Androgen and behavior in the male three-spined stickleback, Gasterosteus aculeatus. II. Castration and 11-ketoandrostenedione effects on courtship and parental care during the nesting cycle.

Courtship declines and ceases while parental care increases in the presence of developing eggs during the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus. Furthermore, circulating 11-ketotestosterone (11KT) levels are higher during the initial "courtship phase" than during the later "parental phase," similar to that found in other paternal fishes. This study aimed to investigate a possible functional relationship between changes in 11KT levels and changes in reproductive behavior during the nesting cycle. To this end, groups of nonspawned and spawned male sticklebacks were sham-operated, castrated, or castrated and treated with 11-ketoandrostenedione (11KA), and the effects of the treatments on courtship and parental care were studied. Castration removed circulating 11KT, while 11KA replacement prevented the natural decline in 11KT during the parental phase (11KA converts to 11KT extratesticularly), as assessed by radioimmunoassay. Regardless of treatment, parental care remained low and courtship was present in all nonspawned males, even at the end of the experiment. However, courtship did eventually decline in castrated nonspawned males compared to the other two nonspawned groups. In all treatments of spawned males there was a drastic decline in courtship and an increase in parental care. In castrated spawned males, however, the decline in courtship came earlier than in the other two spawned groups. 11KA treatment did not prevent the natural decline in courtship/increase in parental care in spawned males, indicating that the natural decline in 11KT is not responsible for the main portion of the rapid changes in these behaviors over the stickleback's nesting cycle. The limited effects of castration also exclude other gonadal hormones from being responsible for most of these changes.

Androgen and behavior in the male three-spined stickleback, Gasterosteus aculeatus I.--changes in 11-ketotestosterone levels during the nesting cycle.

Circulating 11-ketotestosterone (11KT) levels are higher during the courtship phase than during the later parental phase in a number of male teleosts. The present study describes the temporal changes in 11KT levels and their relationships to changes in courtship behavior, after different number of spawnings, over the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus, a small teleost showing pronounced paternal behavior. Plasma 11KT levels, measured by radioimmunoassay, were approximately 34 times higher during the initial courtship phase than at the end of the following parental phase in spawned males. In addition, males that spawned with three or more females on a single day showed an earlier decline in 11KT levels and in courtship behavior compared to males that were only allowed to spawn with a single female. Plasma 11KT levels remained high in males not allowed to spawn.

High levels of antibodies against Clq are associated with disease activity and nephritis but not with other organ manifestations in SLE patients.

OBJECTIVE: Serum concentration of antibodies to C1q (C1qAb) has been reported to be elevated in a high percentage of patients with systemic lupus erythematosus (SLE). The associations of high C1qAb levels with different clinical manifestations and the activity of the disease, however, are not definitely understood. METHODS: We measured the levels of IgG type C1qAb in the sera of 137 patients with SLE using an ELISA method. RESULTS: Serum concentrations of C1qAb were found to be higher (p < 0.0001) in SLE patients than in healthy controls. High titer (> 66 AU/ml) C1qAb was found in 40/137 (29.2%) SLE patients, and 4/192 (2.1%) healthy controls (p < 0.0001). A strong negative correlation (R = -0.4, p < 0.0001) between the age of the patients and the C1qAb titers could be detected. C1qAb levels in clinically active SLE patients significantly (p < 0.0001) exceeded those measured in the sera of patients in the inactive stage of the disease. A significant positive correlation was detected between C1qAb levels and the laboratory activity markers (anti-DNA, low C3 level) of the disease. We found a significant negative correlation between levels of C1qAb and a negative acute phase protein, alpha2-HS-glycoprotein. Renal involvement was present in 11/40 (27.5%) and 11/97 (11%) of the patients with high and low titers of C1qAb, respectively (p = 0.038). The prevalence of other organ manifestations was, however, the same in the patients with or without high titer C1qAb. CONCLUSION: These findings indicate that C1qAb measurement is a useful method for detecting the activity of SLE and predicting renal manifestations, but not other organ involvement in the disease.

[Transient Q-waves in coronary disease]. / Tranziens Q-hullámok coronariabetegségben.

The authors discuss the transient ischemic Q waves in various situations. Five cases are presented. Two patients had exercise-induced Q waves, one patient had right bundle branche block-dependent Q waves, one patient had transient Q waves after thrombolytic therapy and one patient had transient Q waves caused by Prinzmetal angina. Profound ischemia may not result in necrosis but may cause myocardial stunning. Myocardial stunning may be accompanied by deranged electrophysiologic activity with transient loss of electromotive forces.

Sodium-cromoglycate (Cromolyn) selectively increases the binding and phagocytosis of unsensitized target cells by rat peritoneal macrophages.

The influence of sodium-cromoglycate (cromolyn) on the binding and ingestion of sheep erythrocytes (SRBC) by elicited rat peritoneal macrophages (M phi) was studied using unsensitized SRBC. SRBC sensitized by homologous IgG or by IgM and complement as target cells. Preincubation of M phi with the drug (1 nM/1-2 mM/1) markedly enhanced both binding and ingestion of uncoated SRBC. The IgG-related increment in binding and phagocytosis was not significantly influenced by the drug. When target cells were coated by IgM and complement cromolyn pretreatment was ineffective. Preincubation of M phi by bovine brain gangliosides (BBG) diminished the cromolyn-induced enhancement of target cell binding and phagocytosis. When SRBC were pretreated by BBG, an increase of binding and phagocytosis was observed. These data suggest that cromoglycate may enhance the capacity of M phi to bind erythrocytes via ganglioside structures. Coating SRBC by complement components appears to interfere with binding of erythrocytes to M phi ganglioside receptors.

Rat IgG subclasses mediating binding and phagocytosis of target cells by homologous macrophages.

Attachment and ingestion of 51Cr-labelled TNP-SRBC sensitized by rat IgG1, IgG2a or IgG2b-type antibodies by homologous, elicited peritoneal macrophages were studied. IgG1 was found to be the most efficient isotype in mediating these functions. The antibody doses required for a significant attachment were found to differ with the isotype of Ab, while doses needed for a significant phagocytosis and antibody-dependent cellular cytotoxicity (ADCC) varied between 400-700 Ab/SRBC with all the isotypes studied. Both binding and phagocytosis were also influenced by the degree of hapten conjugation when target cells were sensitized by IgG1. Inhibition of these functions by soluble immune complexes and monomeric immunoglobulins suggests the involvement of two Fc gamma R in binding of the three isotypes. Based on the present work and on previous results we conclude that IgG2a interacts with a receptor binding complexed IgG only (Fc gamma RII), IgG2b binds to a different receptor which appears to bind monomeric ligand as well (Fc gamma RI), while IgG1 seems to interact with both types of receptor. We propose that phagocytosis can be mediated by both Fc gamma RI and Fc gamma RII.

Antibody mediated lysis of hapten-conjugated target cells by macrophages and by complement: the influence of IgG subclass, antibody and hapten density.

Lysis of 51Cr-labeled TNP-SRBC sensitised by rat IgG1 or IgG2a type antibodies by homologous, paraffin oil-elicited peritoneal macrophages (ADCC) or by homologous complement was studied. IgG2a was found to be markedly more efficient in mediating both ADCC and complement dependent lysis compared to IgG1. Inhibition of the ADCC pointed to the involvement of separate but partially overlapping interaction sites for the two isotypes. We suggest that FcRII type receptors play a favoured role in both IgG2a and IgG1 mediated ADCC. The threshold amount of bound antibody required for ADCC was lower than that sufficient for complement dependent lysis regardless on the subclass or on hapten density. The extent of lysis (both ways) was found to depend on hapten density using equal amounts of antibody. The results are interpreted as terms of the possible requirements for association of IgG molecules on the target cell surface.

Interaction between rat peritoneal macrophages and sensitised erythrocytes: dependence on IgG subclass, antibody density and the degree of hapten conjugation.

The interaction between macrophages (M phi) and antibody sensitised target cells was studied by the use of rat peritoneal macrophages, TNP hapten conjugated sheep red blood cells (SRBC) and homologous antibodies of subclasses IgG1 and IgG2a. Under optimal conditions, the great majority of the M phi formed rosettes with IgG1-sensitised antibodies while a maximum of 50% was achieved when target cells were sensitised by IgG2a. Using a double rosette technique, the major part of rosette-forming cells was found to bind both of the isotypes. IgG1-mediated rosette formation was observed at very low degrees of sensitisation as opposed to IgG2a-mediated target cell binding. Not only the amount of bound antibody but also the degree of hapten conjugation (epitope density) appear to influence the ratio of rosette-forming cells. IgG1-mediated rosette formation was partially inhibited by monomeric IgG1 and more efficiently by soluble ovalbumin (OVA)-anti-OVA complexes involving IgG1-type antibodies, while IgG2a mediated rosette formation was inhibited by OVA-anti-OVA complexes containing IgG2a type antibodies, and less efficiently by complexes involving IgG1. No inhibition was found by monomeric IgG2a. Based on the present data, we propose that two types of receptors are involved in the interaction of M phi and target cells coated by IgG1 and/or IgG2a type antibodies. One requires a multiple antibody-receptor interaction, binding both subclasses at overlapping binding sites; the other is able to interact with IgG1 and does not depend on the multiplicity of interactions.

Fc-dependent effector functions of idiotype-anti-idiotype immune complexes.

Some effector functions of antigen-antibody and antibody-antibody (idiotype-anti-idiotype) complexes were analyzed. As a model system a monoclonal IgM antibody specific for the hapten NP (antibody B1-8) was reacted either with hapten and hapten-carrier conjugates or with monoclonal anti-idiotope antibodies with specificity for B1-8 idiotopes. The precipitating, C1q-binding, complement-activating and Fc receptor binding properties of these complexes were compared. Binding of both hapten-carrier conjugates and anti-idiotope antibodies to B1-8 results in formation of complexes which depending on the B1-8:ligand ratio precipitate, activate complement, bind C1q and exhibit increased avidity for Fc mu and Fc gamma receptors of mouse spleen cells. In both types of complexes cross-linking of IgM molecules is essential for triggering these Fc-dependent functions, and a functional heterogeneity if idiotype-anti-idiotope complexes based on different idiotype-anti-idiotope ratios could also be observed. The functional similarity of B1-8-hapten-carrier and B1-8-anti-idiotope complexes suggests that regulatory functions so far assigned to antigen-antibody complexes could be carried out also by idiotype-anti-idiotope complexes.

Immunogenicity of antigen complexed with antibody. I. Role of different isotypes.

IgM-, IgG1-, IgG2a-, IgG2b- and IgE-type anti-ovalbumin antibodies were isolated from rat immune sera and the complexes formed by antibodies of a defined isotype and the antigen were compared for antibody avidity and interaction with homologous complement. IgG2a-containing complexes consumed total complement with the greatest efficiency. IgG2a-, IgG2b- and IgM-containing complexes displayed similar activities in activation of the alternative pathway while IgG1 was less efficient. Reduction and alkylation of IgG2a antibodies abolished the capacity of the complex to activate the alternative pathway, but not their total complement consumption. The complement-dependent solubilization was greatest when complexes contained IgG1- or IgM-type antibodies and lowest with IgG2a-containing complexes. Inbred Long Evans rats immunized by complexes containing IgG1 or IgG2b antibodies displayed a markedly lower delayed-type hypersensitivity (DTH) compared with those immunized by antigen alone. Immunization with IgG2a- or IgE-containing complexes resulted in a slightly decreased DTH, while IgM-containing complexes induced DTH like ovalbumin alone. IgG2a-containing complexes elicited an antibody response markedly higher than that found in animals immunized by antigen alone. The same effect was found when complexes contained reduced-alkylated IgG2a. Immunization with complexes containing the other isotypes involved in the study induced an antibody response similar to the antigen in phosphate-buffered saline.

Complement-mediated fragmentation of soluble and insoluble immune complexes containing porcine anti-DNP antibodies.

Complement-mediated release of soluble immune complexes and immune precipitates containing DNP-PSA and precipitating or non-precipitating porcine anti-DNP antibodies was studied. A decrease in the average size of soluble immune complexes indicating their fragmentation was observed during incubation in excess human serum, the extent of the complex release was found to be in direct proportion to the time of incubation. The effect was complement-dependent. In the second part of the study, complement-dependent solubilization of the immune precipitates of the precipitating antibody preparation was compared to the solubilization of the precipitates of the non-precipitating antibody formed in the presence of PEG. Although, both types of precipitates activated complement in about the same extent, complexes of non-precipitating antibody were solubilized much slower than those of the precipitating one. As avidity of both antibody preparations to the antigen was high, the observed differences in the rate of the complex solubilization probably reflected differences in the structure of the two types of complexes.

Classes and subclasses of rat antibodies: reaction with the antigen and interaction of the complex with the complement system.

Antibodies against the dinitroaminophenyl (DNAP) haptenic group were raised in outbred CFY rats using HSA or LPS as carrier. Antibodies isolated by immunoadsorbent techniques were resolved into fractions representing distinct isotypes, and the resulting fractions were tested for avidity. Subclass IgG2a was found to contain antibodies of an avidity index lower than those of other IgG subclasses or IgM. IgG2a was the only isotype detected when DNAP-LPS was used for immunization. Complexes containing defined isotypes were compared for their capacity to activate homologous complement. IgGl type antibody-containing complexes displayed a low complement activating capacity compared with those containing IgG2b, IgG2c or IgG2a. The latter subclass when complexed with antigen can thus induce complement-dependent processes in spite of a low avidity. Insoluble complexes of IgG antibodies were rapidly solubilized in rat serum (CRA phenomenon), except those containing predominantly IgG2c.

Studies of pre- and postimmunization antibodies of sheep.

Serum samples were taken serially from three nonimmunized sheep over a long period of time. Antibodies to human serum albumin (HSA), ovalbumin (OA) and FITC were separated from the samples. Than, two of the animals were injected with HSA+ complete Freund's adjuvant, the third with adjuvant without antigen. Serial postimmunization serum samples were subjected to the same procedures as the pre-immunization ones. The specific antibodies increased in concentration, and only the postimmunization antibody population was able to precipitate. In the presence of the antigen, the postimmunization antibodies bound to the Fc receptors of lymphocytes to an increased degree. There was no difference between pre- and postimmunizaton antibody populations either in complement-activating capacity or in the quantity of antigen necessary for reaching antigen-antibody equivalence. Isoelectro-focusing showed no new bands which would indicate antibodies different from the pre-existing ones. However, changes were observed in the relative participation of the antibodies forming different bands. No sharp limit was observed between pre- and postimmunization antibody populations. The Ig increment demonstrated after immunization was accompanied by a similar increment in the specific antibodies tested in the animal that had not given antigen, but not in the others. The authors attribute a role to humoral antibodies already in the earliest phase of immune response.