Antagonistic effects of interferon-gamma and interleukin-4 on fibroblast cultures

The uptake of the Cd-metallothionein complex (CdMT) into LLC-PK1 cells was investigated and compared with that of CdCl2. The cells were incubated at 37 degrees C for up to 45 min with 1 microM Cd, as either CdMT or CdCl2 at pH 5.5, 6.4, or 7.4. Under all the experimental conditions described below, the accumulation of Cd from CdMT was markedly lower than that from CdCl2. Cd accumulation at pH 7.4 from CdMT increased linearly with the time of incubation, whereas Cd accumulation from CdCl2 was saturable. Metabolic inhibitors, 2,4-dinitrophenol (DNP) and carbonylcyanide p-(trifluoromethoxy)-phenylhydrazone (FCCP), and incubation at low temperature significantly decreased Cd accumulation from both Cd compounds. Coincubation with 30 microM ZnCl2, an antagonist of CdCl2 uptake, slightly decreased Cd accumulation from CdMT, but it markedly decreased that from CdCl2. Cd accumulation from CdMT at pH 5.5 was significantly higher than at pH 6.4 or 7.4, whereas Cd accumulation from CdCl2 at pH 5.5 and 6.4 was significantly lower than at pH 7.4. Although Cd accumulation from CdCl2 at pH 7.4 was significantly decreased by coincubation with 100 microM cysteine or glutathione (GSH), this decrease was not observed at pH 5.5 or 6.4. A small amount of Cd was removed, by the chelating agent EGTA, from the cell membranes after incubation with CdMT at pH 6.4 and 7.4, whereas a considerable amount of Cd was removed by EGTA after incubation with CdMT at pH 5.5 and with CdCl2 at three pHs. It appears that the CdMT complex is taken up into LLC-PK1 cells partially via an energy-dependent process, and the increase in Cd accumulation at low pH is due to the liberation of Cd. High stability and molecular size of the CdMT complex explains why it is not taken up readily into LLC-PK1 cells.     

Circulating levels of interleukin 1 beta and of interleukin 1 receptor antagonist in systemic juvenile chronic arthritis

OBJECTIVE: To measure circulating interleukin 1 beta (IL-1 beta) and IL-1 receptor antagonist (IL-1Ra) levels in patients with systemic juvenile chronic arthritis (JCA) and to evaluate their correlation with disease activity. METHODS: IL-1 beta and IL-1Ra levels were measured by ELISA in 45 patients with JCA (20 systemic, 10 polyarticular and 15 pauciarticular) and in 15 healthy controls. RESULTS: Plasma IL-1 beta levels were undetectable in the majority of patients with systemic JCA, and detectable levels were not associated with different treatments or with parameters of disease severity. Serum IL-1Ra levels were markedly increased in patients with systemic JCA and significantly correlated with the persistence of systemic features, the extent and severity of joint involvement, and with C-reactive protein concentrations. Serum IL-1Ra levels were also significantly correlated with IL-6 levels. CONCLUSION: These results argue against a relevant role of IL-1 in systemic JCA. The increase in IL-1Ra levels does not appear to reflect an increase in IL-1 production, but may rather be induced by IL-6.     

Differential effect of interleukin 10 on interleukin 12- and interleukin 2-mediated killer induction from blood mononuclear cells

Symptomatic tarsal coalition is often considered to be synonymous with peroneal spastic flatfoot. The association of the cavovarus foot type with tarsal coalition is less well established and has been described only in children. This article describes a case of an adult female with symptomatic cavovarus feet with talocalcaneal coalition. The authors theorize about the pathology of muscle spasm and pain in patients with this condition.     

CD81 on B cells promotes interleukin 4 secretion and antibody production during T helper type 2 immune responses

Mice lacking CD81 (TAPA-1), a widely expressed tetraspanin molecule, have impaired antibody responses to protein antigens. This defect is specific to antigens that preferentially stimulate a T helper 2 response (ovalbumin or keyhole limpet hemocyanin in alum) and is only seen with T cell-dependent antigens. Absence of CD81 on B cells is sufficient to cause the defect. Also, antigen-specific interleukin (IL) 4 production is greatly reduced in the spleen and lymph nodes of CD81-null mice compared with heterozygous littermates. Thus, expression of CD81 on B cells is critical for inducing optimal IL-4 and antibody production during T helper 2 responses. These findings suggest that CD81 may interact with a ligand on T cells to signal IL-4 production. By using a soluble form of CD81 as a probe, a putative ligand for CD81 was identified on a subset of B and T cells. Two possible models for the interaction of CD81 on B cells with a potential ligand on either B or T cells are proposed.     

Use of anti-CD3 epsilon F(ab')2 fragments in vivo to modulate graft-versus-host disease without loss of graft-versus-leukemia reactivity after MHC-matched bone marrow transplantation

The use of T cell-specific mAb in vivo for prevention and treatment of graft-vs-host disease (GVHD) and its impact on graft-vs-leukemia (GVL) reactivity was examined in a murine model of MHC-matched bone marrow transplantation (BMT). F(ab')2 fragments of a CD3 epsilon-specific mAb were administered to irradiated AKR (H-2k) hosts after transplantation of BM plus spleen cells from B10.BR donors (BMS chimeras). The effects on GVH and GVL reactivity were Ab dose- and schedule-dependent. A short course of mAb (qe2d, days 0 to 8) prevented clinical evidence of GVHD and mortality. Anti-CD3 F(ab')2 mAb reversed clinical symptoms of acute GVHD when delayed up to 18 days post-transplant. Anti-host (Mls-1a)-specific V beta 6+ cells were absent from the spleens of GVH-negative control mice, but persisted in Ab-treated BMS chimeras despite the absence of GVHD. Leukemic mice given 16.7 micrograms of Ab on days 0, 2, and 4 survived leukemia-free without developing severe GVHD. A longer course of Ab completely prevented GVHD, but led to leukemia relapse in tumor-bearing hosts, despite engraftment of donor T cells. The GVL effect was quantitatively stronger when Ab was used for GVH therapy as compared with GVH prevention. Some Ab-treated, GVH-free chimeras relapsed with lymphomas in unusual sites, suggesting that occult tumor cells may persist in nonlymphoid tissues. These experiments demonstrate that T cell-specific mAb can be used successfully in vivo to avoid severe GVHD, but that excessive or ill-timed administration of Ab may eliminate GVL reactivity.     

Mechanism of action of BCG-tumor cell vaccines in the generation of systemic tumor immunity. II. Influence of the local inflammatory response on immune reactivity

The intradermal injection of a vaccine composed of 10(7) X-irradiated syngeneic hepatocarcinoma line 10 (L10) cells admixed with 10(8) Mycobacterium bovis strain BCG into inbred Sewall Wright strain 2 guinea pigs induces a local inflammatory reaction and effectively immunizes against a contralateral challenge with viable L10 cells. The relationship between the local inflammatory reaction and the generation of tumor immunity was studied. Immunization against L10 was most effective when both L10 cells and BCG were injected into the same site, less effective when they were injected into separate sites with common lymphatic drainage, and not effective when they were injected at totally separate sites. Enzymatic dissociation of dermal vaccination sites revealed that vaccination with BCG and L10 cells combined induced a significantly greater inflammatory response than did vaccination with BCG alone or L10 cells alone; the inflammatory response was also greater than the combination of these individual responses, suggesting that BCG and L10 cells interacted synergistically in the elicitation of an inflammatory response. Sites receiving a combined vaccine of BCG and L10 cells were infiltrated rapidly by inflammatory cells, and surgical excision of these sites as early as 24 hours after vaccine administration did not affect significantly the development of immune responsiveness. However, vaccination sites induced by the injection of BCG and L10 cells at separate but adjacent sites were slowly infiltrated by inflammatory cells, and surgical removal of these sites within 96 hours of vaccination inhibited later immune responsiveness. Quantitative cellular analysis of these inflammatory reactions showed that inflammation was related to tumor-reactive immunity such that the greater the initial inflammatory process, the greater the resistance to tumor challenge.     

Interleukin-2 inhibits graft-versus-host disease-promoting activity of CD4+ cells while preserving CD4- and CD8-mediated graft-versus-leukemia effects

We have recently shown that a short course of high-dose interleukin-2 (IL-2) can markedly inhibit the graft-versus-host disease (GVHD)-promoting activity of donor CD4+ T cells. The difficulty in dissociating GVHD-promoting from graft-versus-leukemia (GVL) effects of alloreactive donor T cells currently prevents clinical bone marrow transplantation (BMT) from fulfilling its full potential. To test the capacity of IL-2 treatment to promote such a dissociation, we have developed a new murine transplantable acute myelogenous leukemia model using a class II major histocompatibility complex-positive BALB/c Moloney murine leukemia virus-induced promonocytic leukemia, 2B-4-2. BALB/c mice receiving 2.5 x 10(5) 2B-4-2 cells intravenously 1 week before irradiation and syngeneic BMT died from leukemia within 2 to 4 weeks after BMT. Administration of syngeneic spleen cells and/or a 2.5-day course of IL-2 treatment alone did not inhibit leukemic mortality. In contrast, administration of non-T-cell-depleted fully allogeneic B10 (H-2b) spleen cells and T-cell-depleted B10 marrow led to a significant delay in leukemic mortality in IL-2-treated mice. In these animals GVHD was inhibited by IL-2 treatment. GVL effects were mediated entirely by donor CD4+ and CD8+ T cells. Remarkably, IL-2 administration did not diminish the magnitude of the GVL effect of either T-cell subset. This was surprising, because CD4-mediated GVHD was inhibited in the same animals in which CD4-mediated GVL effects were not reduced by IL-2 treatment. These results suggest a novel mechanism by which GVHD and GVL effects of a single unprimed alloreactive T-cell subset can be dissociated; different CD4 activities promote GVHD and GVL effects, and the former, but not the latter activities are inhibited by treatment with IL-2.     

The effects of heparinase 1 and protamine on platelet reactivity

BACKGROUND: Protamine is currently the most widely used drug for the reversal of heparin anticoagulation. Heparinase 1 (heparinase) is being evaluated as a possible alternative to protamine for the reversal of heparin anticoagulation. The authors evaluated the effects of equivalent doses of heparinase and protamine on platelet reactivity by measuring agonist-induced P-selectin expression. METHODS: After Institutional Review Board (IRB) approval, informed consent was obtained from 12 healthy volunteers and 8 patients undergoing surgery requiring cardiopulmonary bypass (CPB). Twenty-four ml of blood was obtained from each volunteer; 10 ml of blood was obtained from each patient before the CPB, and another 10 ml was obtained after CPB. Heparin was neutralized using heparinase or protamine. Platelet reactivity was assessed by measuring the expression of P-selectin after stimulation of platelets with increasing concentrations of a thrombin receptor agonist peptide (TRAP). Data were analyzed using analysis of variance. P < 0.05 was considered significant. RESULTS: For the healthy volunteers, the activated coagulation times (ACTs) of the heparinized samples returned to baseline values with heparinase (12.5 U/ml) or protamine (32.5 microg/ml). For the 8 patients, the ACTs returned to baseline with heparinase (20 U/ml) or protamine (50 microg/ml). The authors found no difference in the expression of P-selectin in samples neutralized with heparinase, but samples neutralized with protamine showed a significant decrease in the expression of P-selectin when compared with heparinized samples. CONCLUSIONS: At dosages that reverse the anticoagulant effects of heparin, heparinase has minimal effects on platelets, whereas platelet reactivity was markedly inhibited by protamine.     

Varied effects of polyadenylic-polyuridylic acid on immune responses

The effect of the double-stranded synthetic polynucleotide, poly(A).poly(U), on the immune response of inbred mouse strains to multichain synthetic polypeptides was studied. Poly(A).poly(U) did not affect immune responses controlled by H-2 linked genes. Thus, when either (T,G)-A- -L or (Phe,G)-A--L were injected into high or low responder mice followed by administration of poly(A).poly(U) 24 h after immunization, no increase in the antibody titers was observed. In contrast, poly(A).poly(U) increased significantly the response to polyproline, which is controlled by a non H-2 linked gene, in low responder mice. However, the polyribonucleotide had no effect on the antibody titers of the SJL mice, the high responders to multichain polyproline. When poly(A).poly(U) was injected into DBA/1 mice following immunization with (Phe,G)-Pro- -L, the polynucleotide enhanced the low response to the Pro- -l region at the expense of the anti (Phe,G) response which is normally high in this mouse strain. In this case poly(A).poly(U) caused an intramolecular antigenic competition. The general conclusion of this study is that the chemical nature of the antigenic determinant plays an important role in determining the type of influence exerted by poly(A).poly(U).     

A randomized phase II trial of interleukin 2 and interleukin 2-interferon alpha in advanced renal cancer

An 81 year old right handed woman developed a left alien hand syndrome characterised by involuntary movements of choking and hitting the face, neck, and shoulder. The patient showed multiple disorders of primary sensation, sensory processing, hemispatial attention, and visual association, as well as a combination of sensory, optic, and cerebellar ataxia (triple ataxia) of the left arm in the absence of motor neglect or hemiparesis. Imaging studies disclosed subacute infarction in the right thalamus, hippocampus, inferior temporal lobes, splenium of corpus callosum, and occipital lobe due to right posterior cerebral artery occlusion. This rare syndrome should be considered as a "sensory" or "posterior" form of the alien hand syndrome, to be distinguished from the "motor" or "anterior" form described more commonly.     

T1/ST2 is preferentially expressed on murine Th2 cells, independent of interleukin 4, interleukin 5, and interleukin 10, and important for Th2 effector function

T helper (Th) cells can be categorized according to their cytokine expression. The differential induction of Th cells expressing Th1 and/or Th2 cytokines is key to the regulation of both protective and pathological immune responses. Cytokines are expressed transiently and there is a lack of stably expressed surface molecules, significant for functionally different types of Th cells. Such molecules are of utmost importance for the analysis and selective functional modulation of Th subsets and will provide new therapeutic strategies for the treatment of allergic or autoimmune diseases. To this end, we have identified potential target genes preferentially expressed in Th2 cells, expressing interleukin (IL)-4, IL-5, and/or IL-10, but not interferon-gamma. One such gene, T1/ST2, is expressed stably on both Th2 clones and Th2-polarized cells activated in vivo or in vitro. T1/ST2 expression is independent of induction by IL-4, IL-5, or IL-10. T1/ST2 plays a critical role in Th2 effector function. Administration of either a mAb against T1/ST2 or recombinant T1/ST2 fusion protein attenuates eosinophilic inflammation of the airways and suppresses IL-4 and IL-5 production in vivo following adoptive transfer of Th2 cells.     

State of immune reactivity of persons having occupational contact with Pu-239

We have previously shown that intact plants and cultured plant cells can metabolize and detoxify formaldehyde through the action of a glutathione-dependent formaldehyde dehydrogenase (FDH), followed by C-1 metabolism of the initial metabolite (formic acid). The cloning and heterologous expression of a cDNA for the glutathione-dependent formaldehyde dehydrogenase from Zea mays L. is now described. The functional expression of the maize cDNA in Escherichia coli proved that the cloned enzyme catalyses the NAD(+)- and glutathione (GSH)-dependent oxidation of formaldehyde. The deduced amino acid sequence of 41 kDa was on average 65% identical with class III alcohol dehydrogenase from animals and less than 60% identical with conventional plant alcohol dehydrogenases (ADH) utilizing ethanol. Genomic analysis suggested the existence of a single gene for this cDNA. Phylogenetic analysis supports the convergent evolution of ethanol-consuming ADHs in animals and plants from formaldehyde-detoxifying ancestors. The high structural conservation of present-day glutathione-dependent FDH in microorganisms, plants and animals is consistent with a universal importance of these detoxifying enzymes.     

Naltrexone's effects on reactivity to alcohol cues among alcoholic men.

The mechanisms of naltrexone's effects on urges to drink during abstinence are unclear. Naltrexone may suppress either urges to drink specifically or appetitive responses in general. The effects of naltrexone on cue reactivity to alcoholic and sweet nonalcoholic beverages were investigated. Alcohol-dependent men (N ?=?53) in treatment received naltrexone (50 mg) or placebo. Four hours later, they received baseline assessment, exposure to fruit juice, and exposure to their usual alcoholic beverage in 3-min trials. Naltrexone reduced urge to drink and self-reported attention to the alcohol cues, not at the initial exposure but after repeated exposures to alcohol cues. Naltrexone reduced negative affect across baseline and alcohol trials. No effects of naltrexone on responses to the nonalcoholic appetitive beverage cues were found, suggesting that general appetite suppression does not mediate the effects of naltrexone on urges. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Self-efficacy as a component of active coping: effects on cardiovascular reactivity

Active coping remains a poorly understood construct in cardiovascular reactivity testing. We have shown that active coping comprises two independent effects: the enhanced control and the effort of exercising control. The present study tested the proposition that, with effort left unconstrained, increased self-efficacy will increase cardiovascular response. Forty women were assigned to low or high self-efficacy conditions; self-efficacy was manipulated using false feedback. Subjects then engaged in a video game shape-matching task, while blood pressure and heart rate were monitored. SBP and DBP changes were smaller in the low self-efficacy group, as predicted: 17.9 versus 25.2 mmHG for SBP (p < 0.05); and 8.7 versus 13.0 mmHG for DBP (p = 0.07). Heart rate was similar for the two conditions. We conclude that self-efficacy for a task may be an integral part of the active coping process, indirectly affecting the blood pressure response by acting on the effort involved in the coping response.     

Nitrogenase reactivity: effects of pH on substrate reduction and CO inhibition

Molybdenum nitrogenase is composed of two separately purified proteins designated the iron protein (Fe protein) and the molybdenum-iron protein (MoFe protein), with the latter containing the substrate reduction site which is a metal cluster designated the iron-molybdenum cofactor (FeMo cofactor). In addition to its physiological substrates H+ and N2, nitrogenase reduces a number of nonphysiological substrates (e.g. C2H2 and N3-) and interacts with a number of similar molecules (e.g. CH3NC and CO) that serve as specific inhibitors. Despite their great diversity, all substrates are reduced by multiples of two electrons and require equivalent numbers of electrons and protons. Although the electron donor to a substrate is believed to be FeMo cofactor, the nature of the proton donor is unknown and might be different for different substrates. Here we report a three-component buffer assay system that eliminates variables of buffer type, ionic strength, and ATP and reductant availability and that is compatible with the nitrogenase system in the pH range 5.0-9.8. Preincubated studies and studies of the effects of pH on H2 evolution under Ar, H2 evolution under N2, H2 evolution under CO, and C2H2 reduction show that there is a group with a pK of ca. 6.3 that must be deprotonated for substrate reduction to occur and that there is a group with a pK of ca. 9.0 that must be protonated for substrate reduction to occur.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effects of PEG-interleukin-2 and interleukin-2 on essential hypertension and cellular immune function in the spontaneously hypertensive rat

The effects of recombinant human interleukin-2 covalently linked to polyethylene glycol (PEG-IL-2) or interleukin-2 (IL-2) on hypertension and in vitro suppressor T cell function in the spontaneously hypertensive rats (SHR) were investigated. Male young prehypertensive (4 weeks old) SHRs and adult (10 weeks old) SHRs with established hypertension were injected with low (5,000 units (u)/kg) or high (50,000-100,000 u/kg) dose of PEG-IL-2 or IL-2 as a single bolus or repeated injections. Systolic blood pressure was measured twice weekly using the tail-cuff technique. Systolic blood pressure in the PEG-IL-2 or IL-2 treated animals, irrespective of age, dose, or route of injection, did not differ significantly from that measured in vehicle-treated controls over a 10 week period. Mean arterial pressure measured by intra-arterial catheter was 159 +/- 7 mm Hg 10 weeks after treatment with repeated injections of 5,000 u/kg of PEG-IL-2 and 158 +/- 9 mm Hg in vehicle-treated controls. All rats injected with IL-2 had IL-2-specific IgG antibody in their sera. None of the PEG-IL-2 treated rats had any detectable anti-IL-2 antibodies in their sera. Thus, PEG-IL-2 showed far less immunogenicity than IL-2. Suppressor T (Ts) cells generated from adult SHR spleen cells failed to suppress pokeweed mitogen (PWM)-driven immunoglobulin G (IgG) synthesis. PEG-IL-2 or IL-2 supplementation both in vitro and in vivo restored the ability of adult SHR to generate Ts cells able to inhibit IgG synthesis. Our data suggest that PEG-IL-2 or IL-2 administration does correct a prominent defective Ts cell activity found in adult SHR, but that correction of this immune abnormality is not attended by an attenuation of hypertension.     

Addition of interleukin 12 to low dose interleukin 2 treatment improves antitumor efficacy in vivo

Interleukin 12 (IL-12) enhances lysis mediated by NK- and lymphokine activated killer (LAK) cells. It also causes proliferation of IL-2 stimulated T and NK cells in vitro. For these IL-2 complementing properties murine pulmonary metastases of a coloncarcinoma line were treated with IL-12 and IL-2 or with the individual agents. Results were compared to sham treated controls. IL-2 alone mediated significant tumor reduction but provoked pulmonary edema and concomittand toxicity, graded in three steps. IL-12 combined with an IL-2 dose reduced by 81% still resulted in significant antitumoral activity. Toxicity, however, was not discernable from sham treated controls. IL-12 thus appears as an attractive cytokine for combination with IL-2 in antitumor therapy. Particularly treatment of tumors, like gastrointestinal tract cancers, so far mainly resistant to cell mediated antitumor therapy, might profit from this approach.     

Osteoclasts and effects of interleukin 4 in development of chronic osteomyelitis

The cellular response to trauma and infection was studied in a murine model of posttraumatic osteomyelitis. Osteoclast response differed markedly depending on whether infection with Staphylococcus aureus accompanied the bone trauma. In animals recovering from sterile trauma, osteoclastic activity that was limited to the damaged or dead bone fragments caused rapid elimination of all recognizable dead bone within 1 week. New bone was laid down in an orderly fashion. Animals with superimposed infection had an intense polymorphonuclear leukocyte response develop. Additionally, osteoclasts behaved as acute inflammatory responders with substantial activity at the margins of the infected site and at previously uninjured tibial cortex adjacent to the infection. Despite the exuberant osteoclast response, bony fragments were not resorbed (for at least 4 weeks after the trauma), that is, sequestra developed, and new bone was laid down over morphologically dead bone and on the cortex (involucrum). When the inhibitory cytokine, interleukin 4 was given in a single dose with the bacterial inoculum, the osteoclast response was moderated with almost complete elimination of osteoclast activity at normal tibial cortex adjacent to the infected site. The limitation of osteoclastic activity did not impair the host's containment of bacterial growth.