Immuno Expression Research Articles

PAX Immunoreactivity Patterns in Anaplastic Wilms Tumors

This immunohistochemistry study explores particular differential immuno expression patterns of PAX genes in unfavorable histology Wilms tumor (UH) and favorable histology Wilms tumor (FH). Some unique features in unfavorable histology Wilms tumor were discovered and they may provide insightful clues of this unique and both clinically and biologically important phenomenon.

P5-11-16: CD44 and CD24 Expression in Ductal Invasive Breast Carcinomas, Classified by Molecular Subtypes and Its Association with Prognostic Factors.

Abstract Backgound: Breast carcinomas (BC) exhibit intra tumoral heterogeneity being stratified into several subgroups based in gene expression profiles or histochemical biomarkers. It was suggested that this heterogeneity is derived in part from the transformation of different subsets of cancer stem cells (CSC) in each intrinsic subgroup. The presence of CSC can be evidenced by phenotypic analysis of CD44 e CD24. This study aimed to identify the CD24 and CD44 immunophenotypes within invasive ductal breast carcinoma (IDC) subgroups defined by immunohistochesmistry markers and determine its influence on prognosis as well as its association with the expression of Ki67, citokeratins (CK5 and CK18) and claudin-7. Methods: Immuno expression of CD44 and CD24 alone or in combination was investigated in 95 IDC cases arranged in a tissue microarray (TMA). The association with subgroups defined as luminal A and B;HER2 rich and triple negative, or with the other markers and prognosis was analyzed. Results: CD44+/CD24- and CD44-/CD24+ were respectively present in 8.4% and 16.8% of the tumors, a lack of both proteins was detected in 6.3%, while CD44+/CD24+ was determined in 45.3% of the tumors. Although there was no significant correlation between subgroups and different phenotypes, the CD44+/CD24-phenotype was more common in the basal subgroups but absent in HER2 tumors, whereas luminal tumors are enriched in CD44-/CD24+ and CD44+/CD24+ cells. The frequency of CD44+/CD24- or CD44-/CD24+ have not been associated with clinical characteristics or biological markers. There was also no significant association of these phenotypes with the event free (DFS) and overall survival (OS). Single CD44+ was evident in 57.9% and was marginally associated to grading and not to any other tumor characteristics, while CD24+ was positive in 74.7% of the tumors, showing a significant association with ER, PR and Ki67 and a marginal association with CK18 and claudin-7. Expression of claudin-7 and Ki67 did not associate with the cancer subgroups, while a positive association between CK18 and the luminal subgroups was found (p=0.03). CD44+ was not significantly associated with OS and DFS whereas CD24+ frequency although not significantly associated with OS was associated with a decrease in DFS (p=0.07). CK5, CK18 and Ki67 expression had no influence in OS or DFS, claudin-7 positive was associated with reduced DFS (p=0.05). Conclusions: There was no significant correlation between CD44+/CD24- tumor cells frequency and event-free or OS. However, a tendency toward a favorable prognosis, was noted. Contrariwise the presence of CD44-/CD24+ suggested a worse prognosis. Both single CD24 and claudin-7 positivity were associated with reduced time of reccurence, suggesting a contribution of these markers to aggressiveness. Supported by FAPESP and CNPq. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-11-16.

Regulation of Connective Tissue Growth Factor (CTGF) Ameliorates the Disease Course in a Rodent Model of Reactive Arthritis (RA).

Gene expression of CTGF, a proangiogenic molecule, is up-regulated by serine proteases (FXa) and thrombin (JBC 275, 2000). We have demonstrated that thrombospondin-1 (TSP1) plays a major role in the assembly of the prothrombinase complex on the surface of neutrophils (PMNs) and that PMNs contain the message for CTGF. Rheumatoid arthritis is a chronic inflammatory disease associated with leukocyte adhesion and extravasation with angiogenesis as its hallmark. There is evidence indicating that TSP1 plays a major role in the pathophysiology of RA (JI 171, 2003). In this genetically susceptible rodent model of RA, a single intraperitoneal (i.p.) injection of peptidoglycan-polysaccharide induces an inflammatory immune response characterized by increased joint diameters in a biphasic manner. The acute phase occurs during the first five days with spontaneous reactivation around day 12 resulting in a progressive and irreversible chronic phase. We tested a well-characterized synthetic peptide derived from TSP1-type 3 repeats under three different therapeutic modalities in the experimental model of RA. First, peptide treatment was administered intravenously (i.v.) during the first five days of the experimental protocol representing the acute phase. Second, peptide treatment was administered i.v. during the acute phase but then i.p. every other day during the chronic phase of the experimental protocol. Third, peptide treatment was administered daily i.p. during the acute phase and every other day during the chronic phase. At all times a positive control group was included as well as a negative control group receiving a scrambled peptide. Total RNA was isolated from the ankle joints of the animals and RT-PCR was performed using specific primers for CTGF and G3PDH. Integrated density values were measured and the ratio of CTGF/G3PDH assessed. Immunohistochemical analysis of CTGF was also performed using Bioquant image analysis software to quantitate CTGF immuno expression. In peptide-treated animals CTGF expression in the cells of the synovial lining was significantly decreased when compared to the disease untreated group. Peptide untreated positive control group was associated with upregulation of the CTGF gene expression (2.53±0.33) in the ankle tissue and localized to the synovial membranes as judge by the immunochemical studies (8.7±0.77). In contrast, peptide treatment downregulated CTGF gene expression (0.9±0.41, p <0.03) as well as CTGF protein expression in synovial membranes (3.9±1.79, p <0.005). The group receiving the scrambled TSP1 peptide was no different from positive control group. These molecular changes where associated with other parameters evaluated including macroscopic changes, namely a decrease in joint diameters observed with peptide treatment group. Peptide treatment blunted the increase in the joint diameters both during the acute phase as well as the chronic phase. The i.v. injection was more effective than the i.p. (p <0.05) injection during the acute phase, however the peptide treatment ameliorated the chronic phase even though it was administered i.p. Histological data indicated that peptide treatment decreased neovascularization, neutrophil infiltration and thickening of the synovial lining in the joint when compared to the disease untreated group. For the first time these results demonstrate a link between TSP1 and CTGF expression and the disease course in an experimental animal model of RA.