Genital epithelial cells from HIV-exposed seronegative (HESN)

commercial sex workers express a discrete cytokine/chemokines profile

upon toll-like receptor activationNyla Dil1 (DVM, MSc (Path), PhD)

Joshua Kimani2, Makobu Kimani2, Francis A. Plummer1, T. Blake Ball11Department 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.

• 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?

Question

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

Methods

TLR 7 and TLR 8 not expressed

TLR and other PRR Expression in GECs

IL-8 and IP-10 Production in GECs HESN GECs showlower levels of proinflammatory IL-8 in response to TLR stimulation.

HESN GECs showlower levels of activatedT cell chemoattractant IP-10 in response toTLR stimulation.

Conclusions

Acknowledgements•Dr. Blake Ball

•Dr. Frank Plummer

•Dr. Joshua Kimani

•Dr. Makobu Kimani

•Pumwani CSW Cohort participants

HESN GECs produce lower levels of basal and TLR mediated IL-8 and IP-10 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.