Despite the immune-privileged status of the male genital tract, infection and inflammation of the male genital tract are important etiological factors in male infertility. A common observation in clinical and experimental orchitis as well as in systemic infection and inflammation are decreased levels of testosterone. Emerging data point to an immunosuppressive role of testosterone. In our study, we substituted testosterone levels in experimental autoimmune orchitis (EAO) in rat by s.c. testosterone implants. EAO development was reduced to 17% when animals were treated with low-dose testosterone implants (3 cm long, EAO+T3) and to 33% when rats were supplied with high-dose testosterone implants (24 cm, EAO+T24) compared with 80% of animals developing disease in the EAO control group. In the testis, testosterone replacement in EAO animals prevented the accumulation of macrophages and significantly reduced the number of CD4+ T cells with a strong concomitant increase in the number of regulatory T cells (CD4+CD25+Foxp3+) compared with EAO control. In vitro testosterone treatment of naive T cells led to an expansion of the regulatory T cell subset with suppressive activity and ameliorated MCP-1–stimulated chemotaxis of T lymphocytes in a Transwell assay. Moreover, expression of proinflammatory mediators such as MCP-1, TNF-α, and IL-6 in the testis and secretion of Th1 cytokines such as IFN-γ and IL-2 by mononuclear cells isolated from testicular draining lymph nodes were decreased in the EAO+T3 and EAO+T24 groups. Thus, our study shows an immunomodulatory and protective effect of testosterone substitution in the pathogenesis of EAO and suggests androgens as a new factor in the differentiation of regulatory T cells.

Disorders or differences in sex development (DSD) comprise a heterogeneous group of conditions with an atypical sex development. For optimal diagnosis, highly specialised laboratory analyses are required across European countries. Working group 3 of EU COST (European Cooperation in Science and Technology) Action BM 1303 ‘DSDnet’ ‘Harmonisation of Laboratory Assessment’ has developed recommendations on laboratory assessment for DSD regarding the use of technologies and analytes to be investigated. This position paper on steroid hormone analysis in diagnosis and treatment of DSD was compiled by a group of specialists in DSD and/or hormonal analysis, either from participating European countries or international partner countries. The topics discussed comprised analytical methods (immunoassay/mass spectrometry-based methods), matrices (urine/serum/saliva) and harmonisation of laboratory tests. The following positions were agreed upon: support of the appropriate use of immunoassay- and mass spectrometry-based methods for diagnosis and monitoring of DSD. Serum/plasma and urine are established matrices for analysis. Laboratories performing analyses for DSD need to operate within a quality framework and actively engage in harmonisation processes so that results and their interpretation are the same irrespective of the laboratory they are performed in. Participation in activities of peer comparison such as sample exchange or when available subscribing to a relevant external quality assurance program should be achieved. The ultimate aim of the guidelines is the implementation of clinical standards for diagnosis and appropriate treatment of DSD to achieve the best outcome for patients, no matter where patients are investigated or managed.

In order to further characterize oestrogen production and metabolism during bovine pregnancy, free (f) and conjugated (c) estrone (El), total free and conjugated oestrogens (tfcOe) and total free oestrogens (tfOe) were determined as marker oestrogens in blood plasma respectively in urine and feces of 10 pregnant cows. For the determination of individual oestrogens blood, urine and feces samples of days 240, 200, 160, 100, 60, 30, 10 and 5 prior to parturition were pooled and the free, sulfo (sc)- and glucuconjugated (gc) forms of El, 17β-estradiol (E2β) and 17α-estradiol (E2α) were obtained following differential enzyme hydrolysis and separation by HPLC; hormone assay was by established RIA-procedures. FEl and cEl concentration in blood plasma, tfOe in feces and tfcOe in urine showed a similar pattern. A first rise occurred between days 110 and 120 of pregnancy, an additional overproportional rise commenced at around days 230—250. Highest concentrations were measured in feces (tfOe ca. 500 ng/g 1 day a. p.), followed by urine (tfcOe ca. 3.5 ng/mosmol 2 days a. p.) and blood plasma (fEl ca. 8 nmol/1 and cEl ca. 20 nmol/1 2 days a. p.). Determination of individual oestrogens in blood plasma revealed that fE2β and fE2α could only be found 10 days a. p. while the conjugated forms could already be detected on days 100 and 160 a. p. With 62% El was the dominant oestrogen, followed by E2α (37%) and E2β (1.0%); El occurred predominantly as sulfate, E2α and E2β predominantly as glucuronide. Main metabloite in feces was fE2α (56.7%), followed by fE2β (32%) and fEl (11.3%); conjugated oestrogens were not detected. Main metabolite in urine was scEl followed by gcE2α and gcE2β. ScE2α and scE2β were not detected or were present in small quantities only. Hormonal changes over time were highly significant. Main product of placental oestrogen synthesis is scEl, the concentrations of f and c E2β and E2α in plasma largely result from oestrogen metabolism and enterohepatic circulation.