Characterization of a glycosphingolipid antigen defined by the monoclonal antibody MBr1 expressed in normal and neoplastic epithelial cells of human mammary gland

The antigen defined by a monoclonal antibody, MBr1, was found to be expressed in normal human mammary gland epithelia and human mammary carcinoma cells (Menard, S., Tagliabue, E., Canevari, S., Fossati, G., and Colnaghi, M. I. (1983) Cancer Res. 43, 1295-1300). The antigen has been isolated from breast cancer cell line MCF-7, which was used as immunogen, and its structure was determined by methylation analysis, NMR spectroscopy, direct probe mass spectrometry, and enzymatic degradation as identified below. Fuc alpha 1----2Gal beta 1----3GalNAc beta 1----3Gal alpha 1----4Gal beta 1----4Glc beta 1----1Cer The antibody cross-reacted weakly with fucosylasialo-GM1 (IV2FucGg4), which shares the same terminal sequence, Fuc alpha 1----2Gal beta 1----3GalNAc, with this antigen. However, various other structures, including lacto-series H structure (Fuc alpha 1----2 Gal beta 1----4/or 3GlcNAc beta 1----3Gal), did not show any reactivity with this antibody. Therefore, this antigen represents a blood group H antigen with a globo-series structure which is abundant in human teratocarcinoma (Kannagi, R., Levery, S. B., Ishigami, F., Hakomori, S., Shevinsky, L. H., Knowles, B. B., and Solter, D. (1983) J. Biol. Chem. 258, 8934-8942), although its presence must be limited in normal adult human tissue

Ligand-dependent EGFR activation induces the co-expression of IL-6 and PAI-1 via the NFkB pathway in advanced-stage epithelial ovarian cancer.

The epidermal growth factor receptor (EGFR), a member of the ErbB family of receptor tyrosine kinases, is expressed in up to 70% of epithelial ovarian cancers (EOCs), where it correlates with poor prognosis. The majority of EOCs are diagnosed at an advanced stage, and at least 50% present malignant ascites. High levels of IL-6 have been found in the ascites of EOC patients and correlate with shorter survival. Herein, we investigated the signaling cascade led by EGFR activation in EOC and assessed whether EGFR activation could induce an EOC microenvironment characterized by pro-inflammatory molecules. In vitro analysis of EOC cell lines revealed that ligand-stimulated EGFR activated NFkB-dependent transcription and induced secretion of IL-6 and plasminogen activator inhibitor (PAI-1). IL-6/PAI-1 expression and secretion were strongly inhibited by the tyrosine kinase inhibitor AG1478 and EGFR silencing. A significant reduction of EGF-stimulated IL-6/PAI-1 secretion was also obtained with the NFkB inhibitor dehydroxymethylepoxyquinomicin. Of 23 primary EOC tumors from advanced-stage patients with malignant ascites at surgery, 12 co-expressed membrane EGFR, IL-6 and PAI-1 by immunohistochemistry; both IL-6 and PAI-1 were present in 83% of the corresponding ascites. Analysis of a publicly available gene-expression data set from 204 EOCs confirmed a significant correlation between IL-6 and PAI-1 expression, and patients with the highest IL-6 and PAI-1 co-expression showed a significantly shorter progression-free survival time (P=0.028). This suggests that EGFR/NFkB/IL-6-PAI-1 may have a significant impact on the therapy of a particular subset of EOC, and that IL-6/PAI-1 co-expression may be a novel prognostic marker

Broad-Spectrum Inhibition of HIV-1 by a Monoclonal Antibody Directed against a gp120-Induced Epitope of CD4

To penetrate susceptible cells, HIV-1 sequentially interacts with two highly conserved cellular receptors, CD4 and a chemokine receptor like CCR5 or CXCR4. Monoclonal antibodies (MAbs) directed against such receptors are currently under clinical investigation as potential preventive or therapeutic agents. We immunized Balb/c mice with molecular complexes of the native, trimeric HIV-1 envelope (Env) bound to a soluble form of the human CD4 receptor. Sera from immunized mice were found to contain gp120-CD4 complex-enhanced antibodies and showed broad-spectrum HIV-1-inhibitory activity. A proportion of MAbs derived from these mice preferentially recognized complex-enhanced epitopes. In particular, a CD4-specific MAb designated DB81 (IgG1Κ) was found to preferentially bind to a complex-enhanced epitope on the D2 domain of human CD4. MAb DB81 also recognized chimpanzee CD4, but not baboon or macaque CD4, which exhibit sequence divergence in the D2 domain. Functionally, MAb DB81 displayed broad HIV-1-inhibitory activity, but it did not exert suppressive effects on T-cell activation in vitro. The variable regions of the heavy and light chains of MAb DB81 were sequenced. Due to its broad-spectrum anti-HIV-1 activity and lack of immunosuppressive effects, a humanized derivative of MAb DB81 could provide a useful complement to current preventive or therapeutic strategies against HIV-1

Human Cutaneous Melanomas Lacking MITF and Melanocyte Differentiation Antigens Express a Functional Axl Receptor Kinase

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A functional gene expression analysis in epithelial sinonasal cancer: Biology and clinical relevance behind three histological subtypes

Epithelial sinonasal cancers (SNCs) are rare diseases with overlapping morphological features and a dismal prognosis. We aimed to investigate the expression differences among the histological subtypes for discerning their molecular characteristics. We selected 47 SNCs: (i) 21 nonkeratinizing squamous cell carcinomas (NKSCCs), (ii) 13 sinonasal neuroendocrine cancers (SNECs), and (iii) 13 sinonasal undifferentiated cancers (SNUCs). Gene expression profiling was performed by DASL (cDNA-mediated annealing, selection, extension, and ligation) microarray analysis with internal validation by quantitative RT-PCR (RT-qPCR). Relevant molecular patterns were uncovered by sparse partial-least squares discriminant analysis (sPLS-DA), microenvironment cell type (xCell), CIBERSORT, and gene set enrichment (GSEA) analyses. The first two sPLS-DA components stratified samples by histological subtypes. xCell highlighted increased expression of immune components (CD8 + effector memory cells, in SNUC) and \u201cother cells\u201d: keratinocytes and neurons in NKSCC and SNEC, respectively. Pathway enrichment was observed in NKSCC (six gene sets, proliferation related), SNEC (one gene set, pancreatic \u3b2-cells), and SNUC (twenty gene sets, some of them immune-system related). Major neuroendocrine involvement was observed in all the SNEC samples. Our high-throughput analysis revealed a good diagnostic ability to differentiate NKSCC, SNEC, and SNUC, but indicated that the neuroendocrine pathway, typical and pathognomonic of SNEC is also present at lower expression levels in the other two histological subtypes. The different and specific profiles may be exploited for elucidating their biology and could help to identify prognostic and therapeutic opportunities

A STUDY ON CORRELATION BETWEEN ELECTROMYOGRAPHICAL AND MORPHOLOGICAL FINDINGS OF BACK MUSCLES IN SCOLIOSIS, ESPECIALLY CHANGES IN THE INTRAMUSCULAR NERVE ENDINGS.

3. In the cases of neurofibromatosis with scoliosis, intramuscular nerve endings showed almost normal morphological appearances but, comparative histograms of the diameters of the end-plates in each 5μ. group showed 2 peak formations.. One of these cases recorded fibrillation voltage at the concave side showed abnormal staining of terminal filamints of the end-plates in deep back muscles at the same side. The sensory nerve endings showed perfectly preserved histological appearances. 4. In cases of rachitogenic scoliosis, motor end-plates of the back muscles were observed using the histochemical method of cholinesterase staining and vital staining with methylene blue. Histograms of the diameters of the end-plates showed a noted reduction in size and in number of units, but no apparent degenerative changes in sensory and motor endings or in terminal axons were seen. 5. In a case of discogenic scoliosis, silver impregnation was applied on the back muscles at both sides of the curvature. In the convex side, collateral branching and swelling of the nerve fibers was observed and also the motor end-plates showed an abnormality of staining in terminal arborization, but, fibrillation voltage was not recorded. 6. In congenital scoliosis, electromyographic findings failed to trace fibrillation voltage, but, terminal axons in deep muscles, showed collateral branchig and an increased terminal innervation ratio in a highly affected case. In slightly curved cases, atrophic changes, and intramuscular fibers and endings appeared almost normal. 7. In the so-called idiopathic scoliosis , about 50% had not traced fibrillation voltage in the paravertebral back muscles. Degenerative findings.of the intramuscular nerve fibers and endings were not observed, but comparative histograms of the diameters of the end -plates showed 2 peak formations in the convex side of the back muscles. Innervation ratio was not changed and sensory nerve endings showed normal appearances.8. In idiopathic scoliosis, characterised by fibrillation voltage which was traced in the paravertebral back muscles, intramuscular motor nerve endings were degenerated in deep muscles of the convex side, and disseminated muscle atrophy in the concave side. In deep muscles of the convex side, terminal axons showed collateral branching and motor endplates, club-like swelling, and 2 peak formations of the histograms of the diameters. However, sensory nerve endings were normal in appearance. These findings lead to a conclusion that changes in the motor endings observed in paralytic scoliosis differed from histological changes in the cases of idiopathic scoliosis, in which fibrillation voltage had been traced. In paralytic scoliosis, histological changes of the back muscles showed various kinds of degenerative findings, but, in cases of idiopathic scoliosis with fibrillation voltage they showed collateral branching and degenerative changes of the end-plates, respectively, in deep back muscles especially in the convex side. In other kinds of scoliosis, neuromuscular changes were largely influenced by their own basic disorders such as, degeneration of discs, abnormality of vertebral bodies, nutritional deficiencies and metablic disorders. In the cases of idiopathic scoliosis without fibrillation voltage intramuscular nerve endings were preserved in good condition, but almost all of the cases of non-paralytic scoliosis showed an abnormality of the histograms of the endplates, or atrophic changes in deep muscles. Prophylactic treatment must be employed on the back muscles for the prevention of further deformity and progression of scoliosis.An electromyographical and histological study on 20 cases of various kinds of scoliosis was carried out. In particular, a biopsy of the back muscles at the apex of the primary curve was performed using the methods of gold chloride staining, silver impregnation modified by Seto, vital staining with methylene blue, and the histochemical demonstration of cholinesterase on the subneural apparatuses of the endplates. Specimens were taken from superficial (M. longissimus dorsi) and deep (M. multifidus) back muscles of scoliosis patients and in all cases, an electromyographic fibrillation voltage was picked up under deep general anesthesia with ether. This was done from the paravertebral back muscles and a comparison of histologic and electromyographic findings was performed. The results obtained are summarized as follows : 1. Motor end-plates of normal back muscles, dissected at autopsy from adults were found to be concentrated in band-like narrow zones and situated at the mid-point of the muscle fibers. Through comparative histograms of the diameters of subneural apparatuses and those of the end-plates of gold chloride staining showed a close correlation. 2. Intramuscular endings in the cases of paralytic scoliosis showed remarkable collateral axonic sprouting and various kinds of pathological changes of the end-plates, multiple innervation, large and and small end-plates, thickened terminal filaments, failure to stain, irregular swelling, and abnormal terminal expansion. The motor endings of the musclespindle were also remarkably degenerated but sensory nerve fibers and endings remained undisturbed. In neuromuscular endings, pathological changes of the back muscles at the apex of the primary curve, displayed a distinguishing feature in a lack of uniformity in the degree of alignment and curvature, but, compared with superficial muscles, the deep muscles were significantly involved. In all cases a giant spike and fibrillation voltage was recorded from wide-spread various parts of the back muscles

Gene expression profiling of advanced ovarian cancer: characterization of a molecular signature involving fibroblast growth factor 2

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Whole-transcriptome analysis links trastuzumab sensitivity of breast tumors to both HER2 dependence and immune cell infiltration.

While results thus far demonstrate the clinical benefit of trastuzumab, some patients do not respond to this therapy. To identify a molecular predictor of trastuzumab benefit, we conducted whole-transcriptome analysis of primary HER2+ breast carcinomas obtained from patients treated with trastuzumab-containing therapies and correlated the molecular portrait with treatment benefit. The estimated association between gene expression and relapse-free survival allowed development of a trastuzumab risk model (TRAR), with ERBB2 and ESR1 expression as core elements, able to identify patients with high and low risk of relapse. Application of the TRAR model to 24 HER2+ core biopsies from patients treated with neo-adjuvant trastuzumab indicated that it is predictive of trastuzumab response. Examination of TRAR in available whole-transcriptome datasets indicated that this model stratifies patients according to response to trastuzumab-based neo-adjuvant treatment but not to chemotherapy alone. Pathway analysis revealed that TRAR-low tumors expressed genes of the immune response, with higher numbers of CD8-positive cells detected immunohistochemically compared to TRAR-high tumors. The TRAR model identifies tumors that benefit from trastuzumab-based treatment as those most enriched in CD8-positive immune infiltrating cells and with high ERBB2 and low ESR1 mRNA levels, indicating the requirement for both features in achieving trastuzumab response