A HIV-1 Gp41 Peptide-Liposome Vaccine Elicits Neutralizing Epitope-Targeted Antibody Responses in Healthy Individuals.

Background: HIV-1 vaccine development is a global health priority. Broadly neutralizing antibodies (bnAbs) which target the HIV-1 gp41 membrane-proximal external region (MPER) have some of the highest neutralization breadth. An MPER peptide-liposome vaccine has been found to expand bnAb precursors in monkeys. Methods: The HVTN133 phase 1 clinical trial (NCT03934541) studied the MPER-peptide liposome immunogen in 24 HIV-1 seronegative individuals. Participants were recruited between 15 July 2019 and 18 October 2019 and were randomized in a dose-escalation design to either 500 mcg or 2000 mcg of the MPER-peptide liposome or placebo. Four intramuscular injections were planned at months 0, 2, 6, and 12. Results: The trial was stopped prematurely due to an anaphylaxis reaction in one participant ultimately attributed to vaccine-associated polyethylene glycol. The immunogen induced robust immune responses, including MPER+ serum and blood CD4+ T-cell responses in 95% and 100% of vaccinees, respectively, and 35% (7/20) of vaccine recipients had blood IgG memory B cells with MPER-bnAb binding phenotype. Affinity purification of plasma MPER+ IgG demonstrated tier 2 HIV-1 neutralizing activity in two of five participants after 3 immunizations. Conclusions: MPER-peptide liposomes induced gp41 serum neutralizing epitope-targeted antibodies and memory B-cell responses in humans despite the early termination of the study. These results suggest that the MPER region is a promising target for a candidate HIV vaccine.

Achieving intracellular cytokine staining assay concordance on two continents to assess HIV vaccine-induced T-cell responses.

The HIV Vaccine Trials Network (HVTN) conducts clinical trials on 4 continents in pursuit of a safe and effective HIV vaccine. Cellular immune responses to vaccination that define vaccine immunogenicity and/or immune correlates of protection can be measured using multiparameter intracellular cytokine staining (ICS) assays. The HVTN cellular immunology laboratory, located in Seattle, WA, conducts ICS assays for vaccine trials according to Good Clinical Laboratory Practices (GCLP). In 2013, the HVTN established a second GCLP compliant cellular immunology laboratory in Cape Town, South Africa to assess vaccine immunogenicity for HVTN trials conducted on the African continent. To ensure ICS readouts in the 2 laboratories were directly comparable, we conducted concordance testing using PBMC from healthy controls and vaccine trial participants. Despite standardized procedures and instrumentation, shared quality control measures and quality assurance oversight, several factors impacted our ability to obtain close agreement in T-cell responses measured in the 2 laboratories. One of these was the type of fetal bovine serum (FBS) used in the assay, which impacted lymphocyte cell viability and background responses. In addition, the differences in supernatant removal technique also significantly affected our ability to detect positive responses to vaccine antigens. Standardization of these factors allowed us to achieve and maintain ICS assay concordance across the 2 laboratories over multiple years, accelerating our efforts to evaluate HIV vaccines. The insights gained in this process are valuable for assay transfer efforts by groups of investigators that need to directly compare data generated in different laboratories around the globe.

Subjective Cognitive Complaints: Predictors and Health Outcomes in People Living with HIV.

There is a paucity of research on the prevalence of subjective cognitive complaints in people living with human immunodeficiency virus, along with the predictors and outcomes related to these complaints. We assessed demographics, substance use and psychiatric predictors, and HIV-related outcomes associated with subjective cognitive complaint items from the Cognitive Difficulties Scale. The sample consisted of 889 people living with HIV in the survey-based Florida Cohort. Results of multivariable regression models indicated that age (45-54), hazardous alcohol consumption, more frequent marijuana use and psychiatric symptoms (depression, anxiety, PTSD) were significant predictors of subjective cognitive complaints. Subjective cognitive complaints were associated with lower adherence to antiretroviral therapy in bivariate analyses, but this relationship was no longer significant after controlling for depression, race, alcohol and drug use. Further research into the relationship between depressive and subjective cognitive complaints may provide additional avenues for intervention.

RESUMEN: Existe una escasez de investigación sobre la prevalencia de quejas cognitivas subjetivas en personas que viven con el virus de la inmunodeficiencia humana (VIH), junto con los predictores y los resultados relacionados con estas quejas. Evaluamos la demografía, el uso de sustancias y los predictores psiquiátricos, y los resultados relacionados con el VIH asociados con los ítems de quejas cognitivas subjetivas de la Escala de Dificultades Cognitivas. La muestra consistió en 889 personas que viven con el VIH en la cohorte de Florida basada en la encuesta. Los resultados de los modelos de regresión multivariable indicaron que la edad (45-54), el consumo peligroso de alcohol, el uso más frecuente de marihuana y los síntomas psiquiátricos (depresión, ansiedad, trastorno de estrés postraumático) fueron predictores significativos de quejas cognitivas subjetivas. Las quejas cognitivas subjetivas se asociaron con una menor adherencia a la terapia antirretroviral en los análisis bivariados, pero esta relación dejó de ser significativa después de controlar la depresión, la raza, el alcohol y el consumo de drogas. La investigación adicional sobre la relación entre las quejas cognitivas depresivas y subjetivas puede proporcionar vías adicionales de intervención.

Defining the optimal cut-point of self-reported ART adherence to achieve viral suppression in the era of contemporary HIV therapy: a cross-sectional study.

BACKGROUND: When considering adherence to antiretroviral therapy (ART) for HIV, many different cut-points are used. The primary goals of this study were to identify a level of self-reported medication adherence that best distinguished HIV viral suppression from non-suppression, and to compare the ability of a single-item and a 3-item adherence questionnaire to predict HIV viral suppression. METHODS: This cross-sectional analysis included 380 persons with HIV (PWH) from the Florida Cohort study who completed a self-reported ART adherence measure within 30-days of having an HIV viral load test. We used Receiver Operating Characteristic (ROC) curve analyses and ROCContrast to compare the ability of a single-item and a 3-item self-reported adherence measure to predict HIV viral suppression (defined as ≤ 200 copies/mL). We used the Youden index and chi square statistics to assess specific cut-points, and repeated the analysis with a different definition of HIV viral suppression (≤ 1000 copies/mL). RESULTS: The mean percent adherence was 92.4% using the single-item score and 90.4% using the 3-item score; 81.6% had viral suppression. The areas under the curve for the single-item and 3-item adherence measures were generally poor overall and not significantly different from each other (0.589 and 0.580, p = 0.67). The Youden index identified cut-points of 93% and 89% as maximizing the sensitivity and specificity for the single-item and 3-item measures, respectively, whereas a cut-point of 80% on the single-item measure was best able to discriminate those with viral suppression (58% vs. 84%, p < 0.001). Results were similar with viral suppression defined as ≤ 1000 copies/mL. CONCLUSIONS: In this sample of PWH, a single question on medication adherence was as good as a 3-item questionnaire in predicting HIV viral suppression, although neither had good discriminatory ability. A cut-point close to 90% adherence maximized sensitivity and specificity, although viral suppression was very similar for nearly all measures above 80%.

Factors associated with current and severe pain among people living with HIV: results from a statewide sample.

BACKGROUND: People living with HIV (PLHIV) are more likely to suffer from pain compared to the general public. Pain often clusters with mental health symptoms and substance use. This study sought to evaluate mental health and substance use factors associated with any pain and severe pain intensities among PLHIV. METHODS: Data were derived from HIV+ adults (N = 733) recruited from community health centers across Florida who completed questionnaires regarding demographics, chronic pain, HIV clinical outcomes, mental health symptoms, and substance use information. Pain was assessed using the Brief Pain Inventory (BPI) short form. Multivariate logistic regression analysis was utilized to assess the relationship between selected covariates and pain. RESULTS: Approximately half (45.0%) of participants reported having any current pain while 16.1% reported severe pain. The odds of having any current pain were 2.49 (CI 95% 1.48, 4.18, p <  0.01) times greater among PLHIV reporting anxiety and 1.69 (CI 95% 1.11, 2.57, p = 0.01) times greater among PLHIV reporting PTSD compared to those without those factors. The odds of having severe pain were 2.03 (CI 95% 1.03, 4.01, p = 0.04) times greater among PLHIV reporting anxiety and 2.02 (CI 95% 1.26, 3.24, p <  0.01) times greater among female participants compared to PLHIV without those factors respectively. Factors including depression, alcohol consumption, and marijuana use were not statistically associated with any current pain nor with severe pain. CONCLUSION: The relationship between pain and mental health is complex. Thus, future research is needed to determine if pain treatments may reduce mental health symptoms or if treatments can be targeted to address both issues simultaneously.

Perceived Positive Consequences Are Associated with Unhealthy Alcohol Use Among Persons Living with HIV.

BACKGROUND: Understanding how perceived positive consequences are associated with drinking may help improve effectiveness of alcohol reduction interventions among people living with HIV (PLWH). We aimed to determine whether perceived positive consequence scores varied by sociodemographic, drinking, mental health or substance use variables. METHODS: Perceived positive consequences of drinking were assessed using the PROMIS: Positive Consequences-Short Form. Unhealthy alcohol use was measured using a modified AUDIT-C. We used multiple linear regression to identify factors associated with perceived positive consequence. RESULTS: 328 PLWH who consumed at least one alcoholic beverage in the last 6 months participated in the Florida Cohort study (mean age=46, 69% male, 58% Black). Perceived positive consequence scores ranged from 0 to 28 (mean=16.1, SD=6.9). Perceived positive consequence scores increased by 0.8 points for each 1-point increase in AUDIT-C score. Demographics, thoughts on reducing alcohol use, other substance use, depression, and anxiety were not significantly associated with perceived positive consequences. CONCLUSIONS: Our findings suggest perceived positive consequences are associated with unhealthy alcohol use. Positive consequences should be considered with negative consequences in a decisional balance when intervening on alcohol use among PLWH.

CONTEXTE: Comprendre comment les conséquences positives perçues sont associées à la consommation d'alcool peut aider à améliorer l'efficacité des interventions de réduction de l'alcool chez les personnes vivant avec le VIH (PVVIH). Nous avons cherché à déterminer si les scores des conséquences positives perçues variaient selon les variables sociodémographiques, de consommation d'alcool, de santé mentale ou de consommation de substances. MÉTHODES: Les conséquences positives perçues de la consommation d'alcool ont été évaluées à l'aide du PROMIS: Positive Consquences - Short Form -conséquences positives­version abrégée. La consommation d'alcool malsaine a été mesurée à l'aide d'un AUDIT-C modifié. Nous avons utilisé une régression linéaire multiple pour identifier les facteurs associés aux conséquences positives perçues. RÉSULTATS: 328 PVVIH ayant consommé au moins une boisson alcoolisée au cours des 6 derniers mois ont participé à l'étude de la cohorte de Floride (âge moyen=46 ans, 69% d'hommes, 58% de noirs). Les scores des conséquences positives perçues allaient de 0 à 28 (moyenne=16.1, ET=6.9). Les scores de conséquences positives perçues ont augmenté de 0.8 point pour chaque augmentation de 1 point du score AUDIT-C.Les données démographiques, les réflexions sur la réduction de la consommation d'alcool, la consommation d'autres substances, la dépression et l'anxiété n'étaient pas associées de manière significative aux conséquences positives perçues. CONCLUSIONS: Nos résultats suggèrent que les conséquences positives perçues sont associées à une consommation d'alcool malsaine. Les conséquences positives doivent être considérées avec des conséquences négatives dans le processus décisionnel lors de l'intervention sur la consommation d'alcool chez les PVVIH.

Increasing antioxidant activities of the glutamine-cysteine mixture by the glutaminase from Bacillus amyloliquefaciens.

In this study, we found that glutaminase from Bacillus amyloliquefaciens could increase antioxidant activities of Gln-Cys mixture, demonstrated by the higher superoxide anion and DPPH radical scavenging activities, Fe2+-chelating and reducing power. According to UPLC-Q-TOF-MS/MS results, we identified γ-[Glu](n=1,2,3,4)-Cys in Gln-Cys mixture in the presence of glutaminase. The yields of γ-Glu-Cys (GC) and γ-Glu-γ-Glu-Cys (GGC) were 40.92% and 22.79% respectively, under the established optimum conditions: pH 10, 37 °C, 3 h, 0.1 mol/l Gln: 0.1 mol/l Cys = 1:1, and glutaminase at 0.1% (m/v). The antioxidant properties of GC, GGC, glutathione and Gln-Cys mixture in the presence of glutaminase were further compared and we found GC exhibited the highest superoxide anion and DPPH radical scavenging activities, and Fe2+-chelating and reducing power. Therefore, glutaminase caused the increase of antioxidant activity of Gln-Cys mixture and might be attributed to GC which was synthesized by glutaminase via transpeptidation, and GC has the potential to be used in food and nutraceutical applications as an antioxidant peptide.

The prevalence and patterns of substance use by birth cohort among HIV-positive adults in Florida.

OBJECTIVES: Antiretroviral therapy is affording longer lifespans for people living with HIV (PLWH), yet factors such as substance use play an increasing role in morbidity and mortality in this population. Though previous studies have examined substance use differences between age cohorts of PLWH, no study has examined the influence of birth cohort on current substance use patterns. Thus, this study investigated the prevalence of past 12-month self-reported substance use between four birth cohorts, <1970 (M age = 54.1), 1970s (M age = 41.5), 1980s (M age = 31.3 years old), and 1990s (M age = 23.2 years old) of PLWH in Florida. METHODS: PLWH (N = 934) recruited from community health clinics in Florida completed a questionnaire assessing sociodemographics, health status, and substance use. Multivariate logistic regressions utilizing the <1970 cohort as the referent group examined the relationship between birth cohort and substance use. RESULTS: The 1980s cohort had significantly greater odds of marijuana use compared to the oldest cohort (<1970s), while the three younger cohorts (1970s, 1980s, and 1990s) evidenced a significantly greater odds of ecstasy use compared to the oldest group. Contrastingly, the three younger birth cohorts reported significantly less crack use than the oldest cohort, while the youngest group (1990s) also demonstrated an 80% reduction in injection drug use compared to the oldest group. CONCLUSION: The older cohort evidenced significantly greater crack and injection drug use, while the younger cohorts evidenced greater marijuana and ecstasy use. Therefore, it is important to develop age-specific substance use interventions among PLWH.

The association of therapeutic versus recreational marijuana use and antiretroviral adherence among adults living with HIV in Florida.

PURPOSE: Marijuana use is common among people living with HIV (PLWH), but its association with antiretroviral therapy (ART) adherence is unclear. This study examined the association between reason for marijuana use and ART adherence in a sample of adults living with HIV. PATIENTS AND METHODS: Participants (N=703) recruited from seven community health centers in Florida completed a 45-minute questionnaire assessing demographics, symptoms of anxiety and depression, ART adherence, and substance use, including reasons for marijuana use. ART adherence was defined as the proportion of days in the last 30 days participants did not miss any medication and dichotomized as optimal (≥95%) and suboptimal (<95%). Multivariate logistic regression analysis assessed the association between therapeutic marijuana use to manage HIV symptoms (ie, improve appetite/gain weight, induce sleep, relieve nausea/vomiting, relieve pain, relieve anxiety/depression/stress) versus recreational marijuana use and ART adherence. RESULTS: Approximately one third (33.2%) of the participants reported using marijuana in the past 3 months. Of marijuana users, 21.8% reported using marijuana only for therapeutic purposes to manage HIV-associated medical symptoms, while 78.2% reported recreational use. After controlling for covariates, therapeutic use of marijuana was not associated with ART adherence (AOR =1.19, 95% CI =0.60-2.38, p=0.602) while recreational marijuana users showed significantly greater odds of suboptimal ART adherence compared to nonusers (AOR =1.80, 95% CI =1.18-2.72, p=0.005). CONCLUSION: Our results suggest differences in ART adherence between individuals who report recreational versus therapeutic marijuana use. Continued research examining the health implications of marijuana use among adults living with HIV is important as legalization of recreational and medical marijuana proliferates in the United States.

Protein interactions and regulation of EscA in enterohemorrhagic E. coli.

Infections caused by enterohemorrhagic Escherichia coli (EHEC) can lead to diarrhea with abdominal cramps and sometimes are complicated by severe hemolytic uremic syndrome. EHEC secretes effector proteins into host cells through a type III secretion system that is composed of proteins encoded by a chromosomal island, locus for the enterocyte effacement (LEE). EspA is the major component of the filamentous structure connecting the bacteria and the host's cells. Synthesis and secretion of EspA must be carefully controlled since the protein is prone to polymerize. CesAB, CesA2, and EscL have been identified as being able to interact with EspA. Furthermore, the intracellular level of EspA declines when cesAB, cesA2, and escL are individually deleted. Here, we report a LEE gene named l0033, which also affects the intracellular level of EspA. We renamed l0033 as escA since its counterpart in enteropathogenic E. coli has been recently described. Similar to CesAB, EscL, and CesA2, EscA interacts with EspA and enhances the protein stability of EspA. However, EscA is also able to interact with inner membrane-associated EscL, CesA2, and EscN, but not with cytoplasmic CesAB. In terms of gene organizations, escA locates in LEE3. Expression of EscA is faithfully regulated via Mpc, the first gene product of LEE3. Since Mpc is tightly regulated to low level, we suggest that EscA is highly synchronized and critical to the process of escorting EspA to its final destination.