Non-Surgical Periodontal Therapy's Influence on Alpha-Synuclein and Inflammatory Marker Levels: A Pilot Study.

Background: Microbial dysbiosis may contribute to alpha-synuclein (α-Syn) homeostasis disruption, yet the burden of inflammatory periodontal infection and its treatment have never been studied in this regard. We aimed to compare the cytokine and α-Syn levels in the saliva and blood of patients with periodontitis who underwent non-surgical periodontal therapy (NSPT) and those of their healthy counterparts. Methods: Periodontal examination and saliva and blood sample collection were carried out in incoming patients at a university clinic. The periodontitis group (PG) received NSPT. The sample collection and periodontal observation were repeated 30 days after. IL-6, IL1-ß and total α-Syn were quantified using immunoassay methods. The periodontal inflamed surface area (PISA) was calculated as a proxy for periodontal inflammation. Results: Eleven participants formed the PG, and there were fifteen healthy controls (HC). At baseline, no correlation between salivary and plasma α-Syn was found. The salivary α-Syn levels revealed a tendency to decrease 30 days after, particularly in the PD cases. The variation in PISA and α-Syn showed significant correlation. Salivary α-Syn correlated negatively with salivary IL-6 levels at both timepoints in the total sample (rho = -0.394 and rho = -0.451) and in the HC (rho = -0.632 and rho = -0.561). Variations in plasma IL-6 and α-Syn were negatively correlated (rho = -0.518) in the healthy participants. Baseline plasma IL1-ß negatively correlated with plasmatic α-Syn at 30 days in the HC (rho = -0.581). Conclusions: Salivary and plasma α-Syn bioavailability operate independently, and periodontal diagnosis was not a confounding factor. Salivary α-Syn levels were significantly affected by NSPT, contrary to plasma levels. These results should be confirmed in future larger and prospective studies.

Insights into Candida Colonization in Intensive Care Unit Patients: A Prospective Multicenter Study.

The skin mycobiota plays a significant role in infection risk, pathogen transmission, and personalized medicine approaches in intensive care settings. This prospective multicenter study aimed to enhance our understanding of intensive care units' (ICUs') Candida colonization dynamics, identify modifiable risk factors, and assess their impact on survival risk. Specimens were taken from 675, 203, and 110 patients at the admission (D1), 5th (D5), and 8th (D8) days of ICU stay, respectively. The patient's demographic and clinical data were collected. Candida isolates were identified by conventional culture-based microbiology combined with molecular approaches. Overall, colonization was 184/675 (27.3%), 87/203 (42.8%), and 58/110 (52.7%) on D1, D5, and D8, respectively. Candida colonization dynamics were significantly associated with ICU type (odds ratio (OR) = 2.03, 95% CI 1.22-3.39, p = 0.007), respiratory infection (OR = 1.74, 95% CI 1.17-2.58, p = 0.006), hemodialysis (OR = 2.19, 95% CI 1.17-4.10, p = 0.014), COVID-19 (OR = 0.37, 95% CI 0.14-0.99, p = 0.048), and with a poor 3-month outcome (p = 0.008). Skin Candida spp. colonization can be an early warning tool to generate valuable insights into the epidemiology, risk factors, and survival rates of critically ill patients, and should be considered for epidemiological surveillance.

Antibacterial Effect of Ozone on Cariogenic Bacteria and Its Potential Prejudicial Effect on Dentin Bond Strength-An In Vitro Study.

Ozone is increasingly utilized in dental caries treatment due to its antibacterial properties. In a context of limited studies and no consensus on protocols, this research aims to assess ozone's antibacterial efficacy on cariogenic bacteria and its potential adverse impact on dentin bond strength. Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei, and Actinomyces naeslundii suspensions were exposed to 40 µg/mL of ozone gas and 60 µg/mL of ozonated water (80 s) via a medical ozone generator. Negative and positive control groups (chlorhexidine 2%) were included, and UFC/mL counts were recorded. To examine microtensile bond strength (µTBS), 20 human molars were divided into four groups, and class I cavities were created. After ozone application, samples were restored using an etch-and-rinse and resin composite, then sectioned for testing. The SPSS v. 28 program was used with a significance level of 5%. The µTBS results were evaluated using one-way ANOVA, Tukey HSD, and Games-Howell. Bacterial counts reduced from 106 to 101, but dentin µTBS was significantly impacted by ozone (ANOVA, p < 0.001). Despite ozone's attractive antibacterial activity, this study emphasizes its detrimental effect on dentin adhesion, cautioning against its use before restorative treatments.

Revisiting Oral Antiseptics, Microorganism Targets and Effectiveness.

A good oral health status is mostly dependent on good oral hygiene habits, which knowingly impacts systemic health. Although controversial, chemical oral antiseptics can be useful in adjunct use to mechanical dental plaque control techniques in the prevention and management of local and overall health and well-being. This review aims to revisit, gather and update evidence-based clinical indications for the use of the most popular oral antiseptics, considering different types, microorganism targets and effectiveness in order to establish updated clinical recommendations.

Association of Herpesvirus and Periodontitis: A Clinical and Laboratorial Case-Control Study.

OBJECTIVES: A significant influence of the Herpesviridae family in the progression of periodontal disease has been suggested. The aim of this study was to investigate the potential association of four Herpesviruses (HSV-1, HSV-2, cytomegalovirus [CMV], and Epstein-Barr virus [EBV]) with periodontal disease using a qualitative test for evaluating the presence or absence of viral DNA in crevicular fluid samples of both healthy periodontal patients and periodontal compromised patients. MATERIALS AND METHODS: A case-control study was conducted in 100 participants at a university clinic. A qualitative test was used for evaluating the presence/absence of viral DNA in crevicular fluid samples of both healthy periodontal patients and periodontal compromised patients, and considering the periodontitis staging (stage II, stage III, and stage IV) and grading (grade A, grade B, and grade C). STATISTICAL ANALYSIS: The distribution of the same exposure variables to the periodontitis staging and grading was compared using Chi-square, Fisher's exact, and Gamma tests depending on the variable characteristics. The significance level was set at 5%. The association of the variables: age, sex, diabetes, smoking, alcohol, and oral hygiene was also considered. RESULTS: The prevalence of Herpesviridae family virus DNA was 6% for the periodontal healthy group and 60% for the periodontitis group (roughly 60% on periodontitis stages II, III, and IV, p <0.001; and twofold increase in moderate and rapid progression grades compared with the slow progression grade, p <0.001). HSV1 DNA was prevalent in all periodontitis stages and grades. HSV 2, EBV, and CMV DNA had increasing prevalence rates in more severe stages (stages III and IV, p <0.001); while considering periodontitis grade, HSV2 (p = 0.001), CMV (p = 0.019) and EBV (p <0.001) DNA were prevalent only in grades B and C, with EBV DNA registering a marked prevalence in grade C. CONCLUSION: A significant different distribution of Herpesviridae virus DNA per each stage of disease was registered.

An HIV-1/HIV-2 Chimeric Envelope Glycoprotein Generates Binding and Neutralising Antibodies against HIV-1 and HIV-2 Isolates.

The development of immunogens that elicit broadly reactive neutralising antibodies (bNAbs) is the highest priority for an HIV vaccine. We have shown that a prime-boost vaccination strategy with vaccinia virus expressing the envelope glycoprotein gp120 of HIV-2 and a polypeptide comprising the envelope regions C2, V3 and C3 elicits bNAbs against HIV-2. We hypothesised that a chimeric envelope gp120 containing the C2, V3 and C3 regions of HIV-2 and the remaining parts of HIV-1 would elicit a neutralising response against HIV-1 and HIV-2. This chimeric envelope was synthesised and expressed in vaccinia virus. Balb/c mice primed with the recombinant vaccinia virus and boosted with an HIV-2 C2V3C3 polypeptide or monomeric gp120 from a CRF01_AG HIV-1 isolate produced antibodies that neutralised >60% (serum dilution 1:40) of a primary HIV-2 isolate. Four out of nine mice also produced antibodies that neutralised at least one HIV-1 isolate. Neutralising epitope specificity was assessed using a panel of HIV-1 TRO.11 pseudoviruses with key neutralising epitopes disrupted by alanine substitution (N160A in V2; N278A in the CD4 binding site region; N332A in the high mannose patch). The neutralisation of the mutant pseudoviruses was reduced or abolished in one mouse, suggesting that neutralising antibodies target the three major neutralising epitopes in the HIV-1 envelope gp120. These results provide proof of concept for chimeric HIV-1/HIV-2 envelope glycoproteins as vaccine immunogens that can direct the antibody response against neutralising epitopes in the HIV-1 and HIV-2 surface glycoproteins.

Non-Surgical Periodontal Treatment Impact on Subgingival Microbiome and Intra-Oral Halitosis.

The purpose of this study was to characterize and compare subgingival microbiome before and after periodontal treatment to learn if any changes of the subgingival microbiome were reflected in intra-oral halitosis. We tested the hypothesis that intra-oral halitosis (Volatile sulfur compounds levels) correlates with corresponding subgingival bacterial levels before and after periodontal treatment. Twenty patients with generalized periodontitis completed the study. Subgingival plaque samples were collected at baseline and 6-8 weeks after nonsurgical periodontal therapy. Full-mouth periodontal status assessed probing depth (PD), clinical attachment loss (CAL), gingival recession (REC), bleeding on probing (BoP), PISA and PESA. Halitosis assessment was made using a volatile sulfur compounds (VSC) detector device. Periodontal measures were regressed across VSC values using adjusted multivariate linear analysis. The subgingival microbiome was characterized by sequencing on an Illumina platform. From a sample of 20 patients referred to periodontal treatment, 70% were females (n = 14), with a mean age of 56.6 (±10.3) years; full-mouth records of PD, CAL, BOP (%) allowed to classify the stage and grade of periodontitis, with 45% (n = 9) of the sample having Periodontitis Stage IV grade C and 95% (n = 19) had generalized periodontitis. The correlation of bacterial variation with VSCs measured in the periodontal diagnosis and in the reassessment after treatment were evaluated. Fusobacterium nucleatum, Capnocytophaga gingivalis and Campylobacter showaei showed correlation with the reduction of VSC after periodontal treatment (p-value = 0.044; 0.047 and 0.004, respectively). Capnocytophaga sputigena had a significant reverse correlation between VSCs variation from diagnosis (baseline) and after treatment. Microbial diversity was high in the subgingival plaque on periodontitis and intra-oral halitosis participants of the study. Furthermore, there were correlations between subgingival plaque composition and VSC counting after periodontal treatment. The subgingival microbiome can offer important clues in the investigation of the pathogenesis and treatment of halitosis.

Data of Nebivolol on oxidative stress parameters in hypertensive patients.

Oxidative stress is a key feature in hypertension, since reactive oxygen species are involved in all stages of cardiovascular diseases. Saliva is a body fluid that can be used to investigate alterations in the oxidative system with several specific advantages over blood. Nebivolol is a third-generation selective ß1-adrenergic receptor antagonist that promotes vasodilation and has been shown to reduce oxidative stress in pre-clinical and clinical studies. The use of Nebivolol in different periods of treatment demonstrated that it is an efficient anti-hypertensive drug. We evaluated the oxidative stress biomarkers and the enzymatic and non-enzymatic antioxidant systems in saliva of hypertensive patients before and after the use of anti-hypertensive therapeutic doses of Nebivolol, since saliva can be used as an auxiliary tool to analyze parameters of oxidative stress.

Spiro-ß-lactam BSS-730A Displays Potent Activity against HIV and Plasmodium.

The high burden of malaria and HIV/AIDS prevents economic and social progress in developing countries. A continuing need exists for development of novel drugs and treatment regimens for both diseases in order to address the tolerability and long-term safety concerns associated with current treatment options and the emergence of drug resistance. We describe new spiro-ß-lactam derivatives with potent (nM) activity against HIV and Plasmodium and no activity against bacteria and yeast. The best performing molecule of the series, BSS-730A, inhibited both HIV-1 and HIV-2 replication with an IC50 of 13 ± 9.59 nM and P. berghei hepatic infection with an IC50 of 0.55 ± 0.14 µM with a clear impact on parasite development. BSS-730A was also active against the erythrocytic stages of P. falciparum, with an estimated IC50 of 0.43 ± 0.04 µM. Time-of-addition studies showed that BSS-730A potentially affects all stages of the HIV replicative cycle, suggesting a complex mechanism of action. BSS-730A was active against multidrug-resistant HIV isolates, with a median 2.4-fold higher IC50 relative to control isolates. BSS-730A was equally active against R5 and X4 HIV isolates and displayed strong synergism with the entry inhibitor AMD3100. BSS-730A is a promising candidate for development as a potential therapeutic and/or prophylactic agent against HIV and Plasmodium.

High Hydrostatic Pressure as Sterilization Method for Drug-Loaded Intraocular Lenses.

Sterilization is a key step in the manufacturing of drug-loaded intraocular lenses (IOLs). Two of the most used methods to sterilize commercial IOLs are steam heat and gamma radiation. However, when the IOLs are loaded with drugs, the adequacy of those methods must be questioned because sterilization may affect the activity of the drugs and/or the drug release. Recently, high hydrostatic pressure (HHP), which is increasingly used in the food industry, has been applied in the sterilization of gels for medical applications. The objective of this work was to assess the performance of HHP in the sterilization of a commercial acrylic material used for the production of IOLs, both without and with loaded drugs. Bare samples and samples loaded with an antibiotic and two anti-inflammatories were tested, and the results were compared to those obtained with conventional sterilization methods. HHP not only sterilized highly contaminated samples but also enhanced drug loading and did not affect significantly the hydrogel properties. Gamma radiation degraded the drugs in solution; thus, it is adequate only for dry sample sterilization. Steam heat did not affect the release profiles but cannot be applied to temperature-sensitive drugs. We concluded that HHP may advantageously substitute steam heat and gamma radiation in the sterilization of drug-loaded IOLs.

Inhibitory activity of a green and black tea blend on Streptococcus mutans.

Through the years, tea consumption has been associated with good health, and some publications are related to oral health. The bioactive components of green tea are thought to be able to influence the process of caries formation through inhibition of proliferation of the streptococcal agent, interference with the process of bacterial adhesion to tooth enamel, and inhibition of glucosyl transferase and amylase; however, little is known about black tea and oral health. The aim of the present in-vitro study was to determine the inhibitory activity of a novel, patent-pending and proprietary blend of green and black tea aqueous extracts on Streptococcus mutans, a bacterium widely associated with plaque development and tooth decay. A minimum inhibitory concentration (MIC) of 12.5 mg/mL and a minimum bactericidal concentration (MBC) of 12.5 mg/mL was established against S. mutans, meaning that at concentrations of 12.5 mg/mL and higher, the proprietary tea blend is effective against the growth of S. mutans. This MIC concentration is lower than the ones reported in the literature for alcoholic black tea and green tea extracts tested separately. As a promising natural ingredient for oral health, this finding is a good indicator for the use of this proprietary blend of black and green tea water extracts.

Correction: Evaluation of the fusion inhibitor P3 peptide as a potential microbicide to prevent HIV transmission in women.

[This corrects the article DOI: 10.1371/journal.pone.0195744.].

Evaluation of the fusion inhibitor P3 peptide as a potential microbicide to prevent HIV transmission in women.

Microbicides are an important strategy for preventing the sexual transmission of HIV but, so far, the most advanced tenofovir-based microbicides have had modest efficacy. This has been related to adherence problems and high prevalence of tenofovir-resistant HIV-1 strains. P3 is a new peptide with potent activity against HIV that may be a good microbicide candidate. In this work P3 was formulated in a gel of hydroxyethyl cellulose and its activity, stability and safety profile in Balb/c mice were evaluated. HIV infection was fully blocked by a 1.5% gel containing P3 at the IC90 (366.4 nM) concentration. The antiviral activity did not change at 4°C during 4 months and at 25, 37 and 65°C for 1 week. P3 was stable and fully functional at acidic pH up to 24h, under different concentrations of hydrogen peroxide and in the presence of genital fluids up to 48h. P3 had no antibacterial activity and did not affect sperm motility and vitality. Finally, P3 didn't cause significant alterations in the vaginal epithelium of Balb/c mice at 0.06 (456.8 µM) and 0.2 mg/day (1522.7 µM) doses. These findings indicate that P3 is an excellent candidate for further development as a microbicide gel for the prevention of HIV transmission in women.

Influência do Nebivolol sobre o estresse oxidativo em pacientes hipertensos / Influence of Nebivolol on oxidative stress in hypertensive patients

O Nebivolol (Neb) é um antagonista dos receptores ß1-Adrenérgicos (ß1-RA) de terceira geração que causa vasodilatação efeito mediado pelo óxido nítrico e redução do estresse oxidativo (EO) em modelos pré-clínicos e estudos clínicos de Hipertensão Arterial Sistêmica (HAS). Este estudo avaliou os níveis de biomarcadores de estresse oxidativo e atividade de sistemas antioxidantes enzimáticos e não-enzimáticos na saliva de pacientes hipertensos antes e após o tratamento com doses terapêuticas anti-hipertensivas de Neb. Vinte e quatro pacientes hipertensos foram tratados com 5 mg/dia de Neb por oito semanas consecutivas. Amostras de saliva não estimulada foram coletadas de pacientes com idade entre 25 a 70 anos para análise da capacidade antioxidante total (CAT), a qual foi medida pelo Método FRAP (Ferric Reducing Antioxidant Power), a atividade enzimática da superóxido dismutase (SOD) e quantificação do ácido úrico (AU) antes (NT, grupo não tratado) e após tratamento (T, grupo tratado) com Neb. Adicionalmente, parâmetros clínicos, bioquímicos e eletrocardiográficos foram avaliados em ambos os grupos. Os grupos foram comparados pelo teste t de Student, pareado (p<0,05). Neb não alterou os valores de CAT (NT:1,697 ± 0,433 versus T: 1,674 ± 0,092 mmol/L, p=0,8454), ou a atividade da SOD (EA- NT: 66,56 ± 4,977 versus T: 60,66 ± 6,376 UE/mL, p=0,4886) na saliva. No entanto, o tratamento com Neb aumentou a concentração do AU em pacientes tratados (NT: 24,64 ± 1,686 versus T: 35,56 ± 2,014 mg/mL, p=0,0002). Os parâmetros clínicos, bioquímicos e eletrocardiográficos foram positivamente alterados com o tratamento com Neb. Nosso estudo demonstrou que o tratamento com Neb aumentou o AU na saliva de pacientes hipertensos em contexto de prática clínica real, aumentando a atividade do sistema antioxidante não-enzimático na saliva de pacientes que foram tratados com Neb. Nossos dados sugerem que o Neb pode estender seu potencial terapêutico, elevando a proteção cardiovascular contra o estresse oxidativo em adição ao seu efeito anti-hipertensivo(AU)

Nebivolol (Neb) is third-generation selective ß1-adrenergic receptor antagonist which causes vasodilation (by nitric oxide release) and has been shown to reduce oxidative stress (OS) in pre-clinical models and clinical studies. This study aimed to evaluate the OS levels and the enzymatic and non-enzymatic antioxidant systems in saliva of patients with ESH before and after anti-hypertensive therapeutic doses of Neb. Twenty-four essential hypertensive patients were treated with 5 mg Neb once daily for a total of 8 weeks. Unstimulated saliva samples were collected from 25 to 70 years-old patients for evaluation of total antioxidant capacity (TAC), which were measured by the ferric reducing antioxidant power (FRAP) assay, as well as to assess the activity of enzymatic (superoxide dismutase, SOD) and non-enzymatic (uric acid, UA) antioxidant systems, respectively before (C, control group) and after treatment (T, treated group) with Neb. Additionally, clinical, biochemical and electrocardiographic parameters were evaluated in both groups. Data were analyzed by paired Student's t-test (p < 0.05). Neb did not alter TAC values (UT: 1,697 ± 0,433 versus T: 1,674 ± 0,092 mmol/L, p=0,8454), or SOD activity (EA-UT: 66,56 ± 4,977 versus T: 60,66 ± 6,376 UE/mL, p=0,4886) in saliva. However, Neb treatment increased salivary UA activity in treated patients (UT: 24,64 ± 1,686 versus T: 35,56 ± 2,014 mg/mL, p=0,0002). Clinical, electrocardiographic and biochemical parameters were positively changed with Neb treatment. Our study showed that Neb increased UA, an OS biomarker, in saliva of hypertensive patients in a context of real clinical practice. Therefore this compound could extend its therapeutic potential, providing cardiovascular protection in addition to its ß1-antagonist effect(AU)

Evolution of the human immunodeficiency virus type 2 envelope in the first years of infection is associated with the dynamics of the neutralizing antibody response.

BACKGROUND: Differently from HIV-1, HIV-2 disease progression usually takes decades without antiretroviral therapy and the majority of HIV-2 infected individuals survive as elite controllers with normal CD4⁺ T cell counts and low or undetectable plasma viral load. Neutralizing antibodies (Nabs) are thought to play a central role in HIV-2 evolution and pathogenesis. However, the dynamic of the Nab response and resulting HIV-2 escape during acute infection and their impact in HIV-2 evolution and disease progression remain largely unknown. Our objective was to characterize the Nab response and the molecular and phenotypic evolution of HIV-2 in association with Nab escape in the first years of infection in two children infected at birth. RESULTS: CD4⁺ T cells decreased from about 50% to below 30% in both children in the first five years of infection and the infecting R5 viruses were replaced by X4 viruses within the same period. With antiretroviral therapy, viral load in child 1 decreased to undetectable levels and CD4+ T cells recovered to normal levels, which have been sustained at least until the age of 12. In contrast, viral load increased in child 2 and she progressed to AIDS and death at age 9. Beginning in the first year of life, child 1 raised high titers of antibodies that neutralized primary R5 isolates more effectively than X4 isolates, both autologous and heterologous. Child 2 raised a weak X4-specific Nab response that decreased sharply as disease progressed. Rate of evolution, nucleotide and amino acid diversity, and positive selection, were significantly higher in the envelope of child 1 compared to child 2. Rates of R5-to-X4 tropism switch, of V1 and V3 sequence diversification, and of convergence of V3 to a ß-hairpin structure were related with rate of escape from the neutralizing antibodies. CONCLUSION: Our data suggests that the molecular and phenotypic evolution of the human immunodeficiency virus type 2 envelope are related with the dynamics of the neutralizing antibody response providing further support for a model in which Nabs play an important role in HIV-2 pathogenesis.

An ancestral HIV-2/simian immunodeficiency virus peptide with potent HIV-1 and HIV-2 fusion inhibitor activity.

OBJECTIVES: To produce new fusion inhibitor peptides for HIV-1 and HIV-2 based on ancestral envelope sequences. METHODS: HIV-2/simian immunodeficiency virus (SIV) ancestral transmembrane protein sequences were reconstructed and ancestral peptides were derived from the helical region 2 (HR2). The activity of one ancestral peptide (named P3) was examined against a panel of HIV-1 and HIV-2 primary isolates in TZM-bl cells and peripheral blood mononuclear cells and compared to T-20. Peptide secondary structure was analyzed by circular dichroism. Resistant viruses were selected and resistance mutations were identified by sequencing the env gene. RESULTS: P3 has 34 residues and overlaps the N-terminal pocket-binding region and heptad repeat core of HR2. In contrast to T-20, P3 forms a typical α-helical structure in solution, binds strongly to the transmembrane protein, and potently inhibits both HIV-2 (mean IC50, 63.8 nmol/l) and HIV-1 (11 nmol/l) infection, including T-20-resistant isolates. The N43K mutation in the HR1 region of HIV-1 leads to 120-fold resistance to P3 indicating that the HR1 region in transmembrane glycoprotein is the target of P3. No HIV-2-resistant mutations could be selected by P3 suggesting that the genetic barrier to resistance is higher in HIV-2 than in HIV-1. HIV-1-infected patients presented significantly lower P3-specific antibody reactivity compared to T-20. CONCLUSION: P3 is an HIV-2/SIV ancestral peptide with low antigenicity, high stability, and potent activity against both HIV-1, including variants resistant to T-20, and HIV-2. Similar evolutionary biology strategies should be explored to enhance the production of antiviral peptide drugs, microbicides, and vaccines.

Resistance to antibody neutralization in HIV-2 infection occurs in late stage disease and is associated with X4 tropism.

OBJECTIVES: To characterize the nature and dynamics of the neutralizing antibody (NAb) response and escape in chronically HIV-2 infected patients. METHODS: Twenty-eight chronically infected adults were studied over a period of 1-4 years. The neutralizing activity of plasma immunoglobulin G (IgG) antibodies against autologous and heterologous primary isolates was analyzed using a standard assay in TZM-bl cells. Coreceptor usage was determined in ghost cells. The sequence and predicted three-dimensional structure of the C2V3C3 Env region were determined for all isolates. RESULTS: Only 50% of the patients consistently produced IgG NAbs to autologous and contemporaneous virus isolates. In contrast, 96% of the patients produced IgG antibodies that neutralized at least two isolates of a panel of six heterologous R5 isolates. Breadth and potency of the neutralizing antibodies were positively associated with the number of CD4(+) T cells and with the titer and avidity of C2V3C3-specific binding IgG antibodies. X4 isolates were obtained only from late stage disease patients and were fully resistant to neutralization. The V3 loop of X4 viruses was longer, had a higher net charge, and differed markedly in secondary structure compared to R5 viruses. CONCLUSION: Most HIV-2 patients infected with R5 isolates produce C2V3C3-specific neutralizing antibodies whose potency and breadth decreases as the disease progresses. Resistance to antibody neutralization occurs in late stage disease and is usually associated with X4 viral tropism and major changes in V3 sequence and conformation. Our studies support a model of HIV-2 pathogenesis in which the neutralizing antibodies play a central role and have clear implications for the vaccine field.

Baseline susceptibility of primary HIV-2 to entry inhibitors.

BACKGROUND: The baseline susceptibility of primary HIV-2 to maraviroc (MVC) and other entry inhibitors is currently unknown. METHODS: The susceptibility of 19 HIV-2 isolates obtained from asymptomatic and AIDS patients and seven HIV-1 clinical isolates to the fusion inhibitors enfuvirtide (ENF) and T-1249, and to the coreceptor antagonists AMD3100, TAK-779 and MVC, was measured using a TZM-bl cell-based assay. The 50% inhibitory concentration (IC(50)), 90% inhibitory concentration (IC(90)) and dose-response curve slopes were determined for each drug. RESULTS: ENF and T-1249 were significantly less active on HIV-2 than on HIV-1 (211- and 2-fold, respectively). AMD3100 and TAK-779 inhibited HIV-2 and HIV-1 CXCR4 tropic (X4) and CCR5 tropic (R5) variants with similar IC(50) and IC(90) values. MVC, however, inhibited the replication of R5 HIV-2 variants with significantly higher IC(90) values (42.7 versus 9.7 nM; P<0.0001) and lower slope values (0.7 versus 1.3; P<0.0001) than HIV-1. HIV-2 R5 variants derived from AIDS patients were significantly less sensitive to MVC than variants from asymptomatic patients, this being inversely correlated with the absolute number of CD4(+) T-cells. CONCLUSIONS: T-1249 is a potent inhibitor of HIV-2 replication indicating that new fusion inhibitors might be useful to treat HIV-2 infection. Coreceptor antagonists TAK-779 and AMD3100 are also potent inhibitors of HIV-2 replication. The reduced sensitivity of R5 variants to MVC, especially in severely immunodeficient patients, indicates that the treatment of HIV-2-infected patients with MVC might require higher dosages than those used in HIV-1 patients, and should be adjusted to the disease stage.

Origin and epidemiological history of HIV-1 CRF14_BG.

BACKGROUND: CRF14_BG isolates, originally found in Spain, are characterized by CXCR4 tropism and rapid disease progression. This study aimed to identify the origin of CRF14_BG and reconstruct its epidemiological history based on new isolates from Portugal. METHODOLOGY/PRINCIPAL FINDINGS: C2V3C3 env gene sequences were obtained from 62 samples collected in 1993-1998 from Portuguese HIV-1 patients. Full-length genomic sequences were obtained from three patients. Viral subtypes, diversity, divergence rate and positive selection were investigated by phylogenetic analysis. The molecular structure of the genomes was determined by bootscanning. A relaxed molecular clock model was used to date the origin of CRF14_BG. Geno2pheno was used to predict viral tropism. Subtype B was the most prevalent subtype (45 sequences; 73%) followed by CRF14_BG (8; 13%), G (4; 6%), F1 (2; 3%), C (2; 3%) and CRF02_AG (1; 2%). Three CRF14_BG sequences were derived from 1993 samples. Near full-length genomic sequences were strongly related to the CRF14_BG isolates from Spain. Genetic diversity of the Portuguese isolates was significantly higher than the Spanish isolates (0.044 vs 0.014, P<0.0001). The mean date of origin of the CRF14_BG cluster was estimated to be 1992 (range, 1989 and 1996) based on the subtype G genomic region and 1989 (range, 1984-1993) based on the subtype B genomic region. Most CRF14_BG strains (78.9%) were predicted to be CXCR4. Finally, up to five amino acids were under selective pressure in subtype B V3 loop whereas only one was found in the CRF14_BG cluster. CONCLUSIONS: CRF14_BG emerged in Portugal in the early 1990 s soon after the beginning of the HIV-1 epidemics, spread to Spain in late 1990 s as a consequence of IVDUs migration and then to the rest of Europe. CXCR4 tropism is a general characteristic of this CRF that may have been selected for by escape from neutralizing antibody response.

Evolutionary and structural features of the C2, V3 and C3 envelope regions underlying the differences in HIV-1 and HIV-2 biology and infection.

BACKGROUND: Unlike in HIV-1 infection, the majority of HIV-2 patients produce broadly reactive neutralizing antibodies, control viral replication and survive as elite controllers. The identification of the molecular, structural and evolutionary footprints underlying these very distinct immunological and clinical outcomes may lead to the development of new strategies for the prevention and treatment of HIV infection. METHODOLOGY/PRINCIPAL FINDINGS: We performed a side-by-side molecular, evolutionary and structural comparison of the C2, V3 and C3 envelope regions from HIV-1 and HIV-2. These regions contain major antigenic targets and are important for receptor binding. In HIV-2, these regions also have immune modulatory properties. We found that these regions are significantly more variable in HIV-1 than in HIV-2. Within each virus, C3 is the most entropic region followed by either C2 (HIV-2) or V3 (HIV-1). The C3 region is well exposed in the HIV-2 envelope and is under strong diversifying selection suggesting that, like in HIV-1, it may harbour neutralizing epitopes. Notably, however, extreme diversification of C2 and C3 seems to be deleterious for HIV-2 and prevent its transmission. Computer modelling simulations showed that in HIV-2 the V3 loop is much less exposed than C2 and C3 and has a retractile conformation due to a physical interaction with both C2 and C3. The concealed and conserved nature of V3 in the HIV-2 is consistent with its lack of immunodominancy in vivo and with its role in preventing immune activation. In contrast, HIV-1 had an extended and accessible V3 loop that is consistent with its immunodominant and neutralizing nature. CONCLUSIONS/SIGNIFICANCE: We identify significant structural and functional constrains to the diversification and evolution of C2, V3 and C3 in the HIV-2 envelope but not in HIV-1. These studies highlight fundamental differences in the biology and infection of HIV-1 and HIV-2 and in their mode of interaction with the human immune system and may inform new vaccine and therapeutic interventions against these viruses.