The Mechanism Study About Proliferation Effect Of Estradiol On Regulatory T Cell Derived From Male Ankylosing Spondylitis Patients

Part Ⅰ Immunomagnetic beads separation regulatory T cells and its purity identificationOBJECTIVE: To evaluate the purity of regulatory T cells which separate with Immunomagnetic beads approach.METHODS: Blood collection from male ankylosing spondylitis patients vein.To separate Peripheral Blood Mononuclear Cells from Venous blood with density gradient centrifugation.Take advantage of immunomagnetic beads and MACS separator to get regulatory T cell,to estimate the purity of regulatory T cell with flow cytometry after cell culture in good condition.RESULT: it’s a good way and smart choice to separate regulatory T cell from human PBMC in MACS approach,the Purity of regulatory T cell could be more than 95%.CONCLUSION: We could take MACS approach into consideration to separate regulatory T cell and for further research.Part Ⅱ To explore the proliferation effect of estradiol on regulatory T cells which derived from male ankylosing spondylitis patientsOBJECTIVE: To explore the proliferation effect of estradiol on regulatory T cells which derived from male ankylosing spondylitis patients and the optimum Stimulation concentration of estradiol.METHODS: Blood collection from male ankylosing spondylitis patients vein.To separate Peripheral Blood Mononuclear Cells from Venous blood with density gradient centrifugation and then continue to separate CD4~+T cells,To culture CD4~+T cell with different concentration of estradiol media(0nmol/L,1nmol/L,10nmol/L,100nmol/L)for 72 hours.RESULT: After culture with different concentration estradiol media,we found that the proportion of regulatory T cells among CD4~+T cells increased between 0-10 n M estradiol stimulation but decreased in 100 n M culture media.CONCLUSION: Estradiol could stimulate the proliferation of regulatory T cells between 0-10 n M concentration but a higher concentration maybe limit its proliferation.