Powell B L, Drutz D J, Huppert M, Sun S H
Infect Immun. 1983 May;40(2):478-85. doi: 10.1128/iai.40.2.478-485.1983.
Pregnancy is a major risk factor for coccidioidal dissemination. Because rates of Coccidioides immitis growth and endospore release are stimulated in vitro by levels of unbound progesterone and 17 beta-estradiol that are achievable, in vivo, in the sera of pregnant women (i.e., 10(-9) to 10(-8) M), a specific-hormone-binding system in C. immitis was sought. Fungal cytosols were incubated with tritiated steroids plus or minus radioinert steroids to identify specific binding systems. All five strains of C. immitis tested exhibited specific saturable binding for progestin, estrogen, androgen, and (to a lesser extent) corticosterone and glucocorticoid hormone classes. Only low or inconsistent estrogen or androgen binding was found in Blastomyces dermatitidis and Torulopsis glabrata. Cryptococcus neoformans, Paracoccidioides brasiliensis, and non-albicans Candida species showed no binding. Scatchard analysis of progestin and estrogen binding in C. immitis revealed a high-capacity, low-affinity binding system that was unaffected by RNase and DNase, but 40 to 60% degraded by trypsin or heating. Ammonium sulfate precipitation resolved a high-affinity, low-capacity binding system (Kd = 1.24 X 10(-9) to 3.60 X 10(-8) M; number of binding sites = 0.014 to 0.20 pmol/mg of protein). The Kd of this system is sufficient to compete for unbound hormone in the sera of pregnant women. The high-capacity, low-affinity system may serve as a repository for hormone before its attachment to the specific binder. These studies suggest that the effects of nanomolar concentrations of sex hormones on C. immitis may be mediated by a specific cytosol protein-binding system and that stimulatory events observed in vitro may have relevance for the mechanism of coccidioidal dissemination in pregnancy.
怀孕是球孢子菌播散的主要危险因素。由于在体外,未结合的孕酮和17β-雌二醇的水平(即10^(-9)至10^(-8)M)可刺激粗球孢子菌的生长和内生孢子释放,而在孕妇血清中可达到这些水平,因此研究了粗球孢子菌中的一种特异性激素结合系统。将真菌胞质溶胶与氚标记的类固醇加或不加放射性惰性类固醇一起孵育,以鉴定特异性结合系统。所测试的所有五株粗球孢子菌均表现出对孕激素、雌激素、雄激素以及(程度较轻的)皮质酮和糖皮质激素类别的特异性可饱和结合。在皮炎芽生菌和光滑球拟酵母中仅发现低水平或不一致的雌激素或雄激素结合。新型隐球菌、巴西副球孢子菌和非白色念珠菌属未显示结合。对粗球孢子菌中孕激素和雌激素结合的Scatchard分析揭示了一种高容量、低亲和力的结合系统,该系统不受核糖核酸酶和脱氧核糖核酸酶的影响,但40%至60%会被胰蛋白酶或加热降解。硫酸铵沉淀解析出一种高亲和力、低容量的结合系统(解离常数Kd = 1.24×10^(-9)至3.60×10^(-8)M;结合位点数量 = 0.014至0.20 pmol/mg蛋白质)。该系统的Kd足以与孕妇血清中的未结合激素竞争。高容量、低亲和力系统可能在激素附着于特异性结合剂之前充当激素的储存库。这些研究表明,纳摩尔浓度的性激素对粗球孢子菌的影响可能由一种特异性胞质溶胶蛋白结合系统介导,并且在体外观察到的刺激事件可能与怀孕期间球孢子菌播散的机制相关。