Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Mar;11(2):512-522.
doi: 10.1038/mi.2017.69. Epub 2017 Aug 9.

Vaccine-modified NF-kB and GR Signaling in Cervicovaginal Epithelium Correlates With Protection

Affiliations
Free PMC article

Vaccine-modified NF-kB and GR Signaling in Cervicovaginal Epithelium Correlates With Protection

L Shang et al. Mucosal Immunol. .
Free PMC article

Abstract

Cervicovaginal epithelium plays a critical role in determining the outcome of virus transmission in the female reproductive tract (FRT) by initiating or suppressing transmission-facilitating mucosal immune responses in naïve and SIVmac239Δnef-vaccinated animals, respectively. In this study, we examined the very early responses of cervical epithelium within 24 h after vaginal exposure to SIV in naive and SIVmac239Δnef-vaccinated rhesus macaques. Using both ex vivo and in vivo experimental systems, we found that vaginal exposure to SIV rapidly induces a broad spectrum of pro-inflammatory responses in the epithelium associated with a reciprocal regulation of NF-kB and glucocorticoid receptor (GR) signaling pathways. Conversely, maintenance of high-level GR expression and suppression of NF-kB expression in the epithelium were associated with an immunologically quiescent state in the FRT mucosa and protection against vaginal challenge in SIVmac239Δnef-vaccinated animals. We show that the immunologically quiescent state is induced by FCGR2B-immune complexes interactions that modify the reciprocal regulation of NF-kB and GR signaling pathways. Our results suggest that targeting the balance of NF-kB and GR signaling in early cervicovaginal epithelium responses could moderate mucosal inflammation and target cell availability after vaginal infection, thereby providing a complementary approach to current prevention strategies.

Conflict of interest statement

DISCLOSURE

The authors declared no conflict of interest.

Figures

Figure 1
Figure 1. SIV exposure induces epithelial expression of pro-inflammatory chemokines, cytokines, and other inflammatory mediators in naïve but not SIVΔnef-vaccinated animals
(A) The global transcriptional profiles of cervical explants of rhesus macaques after 8h topical exposure to SIV ex vivo were examined by microarray analysis and compared pairwise between naïve versus SIV-exposed tissues from the same animal. Topical SIV exposure rapidly increased cervical expression of a spectrum of pro-inflammatory chemokines, cytokines, and other inflammatory mediators as described under Results. (B) Log10 fold changes of selected genes detected by microarray analysis were measured by quantitative real time PCR. (C) Images of increased expression ex vivo of some of the chemokines, cytokines and inflammatory mediators in cervical epithelium at 8h shown in 1A in descending order. (D) Increased expression in vivo in cervix of the same chemokines, cytokines, and inflammatory mediators following vaginal challenge in naïve animals but not SIVΔnef-vaccinated animals. (E) Quantification of expression levels by quantitative image analysis of immunohistochemistry-stained in vivo tissues.
Figure 1
Figure 1. SIV exposure induces epithelial expression of pro-inflammatory chemokines, cytokines, and other inflammatory mediators in naïve but not SIVΔnef-vaccinated animals
(A) The global transcriptional profiles of cervical explants of rhesus macaques after 8h topical exposure to SIV ex vivo were examined by microarray analysis and compared pairwise between naïve versus SIV-exposed tissues from the same animal. Topical SIV exposure rapidly increased cervical expression of a spectrum of pro-inflammatory chemokines, cytokines, and other inflammatory mediators as described under Results. (B) Log10 fold changes of selected genes detected by microarray analysis were measured by quantitative real time PCR. (C) Images of increased expression ex vivo of some of the chemokines, cytokines and inflammatory mediators in cervical epithelium at 8h shown in 1A in descending order. (D) Increased expression in vivo in cervix of the same chemokines, cytokines, and inflammatory mediators following vaginal challenge in naïve animals but not SIVΔnef-vaccinated animals. (E) Quantification of expression levels by quantitative image analysis of immunohistochemistry-stained in vivo tissues.
Figure 2
Figure 2. Dynamic changes in NF-kB signaling pathways in early epithelial responses to SIV ex vivo
(A) Microarray analysis ex vivo of cervical explant tissues of rhesus macaques after 8h exposure to topical SIV. Increased expression of NF-kB1p50 and NF-kBp65/RelA and decreases in NF-kB signaling inhibitors, COMMD1, COMMD2, and COMMD10 (yellow boxes); increased expression of IRF1 and cAMP-PKA signaling molecules, ATF3 and CREM (blue boxes). (B) Log10 fold changes of selected genes detected by microarray analysis were measured by quantitative real time PCR. (C) Increased and decreased cervical epithelial expression by immunohistochemical staining of key NF-kB signaling molecules identified by microarray analysis after 8 h exposure to SIV.
Figure 2
Figure 2. Dynamic changes in NF-kB signaling pathways in early epithelial responses to SIV ex vivo
(A) Microarray analysis ex vivo of cervical explant tissues of rhesus macaques after 8h exposure to topical SIV. Increased expression of NF-kB1p50 and NF-kBp65/RelA and decreases in NF-kB signaling inhibitors, COMMD1, COMMD2, and COMMD10 (yellow boxes); increased expression of IRF1 and cAMP-PKA signaling molecules, ATF3 and CREM (blue boxes). (B) Log10 fold changes of selected genes detected by microarray analysis were measured by quantitative real time PCR. (C) Increased and decreased cervical epithelial expression by immunohistochemical staining of key NF-kB signaling molecules identified by microarray analysis after 8 h exposure to SIV.
Figure 3
Figure 3. NF-kB signaling pathways activated in vivo in cervical epithelium of naïve but not SIVΔnef-vaccinated animals
(A) Increased expression of NF-kBp50 and NF-kBp65/RelA in cervical epithelium and decreased expression of COMMD1 in vivo at day 1 and 3 following vaginal exposure to SIV in naïve animals but not SIVΔnef-vaccinated animals (4–5 dpc). Epithelial expression of IRF1, ATF3 and CREM increase in naïve animals following vaginal exposure, as shown ex vivo. In vivo, ATF expression is not increased in vaccinated animals, similar to NF-kB, but IRF1 and CREM expression levels are both elevated in vaccinated animals. (B) Quantification of immunohistochemical epithelial staining.
Figure 4
Figure 4. Epithelial glucocorticoid receptor (GR) expression decreases in response to SIV in naïve but not SIVΔnef-vaccinated animals
(A) Microarray analysis ex vivo of cervical tissue explants of rhesus macaques after 8h topical exposure to SIV. Increases in transcripts for Inhibitors (GRLF1) and repressors (ZNF653) of GR expression and down-regulation of DCAF6 (enhancer of GR expression), TTLL5 (GR-regulated gene), NCOA1/3 and NRIP1 (GR coactivators). (B) In vivo, GR is expressed at a high level mainly in the cervical epithelium of uninfected/unvaccinated animals. Following vaginal challenge, epithelial expression of GR is reduced in unvaccinated animals, but is maintained at high levels in SIVΔnef-vaccinated animals through 7 dpc. (C) Quantitative image analysis of GR expression in the cervical tissues of vaccinated and unvaccinated animals before and after vaginal inoculation.
Figure 5
Figure 5. FCGR2B-Immune complex (I.C.)-mediated suppression of epithelial NF-kB and GR signaling responses to SIV
(A) In ex vivo cervical tissues from naïve rhesus macaques, topical exposure to SIV reduces the epithelial expression of GR and induces epithelial expression NF-kB signaling molecules. Topical application of SIV-specific I.C. onto the cervical epithelium prior to SIV exposure partially suppresses these responses. (B) Quantitative image analysis of immunohistochemical epithelial staining. Fresh cervical tissues from 4 uninfected rhesus macaques were used. Uninfected and infected tissues from the same animal were compared pairwise to measure the effect of SIV-specific I.C.-FCGR2B inhibitory signaling on NF-kB vs. GR signaling pathways. Each dot represents a single animal.

Similar articles

See all similar articles

Cited by 2 articles

References

    1. UNAIDS 2013 Report on the global AIDS epidemic. 2013 http://www.unaids.org/sites/default/files/media_asset/UNAIDS_Global_Report_2013_en_1.pdf.
    1. Quinn TC, Overbaugh J. HIV/AIDS in women: an expanding epidemic. Science. 2005;308:1582–1583. - PubMed
    1. Li Q, et al. Glycerol monolaurate prevents mucosal SIV transmission. Nature. 2009;458:1034–1038. - PMC - PubMed
    1. Li Q, et al. Live simian immunodeficiency virus vaccine correlate of protection: local antibody production and concentration on the path of virus entry. J Immunol. 2014;193:3113–3125. - PMC - PubMed
    1. Miller CJ, et al. Propagation and dissemination of infection after vaginal transmission of simian immunodeficiency virus. J Virol. 2005;79:9217–9227. - PMC - PubMed

Publication types

MeSH terms

Feedback