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1.
Front Reprod Health ; 4: 876729, 2022.
Article in English | MEDLINE | ID: mdl-36303633

ABSTRACT

The role of neutrophils relative to vaginal dysbiosis is unclear. We hypothesize that bacterial vaginosis (BV)-associated bacteria may induce the activation and accumulation of mucosal neutrophils within the female reproductive tract (FRT), resulting in epithelial barrier damage. We collected endocervical cytobrushes from women with and without BV and assessed bacteria community type and frequency/functional phenotypes of neutrophils. We performed in vitro whole blood co-cultures with BV-associated bacteria and healthy vaginal commensals and assessed their impact on epithelial integrity using transepithelial electrical resistance. We demonstrated increased neutrophil frequency (p < 0.0001), activation (p < 0.0001), and prolonged lifespan (p < 0.0001) in the cytobrushes from women with non-Lactobacillus dominant (nLD) communities. Our in vitro co-cultures confirmed these results and identified significant barrier damage in the presence of neutrophils and G. vaginalis. Here, we demonstrate that BV-associated bacteria induce neutrophil activation and increase lifespan, potentially causing accumulation in the FRT and epithelial barrier damage.

2.
Cell ; 184(15): 3899-3914.e16, 2021 07 22.
Article in English | MEDLINE | ID: mdl-34237254

ABSTRACT

The impact of the microbiome on HIV disease is widely acknowledged although the mechanisms downstream of fluctuations in microbial composition remain speculative. We detected rapid, dynamic changes in translocated microbial constituents during two years after cART initiation. An unbiased systems biology approach revealed two distinct pathways driven by changes in the abundance ratio of Serratia to other bacterial genera. Increased CD4 T cell numbers over the first year were associated with high Serratia abundance, pro-inflammatory innate cytokines, and metabolites that drive Th17 gene expression signatures and restoration of mucosal integrity. Subsequently, decreased Serratia abundance and downregulation of innate cytokines allowed re-establishment of systemic T cell homeostasis promoting restoration of Th1 and Th2 gene expression signatures. Analyses of three other geographically distinct cohorts of treated HIV infection established a more generalized principle that changes in diversity and composition of translocated microbial species influence systemic inflammation and consequently CD4 T cell recovery.


Subject(s)
Gastrointestinal Microbiome , HIV Infections/immunology , HIV Infections/microbiology , Antiretroviral Therapy, Highly Active , Biodiversity , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Chemokines/blood , Cohort Studies , Glycolysis , HIV Infections/blood , HIV Infections/drug therapy , Humans , Inflammation/genetics , Inflammation/pathology , Mitochondria/metabolism , Monocytes/metabolism , Nucleic Acids/blood , Principal Component Analysis , Serratia/physiology , Th1 Cells/immunology , Th2 Cells/immunology , Transcription, Genetic , Uganda , Viral Load/immunology
3.
Nat Commun ; 11(1): 6147, 2020 12 01.
Article in English | MEDLINE | ID: mdl-33262350

ABSTRACT

Bacterial vaginosis is a condition associated with adverse reproductive outcomes and characterized by a shift from a Lactobacillus-dominant vaginal microbiota to a polymicrobial microbiota, consistently colonized by strains of Gardnerella vaginalis. Metronidazole is the first-line treatment; however, treatment failure and recurrence rates remain high. To understand complex interactions between Gardnerella vaginalis and Lactobacillus involved in efficacy, here we develop an ordinary differential equation model that predicts bacterial growth as a function of metronidazole uptake, sensitivity, and metabolism. The model shows that a critical factor in efficacy is Lactobacillus sequestration of metronidazole, and efficacy decreases when the relative abundance of Lactobacillus is higher pre-treatment. We validate results in Gardnerella and Lactobacillus co-cultures, and in two clinical cohorts, finding women with recurrence have significantly higher pre-treatment levels of Lactobacillus relative to bacterial vaginosis-associated bacteria. Overall results provide mechanistic insight into how personalized differences in microbial communities influence vaginal antibiotic efficacy.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Metronidazole/administration & dosage , Microbiota , Vaginosis, Bacterial/drug therapy , Bacteria/classification , Bacteria/drug effects , Bacteria/genetics , Bacteria/growth & development , Cohort Studies , Female , Gardnerella vaginalis/drug effects , Gardnerella vaginalis/genetics , Gardnerella vaginalis/growth & development , Humans , Lactobacillus/drug effects , Lactobacillus/genetics , Lactobacillus/growth & development , Microbiota/drug effects , Treatment Outcome , Vagina/drug effects , Vagina/microbiology , Vaginosis, Bacterial/microbiology
4.
PLoS Pathog ; 16(12): e1009024, 2020 12.
Article in English | MEDLINE | ID: mdl-33270801

ABSTRACT

Despite the efficacy of antiretroviral-based pre-exposure prophylactics (PrEP) in men who have sex with men, studies in women have produced widely varying outcomes. Recent evidence demonstrates that vaginal microbial communities are associated with increased HIV acquisition risk and may impact PrEP efficacy. Here, we investigate the mechanisms underlying how vaginal bacteria alter PrEP drug levels and impact HIV infection rates ex vivo. Using cervicovaginal lavages (CVLs) from women with or without bacterial vaginosis (BV), we identified microbial metabolism of PrEP drugs in BV samples through LC-MS/MS analysis of soluble drug levels and metabolite formation in dual T-cell cultures. CVL samples were assessed for microbiome analysis using sequencing of bacterial 16S rRNA genes. We also observed non-Lactobacillus bacteria that are associated with BV may potentially impact PrEP efficacy through increased HIV infection rates in co-cultures containing Lactobacillus or BV bacteria, PrEP drugs, CEM-GFP cells, and HIV-1LAI virus. Finally, we used these data to develop a novel predictive mathematical simulation modeling system to predict these drug interactions for future trials. These studies demonstrate how dysbiotic vaginal microbiota may impact PrEP drugs and provides evidence linking vaginal bacteria to PrEP efficacy in women.


Subject(s)
HIV Infections/transmission , Microbiota/physiology , Pre-Exposure Prophylaxis/methods , Vagina/microbiology , Adult , Anti-HIV Agents/therapeutic use , Anti-Retroviral Agents/therapeutic use , Chromatography, Liquid/methods , Dysbiosis/microbiology , Female , HIV Infections/drug therapy , HIV-1/metabolism , HIV-1/pathogenicity , Humans , Microbiota/genetics , RNA, Ribosomal, 16S/genetics , Tandem Mass Spectrometry/methods , Treatment Outcome , Vagina/drug effects , Vaginosis, Bacterial/complications , Vaginosis, Bacterial/drug therapy
6.
Mucosal Immunol ; 13(3): 471-480, 2020 05.
Article in English | MEDLINE | ID: mdl-31797911

ABSTRACT

The diverse bacterial communities that colonize the gastrointestinal tract play an essential role in maintaining immune homeostasis through the production of critical metabolites such as short-chain fatty acids (SCFAs) and this can be disrupted by antibiotic use. However, few studies have addressed the effects of specific antibiotics longitudinally on the microbiome and immunity. We evaluated the effects of four specific antibiotics: enrofloxacin, cephalexin, paromomycin, and clindamycin, in healthy female rhesus macaques. All antibiotics disrupted the microbiome, including reduced abundances of fermentative bacteria and increased abundances of potentially pathogenic bacteria, including Enterobacteriaceae in the stool, and decreased Helicobacteraceae in the colon. This was associated with decreased SCFAs, indicating altered bacterial metabolism. Importantly, antibiotic use also substantially altered local immune responses, including increased neutrophils and Th17 cells in the colon. Furthermore, we observed increased soluble CD14 in plasma, indicating microbial translocation. These data provide a longitudinal evaluation of antibiotic-induced changes to the composition and function of colonic bacterial communities associated with specific alterations in mucosal and systemic immunity.


Subject(s)
Anti-Bacterial Agents/pharmacology , Colon , Gastrointestinal Microbiome/drug effects , Immunity, Mucosal/drug effects , Intestinal Mucosa/drug effects , Intestinal Mucosa/immunology , Intestinal Mucosa/microbiology , Animals , Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/pharmacokinetics , Bacteria , Biodiversity , Biomarkers , Drug Administration Schedule , Drug Monitoring , Fatty Acids, Volatile/metabolism , Feces/cytology , Feces/microbiology , Gas Chromatography-Mass Spectrometry , Immunophenotyping , Intestinal Mucosa/pathology , Macaca mulatta , Neutrophil Infiltration/drug effects , Neutrophil Infiltration/immunology , T-Lymphocyte Subsets/drug effects , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , Tissue Distribution
7.
Invest Ophthalmol Vis Sci ; 60(6): 2218-2225, 2019 05 01.
Article in English | MEDLINE | ID: mdl-31108552

ABSTRACT

Purpose: To evaluate relationships among retinal vascular caliber and biomarkers of systemic inflammation in patients with AIDS. Methods: A total of 454 participants with AIDS had retinal vascular caliber (central retinal artery equivalent and central retinal vein equivalent) determined from enrollment retinal photographs by reading center graders masked to clinical and biomarker information. Cryopreserved plasma specimens were assayed for inflammatory biomarkers, including C-reactive protein (CRP), IL-6, interferon-γ inducible protein (IP)-10, kynurenine/tryptophan (KT) ratio, and intestinal fatty acid binding protein (I-FABP). Results: In the simple linear regression of retinal vascular caliber on plasma biomarkers, elevated CRP, IL-6, and IP-10 were associated with retinal venular dilation, and elevated KT ratio with retinal arteriolar narrowing. In the multiple linear regression, including baseline characteristics and plasma biomarkers, AMD was associated with dilation of retinal arterioles (mean difference: 9.1 µm; 95% confidence interval [CI] 5.2, 12.9; P < 0.001) and venules (mean difference, 10.9 µm; 95% CI, 5.3, 16.6; P < 0.001), as was black race (P < 0.001). Hyperlipidemia was associated with retinal venular narrowing (mean difference, -7.5 µm; 95% CI, -13.7, -1.2; P = 0.02); cardiovascular disease with arteriolar narrowing (mean difference, -5.2 µm; 95% CI, -10.3, -0.1; P = 0.05); age with arteriolar narrowing (slope, -0.26 µm/year; 95% CI, -0.46, -0.06; P = 0.009); and IL-6 with venular dilation (slope, 5.3 µm/standard deviation log10[plasma IL-6 concentration]; 95% CI, 2.7, 8.0; P < 0.001). Conclusions: These data suggest that retinal vascular caliber is associated with age, race, AMD, hyperlipidemia, cardiovascular disease, and selected biomarkers of systemic inflammation.


Subject(s)
Acquired Immunodeficiency Syndrome/pathology , Inflammation/pathology , Retinal Vessels/pathology , Adult , Age Factors , Arterioles/pathology , Biomarkers , Cardiovascular Diseases/complications , Female , Humans , Linear Models , Male , Middle Aged , Retinal Artery/pathology , Retinal Vein/pathology , Risk Factors , Venules/pathology
8.
PLoS Pathog ; 15(4): e1007672, 2019 04.
Article in English | MEDLINE | ID: mdl-30973942

ABSTRACT

Gastrointestinal (GI) mucosal dysfunction predicts and likely contributes to non-infectious comorbidities and mortality in HIV infection and persists despite antiretroviral therapy. However, the mechanisms underlying this dysfunction remain incompletely understood. Neutrophils are important for containment of pathogens but can also contribute to tissue damage due to their release of reactive oxygen species and other potentially harmful effector molecules. Here we used a flow cytometry approach to investigate increased neutrophil lifespan as a mechanism for GI neutrophil accumulation in chronic, treated HIV infection and a potential role for gastrointestinal dysbiosis. We report that increased neutrophil survival contributes to neutrophil accumulation in colorectal biopsy tissue, thus implicating neutrophil lifespan as a new therapeutic target for mucosal inflammation in HIV infection. Additionally, we characterized the intestinal microbiome of colorectal biopsies using 16S rRNA sequencing. We found that a reduced Lactobacillus: Prevotella ratio associated with neutrophil survival, suggesting that intestinal bacteria may contribute to GI neutrophil accumulation in treated HIV infection. Finally, we provide evidence that Lactobacillus species uniquely decrease neutrophil survival and neutrophil frequency in vitro, which could have important therapeutic implications for reducing neutrophil-driven inflammation in HIV and other chronic inflammatory conditions.


Subject(s)
Colon/immunology , Gastrointestinal Microbiome/immunology , HIV Infections/immunology , HIV-1/immunology , Inflammation/immunology , Neutrophils/immunology , Rectum/immunology , Colon/microbiology , Colon/pathology , Female , HIV Infections/virology , Humans , Inflammation/pathology , Male , Middle Aged , Neutrophils/cytology , Rectum/microbiology , Rectum/pathology
9.
Am J Ophthalmol ; 199: 230-237, 2019 03.
Article in English | MEDLINE | ID: mdl-30552890

ABSTRACT

PURPOSE: To evaluate the relationships among age-related macular degeneration (AMD), mortality, and biomarkers of systemic inflammation in patients with acquired immunodeficiency syndrome (AIDS). DESIGN: Case-control study. METHODS: In participants with intermediate-stage AMD at enrollment in the Longitudinal Study of the Ocular Complications of AIDS (LSOCA) and 2:1 controls matched for age and sex, cryopreserved baseline plasma specimens were assayed for biomarkers of inflammation, including high-sensitivity C-reactive protein (CRP), interleukin (IL)-6, interferon-γ inducible protein (IP)-10, soluble CD14 (sCD14), soluble CD163 (sCD163), kynurenine/tryptophan (KT) ratio, and intestinal fatty acid binding protein (I-FABP). Main outcome measure was mortality. RESULTS: The study included 189 patients with AMD and 385 controls. In the unadjusted analysis, AMD was associated with mortality (hazard ratio [HR] 1.48; 95% confidence interval [CI] 1.02, 2.15; P = .04). In an adjusted analysis, CRP (HR 1.36; 95% CI 1.08, 1.71; P = .009), IL-6 (HR 1.45; 95% CI 1.11, 1.90; P = .006), and IP-10 (HR 1.41; 95% CI 1.08, 1.84; P = .01) were associated with mortality. In a Cox regression analysis adjusted for human immunodeficiency virus load, blood CD4+ T cell level, CRP, IL-6, and IP-10, the association of AMD with mortality was attenuated (HR 1.08; 95% CI 0.73, 1.59; P = .70), primarily by the addition of the inflammatory biomarkers. CONCLUSIONS: These data suggest that the increased mortality observed in patients with AIDS with AMD is, at least in part, a result of systemic inflammation.


Subject(s)
Acquired Immunodeficiency Syndrome/mortality , Biomarkers/blood , Inflammation/blood , Macular Degeneration/mortality , Acquired Immunodeficiency Syndrome/blood , Adult , Antigens, CD/blood , C-Reactive Protein/metabolism , Case-Control Studies , Chemokine CXCL10/blood , Female , Humans , Interleukin-6/blood , Kynurenine/blood , Longitudinal Studies , Macular Degeneration/blood , Male , Middle Aged , Proportional Hazards Models , Prospective Studies , Risk Factors , Tryptophan/blood
10.
Clin Infect Dis ; 66(12): 1872-1882, 2018 06 01.
Article in English | MEDLINE | ID: mdl-29471387

ABSTRACT

Background: Cannabis is a widely used drug in the United States, and the frequency of cannabis use in the human immunodeficiency virus (HIV)-infected population is disproportionately high. Previous human and macaque studies suggest that cannabis may have an impact on plasma viral load; however, the relationship between cannabis use and HIV-associated systemic inflammation and immune activation has not been well defined. Methods: The impact of cannabis use on peripheral immune cell frequency, activation, and function was assessed in 198 HIV-infected, antiretroviral-treated individuals by flow cytometry. Individuals were categorized into heavy, medium, or occasional cannabis users or noncannabis users based on the amount of the cannabis metabolite 11-nor-carboxy-tetrahydrocannabinol (THC-COOH) detected in plasma by mass spectrometry. Results: Heavy cannabis users had decreased frequencies of human leukocyte antigen (HLA)-DR+CD38+CD4+ and CD8+ T-cell frequencies, compared to frequencies of these cells in non-cannabis-using individuals. Heavy cannabis users had decreased frequencies of intermediate and nonclassical monocyte subsets, as well as decreased frequencies of interleukin 23- and tumor necrosis factor-α-producing antigen-presenting cells. Conclusions: While the clinical implications are unclear, our findings suggest that cannabis use is associated with a potentially beneficial reduction in systemic inflammation and immune activation in the context of antiretroviral-treated HIV infection.


Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , Immunity, Innate/drug effects , Lymphocyte Activation/drug effects , Marijuana Abuse/immunology , Antiretroviral Therapy, Highly Active , CD4-Positive T-Lymphocytes/drug effects , CD8-Positive T-Lymphocytes/drug effects , Dronabinol/analogs & derivatives , Dronabinol/blood , Female , Flow Cytometry , Humans , Inflammation , Male , Middle Aged , Monocytes/drug effects , Viral Load/drug effects
11.
Science ; 356(6341): 938-945, 2017 06 02.
Article in English | MEDLINE | ID: mdl-28572388

ABSTRACT

Antiretroviral-based strategies for HIV prevention have shown inconsistent results in women. We investigated whether vaginal microbiota modulated tenofovir gel microbicide efficacy in the CAPRISA (Centre for the AIDS Program of Research in South Africa) 004 trial. Two major vaginal bacterial community types-one dominated by Lactobacillus (59.2%) and the other where Gardnerella vaginalis predominated with other anaerobic bacteria (40.8%)-were identified in 688 women profiled. Tenofovir reduced HIV incidence by 61% (P = 0.013) in Lactobacillus-dominant women but only 18% (P = 0.644) in women with non-Lactobacillus bacteria, a threefold difference in efficacy. Detectible mucosal tenofovir was lower in non-Lactobacillus women, negatively correlating with G. vaginalis and other anaerobic bacteria, which depleted tenofovir by metabolism more rapidly than target cells convert to pharmacologically active drug. This study provides evidence linking vaginal bacteria to microbicide efficacy through tenofovir depletion via bacterial metabolism.


Subject(s)
Bacteria/metabolism , HIV Infections/microbiology , HIV Infections/prevention & control , Microbiota/physiology , Tenofovir/metabolism , Tenofovir/pharmacology , Vagina/microbiology , Adult , Antiviral Agents/metabolism , Antiviral Agents/pharmacology , Bacteria/genetics , Bacteria/isolation & purification , Biodiversity , Female , Gardnerella/metabolism , Humans , Lactobacillus/metabolism , Mass Spectrometry , Microbiota/genetics , Proteome , RNA, Ribosomal, 16S/genetics , South Africa , Tenofovir/analysis
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