Transcript Genital epithelial cells from HIV-exposed seronegative commercial

Genital epithelial cells from HIVexposed seronegative (HESN)
commercial sex workers express a
discrete cytokine/chemokines profile
upon toll-like receptor activation
Nyla Dil1 (DVM, MSc (Path), PhD)
Joshua Kimani2, Makobu Kimani2, Francis A. Plummer1, T. Blake Ball1
1Department of Medical Microbiology and Infectious Diseases, University of Manitoba &
Public Health Agency of Canada, Winnipeg, Manitoba, Canada; 2Department of Medical
Microbiology, University of Nairobi, Nairobi, Kenya
Background
• Most HIV-1 infections acquired through heterosexual
transmission across female genital mucosa.
• Epithelial cells lining genital mucosa (GECs) act as first
line of defense against mucosal infections.
• Immune locale orchestrated by TLR mediated activation
of GECs can be a critical determinant of HIV-1 infection.
Question
• What is the role of GECs and TLR signaling in GECs in
determining the local mucosal cytokine /chemokines milieu in
HIV resistant and susceptible women?
Methods

Endocervical cytobrush samples from Pumwani CSWs cohort ( HESN (n=22), HIV- (n=24) and
HIV infected (n=23).

GECs purified through a series of nylon membrane filtrations

Cytokine levels in 24 h culture (with or without 25 ug/ml Poly(I:C), or 1ug/ml Pam3CSK4,
1ug/ml LPS, 100 ng/ml Flagellin and 100ng/ml FSL-1 combined) supernatants using the
Milliplex MAP multiplex Human Assay.

TLR expression GEC cell lines from different FGT anatomical sites
TLR and other PRR Expression in GECs
TLR 7 and TLR 8 not expressed
IL-8 and IP-10 Production in GECs
HESN GECs show
lower levels of
proinflammatory IL-8
in response to TLR
stimulation.
HESN GECs show
lower levels of activated
T cell chemoattractant
IP-10 in response to
TLR stimulation.
Conclusions
 HESN GECs produce lower levels of basal and TLR mediated IL-8 and IP10 that may be critical at reducing trafficking to this crucial mucosal site and
hence contributing to HIV-1 resistance.
 This distinct pattern of mucosal epithelial derived chemokine/cytokine
expression contributes to the immune quiescence phenotype observed in the
Pumwani HESN cohort.
Acknowledgements
•Dr. Blake Ball
•Dr. Frank Plummer
•Dr. Joshua Kimani
•Dr. Makobu Kimani
•Pumwani CSW Cohort participants