Kaposi's sarcoma-associated herpesvirus infection promotes differentiation and polarization of monocytes into tumor-associated macrophages

<p>Tumor associated macrophages (TAMs) promote angiogenesis, tumor invasion and metastasis, and suppression of anti-tumor immunity. These myeloid cells originate from monocytes, which differentiate into TAMs upon exposure to the local tumor microenvironment. We previously reported that Kaposi's sarcoma-associated herpes virus (KSHV) infection of endothelial cells induces the cytokine angiopoietin-2 (Ang-2) to promote migration of monocytes into tumors. Here we report that KSHV infection of endothelial cells induces additional cytokines including interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-13 (IL-13) that drive monocytes to differentiate and polarize into TAMs. The KSHV-induced TAMs not only express TAM-specific markers such as CD-163 and legumain (LGMN) but also display a gene expression profile with characteristic features of viral infection. More importantly, KSHV-induced TAMs enhance tumor growth in nude mice. These results are consistent with the strong presence of TAMs in Kaposi's sarcoma (KS) tumors. Therefore, KSHV infection of endothelial cells generates a local microenvironment that not only promotes the recruitment of monocytes but also induces their differentiation and polarization into TAMs. These findings reveal a new mechanism of KSHV contribution to KS tumor development.</p

Maintainable systems with a business object approach

The concept of Business Objects (BOs) has been recently promoted as a new way of exploiting object-orientation for achieving large-grain reuse. In this paper, we address the issue of how to effectively re-engineer business software applications using BOs as a reuse technique. To this end, we first identify the reuse features of business objects and then compare them with other reuse techniques. In addition, we show that software re-engineering can be more economical when business objects are used. Our work also provides guidance on how to develop and use a Business Object Architecture (BOA), which is shared by a group of interrelated and interdependent software applications. We argue that such architecture allows for more efficient reuse and better maintainability and it is illustrated by means of a case study in a realistic manufacturing environment

MGO-adducted rhBD-2 shows a concentration-dependent reduction of bactericidal activity, shown as a reduction in CFU vs unadducted peptide.

<p>CFU within a defined area were counted following radial diffusion assays performed with gram-negative, facultative anaerobes <i>Escherichia coli</i> (<i>e</i>.<i>c</i>.) and <i>Pseudomonas aeruginosa</i> (<i>p</i>.<i>a</i>.), and with the gram-positive, facultative anaerobe <i>Staphylococcus aureus</i> (<i>s</i>.<i>a</i>.) exposed to 0.5 μg/5 μl with or without MGO. Wild-type hBD-2 is highly bactericidal against most gram-negative bacterial strains, including <i>E</i>. <i>coli</i>, but this function is dramatically reduced following 72 h incubation of rhBD-2 (0.5 μg/5 μl) with 100 μM MGO at 37°C (inset). MGO (100 <i>μ</i>M) also reduces rhBD-2 bactericidal function against the gram-positive <i>S</i>. <i>aureus</i> strain. Data is presented as the Mean ± S.D. for N = 5 (<i>e</i>.<i>c</i>.) or N = 6 experiments (<i>p</i>.<i>a</i>., <i>s</i>.<i>a</i>.). Graph line for <i>e</i>.<i>c</i>. is offset slightly for clarity. (♦, ◊: p = 0.01; _*: p = 0.05).</p

Effect of MGO adduction to rhBD-2 on chemoattraction for CEM cells.

<p>Inset, untreated rhBD-2 shows a concentration-dependent optimum in chemoattraction. Fluorescence units are proportional to migrated cell number due to labeling of migrating cell DNA by the CyQuant probe. Lower graph, chemoattractive function is reduced by incubating rhBD-2 with 100 uM MGO for 72 h at 37ଌ. SDF-1 is positive control for chemotaxis of CEM cells. Data is presented as the Mean ± S.E.M. for N₁ = 3, N₂ = 3 experiments _{* *}: p = 0.05).</p

Comparison of deconvoluted tandom MS/MS spectra of untreated (a) vs modified (b) rhBD-2 peptide RYKQIGTCGLPGTK (23–36) after trypsin digestion of the rhBD-2 protein.

<p>The modified hBD-2 peptide was previously incubated in 100 μM MGO at 37°C for 72 h. The presence of the b1 ion with a +54 Da mass shift and unmodified doubly protonated y13 shows that modification of this peptide occurred at Arg²³.</p

Proteomic and Bioinformatic Profile of Primary Human Oral Epithelial Cells

Wounding of the oral mucosa occurs frequently in a highly septic environment. Remarkably, these wounds heal quickly and the oral cavity, for the most part, remains healthy. Deciphering the normal human oral epithelial cell (NHOEC) proteome is critical for understanding the mechanism(s) of protection elicited when the mucosal barrier is intact, as well as when it is breached. Combining 2D gel electrophoresis with shotgun proteomics resulted in identification of 1662 NHOEC proteins. Proteome annotations were performed based on protein classes, molecular functions, disease association and membership in canonical and metabolic signaling pathways. Comparing the NHOEC proteome with a database of innate immunity-relevant interactions (InnateDB) identified 64 common proteins associated with innate immunity. Comparison with published salivary proteomes revealed that 738/1662 NHOEC proteins were common, suggesting that significant numbers of salivary proteins are of epithelial origin. Gene ontology analysis showed similarities in the distributions of NHOEC and saliva proteomes with regard to biological processes, and molecular functions. We also assessed the interindividual variability of the NHOEC proteome and observed it to be comparable with other primary cells. The baseline proteome described in this study should serve as a resource for proteome studies of the oral mucosa, especially in relation to disease processes

Proteomic and Bioinformatic Profile of Primary Human Oral Epithelial Cells

Wounding of the oral mucosa occurs frequently in a highly septic environment. Remarkably, these wounds heal quickly and the oral cavity, for the most part, remains healthy. Deciphering the normal human oral epithelial cell (NHOEC) proteome is critical for understanding the mechanism(s) of protection elicited when the mucosal barrier is intact, as well as when it is breached. Combining 2D gel electrophoresis with shotgun proteomics resulted in identification of 1662 NHOEC proteins. Proteome annotations were performed based on protein classes, molecular functions, disease association and membership in canonical and metabolic signaling pathways. Comparing the NHOEC proteome with a database of innate immunity-relevant interactions (InnateDB) identified 64 common proteins associated with innate immunity. Comparison with published salivary proteomes revealed that 738/1662 NHOEC proteins were common, suggesting that significant numbers of salivary proteins are of epithelial origin. Gene ontology analysis showed similarities in the distributions of NHOEC and saliva proteomes with regard to biological processes, and molecular functions. We also assessed the interindividual variability of the NHOEC proteome and observed it to be comparable with other primary cells. The baseline proteome described in this study should serve as a resource for proteome studies of the oral mucosa, especially in relation to disease processes

Proteomic and Bioinformatic Profile of Primary Human Oral Epithelial Cells

Wounding of the oral mucosa occurs frequently in a highly septic environment. Remarkably, these wounds heal quickly and the oral cavity, for the most part, remains healthy. Deciphering the normal human oral epithelial cell (NHOEC) proteome is critical for understanding the mechanism(s) of protection elicited when the mucosal barrier is intact, as well as when it is breached. Combining 2D gel electrophoresis with shotgun proteomics resulted in identification of 1662 NHOEC proteins. Proteome annotations were performed based on protein classes, molecular functions, disease association and membership in canonical and metabolic signaling pathways. Comparing the NHOEC proteome with a database of innate immunity-relevant interactions (InnateDB) identified 64 common proteins associated with innate immunity. Comparison with published salivary proteomes revealed that 738/1662 NHOEC proteins were common, suggesting that significant numbers of salivary proteins are of epithelial origin. Gene ontology analysis showed similarities in the distributions of NHOEC and saliva proteomes with regard to biological processes, and molecular functions. We also assessed the interindividual variability of the NHOEC proteome and observed it to be comparable with other primary cells. The baseline proteome described in this study should serve as a resource for proteome studies of the oral mucosa, especially in relation to disease processes

Primers used in real-time quantitative RT-PCR.

<p>Primers used in real-time quantitative RT-PCR.</p

Chemotactic molecules involved in inflammatory cell trafficking.

<p>RANTES, Regulated upon Activation Normal T-cell Expression and presumably Secreted; C5a, complement 5a; M-CSF, Macrophage Colony-Stimulating Factor; GM-CSF, Granulocyte-Macrophage Colony Stimulating Factor.</p