A simple method to elute cell-free HIV from dried blood spots improves their usefulness for monitoring therapy.

Background: Dried blood spots (DBS) improve access to HIV viral load (VL) testing, but yield increased VL measurements compared to the plasma reference method because of cell-associated viral nucleic acid. In clinical settings, DBS methods may falsely categorize many patients as failing therapy.

Objectives: Description of a simple method, free virus elution (FVE), to preferentially elute plasma-associated virus from DBS samples with phosphate-buffered saline, and an initial HIV VL performance comparison with standard DBS elution methods.

Wrestling with the repertoire: the promise and perils of next generation sequencing for antigen receptors.

Next generation sequencing technologies are revolutionizing the study of immune repertoires. These methods provide a previously unimaginable amount of sequence data, unfortunately accompanied by numerous challenges associated with error correction and interpretation that remain to be solved. For antigen receptors, these challenges will require dedicated solutions beyond those developed for genome sequencing, which may differ depending on the sequencing technology used and the purpose of the experiment.

Blood T-cell receptor diversity decreases during the course of HIV infection, but the potential for a diverse repertoire persists.

HIV infection results in a decrease in circulating CD4(+) T-cell and naive T-cell numbers. If such losses were associated with an erosion of T-cell receptor (TCR) repertoire diversity in the peripheral T-cell pool, this might exacerbate the state of persistent immunodeficiency. Existing methods for the analysis of the TCR repertoire have demonstrated skewed distributions of TCR genes in HIV-infected subjects but cannot directly measure TCR diversity.

Abacavir increases platelet reactivity via competitive inhibition of soluble guanylyl cyclase.

Objective: To provide a molecular mechanism that explains the association of the antiretroviral guanosine analogue, abacavir, with an increased risk of myocardial infarction.

Design: Drug effects were studied with biochemical and cellular assays.

Methods: Human platelets were incubated with nucleoside analogue drugs ex vivo.

Measurement of absolute T cell receptor rearrangement diversity.

T cell receptor (TCR) diversity is critical for adaptive immunity. Existing methods for measuring such diversity are qualitative, expensive, and/or of uncertain accuracy. Here, we describe a method and associated reagents for estimating the absolute number of unique TCR Vβ rearrangements present in a given number of cells or volume of blood.

Design, construction, and validation of a modular library of sequence diversity standards for polymerase chain reaction.

Methods to measure the sequence diversity of polymerase chain reaction (PCR)-amplified DNA lack standards for use as assay calibrators and controls. Here we present a general and economical method for developing customizable DNA standards of known sequence diversity. Standards ranging from 1 to 25,000 sequences were generated by directional ligation of oligonucleotide "words" of standard length and GC content and then amplified by PCR.

Growth hormone enhances thymic function in HIV-1-infected adults.

Growth hormone (GH) is an underappreciated but important regulator of T cell development that can reverse age-related declines in thymopoiesis in rodents. Here, we report findings of a prospective randomized study examining the effects of GH on the immune system of HIV-1-infected adults. GH treatment was associated with increased thymic mass.

C. elegans AP-2 and retromer control Wnt signaling by regulating mig-14/Wntless.

While endocytosis can regulate morphogen distribution, its precise role in shaping these gradients is unclear. Even more enigmatic is the role of retromer, a complex that shuttles proteins between endosomes and the Golgi apparatus, in Wnt gradient formation. Here we report that DPY-23, the C.

Direct measurement of T-cell receptor repertoire diversity with AmpliCot.

Many studies require the measurement of nucleic acid sequence diversity. Here we describe a method, called AmpliCot, that measures the sequence diversity of PCR products on the basis of DNA hybridization kinetics, thereby avoiding the time, expense and biases associated with cloning and sequencing. SYBR Green dye is used to measure DNA hybridization kinetics in a homogeneous, automated fashion.

HLH-14 is a C. elegans achaete-scute protein that promotes neurogenesis through asymmetric cell division.

Achaete-Scute basic helix-loop-helix (bHLH) proteins promote neurogenesis during metazoan development. In this study, we characterize a C. elegans Achaete-Scute homolog, HLH-14.