Immunohistochemical analysis of pathway HER2 4B5 performance in breast carcinoma expressing HER4.

2012 ◽  
Vol 30 (27_suppl) ◽  
pp. 16-16
Author(s):  
Julie I Jay ◽  
Patrick S Brunhoeber ◽  
Margie H Smith ◽  
Ryan R Williams ◽  
Michael Sugarman ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Immunohistochemical Analysis ◽  
Cross Reactivity ◽  
Target Antigen ◽  
Her2 Expression ◽  
Medical Systems ◽  
Hek 293 ◽  
Membrane Staining ◽  
Positive Score ◽  
Her2 Antigen

16 Background: The ability of pharmacodiagnostic antibodies to specifically detect their target antigen is essential to their performance. Non-clinical protein based models for specificity of two human epidermal growth factor 2 (HER2) pharmacodiagnostic antibodies, Oracle CB11 and PATHWAY HER2 4B5 (4B5) (Ventana Medical Systems Inc., Tucson, AZ, USA), have been reported to demonstrate affinity to both HER2 and HER4. To date there has been no evidence showing that affinity of 4B5 for HER4 impacts the IHC clinical HER2 assay. We therefore sought to investigate whether there exists any risk of the 4B5 antibody detecting HER4-positive but HER2-negative cases as falsely positive due to a hypothetical cross reactivity to HER4. Methods: A HER4 IHC assay was developed by screening ten HER4 antibodies for specificity by staining FFPE blocks of recombinant HEK-293 cells expressing HER1, HER2, HER3 and HER4. Specific HER4 antibodies were then evaluated in breast carcinoma to identify a clone that provided distinct membrane staining in tumour cells. This pattern was used as a reference in order to detect any potential interference of HER4 detection in determining the HER2 score. The HER4 antibody E200 (Epitomics) was selected and used to evaluate a cohort of breast carcinoma cases. The HER2 expression in these cases was then assessed using anti-HER2 clone PATHWAY HER2 4B5 and a HER2 reference antibody, clone SP3 (Spring Bioscience). Results: 117 out of 241 (48%) breast tissue cases evaluated were HER4 positive for membrane staining by IHC using E200. The scoring criteria for determining the HER4 score was based on the system used to score HER2, however a positive score indicated any membrane staining and therefore a 1+, 2+ or 3+ score was considered HER4-positive. 4B5 exhibited similar staining patterns as SP3 in co-expressing HER4 and HER2 tissues indicating that 4B5 was detecting only the HER2 antigen. Eighteen cases were HER4-positive by E200 and level 0 by 4B5 including a HER4 level 3+ case. Conclusions: PATHWAY HER2 4B5, a clinically relevant pharmacodiagnostic antibody for detecting HER2 expression in breast carcinoma, does not demonstrate cross reactivity to HER4 in IHC of FFPE clinical tissues.

Epstein-Barr Virus in Breast Carcinoma in Argentina

2005 ◽  
Vol 129 (3) ◽  
pp. 377-381
Author(s):  
María Victoria Preciado ◽  
Paola Andrea Chabay ◽  
Elena Noemí De Matteo ◽  
Pedro Gonzalez ◽  
Saúl Grinstein ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Epstein Barr Virus ◽  
Breast Biopsy ◽  
Immunohistochemical Analysis ◽  
Nuclear Antigen ◽  
Cancer Etiology ◽  
Barr Virus ◽  
Biopsy Specimens ◽  
Ductal Hyperplasia ◽  
Epstein Barr

Abstract Context.—Because the etiology and progression of breast carcinoma remain unclear, novel mechanisms of disease pathogenesis need to be considered. Recent interest has focused on Epstein-Barr virus (EBV), an oncogenic ubiquitous herpesvirus. Investigations of this association could not only broaden understanding of breast cancer etiology but also have implications regarding early detection, treatment, and prevention. Objective.—To assess EBV presence in breast carcinoma in an Argentine series. Design.—Breast biopsy specimens of 69 women with breast carcinoma and fresh tumor tissue of 39 of these women were collected. As controls, 17 biopsy specimens of fibroadenomas, 9 of benign epithelial proliferation, 4 of atypical ductal hyperplasia, and 10 of usual ductal hyperplasia and 8 normal breast tissues from women were studied. The EBV-infected cells were identified by means of immunohistochemical analysis, using a monoclonal antibody against Epstein-Barr virus–encoded nuclear antigen 1 (EBNA-1). Polymerase chain reaction (PCR) was used to amplify EBV DNA, with primers that cover the EBV encoded RNA (EBER) and BamHIW regions. Results.—Nuclear expression of EBNA-1 was observed in tumor epithelial cells in 24 (35%) of the 69 cases. We confirmed both positive and negative immunohistochemical results by PCR in those cases where good quality DNA was also available, detecting amplification fragments of 108 base pairs (bp) from the EBER region and 122 bp from the BamHIW region. Neither immunohistochemical analysis nor PCR detected any positive EBV results in the control samples. Conclusions.—Our results demonstrate the presence and expression of EBV restricted to epithelial tumor cells in a subset of breast carcinomas studied. However, no significant association was observed between EBV expression and worse clinical and pathologic patient characteristics.

scholarly journals IMMUNOHISTOCHEMICAL EXPRESSION OF HER2 IN ADENOCARCINOMA OF THE STOMACH

2015 ◽  
Vol 52 (2) ◽  
pp. 152-155 ◽  
Author(s):  
Diego Michelon DE CARLI ◽  
Marta Pires da ROCHA ◽  
Luis Carlos Moreira ANTUNES ◽  
Renato Borges FAGUNDES
Keyword(s):  
Gastric Cancer ◽  
Gastric Adenocarcinoma ◽  
Immunohistochemical Analysis ◽  
Anatomical Site ◽  
Her2 Expression ◽  
Immunohistochemical Expression ◽  
Her2 Gene ◽  
Cross Sectional ◽  
Dismal Prognosis ◽  
Location Methods

Background Worldwide, gastric cancer is the fourth cancer in incidence and the second most common cause of cancer death. Gastric cancer is asymptomatic in the early stages and very often diagnosed at advanced stages, determining a dismal prognosis. Expression of the HER2 gene has been identified in about 20% of gastric cancer cases, and its hyper-expression is associated with poor prognosis. Objective To investigate HER2 immunohistochemical expression in gastric adenocarcinoma and its relationship to the histological type and anatomic location. Methods A cross-sectional retrospective study analyzed the immunohistochemical expression of HER2 in a sample of 48 specimens of gastric cancer. Immunohistochemical analysis were performed using avidin-biotin-peroxidase method with C-erb B2 (clone EP1045Y), as a primary antibody (Biocare Medical, USA). Standardized gastric adenocarcinoma‘s HER2 expression criteria has been used in the analysis of samples. Results There were seven cases with reactivity for HER2. Five were of intestinal-type while two cases were of mixed-type in which the expression occurred in the intestinal component. It was identified a significant association of HER2 expression in the intestinal subtype of gastric adenocarcinoma (P=0.003). Regarding the anatomical site, HER2 was positive in only one (16.6%) of the six proximal cases and six (14.28%) of the 42 distal cases (P=0.88). Conclusion HER2 immunoexpression was identified in 14.6% of the samples, and the expression was significantly associated to Lauren’s intestinal subtype.

scholarly journals Leptin, Leptin Receptor, KHDRBS1 (KH RNA Binding Domain Containing, Signal Transduction Associated 1), and Adiponectin in Bone Metastasis from Breast Carcinoma: An Immunohistochemical Study

Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 510
Author(s):  
Paola Maroni ◽  
Alessandro Luzzati ◽  
Giuseppe Perrucchini ◽  
Luca Cannavò ◽  
Paola Bendinelli
Keyword(s):  
Signal Transduction ◽  
Bone Metastasis ◽  
Breast Carcinoma ◽  
Leptin Receptor ◽  
Rna Binding ◽  
Immunohistochemical Analysis ◽  
Binding Domain ◽  
Breast Cancer Patients ◽  
Metastatic Cells ◽  
Rna Binding Domain

Breast cancer patients are at a high risk of complications from bone metastasis. Molecular characterization of bone metastases is essential for the discovery of new therapeutic targets. Here, we investigated the expression and the intracellular distribution of KH RNA binding domain containing, signal transduction associated 1 (KHDRBS1), leptin, leptin receptor (LEPR), and adiponectin in bone metastasis from breast carcinoma and looked for correlations between the data. The expression of these proteins is known in breast carcinoma, but it has not been investigated in bone metastatic tissue to date. Immunohistochemical analysis was carried out on bone metastasis specimens, then semiquantitative evaluation of the results and the Pearson test were performed to determine eventual correlations. KHDRBS1 expression was significantly higher in the nuclei than in the cytosol of metastatic cells; LEPR was prevalently observed in the cytosol and the nuclei; leptin and adiponectin were found in metastatic cells and stromal cells; the strongest positive correlation was between nuclear KHDRBS1 and nuclear LEPR expression. Taken together, our findings support the importance of the leptin/LEPR/KHDRBS1 axis and of adiponectin in the progression of bone metastasis and suggest their potential application in pharmacological interventions.

Immunohistochemical analysis of bcl-2 and p53 protein in breast carcinoma

1996 ◽  
Author(s):  
M Baba ◽  
T Hideshima ◽  
T Shinohara ◽  
J Yamashita ◽  
T Shirakusa
Keyword(s):  
Breast Carcinoma ◽  
P53 Protein ◽  
Immunohistochemical Analysis

Retinoblastoma and p53 gene product expression in breast carcinoma: Immunohistochemical analysis and clinicopathologic correlation

1992 ◽  
Vol 23 (12) ◽  
pp. 1388-1394 ◽  
Author(s):  
Michel Trudel ◽  
Lois Mulligan ◽  
Webster Cavenee ◽  
Richard Margolese ◽  
Jean Côté ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Gene Product ◽  
P53 Gene ◽  
Immunohistochemical Analysis ◽  
Clinicopathologic Correlation ◽  
Product Expression

scholarly journals Human Kallikrein 13: Production and Purification of Recombinant Protein and Monoclonal and Polyclonal Antibodies, and Development of a Sensitive and Specific Immunofluorometric Assay

2003 ◽  
Vol 49 (1) ◽  
pp. 77-86 ◽  
Author(s):  
Carl Kapadia ◽  
Albert Chang ◽  
Georgia Sotiropoulou ◽  
George M Yousef ◽  
Linda Grass ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Amniotic Fluid ◽  
Polyclonal Antibodies ◽  
Biological Fluids ◽  
Expression System ◽  
Immunohistochemical Analysis ◽  
Cross Reactivity ◽  
Yeast Expression System ◽  
Tissue Extracts ◽  
Cancer Tissues

Abstract Background: The aims of this study were to develop immunologic reagents and a sensitive and specific immunoassay for human kallikrein 13 (hK13) and to examine the presence of hK13 in human tissues and biological fluids. Methods: Recombinant hK13 protein was produced and purified with use of a Pichia pastoris yeast expression system. The protein was used as an immunogen to generate mouse monoclonal and rabbit polyclonal anti-hK13 antibodies. A sandwich-type immunoassay was developed with these antibodies. The assay was used to measure hK13 in various biological fluids and tissue extracts. Immunohistochemical analysis was also performed on nondiseased and cancerous prostatic sections. Results: The hK13 immunoassay had a detection limit of 0.05 μg/L and showed no cross-reactivity with homologous kallikreins. The assay was linear at 0–20 μg/L, and within-and between-run CVs were <10% (n = 12). hK13 was detected in tissues, including esophagus, tonsil, trachea, lung, cervix, and prostate. hK13 was also found in seminal plasma, amniotic fluid, follicular fluid, ascites of ovarian cancer patients, breast milk, and cytosolic extracts of ovarian cancer tissues. hK13 was immunohistochemically localized in epithelial cells of both nondiseased and cancerous prostate. hK13 appears to be overexpressed in 50% of ovarian cancer tissues compared with healthy ovarian tissues. Recovery of active enzyme added to milk or amniotic fluid was 70–98%, but was <20% when added to serum, suggesting rapid sequestration by protease inhibitors. In fluids and tissue extracts, hK13 was found in its free (∼30 kDa) form. Conclusions: This immunofluorometric assay for hK13 may be used to examine the value of hK13 as a disease biomarker and to further explore the physiologic and pathobiologic role of this enzyme in human disease.

Intense basolateral membrane staining indicates HER2 positivity in invasive micropapillary breast carcinoma

2020 ◽  
Vol 33 (7) ◽  
pp. 1275-1286
Author(s):  
Shuling Zhou ◽  
Fei Yang ◽  
Qianming Bai ◽  
Anqi Li ◽  
Ming Li ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Basolateral Membrane ◽  
Membrane Staining ◽  
Her2 Positivity

scholarly journals Expression of cell-adhesion molecules in embryonic induction. I. Morphogenesis of nestling feathers.

1985 ◽  
Vol 101 (3) ◽  
pp. 1009-1026 ◽  
Author(s):  
C M Chuong ◽  
G M Edelman
Keyword(s):  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecules ◽  
General Pattern ◽  
Immunohistochemical Analysis ◽  
Dermal Papilla ◽  
Cross Reactivity ◽  
Basement Membranes ◽  
Embryonic Induction ◽  
Feather Follicle

The potential relationship of cell adhesion to embryonic induction during feather formation was examined by immunohistochemical analysis of the spatiotemporal distribution of three cell-adhesion molecules (CAMs), neural CAM (N-CAM), liver CAM (L-CAM), and neuron-glia CAM (Ng-CAM), and of substrate molecules (laminin and fibronectin) in embryonic chicken skin. The N-CAM found at sites of embryonic induction in the feather was found to be similar to brain N-CAM as judged by immuno-cross-reactivity, migratory position in PAGE, and the presence of embryonic to adult conversion. In contrast to the N-CAM found in the brain, however, only one polypeptide of Mr 140,000 was seen. N-CAM-positive dermal condensations were distributed periodically under L-CAM-positive feather placodes at those sites where basement membranes are known to be disrupted. After initiation of induction, L-CAM-positive placode cells became transiently N-CAM-positive. N-CAM was asymmetrically concentrated in the dorsal region of the feather bud, while fibronectin was concentrated in the ventral region. During feather follicle formation, N-CAM was expressed in the dermal papilla and was closely apposed to the L-CAM-positive papillar ectoderm, while the dermal papilla showed no evidence of laminin or fibronectin. The collar epithelium was both N-CAM- and L-CAM-positive. During the formation of the feather filament, N-CAM appeared periodically and asymmetrically on basilar cells located in the valleys between adjacent barb ridges. In contrast to the two primary CAMs, Ng-CAM was found only on nerves supplying the feather and the skin. These studies indicate that at each site of induction during feather morphogenesis, a general pattern is repeated in which an epithelial structure linked by L-CAM is confronted with periodically propagating condensations of cells linked by N-CAM.

scholarly journals Regulation of 3-Phosphoinositide-dependent Protein Kinase-1 (PDK1) by Src Involves Tyrosine Phosphorylation of PDK1 and Src Homology 2 Domain Binding

2007 ◽  
Vol 283 (3) ◽  
pp. 1480-1491 ◽  
Author(s):  
Keum-Jin Yang ◽  
Sanghee Shin ◽  
Longzhen Piao ◽  
Eulsoon Shin ◽  
Yuwen Li ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Complex Formation ◽  
Tyrosine Phosphorylation ◽  
Immunohistochemical Analysis ◽  
Sh2 Domain ◽  
Resting Cells ◽  
Dependent Protein Kinase ◽  
Hek 293 ◽  
Dependent Protein

3-Phosphoinositide-dependent protein kinase-1 (PDK1) appears to play a central regulatory role in many cell signalings between phosphoinositide-3 kinase and various intracellular serine/threonine kinases. In resting cells, PDK1 is known to be constitutively active and is further activated by tyrosine phosphorylation (Tyr9 and Tyr373/376) following the treatment of the cell with insulin or pervanadate. However, little is known about the mechanisms for this additional activation of PDK1. Here, we report that the SH2 domain of Src, Crk, and GAP recognized tyrosine-phosphorylated PDK1 in vitro. Destabilization of PDK1 induced by geldanamycin (a Hsp90 inhibitor) was partially blocked in HEK 293 cells expressing PDK1-Y9F. Co-expression of Hsp90 enhanced PDK1-Src complex formation and led to further increased PDK1 activity toward PKB and SGK. Immunohistochemical analysis with anti-phospho-Tyr9 antibodies showed that the level of Tyr9 phosphorylation was markedly increased in tumor samples compared with normal. Taken together, these data suggest that phosphorylation of PDK1 on Tyr9, distinct from Tyr373/376, is important for PDK1/Src complex formation, leading to PDK1 activation. Furthermore, Tyr9 phosphorylation is critical for the stabilization of both PDK1 and the PDK1/Src complex via Hsp90-mediated protection of PDK1 degradation.

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