Functional classification and associated mapped reads deriving from the CDS associated with putative secreted proteins found in the sialotranscriptome of <i>Amblyomma americanum</i>.

<p>Functional classification and associated mapped reads deriving from the CDS associated with putative secreted proteins found in the sialotranscriptome of <i>Amblyomma americanum</i>.</p

Heat map displaying 98 Kunitz domain containing coding sequences (CDS) that are not uniformly expressed in the four libraries and showing a standard deviation of 1 or more regarding the log(10) transformation of the average normalized data.

<p>For more details, see text.</p

Heat map displaying 138 lipocalin coding sequences (CDS) that are not uniformly expressed in the four libraries and showing a standard deviation of 1 or more regarding the log(10) transformation of the average normalized data.

<p>For more details, see text.</p

Clustering of the differential temporal expression of 817 transcripts from the <i>Amblyomma americanum</i> sialotranscriptome, using the Click algorithm of the Expander program [24].

<p>Nineteen transcripts of the 836 shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0131292#pone.0131292.g001" target="_blank">Fig 1</a> were not clustered. Each graph (A-E) represents an identified cluster. The Y axis represent the log(10) of the normalized FPKM row data, where 0 reads were substituted by 1. Symbols and bars represent the average and standard errors. For more details, see text.</p

Human CD117 (cKit)+ Innate Lymphoid Cells Have a Discrete Transcriptional Profile at Homeostasis and Are Expanded during Filarial Infection

<div><p>Since innate lymphoid cells (ILCs) have been found to play a role in the immune response to helminth parasites in rodents, we sought to determine their role in human helminth infection. By developing multicolor flow cytometry-based methods to identify and enumerate circulating ILCs and their subsets, we were able to identify a subset of cKit+ ILCs defined as Lineage (Lin)−/CD45+/cKit+/CD127+ that were significantly expanded in the filarial-infected individuals (p = 0.0473) as were those cKit+ ILCs that produced IL-13. Additionally, the frequency of these cKit+ ILCs correlated with the frequency of IL-17 producing CD4+ T cells (Th17 cells; p = 0.025). To investigate the function of cKit+ ILCs, sorted, highly purified human ILCs were subjected to transcriptional profiling by RNAseq and compared to appropriate control cells. These cKit+ ILCs expressed TLRs, a broad range of cytokines/cytokine receptors and MHC Class II molecules suggesting that these ILCs sense pathogens independent of other cell types. Functional analysis revealed expanded cKit+ ILC-specific transcription and ILC-specific microRNA precursors.</p></div

Cell sorting yields a purified ILCs that produces Th2-, Th17- and Th1-related cytokines.

<p>(A) Representative flow diagram of the strategy used for cell sorting of cKit+ ILCs (B) Sorted cKit+ ILCs (n = 5) cultured with media alone (closed circles-Unst) or with a cytokine cocktail consisting of rhIL-2, IL-7, IL-25 and IL-33 (open circles-Stim) for 3 days.</p

The frequency of cKit+ ILCs in peripheral blood correlates with the frequency of Th17 cells.

<p>Correlation of the frequencies of cKit+ ILCs and the frequencies of Th17 (A), Th2 (B), and Th1 (C) CD4+ T cells in 21 filarial-infected (circles) and 11 filarial-uninfected individuals (triangles). The p and r-values from the Spearman rank correlation analysis are shown. (D) Correlation of the frequencies of IL-13 producing cKit+ ILCs and Th2 CD4+ T cells from 8 filarial-infected and 6 uninfected individuals. The p-value and r-value from the Spearman rank correlation analysis are shown.</p

At homeostasis cKit+ ILCs express transcripts associated with the ILC2 and ILC3 subsets.

<p>Transcripts differentially expressed between ILCs and lineage depleted PBMCs (control cells). The upregulated and downregulated transcripts from the RNASeq analysis were put into Ingenuity Pathway Analysis for graphical illustration. Pathway depicts genes that are upregulated (red) and downregulated (green) in cKit+ILCs in comparison to lineage-depleted PBMCs and their presumed location in the cell. The top of the figure depicts the cell membrane, with the nucleus towards the bottom of the figure.</p