?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F31150%2F&rft.title=Medroxyprogesterone+acetate+has+minimal+impact+on+HIV-1+replication+in+ectocervical+tissue+ex+vivo&rft.creator=Mariko%2C+Hawa&rft.description=A+link+between+use+of+the+contraceptive+depomedroxyprogesterone+acetate+(DMPA)+and+risk+of+HIV-1+acquisition+has+been+demonstrated+in+epidemiologic+studies.+Peak+medroxyprogesterone-17-acetate+(MPA)+levels+occur+4+days+post-injection+with+serum+progestin+levels+of+2+nM.+However%2C+other+studies+have+evaluated+its+in+vitro+effects+at+supraphysiologic+concentrations+resulting+in+immune+modulation+of+epithelial+cell+lines+and+peripheral+blood+mononuclear+cells.+To+formally+evaluate+MPA+effects+on+HIV-1+infection+in+a+more+physiologic+model%2C+we+utilized+our+polarized+ectocervical+tissue+explant+model.+Our+hypothesis+is+physiologic+concentrations+of+MPA+will+not+affect+HIV-1+replication+in+ectocervical+tissue+ex+vivo.%0D%0A%0D%0AHuman+ectocervical+tissue+was+processed+into+polarized+explants.+Explants+were+cultured+alone+(control)+or+with+MPA+in+the+basolateral+compartment+at+physiological+(0.05nM%2C+0.5nM%2C+1nM%2C+5nM)+(n%3D17)+or+higher+concentrations+(500nM%2C+5%CE%BCM%2C+500%CE%BCM)+(n%3D10)+for+48+h%2C+infected%0D%0Awith+either+5x10%5E2+or+5x10%5E3+TCID50+HIV-1BaL+apically+for+24+h%2C+and+then+washed.+Basolateral+supernatants+were+collected+and+replenished+with+fresh+medium+containing+MPA+every+3-4+days+up+to+21+days.+Supernatant+collections+at+the+48+h+time+point+(pre-infection)+were+tested+for+cytokine+expression+while+all+following+supernatant+collections+post-infection+were+analyzed+for+viral+p24gag+concentrations+via+ELISA.+PBMCs+were+cultured+for+72+hours+in+media+containing+MPA+at+5%CE%BCM%2C+50%CE%BCM%2C+and500%CE%BCM+(n%3D5)%2C+including+a+negative+and+activated+control%2C+then+analyzed+for+antibodies+via+flow+cytometry.%0D%0A%0D%0AViral+replication+over+21+days+in+the+presence+of+supraphysiologic+concentrations+of+MPA+indicated+suppression%2C+while+that+in+the+presence+of+physiologic+concentrations+was+not+significantly+different+from+the+control.+Evaluation+of+cytokine+markers+indicated+a+hormone+dose-dependent+relationship+between+decreasing+MPA+concentrations+(500%CE%BCM-5nM)+and+increasing+expression+of+pro-inflammatory+cytokines+approaching+levels+of+the+control%2C+which+were+significant+for+IL-12+(p40)%2C+IL-6%2C+and+IL-8.%0D%0A%0D%0AMPA+does+not+significantly+affect+HIV-1+replication+at+physiologic+concentrations+but+has+an+overall+suppressive+effect+on+replication+at+supraphysiologic+concentrations+in+ectocervical+tissue+in+cervical+tissue+ex+vivo.+%0D%0A%0D%0AThe+public+health+significance+of+DMPA+continuation+requires+balancing+unintended+pregnancy+while+diminishing+HIV-1+acquisition+risk%2C+especially+when+the+risks+of+infection+do+not+outweigh+the+contraceptive+benefits.&rft.date=2017-06-29&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F31150%2F1%2Fmariko_hawa_ETDApril2017.pdf&rft.identifier=++Mariko%2C+Hawa++(2017)+Medroxyprogesterone+acetate+has+minimal+impact+on+HIV-1+replication+in+ectocervical+tissue+ex+vivo.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++