Figure 1.
Sample collection proceeded from the introitus to the cervix. A Dacron swab was rolled 360° along the vaginal lateral wall. A flocked swab was rolled 360° along the opposite vaginal lateral wall. After vaginal swabs, endocervical swabs were taken with first a Dacron swab and then a flocked swab inserted into the cervical os and turned 360°. Finally, the women were randomized to have a CVL collected with 10 ml of saline, Normosol-R, or tap water. Samples were processed as described in the Methods section.
Figure 2.
Protein levels in female genital tract secretions collected by swabs and cervicovaginal lavages (CVL).
Female genital tract secretions were collected by Dacron swabs (DS) and flocked swabs (FS) from the vagina and the endocervix (cervix) and by CVL using Normosol-R, saline, or water. Protein levels were determined using the Bradford assay. Data are presented as box and whisker plots where the median is the horizontal line through the vertical box which represents the 25–75th percentiles. Values within the 10–90th percentiles are represented by the error bars. Outliers are shown by filled circles. Data were log-transformed and significant changes were determined using linear mixed model and discussed in the results section.
Table 1.
Levels of immune mediators and endogenous antimicrobial activity in female genital tract samples by collection sites and methods.
Table 2.
Levels of immune mediators and endogenous antimicrobial activity in female genital tract samples by CVL diluents.
Figure 3.
Levels of mediators in female genital tract secretions collected by swabs and cervicovaginal lavages (CVL).
Female genital tract secretions were collected by Dacron swabs (DS) and flocked swabs (FS) from the vagina and the endocervix (cervix) and by CVL using Normosol-R, saline, or water. (A) Cytokine levels and IL-8 were measured by luminex technology. (B) Antimicrobial protein levels were measured by commercially available ELISA. β-defensins were measured only in CVL due to limited sample volume from the swabs. Data are presented as box and whisker plots where the median is the horizontal line through the vertical box which represents the 25–75th percentiles. Values within the 10–90th percentiles are represented by the error bars. Outliers are shown by filled circles. Data were log-transformed and significant changes were determined using linear mixed model and discussed in the results section.
Figure 4.
Levels of secretory leukocyte protease inhibitor (SLPI) in female genital tract secretions collected by swabs and cervicovaginal lavages (CVL) in women without or with bacterial vaginosis (BV).
Female genital tract secretions were collected by Dacron swabs (DS) and flocked swabs (FS) from the vagina and the endocervix (cervix) and by CVL using Normosol-R, saline, or water. Women with BV (n = 24) had a Nugent score of ≥7. Data are presented as box and whisker plots where the median is the horizontal line through the vertical box which represents the 25–75th percentiles. Values within the 10–90th percentiles are represented by the error bars. Data were log-transformed and significant changes were determined using linear mixed model and discussed in the results section.
Figure 5.
Levels of antimicrobial activity in female genital tract secretions collected by cervicovaginal lavages (CVL) and swabs.
Female genital tract secretions were collected by Dacron swabs (DS) and flocked swabs (FS) from the vagina and the endocervix (cervix) and by CVL using Normosol-R, saline, or water. Anti-E. coli activity was calculated as the % inhibition of bacterial growth compared to an untreated control. Anti-HIV-1 activity was calculated as the % inhibition of infection as compared to an untreated control. CVL/water was not tested (nt) because water would lyse the pathogens or cells providing non-reportable results. Negative values reflect increased E. coli growth or enhanced HIV-1 infection. Data are presented as box and whisker plots where the median is the horizontal line through the vertical box which represents the 25–75th percentiles. Values within the 10–90th percentiles are represented by the error bars. Outliers are shown by filled circles. Significant changes were determined using linear mixed model and discussed in the results section.