CD8+ T cells from SIV elite controller macaques recognize Mamu-B*08-bound epitopes and select for widespread viral variation

Background

It is generally accepted that CD8⁺ T cell responses play an important role in control of immunodeficiency virus replication. The association of HLA-B27 and -B57 with control of viremia supports this conclusion. However, specific correlates of viral control in individuals expressing these alleles have been difficult to define. We recently reported that transient in vivo CD8⁺ cell depletion in simian immunodeficiency virus (SIV)-infected elite controller (EC) macaques resulted in a brief period of viral recrudescence. SIV replication was rapidly controlled with the reappearance of CD8⁺ cells, implicating that these cells actively suppress viral replication in ECs.

Methods and Findings

Here we show that three ECs in that study made at least seven robust CD8⁺ T cell responses directed against novel epitopes in Vif, Rev, and Nef restricted by the MHC class I molecule Mamu-B*08. Two of these Mamu-B*08-positive animals subsequently lost control of SIV replication. Their breakthrough virus harbored substitutions in multiple Mamu-B*08-restricted epitopes. Indeed, we found evidence for selection pressure mediated by Mamu-B*08-restricted CD8⁺ T cells in all of the newly identified epitopes in a cohort of chronically infected macaques.

Conclusions

Together, our data suggest that Mamu-B*08-restricted CD8⁺ T cell responses effectively control replication of pathogenic SIV_mac239. All seven regions encoding Mamu-B*08-restricted CD8⁺ T cell epitopes also exhibit amino acid replacements typically seen only in the presence of Mamu-B*08, suggesting that the variation we observe is indeed selected by CD8⁺ T cell responses. SIV_mac239 infection of Indian rhesus macaques expressing Mamu-B*08 may therefore provide an animal model for understanding CD8⁺ T cell-mediated control of HIV replication in humans.     

Relationships between anthropometric, body composition and bone mineral parameters in 7-8-year-old rhythmic gymnasts compared with controls

The aim of the study was to investigate the relationships between specific anthropometric (9 skinfolds, 13 girths, 8 lengths and 8 breadths), body composition (body fat %, fat free mass [FFM], fat mass [FM]) parameters and bone mineral parameters (bone mineral density [BMD], bone mineral content [BMC) in young rhythmic gymnasts and same age controls. Eighty nine 7-8-year-old girls participated in this study and were divided to the rhythmic gymnast's (n = 46) and control (n = 43) groups. Body composition was determined by dual energy X-ray absorptiometry (FFM, FM, body fat %, BMD and BMC). Body fat % and FM were lower and BMD and BMC values at lumbar spine (L2-L4) and femoral neck were higher in rhythmic gymnasts compared with controls. All measured skinfold thicknesses were thicker in controls. In girths, lengths and widths there were only few significant differences between the groups. Stepwise multiple regression analysis indicated that skinfold thicknesses (supraspinale and medial calf) influenced L2-L4 BMD only in controls 38.2% (R2x100). Supraspinale and iliac crest skinfold thicknesses characterised L2-L4 BMC 43.9% (R2x100). Calf girths influenced BMD in L2-L4 52.3% (R2x100) in controls. BMC in L2-L4 was dependent only on mid-thigh girths 35.9% (R2x100). BMD in L2-L4 was dependent on tibiale-laterale height 30.0% (R2x100). Biiliocristal breadths together with sitting height characterised BMC in L2-L4 BMD 62.3% (R2x100). In conclusion, we found that the relationships between anthropometry, body composition and bone parameters in young rhythmic gymnasts are weak. In control group first of all lower body anthropometric parameters significantly correlated with BMD and BMC in spine.     

Plasma ghrelin is altered after maximal exercise in elite male rowers

The aim of the present investigation was to investigate plasma ghrelin response to acute maximal exercise in elite male rowers. Eight elite male rowers performed a maximal 6000-m rowing ergometer test (mean performance time: 19 mins 52 secs; 1192.1 +/- 16.4 secs), and venous blood samples were obtained before, immediately after, and after 30 mins of recovery. In addition to ghrelin concentration, leptin, insulin, growth hormone, insulin-like growth factor-1 (IGF-1), testosterone, cortisol, and glucose values were measured. Ghrelin was significantly increased immediately after the exercise (+24.4%; P < 0.05) and was not significantly different than baseline after 30 mins of recovery. Leptin was significantly decreased immediately after the exercise (- 15.8%; P < 0.05) and remained significantly decreased after the first 30 mins of recovery. No changes occurred in insulin concentrations. Growth hormone, IGF-1, and testosterone values were significantly increased and decreased to the pre-exercise level immediately after the exercise and after the first 30 mins of recovery, respectively. Cortisol and glucose values were significantly increased immediately after the exercise and remained significantly increased during the first 30 mins of recovery. There were no relationships between plasma ghrelin and other measured blood parameters after the exercise, nor were changes in ghrelin related to changes in other measured blood biochemical values after the exercise. In conclusion, these results suggest that acute negative energy balance induced by specific maximal short-term exercise elicits a metabolic response with opposite changes in ghrelin and leptin concentrations in elite male athletes.     

Physical Activity and Bone Mineral Accrual in Boys with Different Body Mass Parameters during Puberty: A Longitudinal Study

The aim of our longitudinal study was to investigate the relationships between physical activity and bone mass in boys with different body mass status during the years surrounding pubertal growth spurt. Two hundred and six boys entering puberty took part in this study. The subjects were divided into underweight (), normal weight (), overweight () and obese () groups at baseline according to age related categories. Whole-body DXA scans were performed at baseline, after 12 and 24 months to assess body composition (lean body mass, fat mass), and total body (TB), lumbar spine (LS) and femoral neck (FN) bone mineral density (BMD) parameters. Physical activity was measured by 7- day accelerometry. For longitudinal analysis, multilevel fixed effects regression models were constructed. Biological age, height and lean body mass had an effect for explanation of TB BMD, FN BMD and LS BMD. Moderate to vigorous physical activity (MVPA), vigorous physical activity (VPA) and sedentary time (SED) had the significant effect only on FN BMD. Being an underweight boy at the baseline indicated greater chance (p<0.01) to have lower TB BMD in the future (2 years at follow up) development, compared to normal weight (estimates = −0.038), overweight (estimates = −0.061) and obese boys (estimates = −0.106).     

Relationships between legs bone mineral density, anthropometry and jumping height in prepubertal children

The aim of this study was to determine how the legs bone mineral density (BMD) is influenced by anthropometry and vertical jumping height in prepubertal children. In total, 64 8-11-year-old schoolchildren (27 boys and 37 girls) were studied. All children were at Tanner stage 1. The subjects' height and body mass were measured and BMI calculated. The following anthropometric parameters directly connected with leg were measured: skinfolds--front thigh and medial calf girths--gluteal, thigh, mid-thigh, calf and ankle; lengths--iliospinale height, trochanterion height, trochanteriontibiale laterale, tibiale-laterale height and tibiale mediale-spyrion tibiale; and breadths--biiliocristal, foot length and biepicondylar femur. Total body and legs fat mass and fat %, lean body mass (LBM) and both legs BMD were measured by DXA. Maximal jumping height was measured on the contact mat. Stepwise multiple regression analysis indicated that body height in boys (54.6%; R2 x 100) and body mass in girls (57.3%) were the most important basic anthropometric parameters that influenced BMD in legs. From the measured skinfolds, that of the front thigh characterized legs BMD by 24.9-35.6%. From the girths, the most important parameter to characterize legs BMD was that of calf (50.0-59.1%). Tibiale laterale height was the only length parameter which was highly related with legs BMD (51.1-54.5%). Biepicondylar femur was the most important breadth parameter which characterized legs BMD (51.0-54.8%). Femur breadth and tibiale-laterale height were selected (68.7%) in boys, and tibiale-laterale height and front thigh skinfold thickness (66.0%) in girls when all measured leg anthropometric parameters were analyzed together. From the body composition parameters, the most important parameter to characterize legs BMD was legs LBM (48.9-59.5%). Jumping height did not correlate with legs BMD in any studied groups. In summary, the present study demonstrated that legs LBM together with tibiale-laterale height are the main predictors of legs BMD in prepubertal children.     

Nonhuman primate models and the failure of the Merck HIV-1 vaccine in humans

The adenovirus type 5 (Ad5)-based vaccine developed by Merck failed to either prevent HIV-1 infection or suppress viral load in subsequently infected subjects in the STEP human Phase 2b efficacy trial. Analogous vaccines had previously also failed in the simian immunodeficiency virus (SIV) challenge-rhesus macaque model. In contrast, vaccine protection studies that used challenge with a chimeric simian-human immunodeficiency virus (SHIV89.6P) in macaques did not predict the human trial results. Ad5 vector-based vaccines did not protect macaques from infection after SHIV89.6P challenge but did cause a substantial reduction in viral load and a preservation of CD4+ T cell counts after infection, findings that were not reproduced in the human trials. Although the SIV challenge model is incompletely validated, we propose that its expanded use can help facilitate the prioritization of candidate HIV-1 vaccines, ensuring that resources are focused on the most promising candidates. Vaccine designers must now develop T cell vaccine strategies that reduce viral load after heterologous challenge.     

Analysis of the nucleus-encoded and chloroplast-targeted rieske protein by classic and site-directed mutagenesis of Chlamydomonas.

Three mutants of the alga Chlamydomonas reinhardtii affected in the nuclear PETC gene encoding the Rieske iron-sulfur protein 2Fe-2S subunit of the chloroplast cytochrome b(6)f complex have been characterized. One has a stable deletion that eliminates the protein; two others carry substitutions Y87D and W163R that result in low accumulation of the protein. Attenuated expression of the stromal protease ClpP increases accumulation and assembly into b(6)f complexes of the Y87D and W163R mutant Rieske proteins in quantities sufficient for analysis. Electron-transfer kinetics of these complexes were 10- to 20-fold slower than those for the wild type. The deletion mutant was used as a recipient for site-directed mutant petC alleles. Six glycine residues were replaced by alanine residues (6G6A) in the flexible hinge that is critical for domain movement; substitutions were created near the 2Fe-2S cluster (S128 and W163); and seven C-terminal residues were deleted (G171och). Although the 6G6A and G171och mutations affect highly conserved segments in the chloroplast Rieske protein, photosynthesis in the mutants was similar to that of the wild type. These results establish the basis for mutational analysis of the nuclear-encoded and chloroplast-targeted Rieske protein of photosynthesis.     

Crystal structures of a poplar xyloglucan endotransglycosylase reveal details of transglycosylation acceptor binding

Xyloglucan endotransglycosylases (XETs) cleave and religate xyloglucan polymers in plant cell walls via a transglycosylation mechanism. Thus, XET is a key enzyme in all plant processes that require cell wall remodeling. To provide a basis for detailed structure-function studies, the crystal structure of Populus tremula x tremuloides XET16A (PttXET16A), heterologously expressed in Pichia pastoris, has been determined at 1.8-A resolution. Even though the overall structure of PttXET16A is a curved beta-sandwich similar to other enzymes in the glycoside hydrolase family GH16, parts of its substrate binding cleft are more reminiscent of the distantly related family GH7. In addition, XET has a C-terminal extension that packs against the conserved core, providing an additional beta-strand and a short alpha-helix. The structure of XET in complex with a xyloglucan nonasaccharide, XLLG, reveals a very favorable acceptor binding site, which is a necessary but not sufficient prerequisite for transglycosylation. Biochemical data imply that the enzyme requires sugar residues in both acceptor and donor sites to properly orient the glycosidic bond relative to the catalytic residues.