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31.

Background aims

CD1d-restricted invariant natural killer (iNK) T cells are rare regulatory T cells that may contribute to the immune-regulation in allogeneic stem cell transplantation (ASCT). Here, we sought to develop an effective strategy to expand human iNK T cells for use in cell therapy to prevent graft-versus-host disease (GVHD) in ASCT.

Methods

Human iNK T cells were first enriched from peripheral blood mononuclear cells (PBMCs) using magnetic-activated cell sorting separation, then co-cultured with dendritic cells in the presence of agonist glycolipids, alpha-galactosylceramide, for 2 weeks.

Results

The single antigenic stimulation reliably expanded iNK T cells to an average of 2.8?×?107 per 5?×?108 PBMCs in an average purity of 98.8% in 2 weeks (N?=?24). The expanded iNK T cells contained a significantly higher level of CD4+ and central memory phenotype (CD45RA?CD62L+) compared with freshly isolated iNK T cells, and maintained their ability to produce both Th-1 (interferon [IFN]γ and tumor necrosis factor [TNF]α) and Th-2 type cytokines (interleukin [IL]-4, IL-5 and IL-13) upon antigenic stimulation or stimulation with Phorbol 12-myristate 13-acetate/ionomycin. Interestingly, expanded iNK T cells were highly autoreactive and produced a Th-2 polarized cytokine production profile after being co-cultured with dendritic cells alone without exogenous agonist glycolipid antigen. Lastly, expanded iNK T cells suppressed conventional T-cell proliferation and ameliorated xenograft GVHD (hazard ratio, 0.1266; P < 0.0001).

Conclusion

We have demonstrated a feasible approach for obtaining ex vivo expanded, highly enriched human iNK T cells for use in adoptive cell therapy to prevent GVHD in ASCT.  相似文献   
32.
Abstract Exclosures were used to examine the impact of grazing upon the mortality patterns of populations of six indigenous grass species. The experiment compared unfenced areas with areas from which either sheep only or sheep, rabbits and kangaroos were excluded. There were large interspecific differences in mortality patterns, with Stipa nitida and Aristida browniana having relatively high mortality rates and Eragrostis eriopoda having relatively low mortality rates. Grazing-induced mortality was observed in treatment areas that were grazed by sheep, rabbits and kangaroos and in areas grazed only by rabbits and kangaroos. The short-lived S. nitida appears less likely to suffer grazing-induced mortality than species of intermediate longevity such as Monachather paradoxa. These observations help explain the decline in endemic perennial grasses that has taken place in the vegetation of western New South Wales since European settlement. Management of these rangelands to encourage these grasses must consider total grazing pressure and not simply the impact of livestock.  相似文献   
33.
A Potter spray tower was used to determine the susceptibility of adult female European red mites (Panonychus ulmi (Koch)) and two-spotted spider mites (Tetranychus urticae Koch) to suffocation by petroleum spray oils (PSOs). The LC95 values calculated for P. ulmi against C23 Ampol D-C-Tron NR and C21 Caltex Lovis were 104 and 165 g cm-2, respectively. For T. urticae the values were 169 and 207 g cm-2, respectively. The results are consistent with established relationships between PSO efficacy and increasing PSO molecular weight. The results are discussed in relation to previously reported bioassays on spider mites and to concurrent evaluations of PSOs for use in pome fruit integrated pest management programmes in Australia.  相似文献   
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Using susceptible and resistant reference strains and standard bioassay methods a new miticide product was screened for efficacy and cross-resistance against two-spotted spider mite, Tetranychus urticae Koch. Experimental product NN1 850.0 EW (fenpyroximate and propargite 3:10 ratio) was significantly (P<0.05) more toxic than either compound tested alone (co-toxicity index 1029). Non-overlapping 95% CIs at the LC50 and LC99 level suggest that there is no cross-resistance to NN1 850.0 EW from existing organophosphate and pyrethroid resistance. NN1 850.0 EW is worthy of further development for T. urticae control.  相似文献   
36.
A combination of uni- and multiplex PCR assays targeting 58 virulence genes (VGs) associated with Escherichia coli strains causing intestinal and extraintestinal disease in humans and other mammals was used to analyze the VG repertoire of 23 commensal E. coli isolates from healthy pigs and 52 clinical isolates associated with porcine neonatal diarrhea (ND) and postweaning diarrhea (PWD). The relationship between the presence and absence of VGs was interrogated using three statistical methods. According to the generalized linear model, 17 of 58 VGs were found to be significant (P < 0.05) in distinguishing between commensal and clinical isolates. Nine of the 17 genes represented by iha, hlyA, aidA, east1, aah, fimH, iroNE. coli, traT, and saa have not been previously identified as important VGs in clinical porcine isolates in Australia. The remaining eight VGs code for fimbriae (F4, F5, F18, and F41) and toxins (STa, STb, LT, and Stx2), normally associated with porcine enterotoxigenic E. coli. Agglomerative hierarchical algorithm analysis grouped E. coli strains into subclusters based primarily on their serogroup. Multivariate analyses of clonal relationships based on the 17 VGs were collapsed into two-dimensional space by principal coordinate analysis. PWD clones were distributed in two quadrants, separated from ND and commensal clones, which tended to cluster within one quadrant. Clonal subclusters within quadrants were highly correlated with serogroups. These methods of analysis provide different perspectives in our attempts to understand how commensal and clinical porcine enterotoxigenic E. coli strains have evolved and are engaged in the dynamic process of losing or acquiring VGs within the pig population.  相似文献   
37.

Background  

Gene expression microarray technology permits the analysis of global gene expression profiles. The amount of sample needed limits the use of small excision biopsies and/or needle biopsies from human or animal tissues. Linear amplification techniques have been developed to increase the amount of sample derived cDNA. These amplified samples can be hybridised on microarrays. However, little information is available whether microarrays based on amplified and unamplified material yield comparable results.  相似文献   
38.
The resistance status of Tetranychus urticae Koch to the miticides clofentezine and fenbutatin oxide was investigated in deciduous fruit tree orchards in Western Australia. Mites were collected from 30 orchards across three fruit growing regions and subjected to discriminating–dose assays. There was a significant relationship between clofentezine (=;0.64) and fenbutatin oxide (V=;0.44) use and the proportion of resistant mites in each sample. Improved clofentezine models were obtained when the Perth Hills region (V=;0.90) was modelled separately from Donnybrook and Manjimup (V=;0.80). Clofentezine could be expected to fail in the latter regions after four sprays and in the former region after only one spray. The relatively low pest status of T. urticae in Western Australia did not delay the clofentezine resistance. For any given number of spray applications the response of T. urticae to fenbutatin oxide was more variable than it was to clofentezine.  相似文献   
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