Human SIM2 gene expression analysis in various cancers.

<p>SIM2 gene expression data were extracted from the Oncomine Research Edition. Microarray datasets that show a 2-fold change in SIM2 expression between cancer and control groups and a p value<0.01 are highlighted. (<b>A</b>) Comparison of SIM2 gene expression between cancer and control specimens. Red color indicates SIM2 overexpression and the blue color indicates SIM2 down-regulation in cancer. Numbers in the boxes indicate the number of datasets showing statistical significance. Box plots were obtained from the datasets selected in (<b>A</b>) to highlight significant overexpression of SIM2 in Prostate Carcinoma (1. Prostate Gland (n = 23), 2. Prostate Carcinoma (n = 65); <i>P</i> = 2.41×10⁻¹⁴, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093231#pone.0093231-Yu1" target="_blank">[40]</a>) (<b>B</b>); Colon Carcinoma (1. Colon (n = 10), 2. Colon Carcinoma (n = 5); P = 1.65×10⁻¹²<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093231#pone.0093231-Skrzypczak1" target="_blank">[41]</a>). (<b>C</b>); Breast Carcinoma (1. Breast (n = 4), 2. Invasive Breast Carcinoma (n = 154); P = 2.25×10⁻⁴, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093231#pone.0093231-Gluck1" target="_blank">[42]</a>) (<b>D</b>); Oligodendroglioma (1. Brain (n = 23), 2. Oligodendroglioma (n = 50); P = 3.31×10⁻⁹<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093231#pone.0093231-Sun1" target="_blank">[43]</a>) (<b>E</b>); and Pancreatic Carcinoma (1. Pancreas (n = 16), 2. Pancreatic Carcinoma (n = 36); P = 3.01×10⁻⁷<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093231#pone.0093231-Pei1" target="_blank">[44]</a>) (<b>F</b>).</p

IFN-γ production by CD8 T-cells from SIM2-immunized mice.

<p>Mice were immunized with either SIM2₂₃₇, SIM2₂₃₇+HBV₁₂₈, SIM2₂₃₇+SIM2_240–254 or SIM2_230–256. Splenocytes were harvested and incubated overnight with T2 cells loaded with the SIM2₂₃₇ peptide. IFN-γ was measured by flow cytometry. FACS plots show the median IFN-γ production for each group (A) and replicate data obtained from each group (B).</p

Human MHC-II-restricted epitopes predicted from the SIM2 long peptide using IEDB tool.

<p>Only the top epitopes having the lowest percentile score and lowest IC50 are selected. One epitope is shown for each HLA allele out of 137 predicted binders.</p>a<p>Percentile Rank – Percentage of all peptides binding with this efficacy or lower.</p>b<p>CombLib IC40 – Predicted peptide concentration required to bind 50% of MHC molecules.</p

SIM2_230–256 induces an IFN-γ and CD4 IL-2 response.

<p>IFN-γ production by splenocytes in mice immunized with various treatments. Mice were immunized with either the 9aa SIM2₂₃₇ epitope combined with HBV or SIM2_240–254, or the SIM2_230–256 peptide alone. IFN-γ production was measured by ELISPOT. IL-2 production by CD4 T-cells. CD4 T-cells were sorted from the spleens of immunized mice and tested for reactivity to HBV₁₂₈ and SIM2_240–254 by IL-2 ELISPOT.</p

Splenocytes from SIM2_230–256-immunized mice response to PC3-A2.1 cells.

<p>Splenocytes from HHD mice immunized with HBV and various SIM2_230–256 peptides or HBV alone were co-cultured with PC3, PC3-A2.1 (<b>A</b>) or LNCaP (<b>B</b>). Production of IFN-γ by splenocytes in response to these tumor cell lines was assessed by ELISPOT. Data is representative of 2 experiments and shows mean ± standard deviation.</p