Measurement of the transmitted HIV-1 IIIB diversity from serially diluted viral inoculums.

<p>The viral diversity of the viral inoculum is 1.20±0.09%.</p><p>*No significant difference in viral diversity between the inoculum and all the three different transmitted variants.</p

Characteristics of Viral Inoculums Used in the HIV-1 Transmission and Diversity Studies.

<p>Characteristics of Viral Inoculums Used in the HIV-1 Transmission and Diversity Studies.</p

The competition analysis of the transmission efficiency for the transmitted/founder viruses and chronic HIV-1 isolates.

<p>HIV-1 CH040 and CH058 are transmitted/founder viruses and HIV-1 BAL is a chronic isolate.</p><p>*The transmission efficiency was calculated by dividing the viral RNA copies in the inoculum with the viral RNA copies of the transmitted variants from the bottom wells. Real-time RT PCR with specific primers/probes set which could discriminate CH040, CH058 and BAL viruses was employed.</p

Diversity analysis of the six HIV-1 isolates before and after their mucosal transmission.

<p>Diversity analysis of envelope sequences (C2–V5) from different isolates was performed using MEGA5.0 software. Nucleic acid intra-population diversity was calculated based on nucleotide sequences. The differences between the diversity were plotted as intra-population distances as a function of inoculums and transmitted viruses. Statistical significance of the diversity differences between the inoculums and transmitted viruses was performed using two-sample tests <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032539#pone.0032539-Gilbert1" target="_blank">[25]</a>. (A) HIV-1 subtype B, IIIB; (B) HIV-1 subtype B, BAL; (C) HIV-1 subtype B, clinical isolate 074; (D) HIV-1 subtype B, clinical isolate 015, (E) HIV-1 subtype A, RW92008; (F) HIV-1 subtype C isolate, IN93999.</p

Comparison of distance distribution between the inoculums and the transmitted viral sequences.

<p>The distance distribution for the inoculums and transmitted pairs was shown in pairwise distance histogram. For each pair, the distances between every possibility of pair sequences of inoculums and transmitted variants were calculated respectively. (A) HIV-1 subtype B, IIIB; (B) HIV-1 subtype B, BAL; (C) HIV-1 subtype B, clinical isolate 074; (D) HIV-1 subtype B, clinical isolate 015; (E) HIV-1 subtype A, RW92008; (F) HIV-1 subtype C, IN93999. Black needle bars represent the inoculums pairwise viral sequence distance frequency. Red needle bars represented the transmitted pairwise viral sequence distance frequency.</p

Transmission of EIAV and HIV across cervical mucosa in an organ culture.

<p>*Denotes transmission efficiency of HIV or EIAV RNA from a mixture of EIAV and HIV(1∶1) inoculum. Transmission efficiency was calculated as described in the materials and methods section.</p

Phylogenetic analysis of all viral inoculum and transmitted pairs.

<p>C2-V5 of <i>env</i> nucleotide sequences of the inoculum and transmitted HIV-1 species were aligned for all linked inoculum variants (green lines) and transmitted variants (red lines) and a neighbor-joining tree representing different transmission pairs. Branch lengths are drawn to scale. Viral strain names are shown on the branches, and asterisks are indicated on branches with bootstrap values greater than 99%.</p

Neighbor-joining trees of HIV-1 env C2-V5 sequences individually from six inoculum and transmission pairs.

<p>Aligned nucleotide sequences for six inoculums and transmitted variants were used to generate neighbor-joining trees for individual viral pairs. (A) HIV-1 subtype B, IIIB; (B) HIV-1 subtype B, BAL; (C) HIV-1 subtype B, clinical isolate 074; (D) HIV-1 subtype B, clinical isolate 015; (E) HIV-1 subtype A, RW92008; (F) HIV-1 subtype C, IN93999. White circles: the sequences of the inoculum viruses; black circles: the sequences of the transmitted viruses.</p

<i>L. vaginalis</i> Initiates an Innate Immune Response from FRT Epithelia.

<p>Confluent monolayers of epithelial cells were inoculated with bacteria and coincubated for 24 hr. Conditioned media were collected, clarified and ELISA was used to quantify hBD2, IL-6 and IL-8 concentrations. MOI is the same as in Figure Five. Analyte concentrations are shown as fold induction compared to mock condition, and are the average of three independent experiments where one (p<0.05), two (p<0.01), or three (p<0.001) asterisks indicate a significant increase over the mock-treated condition.</p

Identification and Characterization of Bacterial Vaginosis-Associated Pathogens Using a Comprehensive Cervical-Vaginal Epithelial Coculture Assay

<div><p>Bacterial vaginosis (BV) is the most commonly treated female reproductive tract affliction, characterized by the displacement of healthy lactobacilli by an overgrowth of pathogenic bacteria. BV can contribute to pathogenic inflammation, preterm birth, and susceptibility to sexually transmitted infections. As the bacteria responsible for BV pathogenicity and their interactions with host immunity are not understood, we sought to evaluate the effects of BV-associated bacteria on reproductive epithelia. Here we have characterized the interaction between BV-associated bacteria and the female reproductive tract by measuring cytokine and defensin induction in three types of FRT epithelial cells following bacterial inoculation. Four BV-associated bacteria were evaluated alongside six lactobacilli for a comparative assessment. While responses differed between epithelial cell types, our model showed good agreement with clinical BV trends. We observed a distinct cytokine and human β-defensin 2 response to BV-associated bacteria, especially <em>Atopobium vaginae</em>, compared to most lactobacilli. One lactobacillus species, <em>Lactobacillus vaginalis</em>, induced an immune response similar to that elicited by BV-associated bacteria, stimulating significantly higher levels of cytokines and human β-defensin 2 than other lactobacilli. These data provide an important prioritization of BV-associated bacteria and support further characterization of reproductive bacteria and their interactions with host epithelia. Additionally, they demonstrate the distinct immune response potentials of epithelial cells from different locations along the female reproductive tract.</p> </div