Preliminary results on intersexual differences in gene expression of chemokine K203 in mononuclear cells of chicken.

Sex steroid levels increase during sexual maturation and cause alterations in many physiological and morphological traits. Some of these changes may be connected with age-dependent and intersexual differences in the immune system. This topic is still insufficiently understood, especially in avian species, partially due to methodological limitations. In this study we measured the gene expression of proinflammatory cytokines (IL-1ß, IL-6, IL-18) and chemokines [K60 (IL-8-like chicken chemokine - CXCLi1), CAF (IL-8-like chicken chemokine - CXCLi2), and K203] in mononuclear cells isolated from blood and spleen after in vitro stimulation with lipopolysaccharide (LPS). Samples were collected from chickens at two ages (from pullets before sexual maturity and from sexually mature egglaying hens). After LPS stimulation, a substantial increase was recorded in the gene expression of IL-6 and K203. All other measured genes were expressed at low levels in mononuclear cells irrespective of cell sources. We found a trend toward intersexual differences in K203 expression, but the expression of other cytokines and chemokines did not differ between the two sexes. The effect of stimulation was more pronounced in monocytes than in spleen macrophages, mainly in IL-6, IL-1ß and K203 gene expression. Our findings represent a basis for further studies on the effects exerted by sexual hormones on the immune phenotype of birds.

Maternally derived egg hormones, antibodies and antimicrobial proteins: common and different pathways of maternal effects in Japanese quail.

Avian eggs contain a variety of maternally-derived substances that can influence the development and performance of offspring. The levels of these egg compounds vary in relation to environmental and genetic factors, but little is known about whether there are correlative links between maternal substances in the egg underlying common and different pathways of maternal effects. In the present study, we investigated genetically determined variability and mutually adjusted deposition of sex hormones (testosterone-T, androstenedione-A4 and progesterone-P4), antibodies (IgY) and antimicrobial proteins (lysozyme) in eggs of Japanese quail (Coturnix japonica). We used different genetic lines that were independently selected for yolk T concentrations, duration of tonic immobility and social reinstatement behaviour, since both selections for behavioural traits (fearfulness and social motivation, respectively) produced considerable correlative responses in yolk androgen levels. A higher selection potential was found for increased rather than decreased yolk T concentrations, suggesting that there is a physiological minimum in egg T levels. Line differences in yolk IgY concentrations were manifested within each selection experiment, but no consistent inter-line pattern between yolk IgY and T was revealed. On the other hand, a consistent inverse inter-line pattern was recorded between yolk IgY and P4 in both selections for behavioural traits. In addition, selections for contrasting fearfulness and social motivation were associated with changes in albumen lysozyme concentrations and an inverse inter-line pattern between the deposition of yolk IgY and albumen lysozyme was found in lines selected for the level of social motivation. Thus, our results demonstrate genetically-driven changes in deposition of yolk T, P4, antibodies and albumen lysozyme in the egg. This genetic variability can partially explain mutually adjusted maternal deposition of sex hormones and immune-competent molecules but the inconsistent pattern of inter-line differences across all selections indicates that there are other underlying mechanisms, which require further studies.

Immune responsiveness of Japanese quail selected for egg yolk testosterone content under severe protein restriction.

Yolk testosterone concentrations vary in response to environmental conditions and different testosterone contents can subsequently modify the phenotypic traits of offspring. Apart from effects on growth, proactive behaviour and secondary sexual characteristics, the possible negative impacts of maternal testosterone on the immune system are often considered a limitation for its deposition. The effects of maternal testosterone can be modulated by postnatal environmental conditions, such as the availability of food resources. However, the majority of studies considering the effects of maternal testosterone on the immune system have been conducted under optimum conditions. We evaluated the influence of genetic selection for high (HET) and low (LET) egg testosterone content in Japanese quail on immune responsiveness of offspring to phytohaemagglutinin (PHA) and lipopolysaccharide (LPS) stimulation under severe protein restriction. Protein restriction negatively influenced body weight and performance in the PHA-test. We observed an increase in Cort (corticosterone) and He/Ly (heterophil/lymphocyte ratio) after LPS, while no changes occurred in total IgY levels in the protein-restricted group. HET quails showed higher body mass and total IgY levels and lower He/Ly ratio than LET quails, while the PHA index and Cort concentration did not differ between lines. No interactions were found between protein restriction and genetic line. In conclusion, the immune response was not compromised under conditions of severe protein restriction in the faster growing HET line compared with the LET line. We hypothesise that the immune responsiveness of birds with higher yolk testosterone may be linked with other maternally-derived substances in a context-dependent manner.

Growth and innate immunity are not limited by selection for high egg testosterone content in Japanese quail.

The effects of maternal androgens on fitness-related traits of offspring are generally assumed to be epigenetic adaptations to the environment that may be encountered by the next generation. Possible constraints of high yolk androgen transfer are still not understood, although a suppressed immune response in offspring is frequently considered. The aim of our study was to examine the innate immune defence in high (HET) and low egg testosterone (LET) lines of Japanese quail, which differ in the hormonal milieu of their eggs, thus providing a good physiological model for the study of androgen-mediated maternal effects. Acute phase response was induced by a lipopolysaccharide injection in 12-day-old quail and plasma corticosterone and the heterophil:lymphocyte ratio were measured at 1 and 3 h post-treatment. Basal levels of non-specific antibodies (IgY) were determined in the circulation. We found that HET quail were heavier than LET quail from the second week of age, indicating enhanced post-hatching growth. At 1 h post-lipopolysaccharide challenge, plasma corticosterone concentrations increased in the HET but not in the LET line. The heterophil:lymphocyte ratio rose in both lines at 3 h post-immune challenge, with a more pronounced response in HET quail. Moreover, HET chicks displayed higher IgY levels than LET chicks, suggesting either enhanced passive immunoprotection or stimulated endogenous antibody production. In conclusion, our data demonstrate that the genetic selection for high egg testosterone content positively influences growth and, simultaneously, does not limit the acute phase response in young quail.