With over 100 million new infections every year infections with Chlamydia trachomatis are a worldwide health problem. Chlamydiae are mandatory intracellular bacteria with an unique evolution cycle.
Cell culture experiments showed that mechanisms that limit the intracellular growth of chlamydiae are reduced in hypoxic environment. It is also known that the cell homeostasis and the metabolism of the host are directly influenced by the hypoxic-induced factors (HIF-1-3). It is likely that chlamydiae by themselves interact with and stabilize the central regulator of homeostasis under hypoxia through HIF-1α. With that chlamydiae prevent cellular apoptosis and can end their evolution cycle. It is still unclear if and how HIF-1α influences the immune response of the host against intracellular chlamydiae in vivo and can lead to chronic inflammation.
To get a better understanding of this, in this project the role of HIF-1α during Chlamydia infection in vivo will be analyzed.
Our goal is to investigate the role of HIF-1α during the course of an infection with chlamydiae to acquire a better understanding of the mechanisms chlamydiae use to survive in ordert to reduce consequences of infections and optimize the treatment.
To investigate the role of HIF-1α 25 female C57BL/6Rj (wildtype controls) mice and 25 Lyz 2cre HIF-1α floxed/floxed (HIF-1α knockout) mice will be treated with 2.5 mg medroxyprogesterinacetate and seven days later infected with an inoculum concentration of Chlamydia trachomatis that will be derived from previous experiments.
During the course of infection vaginal swabs and stool samples will be taken and the chlamydiae concentration will be determined using the indirect Immunfluorescence test.
Five wildtype control mice and five HIF-1α knockout mice will each be killed on different days during the course of infection. From the killed mice vaginal swabs and stool samples will be taken and the genital tracts and uteri will be dissected.
From the vaginal swabs the chlamydiae concentration will be determined. The microbiota compositions will be analyzed from the vaginal swabs and stool samples by sequencing the V3/V4 region oft he bacterial 16S rRNA gene. The uteri will be weighed and analyzed on the existence of a hydrosalpinx. The genital tracts and uteri will be enzymatically digested and the HIF-1α and inflammatory proteins will be evaluated using Western Blot.