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6: T cell activation in south african HIV-exposed infants linked to ochratoxin exposure
IDSOG Abstracts
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School of Medicine, University of Washington, Seattle, WA, USA, 2Center for Infectious Disease Research Institute, formerly Seattle Biomedical Research Institute, Seattle, WA, USA, 3Divisions of Paediatrics, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 4Seattle Children’s Research Institute, Seattle, Washington, USA
OBJECTIVES: Early introduction of non-breast milk foods is associated with increased morbidity and mortality in the developing world. This is particularly true for HIV-infected infants, who have a more rapid rate of disease progression if they consume non-breast milk foods. Here we describe a potential mechanism to explain how non-breast milk foods might influence HIV disease progression, exposure to food borne toxins, specifically the immunomodulatory mycotoxin, ochratoxin (OTA). METHODS: This study was based in Khayelitsha, South Africa, an informal settlement outside of Cape Town. We enrolled 151 HIVexposed, uninfected infants at birth into a prospective cohort and followed them until 14 weeks of age. Mothers self-selected feeding modes but exclusive feeding was encouraged. Blood and detailed demographic and health data were collected at birth, 2, 6 and 14 weeks of age. Flow cytometry was performed on cryopreserved PBMC to assess T cell activation, plasma cytokines were measured with multiplex assays, and ochratoxin levels were measured using competitive ELISA. RESULTS: Infants that consumed non-breast milk foods had up to 2 fold higher plasma levels of OTA between 6, 8 and 14 weeks of age. OTA plasma levels correlated with expression of activation markers CCR5, CD38, HLADR and Ki67 on CD4 and CD8 T cells as well as plasma levels of the inflammatory markers TNF-alpha and CXCL10. CONCLUSIONS: Infants in South Africa are exposed to the food borne toxin, OTA, in non-breast milk foods, which may induce T cell activation, a known risk factor for HIV disease progression. These findings suggest that the increased rate of HIV disease progression in infants that consume non-breast milk foods may be, in part, mediated by immune activation induced by food borne toxins.
7 Relationships between host innate immune genetic variation, bacterial vaginosis, and bacterial colonization E. dela Cruz1,2, J. Marrazzo3, T. Hawn2, D. Fredricks1,2 1
University of Washington, Seattle, WA, 2Fred Hutchinson Cancer Research Center, Seattle, WA, 3University of Alabama, Birmingham, AL
OBJECTIVES: Our goals were to identify host innate immune
genotypes associated with prevalent bacterial vaginosis (BV) and colonization with BV-associated bacteria (BVAB). Furthermore, we sought to characterize Lactobacillus colonization patterns associated with risk of incident BV. METHODS: We enrolled women with and without BV in a study of the vaginal microbiota. Human genomic DNA was extracted from whole blood and used to genotype a panel of 14 single nucleotide polymorphisms (SNPs) in toll-like receptor-1 (TLR1), TLR2, TLR4, TLR5, TLR6, as well as regulators of TLR signaling, toll-interacting protein (TOLLIP), and CD180. In a cohort of 43 non-Hispanic, Caucasian participants, colonization with 15 bacterial species was assessed by quantitative PCR applied to DNA extracted from vaginal swabs. An independent cohort of 116 women without prevalent BV was followed for a period of approximately 90 days and assessed for
ˇ
1
ˇ
Lianna F. Wood1,2, Heather Jaspan3,4, Don Sodora2
BV by a study clinician at monthly intervals. DNA extracted from vaginal swabs collected at study enrollment was used to assess Lactobacillus colonization by quantitative PCR. RESULTS: We found a relationship between TLR1 deficiency (defined by genotype TLR1 1805GG) and decreased probability of prevalent BV (odds ratio (OR) ¼ 0.12; 95% confidence interval (95% CI)¼ 0.02 to 0.45) as well as decreased colonization with BVAB1 (OR¼0.178; 95% CI¼0.041 to 0.654) and Megasphaera genogroups 1 and 2 (OR¼0.059; 95% CI¼0.003 to 0.353). In 116 study participants without prevalent BV, a subset of 22 women who were colonized with a combination of L. crispatus (10 7 16S rRNA gene copies per swab), L. iners (10 7 copies per swab), and L. gasseri (10 4 copies per swab) did not develop BV over a 90-day follow up period. CONCLUSIONS: These results suggest that variation in Lactobacillus colonization and innate immunity may influence risk of BV, inflammatory response to BVABs, and likelihood of BVAB colonization. We found correlations between TLR1 deficiency and BV status as well as colonization with bacterial species highly sensitive and specific for the presence of BV. TLR1 deficiency segregates with race and is more common among individuals of European descent, and less common among those of African descent. Genetic variation in TLR1 may in part explain increased risk of BV seen in African and African-American women. Additionally, simultaneous colonization with L. crispatus, L. iners, and L. gasseri may be protective against the development of BV. ˇ
6 T cell activation in south african HIV-exposed infants linked to ochratoxin exposure
8 Detection & implications of lactobacillus iners sub-strain diversity J. B. Holm1,2, P. Gajer1, B. Ma1, M. Humphrys1, J. Ravel1,2 1 Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, MD, USA, 2Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, USA
OBJECTIVES: Lactobacillus iners is the most common species in
vaginal microbiota, and is observed in both healthy and dysbiotic states. Sub-strains of Lactobacillus iners may be associated with specific indigenous microbiota and exogenous infection or disease. Here, we sought to determine if sub-strains of Lactobacillus iners were present before and after antibiotic treatment of Chlamydia trachomatis (CT) infection. METHODS: A subset of females was selected from a larger cohort of 129 participants who tested positive for CT at enrollment. Vaginal samples were collected at baseline prior to treatment with a single dose of azithromycin (1 mg) and every 3 months at each of the three subsequent visits. All subjects included in this study tested negative for CT at visits 2, 3, and 4. Vaginal microbiota were characterized via 16S ribosomal RNA gene sequencing and analysis, and the most abundant taxa observed were compared across visits for each subject. Sequences from L. iners were then isolated and evaluated for the presence of sub-strains using “oligotyping” analyses. The Shannon Entropy diversity index of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene was used to detect bacterial sub-strains. The number of L. iners oligotypes (strains) was examined for each subject across all 4 visits. RESULTS: The relative abundance of L. iners increased between visits 1 and 2. Oligotyping analyses revealed 5 SNPs in the 16S rRNA gene with significantly greater diversity indices than all other positions in the alignment, resulting in up to 93 oligotypes (or sub-strains) observed. The number of L. iners strains observed within a subject increased from visit 1 to visit 2, and this diversity was maintained at subsequent visits. DECEMBER 2016 American Journal of Obstetrics & Gynecology
817
ajog.org
School of Medicine, University of Washington, Seattle, WA, USA, 2Center for Infectious Disease Research Institute, formerly Seattle Biomedical Research Institute, Seattle, WA, USA, 3Divisions of Paediatrics, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 4Seattle Children’s Research Institute, Seattle, Washington, USA
OBJECTIVES: Early introduction of non-breast milk foods is associated with increased morbidity and mortality in the developing world. This is particularly true for HIV-infected infants, who have a more rapid rate of disease progression if they consume non-breast milk foods. Here we describe a potential mechanism to explain how non-breast milk foods might influence HIV disease progression, exposure to food borne toxins, specifically the immunomodulatory mycotoxin, ochratoxin (OTA). METHODS: This study was based in Khayelitsha, South Africa, an informal settlement outside of Cape Town. We enrolled 151 HIVexposed, uninfected infants at birth into a prospective cohort and followed them until 14 weeks of age. Mothers self-selected feeding modes but exclusive feeding was encouraged. Blood and detailed demographic and health data were collected at birth, 2, 6 and 14 weeks of age. Flow cytometry was performed on cryopreserved PBMC to assess T cell activation, plasma cytokines were measured with multiplex assays, and ochratoxin levels were measured using competitive ELISA. RESULTS: Infants that consumed non-breast milk foods had up to 2 fold higher plasma levels of OTA between 6, 8 and 14 weeks of age. OTA plasma levels correlated with expression of activation markers CCR5, CD38, HLADR and Ki67 on CD4 and CD8 T cells as well as plasma levels of the inflammatory markers TNF-alpha and CXCL10. CONCLUSIONS: Infants in South Africa are exposed to the food borne toxin, OTA, in non-breast milk foods, which may induce T cell activation, a known risk factor for HIV disease progression. These findings suggest that the increased rate of HIV disease progression in infants that consume non-breast milk foods may be, in part, mediated by immune activation induced by food borne toxins.
7 Relationships between host innate immune genetic variation, bacterial vaginosis, and bacterial colonization E. dela Cruz1,2, J. Marrazzo3, T. Hawn2, D. Fredricks1,2 1
University of Washington, Seattle, WA, 2Fred Hutchinson Cancer Research Center, Seattle, WA, 3University of Alabama, Birmingham, AL
OBJECTIVES: Our goals were to identify host innate immune
genotypes associated with prevalent bacterial vaginosis (BV) and colonization with BV-associated bacteria (BVAB). Furthermore, we sought to characterize Lactobacillus colonization patterns associated with risk of incident BV. METHODS: We enrolled women with and without BV in a study of the vaginal microbiota. Human genomic DNA was extracted from whole blood and used to genotype a panel of 14 single nucleotide polymorphisms (SNPs) in toll-like receptor-1 (TLR1), TLR2, TLR4, TLR5, TLR6, as well as regulators of TLR signaling, toll-interacting protein (TOLLIP), and CD180. In a cohort of 43 non-Hispanic, Caucasian participants, colonization with 15 bacterial species was assessed by quantitative PCR applied to DNA extracted from vaginal swabs. An independent cohort of 116 women without prevalent BV was followed for a period of approximately 90 days and assessed for
ˇ
1
ˇ
Lianna F. Wood1,2, Heather Jaspan3,4, Don Sodora2
BV by a study clinician at monthly intervals. DNA extracted from vaginal swabs collected at study enrollment was used to assess Lactobacillus colonization by quantitative PCR. RESULTS: We found a relationship between TLR1 deficiency (defined by genotype TLR1 1805GG) and decreased probability of prevalent BV (odds ratio (OR) ¼ 0.12; 95% confidence interval (95% CI)¼ 0.02 to 0.45) as well as decreased colonization with BVAB1 (OR¼0.178; 95% CI¼0.041 to 0.654) and Megasphaera genogroups 1 and 2 (OR¼0.059; 95% CI¼0.003 to 0.353). In 116 study participants without prevalent BV, a subset of 22 women who were colonized with a combination of L. crispatus (10 7 16S rRNA gene copies per swab), L. iners (10 7 copies per swab), and L. gasseri (10 4 copies per swab) did not develop BV over a 90-day follow up period. CONCLUSIONS: These results suggest that variation in Lactobacillus colonization and innate immunity may influence risk of BV, inflammatory response to BVABs, and likelihood of BVAB colonization. We found correlations between TLR1 deficiency and BV status as well as colonization with bacterial species highly sensitive and specific for the presence of BV. TLR1 deficiency segregates with race and is more common among individuals of European descent, and less common among those of African descent. Genetic variation in TLR1 may in part explain increased risk of BV seen in African and African-American women. Additionally, simultaneous colonization with L. crispatus, L. iners, and L. gasseri may be protective against the development of BV. ˇ
6 T cell activation in south african HIV-exposed infants linked to ochratoxin exposure
8 Detection & implications of lactobacillus iners sub-strain diversity J. B. Holm1,2, P. Gajer1, B. Ma1, M. Humphrys1, J. Ravel1,2 1 Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, MD, USA, 2Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, USA
OBJECTIVES: Lactobacillus iners is the most common species in
vaginal microbiota, and is observed in both healthy and dysbiotic states. Sub-strains of Lactobacillus iners may be associated with specific indigenous microbiota and exogenous infection or disease. Here, we sought to determine if sub-strains of Lactobacillus iners were present before and after antibiotic treatment of Chlamydia trachomatis (CT) infection. METHODS: A subset of females was selected from a larger cohort of 129 participants who tested positive for CT at enrollment. Vaginal samples were collected at baseline prior to treatment with a single dose of azithromycin (1 mg) and every 3 months at each of the three subsequent visits. All subjects included in this study tested negative for CT at visits 2, 3, and 4. Vaginal microbiota were characterized via 16S ribosomal RNA gene sequencing and analysis, and the most abundant taxa observed were compared across visits for each subject. Sequences from L. iners were then isolated and evaluated for the presence of sub-strains using “oligotyping” analyses. The Shannon Entropy diversity index of single nucleotide polymorphisms (SNPs) in the 16S rRNA gene was used to detect bacterial sub-strains. The number of L. iners oligotypes (strains) was examined for each subject across all 4 visits. RESULTS: The relative abundance of L. iners increased between visits 1 and 2. Oligotyping analyses revealed 5 SNPs in the 16S rRNA gene with significantly greater diversity indices than all other positions in the alignment, resulting in up to 93 oligotypes (or sub-strains) observed. The number of L. iners strains observed within a subject increased from visit 1 to visit 2, and this diversity was maintained at subsequent visits. DECEMBER 2016 American Journal of Obstetrics & Gynecology
817