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Current concept of HIV pathogenesis in the female genital tract
Abstracts / Journal of Reproductive Immunology 94 (2012) 5–130
strated that GM-CSF stimulated mouse myeloid committed cells have increased production of multi-vesicular bodies, which are involved in HIV gag assembly, intracellular budding and virion release. This finding may represent the mechanism by which GM-CSF treatment circumvents the species-specific post-integration block in HIV budding that restricts HIV replication by mouse cells. doi:10.1016/j.jri.2012.03.484 P 183 Current concept of HIV pathogenesis in the female genital tract G. Spear 1 , P. Mirmonsef 1 , P. Gillevet 2 , M.R. Zariffard 1 , D. Gilbert 1 , D. Burgad 1 , A.L. Landay 1,∗ 1
Rush University Medical Center, Department of Immunology/Microbiology, Chicago, United States 2 George Mason University, Department of Environmental Sciences and Policy, Manassas, United States Changes in the bacterial microbiota of the lower genital tract of women are associated with susceptibility to transmission of human immunodeficiency virus (HIV), as well as other STIs. Our laboratory has explored the question of how the microbiome in the female genital tract can modulate HIV acquisition through interaction with the innate and adaptive immune system. Since the types of genital bacteria could vary depending on geography or cultural influences and these differences could in turn influence HIV transmission, our lab studied how the genital microbiome differed between Rwandan and U.S. women using pyrosequencing. The generaLactobacillus, Gardnerella, Prevotella, Atopobiumand Sneathiawere the most predominant genera and were found at similar frequencies in Rwandan and U.S. women. In contrast,Mycoplasmawas significantly more frequent in Rwandan women whileMegasphaerawas more frequent in U.S. women. Since macaques are used as an animal model of HIV vaginal infection of women, we have also studied the genital microbiota of macaques by pyrosequencing. Both rhesus and pigtailed macaques had a genital microbiota that was low inLactobacillusbut high in levels of bacteria found in women with the condition known as bacterial vaginosis (BV). This suggests that the microbiota in macaques could alter infection by retroviruses. Our laboratory has also studied how pro-inflammatory cytokines and other immune mediators in the lower genital tract are altered in disease states such as STDs. Strong correlations were observed between levels of TNF-a, IL1ß and IL-6, between chemokines IP-10 and MIG and between myeloperoxidase, IL-8 and G-CSF. Samples from women with any STD/colonization had significantly higher levels of IL-8, IL-3, IL-7, IL-1ß, lactoferrin and myeloperoxidase. IL-1ß and lactoferrin were significantly increased in gonorrhea, Chlamydia, cervicitis, bacterial vaginosis and trichomoniasis. Our research also shows that toll-like receptor 2- (TLR2) ligands are present in genital fluids of women with BV and may contribute to induction of these
129
cytokines in that condition. This inflammation may contribute to the increased HIV susceptibility that has been observed in BV. While bacteria release TLR-ligands, they also make short chain fatty acids (SCFAs) such as butyrate, propionate and acetate during fermentation. These SCFAs can reach high levels in the genital tract of women with BV (220 mM). We found that SCFAs alone can induce low levels of pro-inflammatory cytokines such as IL-1ß, IL-8 and IL6. However, in combination with TLR2- and TLR7- but not TLR4- ligands SCFAs potently increase cytokine production. These SCFAs may enter cells through plasma membrane transporters called MCTs. These studies highlight the important role that the genital tract microbiome plays in HIV disease and provides potential insights into novel approaches for HIV prevention strategies. doi:10.1016/j.jri.2012.03.485 P 184 Possible importance of the HIV-1 mutations in vertical transmission and frequent anti retroviral drug resistance mutations in1 protease and reverse transcriptase genes in antiretroviral-naïve children S. Hayakawa, D.Q. Trinh ∗ Nihon University,School of Medicine, Department of Microbiology, Tokyo, Viet Nam In the developed countries, newly acquired pediatric HIV infections have been virtually eliminated; however, pediatric HIV infection remains a significant public health problem in developing. countries in Asia and Africa. In the present study, a molecular epidemiological study was conducted on 104 HIV-1 strains isolated from HIV-infected children hospitalized in Children Hospital 1 in Ho Chi Minh City, Vietnam during 2004-2005. Genetic subtyping based on env C2/V3 sequences revealed that CRF01-AE was the sole circulating recombinant form found in this study. Sequence analysis of the V3 loop showed that GPGQ tetramer was the most common V3 loop core motif identified in the HIV-1 strains studied (89.5%). Then we examined the frequency of the resistance for antiretroviral medicines because they have recently become available for the treatment of children infected with HIV in Vietnam. So far genetic background of HIV-1 drug resistance in antiretroviral-naïve children has yet to be studied. Of the 104 HIV-1 CRF01-AE subtype strains that were previously isolated from antiretroviral-naïve children from the provinces of southern Vietnam, 79 strains were used for amplification and sequence analyses of the protease and reverse transcriptase (RT) genes. Minor mutations were found in the protease gene, including L10I, I13 V, G16E, M36I, D60E, I62 V, I64 V, L63P, H69 K, V82I, and I93L. Of these mutations, M36I and H69 K were detected in all of the strains that were studied. However, all of the amino acid changes in the protease gene were considered to be polymorphisms. In the RT gene, 3 major mutations were detected in 6 strains: the V75 M mutation in 1 strain, the
strated that GM-CSF stimulated mouse myeloid committed cells have increased production of multi-vesicular bodies, which are involved in HIV gag assembly, intracellular budding and virion release. This finding may represent the mechanism by which GM-CSF treatment circumvents the species-specific post-integration block in HIV budding that restricts HIV replication by mouse cells. doi:10.1016/j.jri.2012.03.484 P 183 Current concept of HIV pathogenesis in the female genital tract G. Spear 1 , P. Mirmonsef 1 , P. Gillevet 2 , M.R. Zariffard 1 , D. Gilbert 1 , D. Burgad 1 , A.L. Landay 1,∗ 1
Rush University Medical Center, Department of Immunology/Microbiology, Chicago, United States 2 George Mason University, Department of Environmental Sciences and Policy, Manassas, United States Changes in the bacterial microbiota of the lower genital tract of women are associated with susceptibility to transmission of human immunodeficiency virus (HIV), as well as other STIs. Our laboratory has explored the question of how the microbiome in the female genital tract can modulate HIV acquisition through interaction with the innate and adaptive immune system. Since the types of genital bacteria could vary depending on geography or cultural influences and these differences could in turn influence HIV transmission, our lab studied how the genital microbiome differed between Rwandan and U.S. women using pyrosequencing. The generaLactobacillus, Gardnerella, Prevotella, Atopobiumand Sneathiawere the most predominant genera and were found at similar frequencies in Rwandan and U.S. women. In contrast,Mycoplasmawas significantly more frequent in Rwandan women whileMegasphaerawas more frequent in U.S. women. Since macaques are used as an animal model of HIV vaginal infection of women, we have also studied the genital microbiota of macaques by pyrosequencing. Both rhesus and pigtailed macaques had a genital microbiota that was low inLactobacillusbut high in levels of bacteria found in women with the condition known as bacterial vaginosis (BV). This suggests that the microbiota in macaques could alter infection by retroviruses. Our laboratory has also studied how pro-inflammatory cytokines and other immune mediators in the lower genital tract are altered in disease states such as STDs. Strong correlations were observed between levels of TNF-a, IL1ß and IL-6, between chemokines IP-10 and MIG and between myeloperoxidase, IL-8 and G-CSF. Samples from women with any STD/colonization had significantly higher levels of IL-8, IL-3, IL-7, IL-1ß, lactoferrin and myeloperoxidase. IL-1ß and lactoferrin were significantly increased in gonorrhea, Chlamydia, cervicitis, bacterial vaginosis and trichomoniasis. Our research also shows that toll-like receptor 2- (TLR2) ligands are present in genital fluids of women with BV and may contribute to induction of these
129
cytokines in that condition. This inflammation may contribute to the increased HIV susceptibility that has been observed in BV. While bacteria release TLR-ligands, they also make short chain fatty acids (SCFAs) such as butyrate, propionate and acetate during fermentation. These SCFAs can reach high levels in the genital tract of women with BV (220 mM). We found that SCFAs alone can induce low levels of pro-inflammatory cytokines such as IL-1ß, IL-8 and IL6. However, in combination with TLR2- and TLR7- but not TLR4- ligands SCFAs potently increase cytokine production. These SCFAs may enter cells through plasma membrane transporters called MCTs. These studies highlight the important role that the genital tract microbiome plays in HIV disease and provides potential insights into novel approaches for HIV prevention strategies. doi:10.1016/j.jri.2012.03.485 P 184 Possible importance of the HIV-1 mutations in vertical transmission and frequent anti retroviral drug resistance mutations in1 protease and reverse transcriptase genes in antiretroviral-naïve children S. Hayakawa, D.Q. Trinh ∗ Nihon University,School of Medicine, Department of Microbiology, Tokyo, Viet Nam In the developed countries, newly acquired pediatric HIV infections have been virtually eliminated; however, pediatric HIV infection remains a significant public health problem in developing. countries in Asia and Africa. In the present study, a molecular epidemiological study was conducted on 104 HIV-1 strains isolated from HIV-infected children hospitalized in Children Hospital 1 in Ho Chi Minh City, Vietnam during 2004-2005. Genetic subtyping based on env C2/V3 sequences revealed that CRF01-AE was the sole circulating recombinant form found in this study. Sequence analysis of the V3 loop showed that GPGQ tetramer was the most common V3 loop core motif identified in the HIV-1 strains studied (89.5%). Then we examined the frequency of the resistance for antiretroviral medicines because they have recently become available for the treatment of children infected with HIV in Vietnam. So far genetic background of HIV-1 drug resistance in antiretroviral-naïve children has yet to be studied. Of the 104 HIV-1 CRF01-AE subtype strains that were previously isolated from antiretroviral-naïve children from the provinces of southern Vietnam, 79 strains were used for amplification and sequence analyses of the protease and reverse transcriptase (RT) genes. Minor mutations were found in the protease gene, including L10I, I13 V, G16E, M36I, D60E, I62 V, I64 V, L63P, H69 K, V82I, and I93L. Of these mutations, M36I and H69 K were detected in all of the strains that were studied. However, all of the amino acid changes in the protease gene were considered to be polymorphisms. In the RT gene, 3 major mutations were detected in 6 strains: the V75 M mutation in 1 strain, the