Dominance and diversity in the primary human CD4 T cell response to replication-competent vaccinia virus

Vaccination with replication-competent vaccinia protects against heterologous orthopoxvirus challenge. CD4 T cells have essential roles helping functionally important Ab and CD8 antiviral responses, and contribute to the durability of vaccinia-specific memory. Little is known about the specificity, diversity, or dominance hierarchy of orthopoxvirus-specific CD4 T cell responses. We interrogated vaccinia-reactive CD4 in vitro T cell lines with vaccinia protein fragments expressed from an unbiased genomic library, and also with a panel of membrane proteins. CD4 T cells from three primary vaccinees reacted with 44 separate antigenic regions in 35 vaccinia proteins, recognizing 8 to 20 proteins per person. The integrated responses to the Ags that we defined accounted for 49 to 81% of the CD4 reactivity to whole vaccinia Ag. Individual dominant Ags drove up to 30% of the total response. The gene F11L-encoded protein was immunodominant in two of three subjects and is fragmented in a replication-incompetent vaccine candidate. The presence of protein in virions was strongly associated with CD4 antigenicity. These findings are consistent with models in which exogenous Ag drives CD4 immunodominance, and provides tools to investigate the relationship between Ab and CD4 T cell specificity for complex pathogens.     

Continental patterns in the diet of a top predator: Australia's dingo

相似文献   

Single-Cell RNA-Seq Reveals AML Hierarchies Relevant to Disease Progression and Immunity

Download video (12MB)Help with mp4 files     

Escherichia coli HU protein has a role in the repair of abasic sites in DNA

HU is one of the most abundant DNA binding proteins in Escherichia coli. We find that it binds strongly to DNA containing an abasic (AP) site or tetrahydrofuran (THF) (apparent K_d ≈50 nM). It also possesses an AP lyase activity that cleaves at a deoxyribose but not at a THF residue. The binding and cleavage of an AP site was observed only with the HUαβ heterodimer. Site-specific mutations at K3 and R61 residues led to a change in substrate binding and cleavage. Both K3A(α)K3A(β) and R61A(α)R61A(β) mutant HU showed significant reduction in binding to DNA containing AP site; however, only R61A(α)R61A(β) mutant protein exhibited significant loss in AP lyase activity. Both K3A(α)K3A(β) and R61K(α)R61K(β) showed slight reduction in AP lyase activities. The function of HU protein as an AP lyase was confirmed by the ability of hupA or hupB mutations to further reduce the viability of an E. coli dut(Ts) xth mutant, which generates lethal AP sites at 37°C; the hupA and hupB derivatives, respectively, had a 6-fold and a 150-fold lower survival at 37°C than did the parental strain. These data suggest, therefore, that HU protein plays a significant role in the repair of AP sites in E. coli.     

The pathway of US11-dependent degradation of MHC class I heavy chains involves a ubiquitin-conjugated intermediate.

The human cytomegalovirus protein, US11, initiates the destruction of MHC class I heavy chains by targeting them for dislocation from the ER to the cytosol and subsequent degradation by the proteasome. We report the development of a permeabilized cell system that recapitulates US11-dependent degradation of class I heavy chains. We have used this system, in combination with experiments in intact cells, to identify and order intermediates in the US11-dependent degradation pathway. We find that heavy chains are ubiquitinated before they are degraded. Ubiquitination of the cytosolic tail of heavy chain is not required for its dislocation and degradation, suggesting that ubiquitination occurs after at least part of the heavy chain has been dislocated from the ER. Thus, ubiquitination of the heavy chain does not appear to be the signal to start dislocation. Ubiquitinated heavy chains are associated with membrane fractions, suggesting that ubiquitination occurs while the heavy chain is still bound to the ER membrane. Our results support a model in which US11 co-opts the quality control process by which the cell destroys misfolded ER proteins in order to specifically degrade MHC class I heavy chains.     

Amino-terminal sequence of a large form of basic fibroblast growth factor isolated from human benign prostatic hyperplastic tissue

Homogenization of human benign prostatic hyperplastic tissue in high ionic strength alkaline buffer containing protease inhibitors resulted in the isolation of a 17,400 molecular weight growth factor. When tissue was homogenized in ammonium sulfate at pH 4.5 without protease inhibitors a smaller, 16,600 dalton, growth factor was isolated. Both growth factors reacted with antisera against synthetic peptides whose sequences corresponded to the amino-terminal (1-12), Internal (33-43) and carboxyl-terminal (135-145) portions of basic fibroblast growth factor (bFGF). This suggested that the smaller growth factor was not a truncated form of (1-146) bFGF and that the larger growth factor may contain additional sequences. Amino-terminal sequencing showed the larger growth factor to have the sequence: Ala-Ala-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala-Leu-Pro-Glu-Asp-Gly-Gly-Ser-Gly- Ala-Phe-Pro-. These results show that the larger growth factor is an 8 amino acid extended from of (1-146) bFGF and it is likely that the smaller growth factor is a proteolytic cleavage product of the larger growth factor produced during the extraction procedure.     

Bacterial transport in volcanic tuff cores under saturated flow conditions

An antibiotic‐resistant bacterium was tested for transport through volcanic tuff and sandstone cores. Tuff cores were representative of the geology of Rainier Mesa located on the Nevada Test Site (NTS). Rapid bacterial transport occurred in some of the tuff cores and all sandstone cores under the hydraulic heads used (5–500 cm). Hydraulic conductivity of the tuff cores ranged widely, 9.6 × 10^‐5 to 7.2 x 10^‐3 cm h^‐1. A much narrower range was observed for sandstone cores, 1.6 × 10^‐2 to 5.9 X 10^‐2 cm h^‐1, which served as experimental controls. The percentage of the initial bacterial inoculum recovered within 3 pore volumes from tuff and sandstone cores ranged from 9.4 to 54.7% and 0.20 to 2.9%, respectively. Bacterial recovery appeared to be controlled by the structure of the flow paths in rock cores and not by overall hydraulic conductivity. Saturated clay‐infiltrated and unfractured zeolitized tuff cores were impermeable to water flow, and therefore bacterial transport was not detected. Three routes of bacterial transport were discerned in permeable rock cores by comparison of the breakthrough patterns of bacteria and tracer solution (chloride ions) in cores of differing lithologies. In sandstone cores, where water flowed evenly through the matrix, bacteria were transported in a dispersed manner throughout the sandstone, whereas bacteria were transported primarily along preferred flow paths (fractures or macropores) in permeable tuff cores.     

Do Children Eat What They Say? Validity of Intended Food Choices Among Native American School Children

Objective. To examine the agreement between school children's intended food choices and observed food choices. Design. Native American students in the second through fifth grade completed a questionnaire that asked them to select from 10 paired food choices for a given meal or snack. Three weeks later students chose among foods identical to those on the questionnaire as part of their usual school lunch or breakfast over three consecutive days; afternoon snacks were also offered. Results. Agreement between students' intended food choices and observed food choices was examined across 10 food pairs. The composite K coefficient between intended and observed food choices was 0.09 (95% confidence interval 0.06, 0.12), indicating virtually no agreement above that expected by chance. There were no differences in agreement by sex or grade. Conclusions. Intended food choices were not significantly associated with observed food choices. It is unclear whether intended food choices reflect nutrition knowledge, socially desirable responses, food preferences, or some other dimension of eating behavior. Although responsive to school-based nutrition interventions, the interpretation of changes in intended food choices must be clarified in future research.