Inflammation and tissue repair markers distinguish the nodular sclerosis and mixed cellularity subtypes of classical Hodgkin's lymphoma

Background: Classical Hodgkin's lymphoma (cHL), although a malignant disease, has many features in common with an inflammatory condition. The aim of this study was to establish the molecular characteristics of the two most common cHL subtypes, nodular sclerosis (NS) and mixed cellularity (MC), based on molecular profiling and immunohistochemistry, with special reference to the inflammatory microenvironment. Methods: We analysed 44 gene expression profiles of cHL whole tumour tissues, 25 cases of NS and 19 cases of MC, using Affymetrix chip technology and immunohistochemistry. Results: In the NS subtype, 152 genes showed a significantly higher expression, including genes involved in extracellular matrix (ECM) remodelling and ECM deposition similar to wound healing. Among these were SPARC, CTSK and COLI. Immunohistochemistry revealed that the NS-related genes were mainly expressed by macrophages and fibroblasts. Fifty-three genes had a higher expression in the MC subtype, including several inflammation-related genes, such as C1Qα, C1Qβ and CXCL9. In MC tissues, the C1Q subunits were mainly expressed by infiltrating macrophages. Conclusions and interpretations: We suggest that the identified subtype-specific genes could reflect different phases of wound healing. Our study underlines the potential function of infiltrating macrophages in shaping the cHL tumour microenvironment

Olfactory receptor protein expression in mouse and human ovarian granulosa cells.

<p>Immunohistochemical stains of ovaries from wild type (A, C, E) and mutant (B, D, F) mice using either anti-Olfr68 (A-D) or anti-Olfr1508 (E-F). The high magnification images shown in C and D are from the same ovaries shown at lower magnification in A and B, respectively. All mice were littermates and were treated with 5 IU of PMSG 48 hours before being euthanized to mimic the conditions used for the mRNA profiling analyses. Bars: 100 microns. (G) Western blot analyses of protein extracts obtained from 3 different wild type mouse olfactory tubercles (samples 1–3), 3 different wild type mouse ovarian granulosa cells (samples 4–6), and 2 different normal donors of human luteinized granulosa cells undergoing <i>in vitro</i> fertilization procedures (samples 7–8). Cells from the donor from whom sample 7 was derived were also cultured <i>in vitro</i> for 8 days before being analyzed by western blotting (sample 7a). The blot was probed with an antibody against Olfr68, which shows 82% sequence homology to human OR52A5 and 73% homology to human OR52A1, both with molecular sizes of 36 KDa.</p

Cre-mediated recombination in olfactory tubercles of <i>Fshr-Cre</i> transgenic mice.

<p>Olfactory tubercles of R26R reporter mice either carrying (A) or not carrying (B) the <i>Fshr-Cre</i> transgene were stained for LacZ as reported earlier [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139013#pone.0139013.ref004" target="_blank">4</a>]. The blue color indicative of a positive LacZ colorimetric assay, present in olfactory bulbs from the transgenic line, indicates the presence of Cre-mediated recombination driven by the <i>Fshr-Cre</i> transgene in olfactory tubercles of mice harboring this transgene.</p

Increased expression of olfactory receptors in ovarian follicles lacking a functional Brca1.

<p>Two pairs of wild type (WT) and mutant (MT) mice, respectively 8 and 11 months old, were synchronized in the proestrus phase of their estrous cycle. Granulosa cells were isolated from frozen histological sections of their ovaries using laser capture microdissection. Expression profiling analyses were performed using total mRNA from each cell preparation. (A) Heat map illustrating the expression levels of genes showing differential down-regulation (blue) or up-regulation (yellow) of at least 2-fold in mutant mice from both age groups. The arrows indicate genes belonging to the olfactory receptor family. The heat map in (B) shows olfactory receptor genes with at least 2-fold differential expression in both age groups.</p

Correlation between gene expression levels determined from microarray <i>versus</i> RT-PCR analyses.

<p>Ten loci were randomly selected from the list of differentially expressed loci in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139013#pone.0139013.s002" target="_blank">S1 Table</a> and their relative expression levels were measured using quantitative RT-PCR. Relative expression values obtained from the expression profiling studies and from the RT-PCR data are shown in the table on the left and plotted in the graph shown on the right. The Pearson correlation coefficient was calculated to evaluate the correlation between expression array and real time RT-PCR data.</p