Does rapid HIV disease progression prior to combination antiretroviral therapy hinder optimal CD4 + T-cell recovery once HIV-1 suppression is achieved?

Objective: This article compares trends in CD4+ T-cell recovery and proportions achieving optimal restoration (>=500 cells/µl) after viral suppression following combination antiretroviral therapy (cART) initiation between rapid and nonrapid progressors. Methods: We included HIV-1 seroconverters achieving viral suppression within 6 months of cART. Rapid progressors were individuals experiencing at least one CD4+ less than 200 cells/µl within 12 months of seroconverters before cART. We used piecewise linear mixed models and logistic regression for optimal restoration. Results: Of 4024 individuals, 294 (7.3%) were classified as rapid progressors. At the same CD4+ T-cell count at cART start (baseline), rapid progressors experienced faster CD4+ T-cell increases than nonrapid progressors in first month [difference (95% confidence interval) in mean increase/month (square root scale): 1.82 (1.61; 2.04)], which reversed to slightly slower increases in months 1–18 [-0.05 (-0.06; -0.03)] and no significant differences in 18–60 months [-0.003 (-0.01; 0.01)]. Percentage achieving optimal restoration was significantly lower for rapid progressors than nonrapid progressors at months 12 (29.2 vs. 62.5%) and 36 (47.1 vs. 72.4%) but not at month 60 (70.4 vs. 71.8%). These differences disappeared after adjusting for baseline CD4+ T-cell count: odds ratio (95% confidence interval) 0.86 (0.61; 1.20), 0.90 (0.38; 2.17) and 1.56 (0.55; 4.46) at months 12, 36 and 60, respectively. Conclusion: Among people on suppressive antiretroviral therapy, rapid progressors experience faster initial increases of CD4+ T-cell counts than nonrapid progressors, but are less likely to achieve optimal restoration during the first 36 months after cART, mainly because of lower CD4+ T-cell counts at cART initiation

Another Shipment of Six Short-Period Giant Planets from TESS

We present the discovery and characterization of six short-period, transiting giant planets from NASA's Transiting Exoplanet Survey Satellite (TESS) -- TOI-1811 (TIC 376524552), TOI-2025 (TIC 394050135), TOI-2145 (TIC 88992642), TOI-2152 (TIC 395393265), TOI-2154 (TIC 428787891), & TOI-2497 (TIC 97568467). All six planets orbit bright host stars (8.9 <G< 11.8, 7.7 <K< 10.1). Using a combination of time-series photometric and spectroscopic follow-up observations from the TESS Follow-up Observing Program (TFOP) Working Group, we have determined that the planets are Jovian-sized (R$_{P}$ = 1.00-1.45 R$_{J}$), have masses ranging from 0.92 to 5.35 M$_{J}$, and orbit F, G, and K stars (4753 $<$ T$_{eff}$ $<$ 7360 K). We detect a significant orbital eccentricity for the three longest-period systems in our sample: TOI-2025 b (P = 8.872 days, $e$ = $0.220\pm0.053$), TOI-2145 b (P = 10.261 days, $e$ = $0.182^{+0.039}_{-0.049}$), and TOI-2497 b (P = 10.656 days, $e$ = $0.196^{+0.059}_{-0.053}$). TOI-2145 b and TOI-2497 b both orbit subgiant host stars (3.8 $<$ $\log$ g $<$4.0), but these planets show no sign of inflation despite very high levels of irradiation. The lack of inflation may be explained by the high mass of the planets; $5.35^{+0.32}_{-0.35}$ M$_{\rm J}$ (TOI-2145 b) and $5.21\pm0.52$ M$_{\rm J}$ (TOI-2497 b). These six new discoveries contribute to the larger community effort to use {\it TESS} to create a magnitude-complete, self-consistent sample of giant planets with well-determined parameters for future detailed studies.Comment: 20 Pages, 6 Figures, 8 Tables, Accepted by MNRA

The concordance of the limiting antigen and the Bio-Rad avidity assays in persons from Estonia infected mainly with HIV-1 CRF06_cpx

BACKGROUND: Serological assays to determine HIV incidence have contributed to estimates of HIV incidence, monitoring of HIV spread, and evaluation of prevention strategies. Two frequently used incidence assays are the Sedia HIV-1 LAg-Avidity EIA (LAg) and the Bio-Rad avidity incidence (BRAI) assays with a mean duration of recent infection (MDRI) of 130 and 240 days for subtype B infections, respectively. Little is known about how these assays perform with recombinant HIV-1 strains. We evaluated the concordance of these assays in a population infected mainly with HIV-1 CRF06_cpx. MATERIAL/METHODS: Remnant serum samples (n = 288) collected from confirmed, newly-diagnosed HIV-positive persons from Estonia in 2013 were tested. Demographic and clinical data were extracted from clinical databases. LAg was performed according to the manufacturer’s protocol and BRAI testing was done using a validated protocol. Samples with LAg-pending or BRAI-invalid results were reclassified as recent if they were from persons with viral loads <1000 copies/mL or were reclassified as long-term if presenting with AIDS. RESULTS: In total 325 new HIV infections were diagnosed in 2013 in Estonia. Of those 276 persons were tested with both LAg and BRAI. Using assay results only, the recency rate was 44% and 70% by LAg and BRAI, respectively. The majority of samples (92%) recent by LAg were recent by BRAI. Similarly, 89% of samples long-term by BRAI were long-term by LAg. After clinical information was included in the analysis, the recency rate was 44% and 62% for LAg and BRAI, respectively. The majority of samples (86%) recent by LAg were recent by BRAI and 91% of long-term infections by BRAI were long-term by LAg. CONCLUSIONS: Comparison of LAg and BRAI results in this mostly CRF06_cpx-infected population showed good concordance for incidence classification. Our finding of a higher recency rate with BRAI in this population is likely related to the longer MDRI for this assay

Site-Specific, Insertional Inactivation of <i>incA</i> in <i>Chlamydia trachomatis</i> Using a Group II Intron

<div><p><i>Chlamydia trachomatis</i> is an obligate, intracellular bacterial pathogen that has until more recently remained recalcitrant to genetic manipulation. However, the field still remains hindered by the absence of tools to create selectable, targeted chromosomal mutations. Previous work with mobile group II introns demonstrated that they can be retargeted by altering DNA sequences within the intron’s substrate recognition region to create site-specific gene insertions. This platform (marketed as TargeTron™, Sigma) has been successfully employed in a variety of bacteria. We subsequently modified TargeTron™ for use in <i>C. trachomatis</i> and as proof of principle used our system to insertionally inactivate <i>incA</i>, a chromosomal gene encoding a protein required for homotypic fusion of chlamydial inclusions. <i>C. trachomatis incA</i>::GII(<i>bla</i>) mutants were selected with ampicillin and plaque purified clones were then isolated for genotypic and phenotypic analysis. PCR, Southern blotting, and DNA sequencing verified proper GII(<i>bla</i>) insertion, while continuous passaging in the absence of selection demonstrated that the insertion was stable. As seen with naturally occurring IncA⁻ mutants, light and immunofluorescence microscopy confirmed the presence of non-fusogenic inclusions in cells infected with the <i>incA</i>::GII(<i>bla</i>) mutants at a multiplicity of infection greater than one. Lack of IncA production by mutant clones was further confirmed by Western blotting. Ultimately, the ease of retargeting the intron, ability to select for mutants, and intron stability in the absence of selection makes this method a powerful addition to the growing chlamydial molecular toolbox.</p></div

Formation of non-fusogenic inclusions by <i>incA</i>::GII(<i>bla</i>) mutants in a cell culture infection model.

<p>Cells were infected at an MOI of ∼10 (panels A–C), ∼5 (panel J), or ∼0.1 (panels D–I) with ACE051, DFCT3, or DFCT4 and viewed under 400× phase contrast at 24 (panels A–F) or 48 hours (panels G–J) post infection. White arrows designate the location of inclusions. Images are representative of three independent experiments.</p

PCR verification of <i>incA</i>::GII(<i>bla</i>) clones plaque purified with or without ampicillin selection.

<p>PCR was performed using genomic DNA from the wild type ACE051 strain and the plaque purified strains DFCT3 and DFCT4 to assess intron insertion, loss of the intron donor plasmid, and maintenance of the chlamydial cryptic plasmid. Primers were designed to amplify regions of the <i>incA</i> locus, cryptic plasmid, intron, and pDFTT3 (the intron donor plasmid). Wild type and mutant <i>incA</i> locus maps and plasmid maps are shown in panels A–D. Expected PCR product sizes (in kbp) are shown on each map with the PCR reaction and region amplified represented by black bars. The predicted GII(<i>bla</i>) intron insertion site is shown in panel A with brackets. PCR reactions and primers are detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083989#pone.0083989.s009" target="_blank">Tables S2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083989#pone.0083989.s010" target="_blank">S3</a>. PCR results are shown in panel E. Products were separated on 1% agarose gels and stained with ethidium bromide. Gels were visualized using UV transillumination and photographed. Images have been reversed to aid in product visualization. The template used in the PCR reaction is listed at the top of each lane. Reactions performed are shown at the bottom of each gel. Molecular weight markers were loaded in lane 1 of each gel and the marker sizes are listed in kbp to the left of each gel.</p

IncA Western blot analysis of the <i>incA</i>::GII(<i>bla</i>) mutants.

<p>Cells were grown until confluent and then infected at an MOI of ∼10 with ACE051, DFCT3, or DFCT4 to assess the production of IncA. After 24 hours, infected (and mock infected) cells were lysed with Laemmli buffer and loaded onto 12% SDS-PAGE gels. Gels were then either stained with Coomassie Brilliant Blue (A) or transferred to nitrocellulose for anti-MOMP blotting (B) or anti-IncA blotting (C). Expected protein sizes are 42.5 kDa (MOMP) and 30.3 kDa (IncA). Molecular weight markers are shown in lane one in each panel and marker sizes in kDa are listed to the left of panel A. Blots are representative of three independent experiments.</p

Immunofluorescence analysis of <i>incA</i>::GII(<i>bla</i>) mutants.

<p>Cells were infected at an MOI of ∼10 with ACE051, DFCT3, or DFCT4. After 24 hours post infection, cells were fixed for immuno-probing with mouse anti-MOMP antibodies to detect <i>Chlamydia</i>, panels B/E/H (visualized with a goat anti-mouse Texas Red conjugated secondary antibody), and stained with DAPI, panels A/D/G. For image overlay analysis, anti-MOMP images were false colored red and DAPI images were false colored blue. The resulting overlays are shown in panels C/F/I. Blue arrows in panels C/F/I highlight cell nuclei while red arrows highlight inclusions. All images were taken under oil immersion at 630× using a fluorescent microscope. Infections and immuno-processing were performed three times.</p