scholarly journals Ex Vivo Comparison of Microbicide Efficacies for Preventing HIV-1 Genomic Integration in Intraepithelial Vaginal Cells

2009 ◽  
Vol 54 (2) ◽  
pp. 763-772 ◽  
Author(s):  
M. Juliana McElrath ◽  
Lamar Ballweber ◽  
Andrew Terker ◽  
Allison Kreger ◽  
Polachai Sakchalathorn ◽  
...  
Keyword(s):  
Large Scale ◽  
Hiv Transmission ◽  
Ex Vivo ◽  
Integrase Inhibitor ◽  
Water Soluble ◽  
Cellulose Sulfate ◽  
Genomic Integration ◽  
Significant Difference ◽  
Significant Efficacy ◽  
Hiv 1

ABSTRACT Vaginally applied microbicides hold promise as a strategy to prevent sexual HIV transmission. Several nonspecific microbicides, including the polyanion cellulose sulfate, have been evaluated in large-scale clinical trials but have failed to show significant efficacy. These findings have prompted a renewed search for preclinical testing systems that can predict negative outcomes of microbicide trials. Moreover, the pipeline of potential topical microbicides has been expanded to include antiretroviral agents, such as reverse transcriptase, fusion, and integrase inhibitors. Using a novel ex vivo model of vaginal HIV-1 infection, we compared the prophylactic potentials of two forms of the fusion inhibitor T-20, the CCR5 antagonist TAK-778, the integrase inhibitor 118-D-24, and cellulose sulfate (Ushercell). The T-20 peptide with free N- and C-terminal amino acids was the most efficacious compound, causing significantly greater inhibition of viral genomic integration in intraepithelial vaginal leukocytes, measured by an optimized real-time PCR assay, than the more water-soluble N-acetylated T-20 peptide (Fuzeon) (50% inhibitory concentration [IC50], 0.153 μM versus 51.2 μM [0.687 ng/ml versus 230 ng/ml]; P < 0.0001). In contrast, no significant difference in IC50s was noted in peripheral blood cells (IC50, 13.58 μM versus 7.57 μM [61 ng/ml versus 34 ng/ml]; P = 0.0614). Cellulose sulfate was the least effective of all the compounds tested (IC50, 1.8 μg/ml). These results highlight the merit of our model for screening the mucosal efficacies of novel microbicides and their formulations and potentially rank ordering candidates for clinical evaluation.

scholarly journals Preclinical Evaluation of the HIV-1 Fusion Inhibitor L'644 as a Potential Candidate Microbicide

2012 ◽  
Vol 56 (5) ◽  
pp. 2347-2356 ◽  
Author(s):  
Sarah Harman ◽  
Carolina Herrera ◽  
Naomi Armanasco ◽  
Jeremy Nuttall ◽  
Robin J. Shattock
Keyword(s):  
Viral Entry ◽  
Hiv Transmission ◽  
Ex Vivo ◽  
Biological Fluids ◽  
Host Cells ◽  
Viral Escape ◽  
Escape Mutant ◽  
Potential Candidate ◽  
Preclinical Evaluation ◽  
Hiv 1

ABSTRACTTopical blockade of the gp41 fusogenic protein of HIV-1 is one possible strategy by which microbicides could prevent HIV transmission, working early against infection, by inhibiting viral entry into host cells. In this study, we examined the potential of gp41 fusion inhibitors (FIs) as candidate anti-HIV microbicides. Preclinical evaluation of four FIs, C34, T20, T1249, and L'644, was performed using cellular andex vivogenital and colorectal tissue explant models. Increased and sustained activity was detected for L'644, a cholesterol-derivatized version of C34, relative to the other FIs. The higher potency of L'644 was further increased with sustained exposure of cells or tissue to the compound. The activity of L'644 was not affected by biological fluids, and the compound was still active when tissue explants were treated after viral exposure. L'644 was also more active than other FIs against a viral escape mutant resistant to reverse transcriptase inhibitors (RTIs), demonstrating the potential of L'644 to be included as part of a multiactive antiretroviral (ARV) combination-based microbicide. These data support the further development of L'644 for microbicide application.

scholarly journals HIV drug resistance and HIV transmission risk factors among newly diagnosed individuals in Southwest China

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xianwu Pang ◽  
Kailing Tang ◽  
Qin He ◽  
Jinghua Huang ◽  
Ningye Fang ◽  
...  
Keyword(s):  
Risk Factors ◽  
Drug Resistance ◽  
Hiv Transmission ◽  
Multivariate Logistic Regression Analysis ◽  
Transmission Risk ◽  
Transmitted Drug Resistance ◽  
Common Mutation ◽  
Newly Diagnosed ◽  
Significant Difference ◽  
Hiv 1

Abstract Background The widespread use of antiretroviral therapy (ART) has resulted in the development of transmitted drug resistance (TDR), which reduces ART efficacy. We explored TDR prevalence and its associated risk factors in newly diagnosed individuals in Guangxi. Methods We enrolled 1324 participants who were newly diagnosed with HIV-1 and had not received ART at voluntary counselling and testing centres (VCT) in Guangxi, China, who had not received ART. Phylogenetic relationship, transmission cluster, and genotypic drug resistance analyses were performed using HIV-1 pol sequences. We analysed the association of demographic and virological factors with TDR. Results In total, 1151 sequences were sequenced successfully, of which 83 (7.21%) showed evidence of TDR. Multivariate logistic regression analysis revealed that there was significant difference between the prevalence of TDR and unmarried status (adjusted odds ratio (aOR) = 2.41, 95% CI: 1.23–4.71), and CRF08_BC subtype (aOR = 2.03, 95% CI: 1.13–3.64). Most cases of TDR were related to resistance to non-nucleoside reverse transcriptase inhibitors (4.87%) and V179E was the most common mutation detected. We identified a total of 119 HIV transmission clusters (n = 585, 50.8%), of which 18 (15.1%) clusters showed evidence of TDR (36, 41.86%). Three clusters were identified that included drug-resistant individuals having a transmission relationship with each other. The following parameters were associated with TDR transmission risk: Unmarried status, educational level of junior high school or below, and CRF08_BC subtype may be a risk of the transmission of TDR. Conclusions Our findings indicated that moderate TDR prevalence and highlighted the importance of continuous TDR monitoring and designing of strategies for TDR mitigation.

scholarly journals Purging human ovarian cortex of contaminating leukaemic cells by targeting the mitotic catastrophe signalling pathway

2021 ◽  
Author(s):  
Lotte Eijkenboom ◽  
Callista Mulder ◽  
Bert van der Reijden ◽  
Norah van Mello ◽  
Julia van Leersum ◽  
...  
Keyword(s):  
Myeloid Leukaemia ◽  
Large Scale ◽  
Ex Vivo ◽  
Ovarian Tissue ◽  
Signalling Pathway ◽  
Mitotic Catastrophe ◽  
Ovarian Cortex ◽  
Significant Difference ◽  
Leukaemic Cells

Abstract Purpose Is it possible to eliminate metastasised chronic myeloid leukaemia (CML) and acute myeloid leukaemia (AML) cells from ovarian cortex fragments by inhibition of Aurora B/C kinases (AURKB/C) without compromising ovarian tissue or follicles? Methods Human ovarian cortex tissue with experimentally induced tumour foci of CML, AML and primary cells of AML patients were exposed to a 24h treatment with 1 μM GSK1070916, an AURKB/C inhibitor, to eliminate malignant cells by invoking mitotic catastrophe. After treatment, the inhibitor was removed, followed by an additional culture period of 6 days to allow any remaining tumour cells to form new foci. Ovarian tissue integrity after treatment was analysed by four different assays. Appropriate controls were included in all experiments. Results Foci of metastasised CML and AML cells in ovarian cortex tissue were severely affected by a 24h ex vivo treatment with an AURKB/C inhibitor, leading to the formation of multi-nuclear syncytia and large-scale apoptosis. Ovarian tissue morphology and viability was not compromised by the treatment, as no significant difference was observed regarding the percentage of morphologically normal follicles, follicular viability, glucose uptake or in vitro growth of small follicles between ovarian cortex treated with 1 μM GSK1070916 and the control. Conclusion Purging of CML/AML metastases in ovarian cortex is possible by targeting the Mitotic Catastrophe Signalling Pathway using GSK1070916 without affecting the ovarian tissue. This provides a therapeutic strategy to prevent reintroduction of leukaemia and enhances safety of autotransplantation in leukaemia patients currently considered at high risk for ovarian involvement.

scholarly journals Interleukin-7 HIV Transmission to Cervico-Vaginal Tissue in vivo

2013 ◽  
Vol 18 (2) ◽  
pp. 4-16
Author(s):  
A. Introini ◽  
C. Vanpouille ◽  
A. Lisco ◽  
J. -C Grivel ◽  
L. Borisovich Margolis
Keyword(s):  
T Cells ◽  
Hiv Transmission ◽  
Ex Vivo ◽  
B Cell Lymphoma ◽  
Vaginal Mucosa ◽  
Vaginal Tissue ◽  
Ki 67 ◽  
Interleukin 7 ◽  
Hiv 1

The majority of HIV-1 infections in women transmit through vaginal intercourse, in which virus-containing semen is deposited on the cervico-vaginal mucosa. Semen is more than a mere carrier of HIV-1, since it contains many biological factors, in particular cytokines, that may affect HIV-1 transmission. The concentration of interleukin (IL)-7, one of the most prominent cytokines in semen of healthy individuals, is further increased in semen of HIV-1-infected men. Here, we investigated the potential role of IL-7 in HIV-1 vaginal transmission in an ex vivo system of human cervico-vaginal tissue. We simulated an in vivo situation by depositing HIV-1 on cervico-vaginal tissue in combination with IL-7 at concentrations comparable with those measured in semen of HIV-1-infected individuals. We found that IL-7 significantly enhanced virus replication in ex vivo infected cervico-vaginal tissue. Similarly, we observed an enhancement of HIV-1 replication in lymphoid tissue explants. Analysis of T cells isolated from infected tissues showed that IL-7 reduced CD4+ T cell depletion preventing apoptosis, as shown by the decrease in the number of cells expressing the apoptotic marker APO2.7 and the increase in the expression of the anti-apoptotic protein B-cell lymphoma (Bcl)-2. Also, IL-7 increased the fraction of cycling CD4+ T cells, as evidenced by staining for the nuclear factor Ki-67. High levels of seminal IL-7 in vivo may be relevant to the survival of the founder pool of HIV-1-infected cells in the cervico-vaginal mucosa at the initial stage of infection, promoting local expansion and dissemination of HIV infection.

scholarly journals Monocytes but Not Lymphocytes Carrying HIV-1 on Their Surface Transmit Infection to Human TissueEx Vivo

2016 ◽  
Vol 90 (21) ◽  
pp. 9833-9840 ◽  
Author(s):  
Victor Barreto-de-Souza ◽  
Anush Arakelyan ◽  
Sonia Zicari ◽  
Leonid Margolis ◽  
Christophe Vanpouille
Keyword(s):  
Human Tissue ◽  
Hiv Transmission ◽  
Ex Vivo ◽  
Sexual Transmission ◽  
Hiv Infections ◽  
Productive Infection ◽  
Unprotected Sexual Intercourse ◽  
Hiv 1 ◽  
Monocytes And Lymphocytes

ABSTRACTUnprotected sexual intercourse with HIV-infected men is the major cause of new infections. HIV virions are released into semen by various cells of the male genital tract, as well as by infected monocytes and lymphocytes present in semen. Some of these virions may attach to the surfaces of cells, infected or uninfected. We investigated whether cells carrying attached HIV on their surfaces can transmit infection. We addressed this question in a model system of human tissue exposedex vivoto monocytes and lymphocytes carrying HIV on their surfaces. We gamma irradiated the cells to prevent their productive infection. In spite of comparable amounts of HIV attached to monocytes and lymphocytes, only monocytes were capable of transmitting infection and triggering productive infection in tissue. This HIV-1 transmission was mediated by cell-cell contacts. Our experiments suggest thatin vivo, HIV attached to infected or uninfected monocytes, which far outnumber lymphocytes in HIV-infected semen, may contribute to sexual transmission of HIV from men to their partners.IMPORTANCEThe vast majority of new HIV infections occur through sexual transmission, in which HIV is transferred from the semen of an infected male to an uninfected partner. In semen, HIV-1 particles may exist as free-floating virions; inside infected cells; or attached to the surfaces of cells, whether they are infected or not. Here, we investigated whether HIV attached to the surfaces of monocytes or lymphocytes could transmit infection to human tissue. Incubation of human tissue with monocyte-attached HIV resulted in productive tissue infection. In contrast, there was no infection of tissues when they were incubated with lymphocyte-attached HIV-1. Our results highlight the important role that seminal monocytes may play in HIV transmissionin vivo, especially since monocytes far outnumber lymphocytes in the semen of HIV-infected individuals.

scholarly journals Feasibility of Repurposing the Polyanionic Microbicide, PPCM, for Prophylaxis against HIV Transmission during ART

2011 ◽  
Vol 2011 ◽  
pp. 1-13 ◽  
Author(s):  
Robert A. Anderson ◽  
David Brown ◽  
Erin M. Jackson ◽  
Kenneth A. Feathergill ◽  
James W. Bremer ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Viral Antigen ◽  
Hiv Transmission ◽  
Ex Vivo ◽  
Dna Integrity ◽  
Sperm Dna Integrity ◽  
Hiv Serodiscordant Couples ◽  
Sperm Dna ◽  
Sperm Washing ◽  
Hiv 1

HIV-serodiscordant couples wishing to conceive often seek assisted reproduction, during which spermatozoa from infected men are washed to minimize the risk of HIV transmission to partner and fetus. We sought to improve this method by adding a microbicide, PPCM, as an HIV prophylactic. HIV-1 (BaL) inhibition by PPCM appears irreversible and independent of added Ca2+. Without added Ca2+, PPCM (≤10 mg/mL, ≤90 min), a stimulus of Ca2+-dependent acrosomal loss, has no effect on sperm motility, forward progression, or acrosomal status. PPCM-treated (10 mg/mL) sperm retain their ability to acrosome react when Ca2+ is added. Sperm DNA integrity/function is unaffected by PPCM (≤10 mg/mL). Adding PPCM (5 mg/mL, 30 min) to washing media reduces infectivity (viral antigen p24 and RNA) of ex-vivo HIV-infected semen by 3-4 Logs compared with washing alone. Sperm washing with appropriate extracellular Ca2+ levels and PPCM is significantly more effective than washing alone at reducing HIV infectivity.

Role of vitamin B12 in treating recurrent aphthous stomatitis: A review

2020 ◽  
pp. 1-8
Author(s):  
Rim Taleb ◽  
Bassel Hafez ◽  
Nadim El Kassir ◽  
Hani El Achkar ◽  
Mohamad Mourad
Keyword(s):  
Vitamin B12 ◽  
Large Scale ◽  
Active Form ◽  
Vital Role ◽  
Recurrent Aphthous Stomatitis ◽  
Water Soluble ◽  
Aphthous Stomatitis ◽  
Hematopoietic Stem ◽  
Inactive Form ◽  
Significant Difference

Abstract. Vitamin B12, a water-soluble vitamin, plays a vital role in the formation of hematopoietic stem cells and has been associated with oral mucosal diseases, mainly recurrent aphthous stomatitis (RAS). The latter is a debilitating condition, and B12 was proposed as a potential treatment given its role in regenerating oral mucosal tissue. There is conflicting evidence that B12 deficiency causes RAS. Five of the seven randomized controlled trials reviewed used the inactive form of B12 (cyanocobalamin) as intervention, while the other two used the active form (methylcobalamin). Of the latter two, buccal discs (500 μg B12) showed significant improvement and reduced perceived pain in 77% of the subjects, and submucosal injections showed a significant difference in pain, starting from the second day. Moreover, three studies administered vitamin B12 sublingually with different dosages, which revealed that the higher dose (1000 μg) achieved a significant reduction in outbreaks, number, and duration of ulcers, especially after six months. Multivitamins showed no difference in new RAS episodes and duration. Injectable B12 was compared with the oral form, and nearly 50% of the injection group reported a desired response by the eighth week. An ointment form (500 μg) showed a significant reduction in pain levels after two days of treatment. Based on the available literature, we suggest that a daily dose of 1000 μg of vitamin B12 sublingually for six months can be used to treat RAS. Nevertheless, this conclusion should be considered tentative due to the lack of high quality, large scale studies.

Large-Scale Bicortical Skull Bone Regeneration Using Ex vivo Replication-Defective Adenoviral-Mediated Bone Morphogenetic Protein—2 Gene—Transferred Bone Marrow Stromal Cells and Composite Biomaterials

2009 ◽  
Vol 65 (suppl_6) ◽  
pp. ons75-ons83 ◽  
Author(s):  
Sophia Chia-Ning Chang ◽  
Tsung-Min Lin ◽  
Hui-Ying Chung ◽  
Philip Kuo-Ting Chen ◽  
Feng-Huei Lin ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Large Scale ◽  
Ex Vivo ◽  
Marrow Stromal Cells ◽  
Skull Defect ◽  
Morphogenetic Protein ◽  
Significant Difference ◽  
Group 2 ◽  
Group 1

Abstract Objective: Bone marrow stromal cells (BMSCs) have great potential in bone repair. We developed an animal model to test the hypothesis that ex vivo gene transfer of human bone morphogenetic protein (BMP)-2 to BMSCs via a replication-defective (E1A-deleted) adenovirus vector (AdV) with appropriate biopolymers would enhance autologous bone formation during repair of a large-scale skull defect. Methods: Eighteen miniature swine were treated with AdV BMP-2-transduced BMSCs in biopolymer (group 1), BMSCs in biopolymer (group 2), or biopolymer alone (group 3). After 6 months, the swine were killed, and the skull repair was examined by gross pictures, histology, 3-dimensional computed tomography, and biomechanical study. Results: Group 1 showed complete solid bone formation after 6 months, and hema-toxylin and eosin staining demonstrated the presence of mature, woven, well-mineralized bone. Computed tomography showed wholesome repair of the skull defect. Statistical analysis demonstrated a significant difference in bone thickness between groups 1 and 2. Biomechanical testing showed a statistically significant difference in the stiffness of new bone formed in group 1 compared with group 2. Conclusion: The Ad5 E1A-deleted AdV may be the optimal starting vector in ex vivo gene therapy for benign skeletal diseases. Additionally, the use of the gelatin/tricalcium phosphate ceramic/glutaraldehyde biopolymer with AdV BMP-2 gene transfer strongly enhances the bony healing of critical-size bicortical craniofacial defects. This method can be used by modifying the delivery of constructs to malunion treatment, in regional osteoporosis therapy, and spinal fusion.

Aerosol generation through pars plana vitrectomy

2020 ◽  
pp. bjophthalmol-2020-317214
Author(s):  
Hasan Naveed ◽  
Fong May Chew ◽  
Hanbin Lee ◽  
Edward Hughes ◽  
Mayank A Nanavaty
Keyword(s):  
Pars Plana Vitrectomy ◽  
Ex Vivo ◽  
Dynamic Changes ◽  
Aerosol Generation ◽  
Significant Difference ◽  
Optical Particle Counter ◽  
Pars Plana ◽  
23 Gauge ◽  
Set Up ◽  
Ex Vivo Study

PurposeTo assess whether pars plana vitrectomy (PPV) is an aerosol-generating procedure (AGP) in an ex vivo experimental model.MethodsIn this ex vivo study on 10 porcine eyes, optical particle counter was used to measure particles ≤10 μm using cumulative mode in the six in-built channels: 0.3 μm, 0.5 μm, 1 μm, 2.5 μm, 5 μm and 10 μm aerosols during PPV. Two parts of the study were as follows: (1) to assess the pre-experimental baseline aerosol count in the theatre environment where there are dynamic changes in temperature and humidity and (2) to measure aerosol generation with 23-gauge and 25-gauge set-up. For each porcine eye, five measurements were taken for each consecutive step in the experiment including pre-PPV, during PPV, fluid–air exchange (FAX) and venting using a flute with 23-gauge set-up and a chimney with 25-gauge set-up. Therefore, a total of 200 measurements were recorded.ResultsWith 23-gauge and 25-gauge PPV, there was no significant difference in aerosol generation in all six channels comparing pre-PPV versus PPV or pre-PPV versus FAX. Venting using flute with 23-gauge PPV showed significant reduction of aerosol ≤1 μm. Air venting using chimney with 25-gauge set-up showed no significant difference in aerosol of ≤1 μm. For cumulative aerosol counts of all particles measuring ≤5 μm, compared with pre-PPV, PPV or FAX, flute venting in 23-gauge set-up showed significant reduction unlike the same comparison for chimney venting in 25-gauge set-up.ConclusionPPV and its associate steps do not generate aerosols ≤10 μm with 23-gauge and 25-gauge set-ups.

scholarly journals Limited Sustained Local Transmission of HIV-1 CRF01_AE in New South Wales, Australia

Viruses ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 482 ◽  
Author(s):  
Francesca Di Giallonardo ◽  
Angie N. Pinto ◽  
Phillip Keen ◽  
Ansari Shaik ◽  
Alex Carrera ◽  
...  
Keyword(s):  
Hiv Transmission ◽  
New South ◽  
New South Wales ◽  
Incidence Rates ◽  
Effective Control ◽  
South Wales ◽  
Immunodeficiency Virus ◽  
Hiv 1 ◽  
Local Transmission

Australia’s response to the human immunodeficiency virus type 1 (HIV-1) pandemic led to effective control of HIV transmission and one of the world’s lowest HIV incidence rates—0.14%. Although there has been a recent decline in new HIV diagnoses in New South Wales (NSW), the most populous state in Australia, there has been a concomitant increase with non-B subtype infections, particularly for the HIV-1 circulating recombinant form CRF01_AE. This aforementioned CRF01_AE sampled in NSW, were combined with those sampled globally to identify NSW-specific viral clades. The population growth of these clades was assessed in two-year period intervals from 2009 to 2017. Overall, 109 NSW-specific clades were identified, most comprising pairs of sequences; however, five large clades comprising ≥10 sequences were also found. Forty-four clades grew over time with one or two sequences added to each in different two-year periods. Importantly, while 10 of these clades have seemingly discontinued, the remaining 34 were still active in 2016/2017. Seven such clades each comprised ≥10 sequences, and are representative of individual sub-epidemics in NSW. Thus, although the majority of new CRF01_AE infections were associated with small clades that rarely establish ongoing chains of local transmission, individual sub-epidemics are present and should be closely monitored.

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