Abstract Background Human papillomavirus (HPV) is the main cause of cervical cancer. HPV-vaccines have led to a significant decrease in HPV-infections and related cancer cases. The estimation of the current HPV-prevalence and distribution of different HPV-types among women with cervical dysplasia is important for the future vaccination strategy. Methods By using a multiplexed bead-based immunoassay, we revealed the prevalence of 27 HPV-types in 168 dysplasia women aged 21–70 from Uppsala University hospital, Sweden. Results The prevalence of HPV in low-and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively) were 56.3% and 76.7%, respectively. The oncogenic HPV-types constituted 80.0%, and 97.1% among the HPV-positive LSIL and HSIL-groups, respectively, with HPV16 as the most prevalent type. We found a reduction in oncogenic HPV-types covered by the bi- and quadrivalent vaccines in the vaccinated HSIL-group, suggesting the effectiveness of the HPV-vaccine in preventing dysplasia caused by the covered HPV-types. Oncogenic HPV-types 39 and 59, not covered by any current vaccine have an important prevalence among patients with cervical dysplasia. Conclusions Oncogenic-HPV-types are highly prevalent among women with HSIL. The current vaccine presents effectiveness for reducing the covered HPV-types among dysplasia patients.
Abstract Natural-product derived lectins can function as potent viral inhibitors with minimal toxicity as shown in vitro and in small animal models. We here assessed the effect of rectal application of an anti-HIV lectin-based microbicide Q-Griffithsin (Q-GRFT) in rectal tissue samples from rhesus macaques. E-cadherin + cells, CD4 + cells and total mucosal cells were assessed using in situ staining combined with a novel customized digital image analysis platform. Variations in cell numbers between baseline, placebo and Q-GRFT treated samples were analyzed using random intercept linear mixed effect models. The frequencies of rectal E-cadherin + cells remained stable despite multiple tissue samplings and Q-GRFT gel (0.1%, 0.3% and 1%, respectively) treatment. Whereas single dose application of Q-GRFT did not affect the frequencies of rectal CD4 + cells, multi-dose Q-GRFT caused a small, but significant increase of the frequencies of intra-epithelial CD4 + cells (placebo: median 4%; 1% Q-GRFT: median 7%) and of the CD4 + lamina propria cells (placebo: median 30%; 0.1–1% Q-GRFT: median 36–39%). The resting time between sampling points were further associated with minor changes in the total and CD4 + rectal mucosal cell levels. The results add to general knowledge of in vivo evaluation of anti-HIV microbicide application concerning cellular effects in rectal mucosa.
Abstract Alterations in the vaginal microbiota, including both species composition and functional pathways, have been associated with HPV infection and progression of dysplasia to cervical cancer. To further explore this, shotgun metagenomic sequencing was used to taxonomically and functionally characterize the vaginal microbiota of women with and without cervical dysplasia. Women with histologically verified dysplasia (n = 177; low grade dysplasia (LSIL) n = 81, high-grade dysplasia (HSIL) n = 94, cancer n = 2) were compared with healthy controls recruited from the cervical screening programme (n = 177). Women with dysplasia had a higher vaginal microbial diversity, and higher abundances of Gardnerella vaginalis, Aerococcus christensenii, Peptoniphilus lacrimalis and Fannyhessea vaginae , while healthy controls had higher relative abundance of Lactobacillus crispatus . Genes involved in e.g. nucleotide biosynthesis and peptidoglycan biosynthesis were more abundant in women with dysplasia. Healthy controls showed higher abundance of genes important for e.g. amino acid biosynthesis, (especially L-lysine) and sugar degradation. These findings suggest that the microbiota may have a role in creating a pro-oncogenic environment in women with dysplasia. Its role and potential interactions with other components in the microenvironment deserve further exploration.
Genital mucosa is the main portal of entry for various incoming pathogens, including human immunodeficiency virus (HIV), hence it is an important site for host immune defenses. Tissue-resident memory T (TRM) cells defend tissue barriers against infections and are characterized by expression of CD103 and CD69. In this study, we describe the composition of CD8+ TRM cells in the ectocervix of healthy and HIV-infected women. Study samples were collected from healthy Swedish and Kenyan HIV-infected and uninfected women. Customized computerized image-based in situ analysis was developed to assess the ectocervical biopsies. Genital mucosa and blood samples were assessed by flow cytometry. Although the ectocervical epithelium of healthy women was populated with bona fide CD8+ TRM cells (CD103+CD69+), women infected with HIV displayed a high frequency of CD103−CD8+ cells residing close to their epithelial basal membrane. Accumulation of CD103−CD8+ cells was associated with chemokine expression in the ectocervix and HIV viral load. CD103+CD8+ and CD103−CD8+ T cells expressed cytotoxic effector molecules in the ectocervical epithelium of healthy and HIV-infected women. In addition, women infected with HIV had decreased frequencies of circulating CD103+CD8+ T cells. Our data provide insight into the distribution of CD8+ TRM cells in human genital mucosa, a critically important location for immune defense against pathogens, including HIV.
Abstract Background: There is increasing evidence that the vaginal microbiome influences women’s health. V agina l dysbiosis with a low abundance of L actobacillus h as been connected to gynaecological and obstetric complications. However, the interplay between the vaginal microbiome and the microbiome in other body sites is not yet described. In addition, fluctuating endogenous sex hormones may exert an effect on microbiome composition and are markedly changed by hormonal contraception . This study includes a cohort of 160 healthy young Caucasian women using three different contraceptive regimens: non-hormonal methods, combined oral contraceptive (COC) or levonorgestrel intra-uterine system (LNG-IUS ) . The oral, vaginal, rectal and faecal microbiome s are characterized by shot gun sequencing during each phase of the menstrual cycle (menses, follicular and luteal phases). Results: The use of COC and LNG-IUS do not affect the microbiome composition or diversity . However , an increased diversity in the vaginal microbiome is observed during menses , followed by a subsequent expansion of Lactobacillus during the follicular and luteal phase s which correlate s with measured serum oestradiol levels (r = 0.11, p<0.001) . During menses 89 women (58%) ha ve a dysbiotic vaginal microbiome with less than 60% Lactobacillus spp . This decline s to 49 (32%) in the follicular phase (p = 1.3E-5) and 44 (29%) in the luteal phase (p = 4.6E-7) . During menses, bacterial richness and diversity in saliva reach its lowest while no difference is observed in the faecal microbiome . T he microbiome in different body sites is on average more similar within the same individual than between individuals , despite phase or hormonal treatment . Only the vagina present s a clear cluster structure with dominance of either Lactobacillus crispatus , Lactobacillus iners , Gardnerella vaginalis or Prevotella spp. Conclusions: The use of COC or LNG-IUS is not associated with changes of the healthy female microbiome except for increased stability in the vagina in LNG-IUS users . Body site is the main driver of the microbiome, including a clear difference between faecal and rectal samples. The menstrual cycle is a confounding factor for microbiome composition in both saliva and the vagina and should be considered when analysing the microbiome in women of reproductive age.
Chronic systemic immune activation significantly influences human immunodeficiency virus (HIV) disease progression. Despite evidence of a pro-inflammatory environment in the genital tract of HIV-infected women, comprehensive investigations into cervical tissue from this region remain limited. Similarly, the consequences of chronic HIV infection on the integrity of the female genital epithelium are poorly understood, despite its importance in HIV transmission and replication. Ectocervical biopsies were obtained from HIV-seropositive (n = 14) and HIV-seronegative (n = 47) female Kenyan sex workers. RNA sequencing and bioimage analysis of epithelial junction proteins (E-cadherin, desmoglein-1, claudin-1, and zonula occludens-1) were conducted, along with CD4 staining. RNA sequencing revealed upregulation of immunoregulatory genes in HIV-seropositive women, primarily associated with heightened T cell activity and interferon signaling, which further correlated with plasma viral load. Transcription factor analysis confirmed the upregulation of pro-inflammatory transcription factors, such as RELA , NFKB1 , and IKZF3 , which facilitates HIV persistence in T cells. Conversely, genes and pathways associated with epithelial barrier function and structure were downregulated in the context of HIV. Digital bioimage analysis corroborated these findings, revealing significant disruption of various epithelial junction proteins in ectocervical tissues of the HIV-seropositive women. Thus, chronic HIV infection associated with ectocervical inflammation, characterized by induced T cell responses and interferon signaling, coupled with epithelial disruption. These alterations may influence HIV transmission and heighten susceptibility to other sexually transmitted infections. These findings prompt exploration of therapeutic interventions to address HIV-related complications and mitigate the risk of sexually transmitted infection transmission.
Certain compositions of vaginal microbiota, and specific bacterial species, seem to be associated with HPV infection and the subsequent development of cervical dysplasia and cancer. In order to better understand the association between vaginal microbiota, HPV-infection and dysplasia, we performed shotgun metagenomic sequencing to taxonomically and functionally characterize the composition of the vaginal microbiota of women with and without cervical dysplasia. The HPV status for all study persons was also analysed.
Methodology
Women with histologically verified cervical dysplasia (n = 161; low grade dysplasia (LSIL) n=73, high-grade dysplasia (HSIL) n= 88) were recruited at Uppsala University hospital, Sweden. Women with two normal consecutive cervical screening tests were included as controls (n= 175) Samples were sequenced using shotgun metagenomics, ALDEx2 was used for differential abundance analysis of metagenomic data, Kraken and Optivag databases for taxonomic data, and metaphlan3 and Humann3 for functional data. All samples were analysed for HPV using Luminex.
Results
A total of 336 women were recruited between 2017–2020. The vaginal microbiota diversity increased with increasing severity of the dysplasia (alpha-diversity measures, Shannon diversity median values: normal =0.771, LSIL= 1.027, HSIL=1.150, and inverse Simpson diversity: normal=1.486, LSIL=1.837, HSIL =2.216). There was a significant difference in diversity when comparing normal to HSIL group (Shannon p < .0001, Inverse Simpson: p < .0001), Figure 1.The relative abundance of Lactobacilli species decreased with increased severity of dysplasia, especially L crispatus. L iners and G vaginalis were more common among LSIL and the vaginal microbiota of the high grade dysplasia were characterized by mainly non-lactobacilli species, for example Fecalibacterium prausnitzii, Eubacterium rectale, Bacteroides uniformis, Blautia obeum and Rumiinococcus bromii.
Conclusion
The vaginal microbiota diversity increased with increasing severity of dysplasia. Further, LSIL and HSIL were characterized by different vaginal microbiota compositions.
Abstract Background: There is increasing evidence that the vaginal microbiome influences women’s health. V agina l dysbiosis with a low abundance of L actobacillus h as been connected to gynaecological and obstetric complications. However, the interplay between the vaginal microbiome and the microbiome in other body sites is not yet described. In addition, fluctuating endogenous sex hormones may exert an effect on microbiome composition and are markedly changed by hormonal contraception . This study includes a cohort of 160 healthy young Caucasian women using three different contraceptive regimens: non-hormonal methods, combined oral contraceptive (COC) or levonorgestrel intra-uterine system (LNG-IUS ) . The oral, vaginal, rectal and faecal microbiome s are characterized by shot gun sequencing during each phase of the menstrual cycle (menses, follicular and luteal phases). Results: The use of COC and LNG-IUS do not affect the microbiome composition or diversity . However , an increased diversity in the vaginal microbiome is observed during menses , followed by a subsequent expansion of Lactobacillus during the follicular and luteal phase s which correlate s with measured serum oestradiol levels (r = 0.11, p<0.001) . During menses 89 women (58%) ha ve a dysbiotic vaginal microbiome with less than 60% Lactobacillus spp . This decline s to 49 (32%) in the follicular phase (p = 1.3E-5) and 44 (29%) in the luteal phase (p = 4.6E-7) . During menses, bacterial richness and diversity in saliva reach its lowest while no difference is observed in the faecal microbiome . T he microbiome in different body sites is on average more similar within the same individual than between individuals , despite phase or hormonal treatment . Only the vagina present s a clear cluster structure with dominance of either Lactobacillus crispatus , Lactobacillus iners , Gardnerella vaginalis or Prevotella spp. Conclusions: The use of COC or LNG-IUS is not associated with changes of the healthy female microbiome except for increased stability in the vagina in LNG-IUS users . Body site is the main driver of the microbiome, including a clear difference between faecal and rectal samples. The menstrual cycle is a confounding factor for microbiome composition in both saliva and the vagina and should be considered when analysing the microbiome in women of reproductive age.
Background Despite mounting evidence of gut-brain involvement in psychiatric conditions, functional data remain limited, and analyses of other microbial niches, such as the vaginal microbiota, are lacking in relation to mental health. This aim of this study was to investigate if the connections between the gut microbiome and mental health observed in populations with a clinical diagnosis of mental illness extend to healthy women experiencing stress and depressive symptoms. Additionally, this study examined the functional pathways of the gut microbiota according to the levels of psychological symptoms. Furthermore, the study aimed to explore potential correlations between the vaginal microbiome and mental health parameters in young women without psychiatric diagnoses. Methods In this cross-sectional study, 160 healthy Danish women (aged 18-40 years) filled out questionnaires with validated scales measuring symptoms of stress and depression and frequency of dietary intake. Fecal and vaginal microbiota samples were collected at the beginning of the menstrual cycle and vaginal samples were also collected at cycle day 8-12 and 18-22. Shotgun metagenomic profiling of the gut and vaginal microbiome was performed. The Kyoto Encyclopedia of Genes and Genomes (KEGG) was used for functional profiling and 56 Gut Brain Modules were analyzed in the fecal samples. Results The relative abundance in the gut of the genera Escherichia , Parabacteroides , and Shigella was higher in women with elevated depressive symptoms. Women with high perceived stress showed a tendency of increased abundance of Escherichia , Shigella , and Blautia . Amongst others, the potentially pathogenic genera, Escherichia and Shigella correlate with alterations in the neuroactive pathways such as the glutamatergic, GABAeric, dopaminergic, and Kynurenine pathways. Vaginosis symptoms were more prevalent in women reporting high levels of stress and depressive symptoms. Conclusions The findings of this study support the concept of a microbiota-associated effect on the neuroactive pathways even in healthy young women. This suggest, that targeting the gut microbiome could be a promising approach for future psychiatric interventions.
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