Effects of the antimicrobic tiamulin on seven gram-negative bacterial fish pathogens

In vitro and in vivo test were carried out with tiamulin and gram-negative bacterial pathogens of fish. Determination of minimum inhibitory concentration for 51 strains of seven species of gram-negative bacterial pathogens showed that only strains of Vibrio anguillarum were sensitive at 1.6-6.25 ppm, while the rest of test strains required 25- greater than 100 ppm. Control of infection was not achieved when tiamulin was fed for 14 days at 5 or 50 mg/kg to rainbow trout (Salmo gairdneri) experimentally infected with Yersinia ruckeri.     

Effects of cyclooxygenase inhibitors and prostaglandin E2 on macrophage activation in vitro

The effect of the cyclooxygenase inhibitors, indomethacin and diclofenac, and of PGE₂ on either resting or stimulated macrophages was investigated. Peritoneal macrophages were obtained from untreated mice and cultured for 10 days. Macrophage activation was induced by zymosan phagocytosis and was monitored by testing for plasminogen activator secretion and the cellular levels of lactate dehydrogenase, β-glucoronidase and alkaline phosphodiesterase I.It was found that cyclooxygenase inhibitors activate resting macrophages and enhance the degree of activation obtained after zymosan phagocytosis. Addition of exogenous PGE₂, on the other hand, had the opposite effect, it suppressed activation induced either by cyclooxygenase inhibitors, phagocytosis or a combination of both. Cyclooxygenase inhibitors and PGE₂ did not affect the hexose monophosphate shunt activity of resting macrophages and had only a minor effect on the respiratory on the respiratory macrophages and had only a minor effect on the respiratory burst occuring during zymosan phagocytosis. It appears, therefore, that the observed changes in the state of activation of the machrophages are not related to hexose monophosphate shunt activity.The described effects suggest that PGE₂ and possibly other cyclooxygenase products may function as inhibitory feed-back regulators of macrophage activation.     

Modulation of nonspecific defence mechanisms and specific immune responses after suppression induced by xenobiotics

Summary In the present study the possible modulation of nonspecific defence mechanisms and specific immune responses after suppression induced by oxytetracycline, oxolinic acid, and lindane – an organochlorine pesticide, were analysed in carp. Modulation was attempted using a natural immunostimulant, dimerized lysozyme (KLP-602, Nika Health Products, USA). Five groups of fish were given intraperitoneal injections of selected doses of the drugs on days 7, 6 and 4 before immunization with Yersinia ruckeri vaccine. At each injection, group I was also given 10 mg kg⁻¹ oxytetracycline, group II 10 mg kg⁻¹ oxolinic acid, group III 10 mg kg⁻¹ lindane, group IV was only immunized, and group V was injected with PBS. The immunostimulant was added in food before and after immunization. The study showed the immunotoxic effects of oxytetracycline, oxolinic acid and lindane; dimerized lysozyme corrected the defence mechanisms suppressed by these drugs and chemicals. The immunostimulatory effects of dimerized lysozyme were better when added before rather than after fish immunization.     

Relationship of prostaglandin secretion by rabbit alveolar macrophages to phagocytosis and lysosomal enzyme release

The phospholipids of rabbit alveolar macrophages were pulse-labelled with [(14)C]-arachidonic acid, and the subsequent release of labelled prostaglandins was measured. Resting macrophages released measurable amounts of arachidonic acid, the prostaglandins E(2), D(2) and F(2alpha) and 6-oxoprostaglandin F(1alpha). Phagocytosis of zymosan increased the release of arachidonic acid and prostaglandins to 2.5 times the control value. In contrast, phagocytosis of inert latex particles had no effect on prostaglandin release. Indomethacin inhibited the release of prostaglandin, and, at high doses (20mug/ml), increased arachidonic acid release. Analysis of the cellular lipids showed that after zymosan stimulation the proportion of label was decreased in phosphatidylcholine, but not in other phospholipids or neutral lipids. Cytochalasin B, at a dose of 2mug/ml, inhibited the phagocytosis induced by zymosan but increased prostaglandin synthesis to 3.4 times the control. These data suggest that the stimulation of prostaglandin synthesis by zymosan is not dependent on phagocytosis. Exposure to zymosan also resulted in the release of the lysosomal enzyme, acid phosphatase. Furthermore, cytochalasin B augmented the zymosan-stimulated release of acid phosphatase at the same dose that stimulated prostaglandin synthesis. However, indomethacin, at a dose that completely inhibited prostaglandin synthesis, failed to block the lysosomal enzyme release. Thus despite some parallels between the release of prostaglandins and lysosomal enzymes, endogenous prostaglandins do not appear to mediate the release of lysosomal enzymes. The prostaglandins released from the macrophages may function as humoral substances affecting other cells.     

Species-dependent effects of dietary lindane and/or zineb on the activation of aflatoxin B1 into mutagenic derivatives

Sprague-Dawley rats and Swiss mice were given diets containing lindane, 125 ppm, or zineb, 5200 ppm, or a mixture of both at the above-mentioned concentrations for 2 and 4 weeks. The effect of pesticide ingestion on the ability of liver S9 to metabolize aflatoxin B1 (AFB1) into mutagenic derivatives was tested by the Salmonella (TA100)/microsome test according to Ames. Control mouse-liver S9 was less efficient (13%) than the corresponding preparation from control rat liver. The ingestion of lindane produced a similar increase in the activities of both rat- (68%) and mouse-liver S9 (62%). Pretreatment with zineb inhibited (46%) rat-liver S9 but caused a marked increase (400%) in the activity of mouse-liver S9. Concomitant exposure to both pesticides showed that lindane released the inhibitory action of zineb on rat-liver S9 and reduced the stimulatory effect of zineb on mouse-liver S9. The inducing action of zineb in mice was a function of the dietary concentration of the pesticide. No effect was observed at dietary concentrations of zineb up to and including 500 ppm.     

A beta-glucan inhibitable receptor on human monocytes: its identity with the phagocytic receptor for particulate activators of the alternative complement pathway

The ligand specificity of the human monocyte receptor that mediates phagocytosis of particulate activators of the human alternative complement pathway was defined by inhibiting the phagocytic response with glycans known to be present in zymosan. When monocytes in monolayers were preincubated with 100 micrograms/ml of beta-glucan and then incubated with 1.25 to 2.5 X 10(6) zymosan particles, the percentage of cells that exhibited phagocytosis was inhibited in a time-dependent manner; maximal inhibition occurred within 20 min of preincubation. beta-Glucan inhibited monocyte phagocytosis of zymosan and rabbit erythrocytes (Er) in a similar dose-dependent fashion and at 100 micrograms/ml reduced monocyte ingestion of 5 X 10(6)/ml zymosan and 2 X 10(8)/ml Er by 63 +/- 8% and 68 +/- 16% (mean +/- SD, n = 3), respectively. The other glycan constituent of zymosan, mannan, was less than 1% as active, and 10 mg/ml of mannan reduced the number of monocytes ingesting zymosan and Er by 56 +/- 12% and 26 +/- 11%, respectively. At concentrations as high as 500 micrograms/ml, beta-glucan had no effect on monocyte Fc, C3b, or fibronectin receptor-mediated functions. Enzymatic hydrolysis of beta-glucan and alpha-mannan with beta-glucosidase or beta-glucanase before their incubation with monocytes abrogated their inhibitory capacity, whereas hydrolysis with alpha-mannosidase or alpha-glucosidase did not. Neither of the two alpha-glucans tested (dextran T-70 and nigeran) affected monocyte ingestion of zymosan particles or sheep erythrocytes (Es) sensitized with rabbit 7S anti-Es (EsIgG) at concentrations as high as 2 mg/ml. In contrast, a number of beta-glucans were active against zymosan but not EsIgG ingestion with a 75% reduction in the number of monocytes ingesting zymosan occurring with 100 micrograms/ml laminarin, 500 micrograms/ml soluble pachyman, and 900 micrograms/ml of soluble pustulan. The galactan, agarose, either in suspensions at 2 mg/ml or in a soluble portion at 600 micrograms/ml failed to affect monocyte ingestion of zymosan particles or Er. Thus, the monocyte receptor for particulate activators that is specifically inhibited by beta-glucan at a rate compatible with a phagocytic process and that recognizes beta-glucans but not alpha-glucans, mannan, or galactan is a beta-glucan receptor.     

Release of beta-hexosaminidase after administration of different agents affecting the reticuloendothelial system. An experimental study in rats

Plasma levels of a lysosomal enzyme, beta-hexosaminidase (beta-N-acetylglucosaminidase, EC 3.2.1.30) were studied in Wistar rats after administration of 99mTc -sulfur colloid, 198Au colloid, gelatine (Haemaccel), alcohol, methylpalmitate and zymosan. The activity of beta-hexosaminidase was increased 10, 30 and 60 min after the zymosan injection. After 24 and 48 h, enzyme levels had returned to those at outset. The transient release of beta-hexosaminidase probably occurred only during the phagocytosis of zymosan which was evaluated by histological examination of lung, liver and spleen. After the injection of all other agents tested, no significant aberration of beta-hexosaminidase levels was seen. Activity distribution of the radio-labeled colloids revealed differences in organ uptake which were attributed to a difference in colloid particle size. Although the colloids tested have been used extensively for determination of reticuloendothelial function and histological studies suggest phagocytosis of the particles, their administration did not affect plasma beta-hexosaminidase levels. Since lysosomal enzymes are cleared from the blood predominantly by liver macrophages, the primary location of particle phagocytosis may explain the present findings.     

Phagocytic and macropinocytic activity in MARCKS-deficient macrophages and fibroblasts

Macrophages express high levels of the myristoylated,alanine-rich, C kinase substrate (MARCKS), an actin cross-linkingprotein. To investigate a possible role of MARCKS in macrophagefunction, fetal liver-derived macrophages were generated from wild-type and MARCKS knockout mouse embryos. No differences between the wild-typeand MARCKS-deficient macrophages with respect to morphology (Wright'sstain) or actin distribution (staining with rhodamine-phalloidin, underbasal conditions or after treatment with phorbol esters, lipopolysaccharide, or both) were observed. We then evaluated phagocytosis mediated by different receptors: Fc receptors tested withIgG-coated sheep red blood cells, complement C3b receptors tested withC3b-coated yeast, mannose receptors tested with unopsonized zymosan,and nonspecific phagocytosis tested with latex beads. We also studiedfluid phase endocytosis in macrophages and mouse embryo fibroblasts byusing FITC-dextran to quantitate this process. In most cases, therewere no differences between the cells derived from wild-type andMARCKS-deficient mice. However, a minor but significant andreproducible difference in rates of zymosan phagocytosis at 45-60min was observed, with lower rates of phagocytosis in theMARCKS-deficient cells. Our data indicate that MARCKS deficiency maylead to slightly decreased rates of zymosan phagocytosis.

     

Involvement of cannabinoid receptor CB2 in dectin-1-mediated macrophage phagocytosis

Cannabinoid receptors are expressed in macrophages, but little is known of their roles. We here examined their involvement in phagocytosis. The presence of 2-arachidonylglycerol, an endocannabinoid, augmented the phagocytosis of zymosan by mouse macrophages, while the phagocytosis of Escherichia coli, Staphylococcus aureus, apoptotic cells or latex beads remained unaffected. An agonist of the cannabinoid receptors CB1 and CB2 also stimulated the phagocytosis of zymosan. The stimulatory effect of 2-arachidonylglycerol was abolished when phagocytosis reactions were carried out in the presence of an antagonist of CB2 but not of CB1. Furthermore, the phagocytosis of zymosan in the presence of 2-arachidonylglycerol was severely inhibited by the addition of a beta-glucan-containing carbohydrate or antibody neutralizing dectin-1, a beta-glucan-recognizing phagocytosis receptor. These results suggested that the activation of CB2 in macrophages leads to the stimulation of dectin-1-mediated phagocytosis.     

Cellular basis of allograft rejection in vivo. V. Examination of the mechanisms responsible for the differing efficacy of monoclonal antibody to CD4+ T cell subsets in low- and high-responder rat strains

MRC OX35, an anti-CD4 mAb, was used to treat high responder Wistar Furth (W/F) (RT1u) and low responder DA (RT1a) rats which had been grafted with directly vascularized hearts from PVG (RT1c) rats across a full MHC plus non-MHC incompatibility. Four doses of mAb at 7 mg/kg given in the first 2 wk postgrafting induced indefinite graft survival (greater than 150 days) in DA hosts, but only delayed rejection to 18 to 42 days in W/F as compared to rejection times of 6 to 8 days in untreated rats. The extension of MRC OX35 treatment to 6 wk in W/F rats induced indefinite graft survival in three of six rats. During treatment MRC OX35 therapy only partially depleted CD4+ cells, and all circulating CD4+ cells were coated with MRC OX35. The capacity of naive CD4+ and CD8+ cells from W/F and DA to be activated to PVG alloantigen was compared both in vitro in an MLC assay and in vivo by an adoptive transfer assay of their capacity to restore rejection of PVG heart grafts in irradiated syngeneic hosts. CD4+ cells from both W/F and DA proliferated in MLC and restored graft rejection. W/F CD8+ cells both proliferated in MLC and restored rejection, but DA CD8+ cells neither proliferated nor reconstituted rejection. Examination of lymphocytes from MRC OX35 treated hosts with long-surviving grafts showed that they were neither depleted of CD4+ T cells nor did they lack the capacity to proliferate to PVG Ag in MLC, this response being similar to that to third-party Ag or by naive lymphocytes. Compared to first-set rejection, PVG skin graft rejection was delayed 2 to 3 days in W/F and 10 to 12 days in DA rats with long-surviving grafts after MRC OX35 therapy, whereas they rejected third-party skin grafts in first-set tempo. These studies show that differences in graft survival in anti-CD4 treated low and high responder strains may be due to the inherent capacity of CD8+ cells to be activated to effect rejection independent of CD4+ cells in W/F but not in DA. In those hosts that accept grafts, there is no evidence of clonal deletion, but there appears to be a form of unresponsiveness akin to that induced in adult rats by other immunosuppressive therapies that protects the graft from rejection.     

阿特拉津和林丹对紫贻贝生长与繁殖净能的影响

本研究评估了亚致死浓度的林丹(1/2的60dLC50,0.935mg/L)和阿特拉津(1/2的60dLC50,3.585mg/L)对海洋性贝类影响的生理学反应。林丹和阿特拉津是已被公认的环境污染物质。56d的实验表明,这两种农药在贝类体内具有积累作用,而且在体内各器官的积累程度与其作用的靶器官相一致。林丹在体内的积累浓度(每克干重372μg)比阿特拉津(每克干重137μg)更高。该贝类在林丹中暴露56d,其氧的消耗比对照组降低10%,而在阿特拉津中则比对照组提高了29%。经林丹和阿特拉津暴露后,增加了氨排泄物。但是,两种农药均使贝类的取食率和吸收率下降。林丹会降低贝类的取食率、氧消耗、氨排泄、食物吸收率以及生长范围。在形式上,阿特拉津对贝类的影响与林丹的影响有不同之处,它会降低贝类的取食率和食物吸收率,但与林丹不同的是,它会提高贝类的氧消耗和氨排泄。总之,阿特拉津会明显地减小贝类繁殖净能。因此,阿特拉津暴露与林丹暴露的毒性症状是一致的。结合组织化学分析,贝类的生理学反应可作为一种理想的环境监测工具。研究结果表明,在二分之一的半致死浓度下暴露两个月,不仅能有效地证明林丹和阿特拉津在贝类组织内具有积累作用,而且对这些积累能产生生理反应。     

Residues of lindane in adipose tissue of lambs and sheep from islands Krk and Cres

Concentrations of lindane from adipose tissue i.e. omentum of sheep from homesteads of island Krk that were treated with lindane, were compared with the concentrations of lindane measured in adipose tissue of non treated sheep from husbandry cooperative on island Cres. Evaluation of food safety for this samples were estimated by comparing obtained results and maximum tolerance concentration (MTC) for lindane (2.0 and 0.2 mg/kg). In treated lambs from Krk measured concentrations of lindane were in range from 0.0038 to 1.8644 mg/kg, and in treated sheep from 0.0094 to 1.646 mg/kg. In control group of lambs and sheep concentration of lindane was < or = 0.0051 mg/kg. Estimation of food safety based on the results from this research are discussable, regarding the fact that MTC prescribed before year 2007 was 2.0 mg/kg. When the new law Regulations of MTC for residues of pesticides in food and animal feed (NN 119/07) came into the force prescribed MTC was 0.02 mg/kg. If food safety of sheep meat from Krk would be estimated based on latter, then 73.4% of samples of lamb meat and 80% of sheep meat could be proclaimed unsafe for human consumption. It is encouraging that after completion of using lindane in year 2005, levels of lindane in adipose tissue of lamb from Krk in year 2006 were under the concentration set by new legislate (< 0.02 mg/kg).     

Yeast mannans inhibit binding and phagocytosis of zymosan by mouse peritoneal macrophages

We have examined the effects of various mannans, glycoproteins, oligosaccharides, monosaccharides, and sugar phosphates on the binding and phagocytosis of yeast cell walls (zymosan) by mouse peritoneal macrophages. A phosphonomannan (PO(4):mannose ratio = 1:8:6) from kloeckera brevis was the most potent inhibitor tested; it inhibited binding and phagocytosis by 50 percent at concentrations of approximately 3-5 μg/ml and 10 μg/ml, respectively. Removal of the phosphate from this mannan by mild acid and alkaline phosphatase treatment did not appreciably reduce its capacity to inhibit zymosan phagocytosis. The mannan from saccharomyces cerevisiae mutant LB301 inhibits phagocytosis by 50 percent at 0.3 mg/ml, and a neutral exocellular glucomannan from pichia pinus inhibited phagocytosis by 50 percent at 1 mg/ml. Cell wall mannans from wild type S. cervisiae X2180, its mnn2 mutant which contains mannan with predominantly 1(arrow)6- linked mannose residues, yeast exocellular mannans and O-phosphonomannans were less efficient inhibitors requiring concentrations of 1-5 mg/ml to achieve 50 percent reduction in phagocytosis. Horseradish peroxidase, which contains high-mannose type oligosaccharides, was also inhibitory. Mannan is a specific inhibitor of zymosan binding and phagocytosis. The binding and ingestion of zymosan but not of IgG- or complement-coated erythrocytes can be obliterated by plating macrophages on substrates coated with poly-L-lysin (PLL)-mannan. Zymosan uptake was completely abolished by trypsin treatment of the macrophages and reduced by 50-60 percent in the presence of 10 mM EGTA. Pretreatment of the macrophages with chloroquine inhibited zymosan binding and ingestion. These results support the proposal that the macrophage mannose/N-acetylglucosamine receptor (P. Stahl, J.S. Rodman, M.J. Miller, and P.H. Schlesinger, 1978, Proc. Natl. Acad. Sci. U.S.A. 75:1399-1403, mediates the phagocytosis of zymosan particles.     

Towards an understanding of human impact upon the hydrology of Lake Naivasha,Kenya

The growth of a strain of Moina macrocopa (Straus 1820) isolated from an experimental stabilization pond in Marrakesh, was examined at seven concentrations of algae (6.25–6.25 × 10⁵ cells ml^–1 and at 5 different temperatures (15–30 °C)). Feeding conditions influenced the growth rate as well as the maximum size that reached 1.8 mm at 25 °C and at the highest algal concentration (6.25 × 10⁵ cell ml^–1). The life span and number of moltings reached a maximum (17.4 days and 13 moltings) at average nutrient concentrations (6.25 × 10⁵ cell ml^–1). Juvenile stages varied from 1 to 3 and adult ones from 6 to 8. In the temperature interval tested, growth rate increased with temperature while longevity decreased. Temperature had less effect on maximal size than nutrient availability. Population density (but not crowding) influenced longevity and survival but had no effect on growth.     

Phagocytic response to fully controlled plural stimulation of antigens on macrophage using on-chip microcultivation system

To understand the control mechanism of innate immune response in macrophages, a series of phagocytic responses to plural stimulation of antigens on identical cells was observed. Two zymosan particles, which were used as antigens, were put on different surfaces of a macrophage using optical tweezers in an on-chip single-cell cultivation system, which maintains isolated conditions of each macrophage during their cultivation. When the two zymosan particles were attached to the macrophage simultaneously, the macrophage responded and phagocytosed both of the antigens simultaneously. In contrast, when the second antigen was attached to the surface after the first phagocytosis had started, the macrophage did not respond to the second stimulation during the first phagocytosis; the second phagocytosis started only after the first process had finished. These results indicate that (i) phagocytosis in a macrophage is not an independent process when there are plural stimulations; (ii) the response of the macrophage to the second stimulation is related to the time" delay from the first stimulation. Stimulations that occur at short time intervals resulted in simultaneous phagocytosis, while a second stimulation that is delayed long enough might be neglected until the completion of the first phagocytic process.     

Release of leukotrienes by human monocytes on stimulation of their phagocytic receptor for particulate activators

The respective capacities of adherent human monocytes to metabolize endogenous arachidonic acid into leukotrienes C4 (LTC4) and B4 (LTB4) in response to activation with an ionophore, A23187, or to phagocytosis of unopsonized zymosan particles and IgG-sensitized sheep erythrocytes ( EsIgG ) were compared under optimal conditions for each stimulus. Resolution of the cellfree supernatant, after ionophore activation, by reverse-phase high performance liquid chromatography (RP-HPLC) identified only two products which eluted at the retention times of LTC4 and LTB4. There was correspondence between their quantitation by integrated optical density and radioimmunoassay, and the recoveries from the initial supernatant were 80% by radioimmunoassay. Activation of adherent monocytes from 12 donors by ionophore and by zymosan particles released 68.1 ng and 10.0 ng LTB4 and 29.5 ng and 2.1 ng LTC4, respectively. With trypsin pretreatment, the monocytes responded fully to ionophore activation but were inhibited in their response to zymosan particles as assessed by phagocytosis and leukotriene release, indicating that the zymosan stimulus acted through a trypsin-sensitive membrane receptor. When the response of adherent monocytes from nine donors to zymosan particles and to EsIgG were compared at identical particle concentrations and with similar numbers of ingesting monocytes, zymosan elicited LTB4 release (mean 6.7 ng) from all and LTC4 (mean 1.5 ng) from eight donors, while EsIgG caused low level release of LTB4 (mean 0.7 ng) from six and LTC4 from only one of the donors. Neither zymosan nor ionophore stimulation led to the metabolism of exogenously added [3H]LTB4 or [3H]LTC4 as assessed by RP-HPLC of the cellfree supernatants and by quantitation of the eluted labeled products. Thus, transmembrane activation of adherent monocytes by their receptor for particulate activators, in contrast to stimulation of their IgG-Fc receptor, reproducibly releases substantial quantities of LTB4 and LTC4, and may represent an important mechanism for regulating the microenvironment in the nonimmune host.     

Effects of in vivo and in vitro lindane treatment on rat ventral prostate lipids

The rat ventral prostate accumulated lindane (gamma-hexachlorocyclohexane) (0.59 +/- 0.07 ppm) when this liposoluble toxicant was injected subcutaneously at a concentration of 1 mg of 100 g body weight for 12 days. Total lipids and phospholipids (especially phosphatidylcholine) amounts were augmented in treated rats. Lindane had no significant influence upon cholesterol mass content in the ventral prostate. Using [1-14C]acetate as radioactive precursor, it was possible to conclude that the mass lipid variations caused by lindane treatment were due, at least in part, to a modification of the endogenous biosynthesis of these lipids. No changes were found in the acetate oxidation to CO2 when control rats and lindane-treated rats were compared.     

T cell subsets in allograft rejection. In situ characterization of T cell subsets in human skin allografts by the use of monoclonal antibodies

We have provided evidence that both major T cell subsets, T4-positive (helper/inducer) and T8-positive (cytotoxic/suppressor), infiltrate human skin allografts. Overall, and in the graft dermis and graft bed, T4-positive cells were predominant (1.5 to 3 times more numerous than T8-positive cells). In contrast, T8-positive cells were relatively more numerous in the epidermis and hair follicles. Rejection probably proceeded by two apparently independent pathways: 1) direct contact killing of graft epithelial cells, presumably by immunologically specific T8-positive cytotoxic cells, and 2) injury of microvascular endothelium of both the graft and graft bed with secondary graft infarction. Although important in first set skin allograft rejection, the mechanism of the second type of killing is uncertain. T4-positive cells were probably involved, as evidenced by their greater numbers; furthermore, studies in mice have shown that transfused helper/inducer cells are able to effect first-set skin graft rejection. It remains to be determined whether T4-positive cells act alone or cooperate with other cells to destroy vessels and bring about graft rejection. Langerhans cells were recognized in epithelial and dermal compartments of both allografts and autografts by their reactivity with anti-T6 and anti-Ia antibodies. We could not determine whether such cells in allografts were of host or donor origin.     

Prolongation of alloskin graft survival by catalytic scavengers of reactive oxygen species

We tested the effects of salen manganese (Salen-Mn) complexes, which are scavengers of reactive oxygen species exhibiting superoxide dismutase and catalase activities on the rejection of and alloresponse to fully allogeneic skin grafts in mice. We showed that pre-transplant treatment of C57Bl/6 donor skin or of BALB/c recipients with Salen-Mn complexes significantly delayed allograft rejection. ELISPOT analysis of alloimmune response of treated mice revealed a significant reduction of the frequency of type 1 cytokine (pro-inflammatory) producing T-cells, while the number of activated T-cells producing type 2 cytokines was elevated. In addition, anti-oxidative treatment of graft recipients resulted in a profound inhibition of their donor-specific cytotoxic T-cell response. Our results indicate that salen manganese complexes mediate their effect on graft rejection both by reducing the susceptibility of graft tissue to ROS-mediated injury and by exerting an anti-inflammatory effect in recipients.     

Host response to tissues placed in the anterior chamber of the eye: demonstration of migration inhibition factor and serum blocking activity.

Allograft immunity of rats to transplantation antigens was demonstrated by in vitro migration inhibition procedures. Spleen cell suspensions from rats sensitized to histocompatibility antigens by skin graft or anterior chamber implants failed to migrate normally in vitro when incubated in the presence of appropriate donor tissue extracts. Tissue grafts transplanted across both major and minor histocompatibility barriers had prolonged survival within the anterior chamber. However, 2 weeks after implantation, the recipient rats showed detectable sensitization to their implants. Serum from 14- to 32-day implant-bearing rats blocked the migration inhibition found in the implanted rats in the presence of corresponding tissue antigen. Such blocking activity was undetectable in the serum from rats which had implants for 7 days or after rejection of the implant was evident. MIF production in implanted and skin-grafted rats was evident at significantly different times in relation to the graft rejection. The asynchrony of MIF synthesis observed in these experimental animals leads us to postulate that the graft rejection and MIF production may be mediated by distinct lymphocyte populations. In addition, the route of the antigen presentation may account in part for the observed differences.