Cutting edge: Experimentally induced immune activation in natural hosts of simian immunodeficiency virus induces significant increases in viral replication and CD4+ T cell depletion.

Chronically SIVagm-infected African green monkeys (AGMs) have a remarkably stable nonpathogenic disease course, with levels of immune activation in chronic SIVagm infection similar to those observed in uninfected monkeys and with stable viral loads for long periods of time. In vivo administration of LPS or an IL-2/diphtheria toxin fusion protein (Ontak) to chronically SIVagm-infected AGMs triggered increases in immune activation and subsequently of viral replication and depletion of intestinal CD4(+) T cells. Our study indicates that circulating microbial products can increase viral replication by inducing immune activation and increasing the number of viral target cells, thus demonstrating that immune activation and T cell proliferation are key factors in AIDS pathogenesis.

The effect of m-xylene on cytotoxicity and cellular antioxidant status in rat dermal equivalents.

Exposure of the skin to volatile organic chemicals (VOCs) can lead to irritation, inflammation and cytotoxicity. Since VOCs are used in industrial, commercial and military applications, concern is mounting with respect to VOC safe exposure limits. Although traditional toxicological assessment of VOCs has utilized animal models, the use of alternative in vitro models is becoming more widespread. We have previously developed a sealed exposure system that prevents chemical loss through evaporation and enables calculation of target cell chemical dose. The present study utilized this in vitro exposure method to assess m-xylene-induced cytotoxicity and antioxidant status in dermal equivalents (dermal fibroblasts in a collagen matrix). At the end of a 1- or 4-h exposure, cytotoxicity was measured using the MTT assay and the EC50 values determined were 1481 +/- 88 and 930 +/- 33, respectively. Decreases in cellular thiols and catalase activity were observed, which occurred in a time and dose-dependent manner. Treatment of dermal equivalents with the antioxidants N-acetylcysteine (NAC) and catalase provided some protection against m-xylene-induced cytotoxicity. When compared to m-xylene exposures, treatment with either 1.0 or 5.0 mM NAC led to increases in the EC50 values at 1 and 4 h. Increases in these EC50 values ranged from 1.22- to 1.32-fold at 1 h and 1.27- to 1.54-fold at 4 h. Although treatment with catalase (1000 U/ml) led to a 1.35-fold increase in cell viability at 1 h, no significant differences were observed at either 1 or 4 h when compared to dermal equivalents exposed to m-xylene alone. These results suggest that exposure to m-xylene leads to a time- and dose-dependent decrease in cellular antioxidants and that cellular thiols may provide protection against the cytotoxic properties of m-xylene.

Association of eleven common, low-penetrance colorectal cancer susceptibility genetic variants at six risk loci with clinical outcome.

BACKGROUND: Low-penetrance genetic variants have been increasingly recognized to influence the risk of tumor development. Risk variants for colorectal cancer (CRC) have been mapped to chromosome positions 8q23.3, 8q24, 9p24.1, 10p14, 11q23, 14q22.2, 15q13, 16q22.1, 18q21, 19q13.1 and 20p12.3. In particular, the 8q24 single nucleotide polymorphism (SNP), rs6983267, has reproducibly been associated with the risk of developing CRC. As the CRC risk SNPs may also influence disease outcome, thus in this study, we evaluated whether they influence patient survival. METHODOLOGY/PRINCIPAL FINDINGS: DNA samples from 583 CRC patients enrolled in the prospective, North Carolina Cancer Care Outcomes Research and Surveillance Consortium Study (NC CanCORS) were genotyped for 11 CRC susceptibility SNPs at 6 CRC risk loci. Relationships between genotypes and patient survival were examined using Cox regression analysis. In multivariate analysis, patients homozygous for the CRC risk allele of rs7013278 or rs7014346 (both at 8 q24) were only nominally significant for poorer overall survival compared to patients homozygous for the protective allele (hazard ratio = 2.20 and 1.96, respectively; P<0.05). None of these associations, however, remained statistically significant after correction for multiple testing. The other nine susceptibility SNPs tested were not significantly associated with survival. CONCLUSIONS/SIGNIFICANCE: We did not find evidence of association of CRC risk variants with patient survival.

Island biogeography reveals the deep history of SIV.

Simian immunodeficiency virus (SIV) lineages have been identified that are endemic to Bioko Island. The time the island formed offers a geological time scale calibration point for dating the most recent common ancestor of SIV. The Bioko viruses cover the whole range of SIV genetic diversity, and each Bioko SIV clade is most closely related to viruses circulating in hosts of the same genus on the African mainland rather than to SIVs of other Bioko species. Our phylogeographic approach establishes that SIV is ancient and at least 32,000 years old. Our conservative calibration point and analyses of gene sequence saturation and dating bias suggest it may be much older.

Cloning and transfer of the Salmonella pathogenicity island 2 type III secretion system for studies of a range of gram-negative genera.

The engineering of bacterial strains with specific phenotypes frequently requires the use of blocks or "cassettes" of genes that act together to perform a desired function. The potential benefits of utilizing type III secretion systems in this regard are becoming increasingly realized since these systems can be used to direct interactions with host cells for beneficial purposes such as vaccine development, anticancer therapies, and targeted protein delivery. However, convenient methods to clone and transfer type III secretion systems for studies of a range of different types of bacteria are lacking. In addition to functional applications, such methods would also reveal important information about the evolution of a given type III secretion system, such as its ability to be expressed and functional outside of the strain of origin. We describe here the cloning of the Salmonella enterica serovar Typhimurium pathogenicity island 2 (SPI-2) type III secretion system onto a vector that can be easily transferred to a range of gram-negative bacterial genera. We found that expression of the cloned SPI-2 system in different Gammaproteobacteria and Alphaproteobacteria (as monitored by SseB protein levels) is dependent on the bacterial strain and growth medium. We also demonstrate that the cloned system is functional for secretion, can direct interactions with macrophages, and can be used as a novel tool to analyze the predicted interaction of SseB with host cells. This work provides a foundation for future applications where the cloned SPI-2 region (or other cloned type III systems) can provide a desired function to an engineered gram-negative strain.

T regulatory cells: aid or hindrance in the clearance of disease?

CD4+ CD25+ T regulatory cells (Tregs) are classified as a subset of T cells whose role is the suppression and regulation of immune responses to self and non-self. Since their discovery in the early 1970s, the role of CD4+ CD25+ Tregs in both autoimmune and infectious disease has continued to expand. This review examines the recent advances on the role CD4+ CD25+ Tregs may be playing in various diseases regarding progression or protection. In addition, advances made in the purification and manipulation of CD4+ CD25+ Tregs using new cell markers, techniques and antibodies are discussed. Ultimately, an overall understanding of the exact mechanism which CD4+ CD25+ Tregs implement during disease progression will enhance our ability to manipulate CD4+ CD25+ Tregs in a clinically beneficial manner.