Differential regulation of the immune response to SRBC by monoclonal antibodies to interferon-gamma

Three monoclonal antibodies against different epitopes of interferon-gamma (IFN-gamma) were used to assess its role as a normal immunomodulatory molecule. Two of these antibodies were able to reduce significantly the primary antibody response to sheep erythrocytes in an in vitro culture system. One of these two antibodies has been reported to suppress both the antiviral and macrophage activation factor activities of IFN-gamma by binding to its carboxyl terminus. These findings indicate that IFN-gamma is an important lymphokine for the maximum expression of the immune response and that it acts via the carboxyl terminus of the molecule. This antibody suppressed the immune response only when added at the initiation of culture, suggesting that the action of IFN-gamma is on an early component of the response. The third monoclonal antibody, which binds to the amino end of IFN-gamma, did not suppress the in vitro response. However, it was able to block the effects exerted by an immunosuppressive dosage of exogenous IFN-gamma on in vitro antibody production. These results indicate that the immunosuppression vitro antibody production. These results indicate that the immunosuppression induced by the addition of IFN-gamma to a primary antibody response and the role that it plays in that response are mediated through different sites on the molecule and, therefore, probably by different mechanisms.     

Japanese quail selected for high plasma corticosterone response deposit high levels of corticosterone in their eggs

Poor habitat quality or body condition often correlates with high responsiveness of the hypothalamo-pituitary-adrenal (HPA) axis rather than with elevated baseline levels of glucocorticoids. We hypothesized that, for egg-laying vertebrates, high responsiveness of the HPA axis would correspond to high concentrations of corticosterone in yolk. We tested the prediction that Japanese quail (Coturnix coturnix japonica) selected for high plasma corticosterone response to brief immobilization (HS quail) would lay eggs with higher yolk corticosterone concentrations than birds selected for low response (LS quail). Quail from both lines were left undisturbed, outside of the stressors associated with daily management, before a first round of egg collection. In a second experiment, quail of both lines were experimentally stressed during the week before egg collection. In both cases we found quail from the HS line to lay eggs with significantly higher yolk corticosterone concentrations than quail of the LS line. After exposure to added experimental stressors, the line difference was more pronounced (increasing from 62% to 96%). There was no line difference in concentrations of yolk testosterone. Our results suggest that (1) genetic differences underly differences in the transfer of maternal corticosterone to yolk and (2) females may be able to control deposition of corticosterone into yolk through a mechanism independent of baseline corticosterone titers.     

Formation of endogenous pyrogen by mononuclear phagocytes

Production of endogenous pyrogen by human and rabbit blood monocytes in response to stimulation with agents of different origin was studied by inhibitory analysis under comparable conditions. Actinomycin D and cytochalasin B were applied. New evidence was obtained about an important role in the mechanism of activation of mononuclear phagocytes of initial interaction between a stimulating agent and the leukocyte membrane and of the biphasic process of endogenous pyrogen production.     

Suppression of the humoral immune response by plasmacytomas: mediation by adherent mononuclear cells.

Mice bearing plasmacytomas have a severely impaired ability to mount a primary immune response; T cells from these mice, however, appear by both in vivo and in vitro criteria to function normally. This unusual pattern of immunodeficiency appears to be mediated by a regulatory cell found in the spleens and peritoneal cavities but not in the lymph nodes or thymuses of mice bearing plasmacytomas. The number of cells with suppressor activity in the spleens of plasmacytoma-bearing mice is directly proportional to the size of the subcutaneous tumor borne by the host. These cells are capable of suppressing antibody production in in vitro cultures of normal spleen cells but have no demonstrable effect on the ability of normal spleen cells to proliferate in vitro in response to phytohemagglutinin or 8-Br-guanosine-3', 5'-monophosphate (T and B cell mitogens, respectively). Characterization of the suppressor cell population on the basis of its cell surface properties, its radioresistance, its morphology, and its ability to adhere to various solid matrices suggest that these cells are adherent mononuclear cells. These data support the concept that plasma cell tumors indirectly induce an impairment in the humoral immune response of their hosts by stimulating the expression of regulatory functions in a population of splenic and peritoneal macrophages.     

Differential antimicrobial activity of human mononuclear phagocytes against the human biovars of Chlamydia trachomatis

The antimicrobial activities of human mononuclear phagocytes against Chlamydia trachomatis were investigated. Phagocytes cultured for 7 days or less were efficiently microbicidal. Almost complete inactivation of organisms from both human biovars was observed after 48 hr of incubation. However, organisms from the lymphogranuloma venereum (LGV) biovar survived in mononuclear phagocytes infected after 8 days or more in culture, whereas those from the trachoma biovar continued to be killed by such cells. Phagocytes cultured as long as 21 days killed the trachoma organisms with the same effectiveness as those cultured for 7 days or less. An ultrastructural study of inoculated phagocytes illustrated phagolysosomal fusion with degradation of organisms from either biovar in phagocytes which had been cultured for 24 hr before infection. Phagolysosomal fusion was not observed in cells which had been cultured for 8 days or more and then infected with LGV. The addition of interferon-gamma to these macrophages partially restored the phagocytes' microbicidal activity for LGV. Furthermore, a synergistic effect was observed when eosinophil peroxidase was added with interferon. Specific antibody failed to neutralize the infectivity of LGV organisms in 8-day or older mononuclear phagocytes. The findings may reflect the differences in disease syndromes between the two biovars, with the trachoma biovar causing more peripheral diseases and the LGV biovar causing a more systemic disease, with lymph node involvement as its main syndrome.     

The effect of althesin on plasma corticosterone levels

Althesin in doses which produced anesthesia (4 and 6 ml kg-1, i.p.) produced biphasic changes in plasma corticosterone levels. Plasma corticosterone showed an increase (p less than 0.05) due to the stress of injection but returned to basal levels by 30 min. Subsequent to the anesthetic effect (approximately 30 min) corticosterone levels increased markedly (p less than 0.01). Althesin's effectiveness showed time of day effects, i.e., Althesin was more effective in the A.M. Rats given 6 ml kg-1 Althesin showed graded plasma corticosterone responses to stresses of varying intensity. Blood withdrawal and surgical stress evoked significant increases in plasma corticosterone but a 2-min holding stress had no effect on plasma corticosterone levels. Instrumented rats receiving supplemental injections (i.p.) presented patterns of plasma corticosterone which were different from those receiving supplemental infusions (i.a.). Whereas plasma corticosterone levels of rats receiving the continuous infusion of Althesin remained relatively constant, corticosterone levels of those which received supplemental injections tended to increase. Collectively, these data suggest that Althesins usefulness as an experimental anesthetic is limited to those studies which are not compromised by stress-induced pituitary-adrenal activity.     

Stimulation of primary immune response by the activation of autoimmune processes]

Experiments were conducted on 220 female mice weighing 20--24 g, with the use of 3 types of antigens (sheep erythrocytes, vaccines from the intestinal and paratyphoid bacilli). There proved to be an increase on the 7th and the 14th day of the formation of specific antibodies under the effect of subcutaneous injection of a homologous blood (0.1--0.3 ml per mouse) 2 hours after the antigen immunization. Hemostimulation not only intensified the antibody-genesis, but also increased the resistance to the infection with the living microbial culture. The stimulating action of the blood injection persisted in irradiation of the mice with gamma-rays in a dose of 300 r. Hemostimulation produced an activation of the normal autoantibody system capable of influencing the function of cells necessary for the antigen assimilation.     

Stimulation of the immune response during blockade of serotonin receptors by cyproheptadine

The role of serotonin receptors in the inhibitory effect of serotoninergic system on immunogenesis was studied using cyproheptadine, a specific blocker of 5-HT2 receptors. It was shown that cyproheptadine administration to CBA mice stimulated the immune response, which was dopamine-dependent and was realized via thymus. With the pituitary stalk destruction, the stimulatory effect of cyproheptadine was not observed, which suggests the participation of 5-HT2 brain receptors in immunogenesis.     

The effect of transdermal corticosterone application on plasma corticosterone levels in pregnant Lacerta vivipara

Relationships between hormones and behaviour can be explored by altering endogenous hormone levels, often through implantation of silastic tubing or osmotic pumps filled with a hormone or its agonists or antagonists. However, organisms in sensitive life-history stages (such as pregnancy) may be adversely affected by the surgical procedures associated with these manipulations, necessitating use of non-invasive techniques. We demonstrate that the application of a sesame oil-corticosterone mixture to the skin of pregnant female common lizards (Lacerta vivipara) elevates plasma levels of the hormone. Pregnant female L. vivipara were captured and treated daily for 1-20 days with the sesame oil-corticosterone mixture (experimental group) or with vehicle only (control group). Blood samples were collected and analyzed for corticosterone by radioimmunoassay. Baseline plasma corticosterone levels were elevated within 1 h in the experimental group. Similar levels ( approximately 145 ng/ml) were found over the subsequent 2 days, and by day 5 had risen significantly higher ( approximately 281.9 ng/ml), where they remained for the duration of the experiment. These increases are comparable to those found in other species using related techniques. No significant changes in plasma corticosterone levels occurred in the control group. Finally, corticosterone levels also were determined for untreated females that were captured, held overnight, sampled, and released to access to the natural range of basal corticosterone levels. Basal plasma levels of corticosterone in pregnant females varied among individuals independently of female body size or corpulence.     

C-reactive protein collaborates with plasma lectins to boost immune response against bacteria

Although human C-reactive protein (CRP) becomes upregulated during septicemia, its role remains unclear, since purified CRP showed no binding to many common pathogens. Contrary to previous findings, we show that purified human CRP (hCRP) binds to Salmonella enterica, and that binding is enhanced in the presence of plasma factors. In the horseshoe crab, Carcinoscorpius rotundicauda, CRP is a major hemolymph protein. Incubation of hemolymph with a range of bacteria resulted in CRP binding to all the bacteria tested. Lipopolysaccharide-affinity chromatography of the hemolymph co-purified CRP, galactose-binding protein (GBP) and carcinolectin-5 (CL5). Yeast two-hybrid and pull-down assays suggested that these pattern recognition receptors (PRRs) form pathogen recognition complexes. We show the conservation of PRR crosstalk in humans, whereby hCRP interacts with ficolin (CL5 homologue). This interaction stabilizes CRP binding to bacteria and activates the lectin-mediated complement pathway. We propose that CRP does not act alone but collaborates with other plasma PRRs to form stable pathogen recognition complexes when targeting a wide range of bacteria for destruction.     

Comparison of plasma ACTH and corticosterone levels after embryo bursectomy

Plasma levels of both adrenocorticotropic hormone (ACTH) and corticosterone (B) were determined in embryos (day 15 of incubation), chicks (day 3 after hatch) and young chickens (8 weeks). Experimental animals were bursectomized at 80 hr of incubation, i.e., before any anlage of the bursa of Fabricius could develop. Bursectomized (BFX) animals were compared to sham-operated controls (T), in basal, resting condition and 7 (ACTH) or 14 min (B) after ether stress was delivered for 30 sec. Basal B and ACTH levels seemed not to be significantly modified in BFX embryos, chicks and chickens. Hypophysial and adrenocortical response to stress appeared more precociously in BFX embryos (day 15 of incubation) than in intact ones (day 19). The non stress-responsive period that was observed for one week after hatch of T birds did not appear in 3-day-old BFX chicks whose both B and ACTH stress-induced levels were as high as in intact adults. In contrast, adrenocortical and pituitary corticotropic responses to stress were markedly impaired (by 50%) in adult BFX chickens as compared to intact controls.     

The role of mononuclear phagocytes in cardiac allograft rejection in the rat: II. Characterization of mononuclear phagocytes extracted from rat cardiac allografts

A method was developed for the extraction of leukocytes infiltrating rat cardiac allografts. Mononuclear phagocytes (MNP) comprised 52.4 ± 5.5% of the cells extracted from allografts at the time of rejection (Day 7). Day 4 allografts and Day 7 syngeneic grafts yielded considerably fewer MNP although numbers of lymphoid cells were similar in all three groups. Allograft MNP were phagocytic for latex particles but only very low numbers were adherent to a variety of surfaces. About 50% were positive for attachment and internalization of opsonized sheep red cells via the Fc receptor. However, fewer cells were able to internalize sheep red cells than were able to bind them when complement receptor-mediated phagocytosis was investigated. Large amounts of plasminogen activator were secreted by allograft MNP while cells from syngeneic grafts produced very little. The possible participation of MNP in the effector phase of a mechanism for allograft rejection similar to delayed-type hypersensitivity is discussed.     

Factors modifying the effect of diazepam on plasma corticosterone levels in rats

We have examined factors that alter the effect of diazepam (DZ) on plasma corticosterone (CS) in rats. DZ had a biphasic effect on plasma CS levels: CS decreased with doses below 5 mg/kg and increased with higher doses. Peak response occurred 90 minutes post injection in both sexes. Plasma DZ levels were significantly higher in females than in males and peak at 10 and 30 minutes post injection in males and females, respectively. There was also a sex difference in the pattern of DZ metabolites. An acute stressor (30 minutes of immobilization) did not affect plasma CS levels in rats injected with a 5 mg/kg dose of DZ. Prenatally stressed animals did not differ in basal CS levels or in their response to 5 mg/kg of DZ compared to prenatally non-stressed animals. These two groups of animals also did not differ in plasma levels of DZ or of its metabolites. By contrast, the 5 mg/kg dose of DZ had no effect on plasma testosterone levels in control animals, but increased it in prenatally stressed animals. Furthermore, compared to non-stressed controls, prenatally stressed animals had lower baseline plasma testosterone levels. These results indicate that the effect of DZ on plasma CS is influenced by endogenous as well as exogenous factors and that these effects vary with the particular biochemical parameter under examination.