosome granulomas or normal spleens are apparently phenotypically indistinguishable and equally capable, in extremely small quantities, of inducing NP-specific DTH, Ts1, and Ts3 immune responses.
Phorbol ester mediates reversible reduction of cloned T lymphocyte cytolysis.
Authors: Orosz C G , Roopenian D C , Bach F H
Published: December 2022

Prolonged contact with nanomole to micromole concentrations of the tumor promoter phorbol myristate acetate causes a significant reduction in the lytic activity of cloned cytolytic T cells. Diminished lysis is apparent even in the presence of agglutinating lectins. PMA does not exert this effect by minimizing clone viability. In fact, these concentrations of PMA cause significant potentiation of antigen-driven proliferation for many of these same clones. The PMA-mediated loss of cytolysis is reversible. Although contact with antigen does not induce or enhance reexpression of cytolysis, PMA-treated cytolytic T cell clones display normal cytolytic activity after contact with lymphokines.
Interleukin 2-dependent natural killer (NK) cell lines from patients with primary T cell immunodeficiencies.
Authors: Flomenberg N , Welte K , Mertelsmann R , O'Reilly R , Dupont B
Published: December 2022

Bulk cultured cell lines with natural killer (NK) activity were derived by in vitro culture with interleukin 2-containing conditioned medium (IL 2-CM) of peripheral blood mononuclear leukocytes (PBL) from patients with primary T cell deficiencies. Lines were developed from three patients with severe combined immunodeficiency (SCID) and one patient with Nezelof's syndrome and contained several populations of cells with distinct phenotypes. All lines contained a cell population expressing the Leu-5 (50K) (sheep red blood cell receptor), 3A1 (40K), and OKT10 antigens, but lacking the pan T cell antigens Leu-1 (67K) and Leu-4 (19K) as well as the markers of T cell subsets Leu-2a (32K) and Leu-3a (56K). These cells failed to express the Leu-7 antigen and only weakly expressed OKM1. In addition, one line contained a population of Leu-5+, 3A1+, OKT10+, Leu-2a+, Leu 1-, and Leu 4- cells. Three of the lines also contained populations with classic T cell (Leu-1 and-Leu 4+) phenotypes. The lines were enriched in NK activity compared with the PBL from which they were derived. Their growth was strictly dependent on IL 2-CM. Highly purified IL 2, lacking any other detectable protein contaminants or lymphokine activities, was capable of supporting the growth of the Leu-5+, 3A1+ "null" cell populations from these lines without alteration in their functional activity or phenotype. Thus, studies of in vitro expanded cell lines from patients with severe disorders of T cell function and thymic involution indicate that this "null" cell population does not require thymic maturation to develop its effector function. This "null" cell population can be maintained in vitro in the presence of IL 2. This finding is analogous to the data obtained from study of NK cells in athymic (nude) mice.
Defective production of interleukin 1 and interleukin 2 in patients with systemic lupus erythematosus (SLE).
Authors: Linker-Israeli M , Bakke A C , Kitridou R C , Gendler S , Gillis S , Horwitz D A
Published: December 2022

The objective of this study was to define some causes of the immunologic impairment characteristic of systemic lupus erythematosus. Blood mononuclear cells from 19 patients were stimulated to produce interleukin 1 and interleukin 2 and compared with controls. A severe defect in both IL 1 and IL 2 activity was observed. The low cytokine levels did not correlate with clinical or serologic activity of disease. These defects could not be explained by concurrent corticosteroid therapy. There was no correlation between a lower number of OKT4+ cells observed in these patients and the levels of IL 2 production, nor did removal of monocytes bring IL 2 to normal. Impaired IL 2 production could not be restored to normal by IL 1. The observed deficiency in these regulatory cytokines may therefore be a primary defect that is important in the pathogenesis of this disorder.
Immunopathology of murine experimental allergic orchitis.
Authors: Kohno S , Munoz J A , Williams T M , Teuscher C , Bernard C C , Tung K S
Published: December 2022

Experimental allergic orchitis (EAO) was induced consistently in BALB/c mice by immunization with homologous testicular tissue homogenate emulsified in complete Freund's adjuvant (CFA) providing that the animals had received simultaneously at least 1 microgram of an extract of Bordetella pertussis rich in pertussigen. All animals thus treated developed orchitis and serum antibody to testicular antigens within 20 days after immunization. The lesions were located in testis (100%), rete testis (37%), cauda epididymis (21%), and vas deferens (37%). Ductus efferentes and caput epididymis were only rarely affected. Early lesions in the seminiferous tubules were characterized by peritubular and/or intratubular accumulation of eosinophils, neutrophils, lymphocytes, and macrophages. This was followed by aspermatogenesis. Late lesions included massive necrosis and extensive fibrosis of the seminiferous tubules. Disruption of blood-testis barrier on day 20 was evidenced by the detection of 1) perfused lanthanum deposits between Sertoli cells and surrounding inflammatory cells inside the seminiferous tubules, 2) deposits of endogenous mouse IgG in germinal epithelium, and 3) probable immune complexes (granular C3) surrounding seminiferous tubules. Murine EAO differed from that of the guinea pig in the lack of involvement of the ductus efferentes, the extensive necrosis, the abundant polymorphonuclear eosinophils in the lesion, and the exquisite requirement of concomitant injection of B. pertussis extract.
Attempts at standardization of lupus-like graft-vs-host disease: inadvertent repopulation by DBA/2 spleen cells of H-2-different nonirradiated F1 mice.
Authors: Van Rappard-Van Der Veen F M , Radaszkiewicz T , Terraneo L , Gleichmann E
Published: December 2022

By induction of a suitable graft-vs-host reaction (GVHR) in nonirradiated, H-2-incompatible F1 mice, one can induce a syndrome strongly resembling systemic lupus erythematosus (SLE). The aim of the present study was to standardize the kind and number of DBA/2 donor cells required for optimal induction of this SLE-like GVH disease (GVHD). Groups of adult (C57BL/10 x DBA/2)F1 (BDF1) mice were injected i.v. with increasing numbers of DBA/2 spleen and lymph-node cells. We found that doses of 100 x 10(6) to 180 x 10(6) spleen and lymph-node cells provided a suitable donor-cell inoculum, whereas doses of donor cells below 100 x 10(6) were suboptimal and doses higher than 180 x 10(6) cells were supraoptimal for the induction of SLE-like GVHD. In a number of those F1 recipients that had received the donor-cell inocula composed of spleen cells, the duration of autoantibody formation was surprisingly brief. This appeared to be due to the fact that these GVH F1 mice were rapidly repopulated by lympho-hemopoietic donor cells. Even after the highest doses of DBA/2 spleen and lymph-node cells administered, this repopulation was not preceded by symptoms of acute GVH disease. Repopulation was avoided and a severe SLE-like disease induced when a mixture deficient in hemopoietic cells, i.e., 280 x 10(6) DBA/2 lymph-node and thymus cells, was used as donor-cell inoculum. Taken together, we reached three conclusions. First, the induction of full-blown SLE-like GVHD depends not only on the injection of a sufficient number of donor T cells but also on the continuous presence of F1 lymphoid cells, which seem to serve as stimulator cells. Second, in addition to the lymphoid hyperplasia and immune-complex glomerulonephritis (ICGN) described previously, the GVHR-induced lesions presenting themselves in the context of SLE-like autoimmunity include a Sjögren-like lymphoid infiltration of the salivary gland, a ubiquitous periarteritis, and a lymphoid infiltration of the bile ducts that is reminiscent of primary biliary cirrhosis. Third, the repopulation by donor cells of nonirradiated, H-2-incompatible hosts need not be accompanied by GVH mortality or symptoms of acute GVHD.
Dependency of human T lymphocyte colony formation on soluble factors produced by accessory or tumor cells.
Authors: Winkelstein A
Published: December 2022

When plated in agar, PHA-stimulated Ficoll-Hypaque separated human mononuclear cells readily form discrete T lymphocyte colonies. Separation of these mononuclear cells on Sephadex G-10 columns yields enriched populations of T lymphocytes (greater than 95% E rosette-positive); these purified cells do not respond to this nonspecific mitogen by undergoing clonal expansion. Colony growth, however, can be restored by the addition of irradiated adherent cells. T cell responses can also be reconstituted by incubating the Sephadex-nonadherent fraction with irradiated cultured human tumor cells of monocytic (U937 and HL60) and B lymphocytic (Raji and Daudi cells) lines, but not from a T cell clone (MOLT-4) or erythroleukemic (K562) cells. Growth can also be induced in cultures containing T cells by adding cells from patients with either acute myeloblastic or B cell chronic lymphocytic leukemia or by the addition of B cell-enriched, monocyte-depleted normal mononuclear cells. Soluble factors prepared from Sephadex-adherent cells or active tumor cell lines are effective in promoting T cell colony formation. These results indicate that accessory cells are required for PHA responses of cloned human T cells. This function can be supplied by both monocytic and B lymphocytic tumor cell lines and is due to the release of soluble factors that act, in conjunction with the mitogen, to activate responsive lymphocytes.
Suppressor cell induction factor: a new mediator released by stimulated human lymphocytes and distinct from previously described lymphokines.
Authors: Kasakura S , Taguchi M , Watanabe Y , Okubo T , Murachi T , Uchino H , Hanaoka M
Published: December 2022

Suppressor cell induction factor (SIF) was produced by alloantigen-stimulated human peripheral blood lymphocytes, and it activated human T cells to become effective suppressors of the mixed lymphocyte reaction (MLR). The activity of SIF was resistant to 56 degrees C and to pH 2, and was precipitated by 50 to 80% saturated ammonium sulfate. SIF had a m.w. range, as determined by gel filtration, of 18,000 to 29,000; it did not bind to DEAE cellulose columns; and it was recovered in the pH range from 6.9 to 7.3 on isoelectric focusing. SIF was biochemically separable from IL 2, BF, IFN-gamma, and CSF. Furthermore, IL 2 activity was completely removed by absorption of MLC supernatants by murine cytotoxic T lymphocyte line (CTLL) cells, whereas SIF activity was unabsorbable, thus distinguishing SIF from IL 2. In addition, antiviral activity of MLC supernatants was completely abolished by anti-human IFN-gamma serum, whereas SIF activity was unaffected by this antiserum, thus distinguishing SIF from IFN-gamma. Since treatment of these supernatants with antiserum against human lymphoblastoid cell IFN(alpha/beta) had no effect on either antiviral or SIF activities in these supernatants, SIF was also distinguishable from IFN alpha/beta. These results indicate that SIF is a distinct new lymphokine with the ability to induce suppressor function in human T cells.