<i>Tmprss2^−/−</i> mutant mice do not exhibit drop in blood oxygen saturation.

<p><i>Tmprss2^−/−</i> and wild type mice were infected with 2×10⁵ FFU PR8M virus and oxygen saturation in the peripheral blood was measured until day 14 p.i. <i>Tmprss2^−/−</i> mice showed only a very mild drop in oxygen saturation whereas wild type mice exhibited a significant decrease that peaked at day 8 p.i.</p

<i>Tmprss2^−/−</i> mutant mice do not exhibit drop in blood oxygen saturation.

<p><i>Tmprss2^−/−</i> and wild type mice were infected with 2×10⁵ FFU PR8M virus and oxygen saturation in the peripheral blood was measured until day 14 p.i. <i>Tmprss2^−/−</i> mice showed only a very mild drop in oxygen saturation whereas wild type mice exhibited a significant decrease that peaked at day 8 p.i.</p

Murine Tmprss2 is not required for spread of H7N7 influenza A virus.

<p>Eight to eleven weeks old female mice were infected with 2×10⁴ FFU mouse-adapted SC35M (H7N7) influenza virus by intra-nasal application and bodyweight (A) and survival (B) was monitored until day 14 p.i. In addition to mice that were found dead, mice with a weight loss of more than 30% of the starting bodyweight were euthanized and recorded as dead. No significant differences were observed in survival between heterozygous and homozygous mutant mice after H7N7 infections (using the log rank test).</p

Viral load in the lungs of <i>Tmprss2^−/−</i> mice after infection with H1N1 (PR8M) influenza A virus.

<p>Eight to eleven weeks old female mice were infected with 2×10⁵ FFU of the PR8M virus. Infectious virus particles were determined in lung homogenates. Viral load was higher in infected wild type mice compared to infected homozygous mutant mice at days 1, 2 and 3 p.i. Individual values, mean and SEM are presented. Detection limit of the assay is at 80 infectious particles per lung indicated by the blue line. Day 1 p.i. n = 9 for <i>Tmprss2^−/−</i>, n = 11 for <i>Tmprss2^+/+</i>, day 2 p.i. n = 6 for <i>Tmprss2^−/−</i>, n = 5 for <i>Tmprss2^+/+</i>, day 3 p.i. n = 7 for <i>Tmprss2^−/−</i>, n = 7 for <i>Tmprss2^+/+</i>.</p

The hemagglutinin of H1N1 PR8M influenza virus is not processed in <i>Tmprss2</i> knock-out mice.

<p>BAL from infected wild type and <i>Tmprss2^−/−</i> male mice was harvested at day 1 after infection with 2×10⁵ FFU PR8M and viral particles were concentrated by centrifugation through a sucrose cushion. As control, BAL from non-infected wild type and <i>Tmprss2^−/−</i> was analyzed. Each sample was loaded undiluted (first lane), and in two dilutions (second lane 1∶1.33, third lane 1∶2). The virus-containing pellets were then analyzed for HA cleavage by Western blots. As loading control, the stripped membranes were incubated with anti-influenza A virus antibody confirming that equal amounts of proteins were loaded for respective undiluted and diluted samples.</p

<i>Tmprss2</i> knock-out mice show reduced mortality after infection with high dose H3N2 influenza A virus infections.

<p>Eight to eleven weeks old female mice were infected with 2×10³ FFU mouse-adapted H3N2 influenza virus by intra-nasal application and survival (A) was monitored until day 14 p.i. In addition to mice that were found dead, mice with a weight loss of more than 30% of the starting body weight were euthanized and recorded as dead. Infectious viral particles were determined in lung homogenates (B). Individual values, mean and SEM are presented. Detection limit of the assay is at 80 infectious particles per lung indicated by the blue line. Homozygous <i>Tmprss2</i> knock-out mice showed significantly reduced mortality compared to wild type and heterozygote mice (p<0.0001 and p = 0.0032, respectively, using the log rank test). Viral load was not significantly different in infected wild type mice compared to infected homozygous mutant mice at days 1 to 3 p.i.</p

Alignment of amino acid sequences of the protease loop region from H1N1 and H3N2 influenza A viruses.

<p>Pathology is strongly reduced in <i>Tmprss2^−/−</i> mice after infection with H1N1 virus and diminished after infection with H3N2 virus.</p

Mild lung pathology and reduced viral spread is observed in <i>Tmprss2^−/−</i> mutant mice.

<p>Eight to twelve weeks old mice were infected intra-nasally with 2×10⁵ FFU of PR8M virus. Serial lung sections were stained at day 1 and day 3 p.i. with anti-influenza antibody and haematoxylin (A–H) or with haematoxylin/eosin (I–P). The overall lung tissues were more densely consolidated with larger numbers of infiltrating immune cells in wild type (I–L) compared to <i>Tmprss2^−/−</i> mice (M–P). In addition, the airways of <i>Tmprss2^+/+</i> mice were surrounded by higher numbers neutrophils and macrophages (I–L) whereas airways of <i>Tmprss2^−/−</i> mice showed lower numbers of immune cell infiltrations (M–P). Virus-infected cells at day 1 p.i. were observed mainly in bronchiolar regions in the lungs of both mice (A, B, E, F). Both wild type and mutant mice showed influenza-positive cells at day 3 p.i. (C, G). However, the overall number of infected cells was lower in <i>Tmprss2^−/−</i> compared to wild type mice. Furthermore, infected cells were mostly limited to bronchiolar regions in <i>Tmprss2^−/−</i> whereas in wild type mice the virus also spread significantly into the alveolar regions (D, H).</p

AR staining profiles of normal prostate, primary PCa and CRPC.

<p>(<b>A</b>) IHC staining for N- and C-terminal AR in normal prostate (NP) (a and b), hyperplastic prostate (HP) (c and d) and primary PCa (e-h) (magnification x200). (<b>B</b>) Comparison of AR staining profiles among normal prostate, hyperplastic prostate and primary PCa. (<b>C</b>) Comparison of AR staining profiles between primary PCa and metastatic CRPC.</p

Integrative Analysis of N-Linked Human Glycoproteomic Data Sets Reveals PTPRF Ectodomain as a Novel Plasma Biomarker Candidate for Prostate Cancer

In an attempt to identify prostate cancer biomarkers with greater diagnostic and prognostic capabilities, we have developed an integrative proteomic discovery workflow focused on N-linked glycoproteins that refines the target selection process. In this work, hydrazide-based chemistry was used to identify N-linked glycopeptides from 22Rv1 prostate cancer cells cultured in vitro, which were compared with glycopeptides identified from explanted 22Rv1 murine tumor xenografts. One hundred and four human glycoproteins were identified in the former analysis and 75 in the latter, with 40 proteins overlapping between data sets. Of the 40 overlapping proteins, 80% have multiple literature references to the neoplastic process and ∼40% to <i>prostatic</i> neoplasms. These include a number of well-known prostate cancer-associated biomarkers, such as prostate-specific membrane antigen (PSMA). By integrating gene expression data and available literature, we identified members of the overlap data set that deserve consideration as potential prostate cancer biomarkers. Specifically, the identification of the extracellular domain of protein tyrosine phosphatase receptor type F (PTPRF) was of particular interest due to the direct involvement of PTPRF in the control of β-catenin signaling, as well as dramatically elevated gene expression levels in the prostate compared to other tissues. In this investigation, we demonstrate that the PTPRF E-subunit is more abundant in human prostate tumor tissue compared to normal control and also detectable in murine plasma by immunoblot and ELISA. Specifically, PTPRF distinguishes between animals xenografted with the 22Rv1 cells and control animals as early as 14 days after implantation. This result suggests that the ectodomain of PTPRF has the potential to function as a novel plasma or tissue-based biomarker for prostate cancer. The workflow described adds to the literature of potential biomarker candidates for prostate cancer and demonstrates a pathway to developing new diagnostic assays