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Background  A  Lactobacillus -dominated cervicovaginal microbiota (VMB) protects women from adverse reproductive health outcomes, but the role of  L .  iners  in the VMB is poorly understood. Our aim was to explore the association between the cervicovaginal  L .  iners  and  L .  crispatus  proteomes and VMB composition.    Methods  The vaginal proteomes of 50 Rwandan women at high HIV risk, grouped into four VMB groups (based on 16S rDNA microarray results), were investigated by mass spectrometry using cervicovaginal lavage (CVL) samples. Only samples with positive 16S results for  L .  iners  and/or  L .  crispatus  within each group were included in subsequent comparative protein analyses:  Lactobacillus crispatus -dominated VMB cluster (with 16S-proven  L .  iners  (n i ) = 0, and with 16S-proven  L .  crispatus  (n c ) = 5),  L .  iners -dominated VMB cluster (n i  = 11, n c  = 4), moderate dysbiosis (n i  = 12, n c  = 2); and severe dysbiosis (n i  = 8, n c  = 2). The relative abundances of proteins that were considered specific for  L .  iners  and  L .  crispatus  were compared among VMB groups.    Results  Forty  Lactobacillus  proteins were identified of which 7 were specific for  L .  iners  and 11 for  L .  crispatus . The relative abundances of  L .  iners  DNA starvation/stationary phase protection protein (DPS), and the glycolysis enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and glucose-6-phosphate isomerase (GPI), were significantly decreased in women with  L .  iners -containing dysbiosis compared to women with a  L .  iners -dominated VMB, independent of vaginal pH and  L .  iners  abundance. Furthermore,  L .  iners  DPS, GAPDH, GPI, and fructose-bisphosphate aldolase (ALDO) were significantly negatively associated with vaginal pH. Glycolysis enzymes of  L .  crispatus  showed a similar negative, but nonsignificant, trend related to dysbiosis.    Conclusions  Most identified  Lactobacillus  proteins had conserved intracellular functions, but their high abundance in CVL supernatant might imply an additional extracellular (moonlighting) role. Our findings suggest that these proteins can be important in maintaining a  Lactobacillus -dominated VMB. Functional studies are needed to investigate their roles in vaginal bacterial communities and whether they can be used to prevent vaginal dysbiosis.

Unique Insights in the Cervicovaginal Lactobacillus iners and L. crispatus Proteomes and Their Associations with Microbiota Dysbiosis