Additional file 1: of Serum micro-rna profiles in patients with autosomal dominant polycystic kidney disease according to hypertension and renal function

List of miRNAs included in the Qiagen miScript miRNA PCR Array Human Serum & Plasma 384HC in Excel format. (XLSX 12.5 KB

An LF epitope based vaccine which stimulates HLA restricted T cell immune responses may confer protection against <i>B. anthracis</i> challenge.

<p>Groups of HLA-DQ8 transgenic mice were immunised 3 times, on days 0, 14 and 35 by the intra-peritoneal route, with either an LF fusion construct comprising a tetanus toxin helper domain (aa 865–1120) and 12 confirmed HLA-restricted LF epitopes (n = 6, black diamonds), or a peptide pool of the LF epitopes expressed in the fusion protein (n = 7, black triangles), a control, sham immunised, group was also included in the experiment (n = 6, black squares). All groups were challenged with 10⁶ cfu <i>B. anthracis</i> STI by the intra-peritoneal route, on day 77, and monitored daily for survival (A). The impact of infection upon survival was described using Kaplan Meier estimation (A). Spleens were recovered from surviving mice at day 21, (LF fusion protein (n = 6, black diamonds), peptide pool (n = 7, black triangles) and sham immunized mice (n = 2, black squares)), and a mean bacterial count per spleen determined following culture of <i>B. anthracis</i> for 24 hours (B). No statistically significant difference was seen between the groups in terms of bacterial burden.</p

Summary detailing the immunogenicity of the promiscuous dominant epitopes identified within this work.

<p>Summary detailing the immunogenicity of the promiscuous dominant epitopes identified within this work.</p

Frequent, large CD4 T cell epitope responses to anthrax LF domain I–III peptide panel in immune human donors.

<p>Frequent, large CD4 T cell epitope responses to anthrax LF domain I-III peptide panel in immune human donors. Table indicates positive T cell IFNγ ELIspot responses that were seen in 3 or more donors from the human donor cohort described in the Methods, comprising a total of 9 donors in the cutaneous anthrax (Kayseri) group and 10 donors in the AVP vaccinees (UK) group.</p

The immunodominant LF epitopes, identified in transgenic mouse strains, show relatively broad binding to common HLA-DR alleles.

<p>The relative binding affinity of peptides to HLA-DR molecules were expressed as a relative activity (ratio of the IC₅₀ of the peptide to the IC₅₀ of the reference peptide which binds strongly to the individual HLA II molecule). Peptides with a high relative binding affinity of <10 are indicated in bold. Means were calculated from at least three independent experiments.</p

Regions HLA-DR and DQ-presented anthrax LF epitopes mapped onto the LF protein structure reveals clustering of immunogenic epitopes.

<p>The structural domains of LF protein are indicated in Roman numerals (A). Immunodominant epitopes identified in this study from mice transgenic for DRB1*0101 (B), DRB1*0401 (C), and DQB1*0302 (D) are superimposed on the LF crystal structure (Protein Data Bank accession code 1J7N). Roman numerals indicate the structural domains. Ribbon diagrams were generated using the Accelrys discovery studio client 2.5 program.</p

Epitope-rich, immunodominant regions of LF and epitopes common to diverse HLA polymorphisms.

<p>Popliteal lymph nodes from mice transgenic for DRB1*0101 (A), DRB1*0401 (B), DRB1*1501 (C), DQB1*0302 (D) and DQB1*0602 (E) (n = 5) were harvested 10 days after immunization with 25 µg LF adjuvanted with Titermax Gold and stimulated with 25 µg of each 10mer peptide in the LF peptide library (LF_31–809). Responses were considered positive if the response was ≥2 SD above the cells plus medium control. Data is represented as scatter plots, showing the responses of individual mice as the stimulation index (SI) calculated as the mean cpm or IFNγ production of triplicate wells in the presence of peptide divided by the mean cpm or IFNγ production in the absence of antigen. Results are given as the mean ± SD/SEM.</p

Overview of allele specific and promiscuous epitopes identified by binding affinity, and immunogenicity in HLA transgenic mice and human subjects.

<p>The overlapping relationships of the epitopes identified in the HLA transgenic responses, HLA-DR binding affinity studies, and in cohorts of vaccinated and infected humans, were demonstrated in a Euler diagram (A). The HLA-DR restricted epitopes identified in (B) HLA transgenic mice and (C) HLA binding affinity studies were visualised as Venn diagrams, to show allele specific and promiscuous epitopes.</p