Characterization of the Primo-Vascular System in the Abdominal Cavity of Lung Cancer Mouse Model and Its Differences from the Lymphatic System

Cancer growth and dissemination have been extensively studied for a long time. Nevertheless, many new observations on anatomy and histopathology of cancer events are still reported such as formation of a vasculogenic-like network inside aggressive tumors. In this research, new kinds of micro-conduits, named primo-vessels, were found inside the abdominal cavity of NCI-H460 lung cancer murine xenograft models. These vascular threads were largely distributed on the surfaces of various organs and were often connected to peritoneal tumor nodules. Histological and immunofluorescent investigations showed that the primo-vessels had characteristic features that were distinctively different from those of similar-looking lymphatic vessels. They had multiple channels surrounded with loose collageneous matrices, which is in contrast to the single-channel structure of other vascular systems. The rod-shaped nuclei aligned longitudinally along the channels were assumed to be the endothelial cells of the primo-vessels, but LYVE-1, a specific marker of lymphatics, was not expressed, which indicates a clear difference from lymphatic endothelial cells. Taken together these findings on and characterization of the novel threadlike vascular structures in cancer models may have important implications for cancer prognosis and for therapy

Sub-Telomere Directed Gene Expression during Initiation of Invasive Aspergillosis

Aspergillus fumigatus is a common mould whose spores are a component of the normal airborne flora. Immune dysfunction permits developmental growth of inhaled spores in the human lung causing aspergillosis, a significant threat to human health in the form of allergic, and life-threatening invasive infections. The success of A. fumigatus as a pathogen is unique among close phylogenetic relatives and is poorly characterised at the molecular level. Recent genome sequencing of several Aspergillus species provides an exceptional opportunity to analyse fungal virulence attributes within a genomic and evolutionary context. To identify genes preferentially expressed during adaptation to the mammalian host niche, we generated multiple gene expression profiles from minute samplings of A. fumigatus germlings during initiation of murine infection. They reveal a highly co-ordinated A. fumigatus gene expression programme, governing metabolic and physiological adaptation, which allows the organism to prosper within the mammalian niche. As functions of phylogenetic conservation and genetic locus, 28% and 30%, respectively, of the A. fumigatus subtelomeric and lineage-specific gene repertoires are induced relative to laboratory culture, and physically clustered genes including loci directing pseurotin, gliotoxin and siderophore biosyntheses are a prominent feature. Locationally biased A. fumigatus gene expression is not prompted by in vitro iron limitation, acid, alkaline, anaerobic or oxidative stress. However, subtelomeric gene expression is favoured following ex vivo neutrophil exposure and in comparative analyses of richly and poorly nourished laboratory cultured germlings. We found remarkable concordance between the A. fumigatus host-adaptation transcriptome and those resulting from in vitro iron depletion, alkaline shift, nitrogen starvation and loss of the methyltransferase LaeA. This first transcriptional snapshot of a fungal genome during initiation of mammalian infection provides the global perspective required to direct much-needed diagnostic and therapeutic strategies and reveals genome organisation and subtelomeric diversity as potential driving forces in the evolution of pathogenicity in the genus Aspergillus

MOUSE CYTOKINE-INDUCED KILLER CELLS DEVELOPED FROM DIFFERENT SOURCES

Introduction: In Kazakhstan, death from colorectal cancer is on the leading position among cancer-related deaths in the population, and since 2013 colorectal cancer is one of the three cancer diseases subject to the National Screening Program. The treatment protocol used for colorectal cancer therapy with metastases has very low efficacy. Another strategy in cancer therapy is immunotherapy with cytokine-induced killer cells (CIK cells). Human CIK cells are isolated from peripheral blood mononuclear cell fraction using IFN-γ, IL-2cytokine and anti-CD3 monoclonal antibodies. As a result, a heterogenous population which consists mainly of CD3+CD56-, CD3+CD56+ cells and of a small population of CD3-CD56+cells is obtained. Among the killer cells obtained, CD3+CD56+ have the greatest cytotoxic activity. For developed preclinical studies of CIK cells in murine model we search the best source of CIK cells within spleen, lymph nodes, bone marrow. Methods: CIK cells will be proliferated from mouse spleen, lymph nodes, bone marrow cells. Spleen, lymph nodes, bone marrow cells without monocytes and erythrocytes expanded with IFN-γ, IL-2cytokines and anti-CD3 monoclonal antibodies for 14 days. Positive selection of CIK cells against NK1.1and DX5 will be performed on immune beads (Miltenyi biotech). Results: CIK cells are characterized by both MHC-restricted and MHC-unrestricted anti-tumor cytotoxicity against a broad range of cancer cells. Mouse CIK cells have distinct phenotype from human CIK cells. NK1.1and DX5 are murine natural killer markers. According to literature data after culturing spleen cells for 21day NK1.1+ and DX5+ of TCRαβ+ CD3+ CD8+ T cells have the greatest cytotoxicity. We evaluated NK1.1+ and DX5+ cells after culturing cells isolated from spleen, lymph nodes and bone marrow for 14 days. NK1.1positive cells were 53,3% and DX5+ were 5% from bone marrow cells, but bone marrow cells showed low amounts of expanded cells. 21,8% of spleen cells showed NK1.1+ phenotype, 20% of DX5 (CD49b). Lymph nodes gave rise to 12,3% NK1.1+ cells. According to proliferation potential and portion of NK1.1+ and DX5+, spleen and lymph nodes are prospective sources of CIK cells. Conclusion: Spleen and lymph node cells may be sources for expansion of mouse CIK cells