Nef enhances HIV-1 replication and infectivity independently of SERINC5 in CEM T cells.

A primary function of HIV-1 Nef is the enhancement of viral infectivity and replication. Whether counteraction of the antiretroviral proteins SERINC3 and SERINC5 is the cause of this positive influence on viral growth-rate and infectivity remains unclear. Here, we utilized CRISPR/Cas9 to knockout SERINC3 and SERINC5 in a leukemic CD4-positive T cell line (CEM) that displays nef-related infectivity and growth-rate phenotypes. Viral replication was attenuated in CEM cells infected with HIV-1 lacking Nef (HIV-1ΔNef). This attenuated growth-rate phenotype was observed regardless of whether the coding regions of the serinc3 or serinc5 genes were intact. Moreover, knockout of serinc5 alone or of both serinc5 and serinc3 together failed to restore the infectivity of HIV1ΔNef virions produced from infected CEM cells. Our results corroborate a similar study using another T-lymphoid cell line (MOLT-3) and indicate that the antagonism of SERINC3 and SERINC5 does not fully explain the virology of HIV-1 lacking Nef.

Demographic and clinical factors associated with same-day discharge and unplanned readmission following shoulder arthroplasty: a retrospective cohort study.

BACKGROUND: Same-day discharge, defined as discharge from the hospital within 24 h of surgery, has been shown to be safe for joint arthroplasty. We examined demographic and clinical factors associated with same-day discharge and unplanned readmission following shoulder arthroplasty in adult patients. METHODS: Utilizing data from the American College of Surgeons National Surgical Quality Improvement Program database, we extracted information of all patients that underwent shoulder arthroplasty. The primary and secondary outcome of interest was same-day discharge and 30-day unplanned readmission, respectively. We utilized multivariable logistic regression to identify covariates associated with these outcomes. RESULTS: There were 17,011 patients analyzed when identifying predictors for same-day discharge. There was an increase in same-day discharge from 2007 to 2016. The odds of same-day discharge were significantly better for males (P < 0.001). The odds of same-day discharge was significantly decreased for every 10-year increase in age and for patients with insulin dependent diabetes, poor functional status, chronic obstructive pulmonary disease, congestive heart failure, bleeding disorder, and comorbidity burden (all P < 0.001). There were 14,276 patients analyzed for hospital readmission. The odds of unplanned readmission were significantly higher for every 10-year increase in age and for patients with poor functional status, congestive heart failure, bleeding disorder, and higher comorbidity burden (all P < 0.001). CONCLUSIONS: The results of this study show that preoperative comorbidities and advanced age reduce the odds of same-day discharge. Risk stratification, preoperative optimization, and coordinated care after surgery may be helpful to optimize patients for same-day discharge.

Inconsistent reversal of HIV-1 latency ex vivo by antigens of HIV-1, CMV, and other infectious agents.

BACKGROUND: A reservoir of replication-competent but latent virus is the main obstacle to a cure for HIV-1 infection. Much of this reservoir resides in memory CD4 T cells. We hypothesized that these cells can be reactivated with antigens from HIV-1 and other common pathogens to reverse latency. RESULTS: We obtained mononuclear cells from the peripheral blood of antiretroviral-treated patients with suppressed viremia. We tested pools of peptides and proteins derived from HIV-1 and from other pathogens including CMV for their ability to reverse latency ex vivo by activation of memory responses. We assessed activation of the CD4 T cells by measuring the up-regulation of cell-surface CD69. We assessed HIV-1 expression using two assays: a real-time PCR assay for virion-associated viral RNA and a droplet digital PCR assay for cell-associated, multiply spliced viral mRNA. Reversal of latency occurred in a minority of cells from some participants, but no single antigen induced HIV-1 expression ex vivo consistently. When reversal of latency was induced by a specific peptide pool or protein, the extent was proportionally greater than that of T cell activation. CONCLUSIONS: In this group of patients in whom antiretroviral therapy was started during chronic infection, the latent reservoir does not appear to consistently reside in CD4 T cells of a predominant antigen-specificity. Peptide-antigens reversed HIV-1 latency ex vivo with modest and variable activity. When latency was reversed by specific peptides or proteins, it was proportionally greater than the extent of T cell activation, suggesting partial enrichment of the latent reservoir in cells of specific antigen-reactivity.

Sensitivity to monoclonal antibody 447-52D and an open env trimer conformation correlate poorly with inhibition of HIV-1 infectivity by SERINC5.

The host protein SERINC5 inhibits the infectivity of HIV-1 virions in an Env-dependent manner and is counteracted by Nef. The conformation of the Env trimer reportedly correlates with sensitivity to SERINC5. Here, we tested the hypothesis that the "open" conformation of the Env trimer revealed by sensitivity to the V3-loop specific antibody 447-52D directly correlates with sensitivity to SERINC5. Of five Envs tested, SF162 was the most sensitive to neutralization by 447-52D, but it was not the most sensitive to SERINC5; instead the Env of LAI was substantially more sensitive to SERINC5 than all the other Envs. Mutational opening of the trimer by substitution of two tyrosines that mediate interaction between the V2 and V3 loops sensitized the Envs of JRFL and LAI to 447-52D as previously reported, but only BaL was sensitized to SERINC5. These data suggest that trimer "openness" is not sufficient for sensitivity to SERINC5.