Effect of ovarian hormones on maturation of dendritic cells from peripheral blood monocytes in dogs     

Viskam WIJEWARDANA  Kikuya SUGIURA  Daluthgamage Patsy H. WIJESEKERA  Shingo HATOYA  Toshiya NISHIMURA  Ryoji KANEGI  Takahiro USHIGUSA  Toshio INABA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(7):771-775

Previously, we reported that ovarian hormones affect the immune response against E. coli isolated from the dogs affected with pyometra. In order to investigate mechanisms underlying the immune modulation, we examined the effects of ovarian hormones on the generation of dendritic cells (DCs), the most potent antigen presenting cell. DCs were differentiated from peripheral blood monocytes (PBMOs) using a cytokine cocktail. Both estrogen receptor and progesterone receptors were expressed by the PBMOs and immature DCs. When various ovarian hormones were added to the culture for the DC differentiation, progesterone significantly decreased the expression of DC maturation markers, such as CD1a, CD80 and CD86, on mature DCs. Conversely, the addition of estrogen to the cultures increased the expression of CD86, but not other maturation makers. Furthermore, DCs differentiated in the presence of progesterone did not stimulate allogeneic mononuclear cells in PB. Taken together, these results indicate that progesterone diminishes the maturation of DCs, leading to decreased immune responses against invading pathogens.  相似文献   

Construction of an expression vector for improved secretion of canine IL-18     

Sugiura K  Akazawa T  Fujimoto M  Wijewardana V  Mito K  Hatoya S  Taketani S  Komori M  Inoue N  Inaba T 《Veterinary immunology and immunopathology》2008,126(3-4):388-391

To construct a vector for caspase-1 independent expression of canine IL-18, the signal sequence of canine IL-12p40 was fused to the sequence of mature IL-18 on the NdeI restriction site which is located at the 3' end of the signal sequence. The resulting vector expressed coding protein from transfected mammalian cells. The expressed protein was shown to have IL-18 bioactivity in a INF-gamma-inducing assay. These results suggest that the expression vector is the desired tool for advancement of dendritic cell (DC)-based cancer therapy, provided that the vector can successfully be transfected into dendritic cells. We propose a simple and widely applicable method for providing the signal sequence.  相似文献   

Generation of canine dendritic cells from peripheral blood monocytes without using purified cytokines   总被引：1，自引：0，他引：1  

Wijewardana V  Sugiura K  Oichi T  Fujimoto M  Akazawa T  Hatoya S  Inaba M  Ikehara S  Jayaweera TS  Inaba T 《Veterinary immunology and immunopathology》2006,114(1-2):37-48

Dendritic cells (DCs), which differentiate in vitro from peripheral blood monocytes (PBMOs) or bone marrow precursors, are a promising candidate for immunotherapy against cancer. The dog, which suffers common types of cancers along with humans, make an ideal large animal model for cancer studies. Monocyte-derived DCs in the dog have not been well characterized, however, since the appropriate condition for in vitro differentiation has not been established. To tackle this problem, we have developed a conditioned media by culturing T cells with immobilized anti-canine CD3 antibody, and sought to induce differentiation of DCs from PBMOs. When purified CD14+ PBMOs were cultured in the presence of 25% T cell conditioned medium (TCCM), the PBMOs increased size and had extended dendritic processes by day 12 of the culture. The cultured PBMOs were found to increase the expression of MHC class II and CD1a molecules, and significantly increased stimulatory activity for allogeneic T cells in the mixed leukocyte reaction. Moreover, the cells significantly increased their expression of IL-18 and IFN-gamma when stimulated with polyinosinic-polycytidylic acid (Poly (I:C)). The cells have a reduced phagocytic activity, which is a common defect in mature DCs. It follows from these results that TCCM does induce the differentiation of DCs from PBMOs.  相似文献   

Isolation and characterization of hematopoietic progenitor cells in canine bone marrow     

Wijewardana V  Sugiura K  Shigeyama N  Moriguchi M  Tsunoda S  Ikehara S  Inaba T 《Veterinary immunology and immunopathology》2007,115(3-4):230-238

For ultimate diagnoses of canine leukemia or malignant lymphoma, we sought to isolate hematopoietic progenitor cells (HPCs) from canine bone marrow (BM) using physiological phenotypes. Canine BM cells were separated by equilibrium discontinued density centrifugation, and HPCs, detected by in vitro colony formation, were significantly enriched in the relatively low density (LD) fraction. In flow cytometry, many CD34 or MHC class II expressing cells were detected in the LD fraction, but these were not significantly enriched. When the LD cells were separated, using a cell-sorting method, into cells with high affinity of wheat germ agglutinin (WGAhigh) and cells with WGAlow, almost all multipotent HPCs (MHPCs) and HPCs committed to myeloid lineage were found in the WGAhigh population. When the WGAhigh population was further stained for rhodamin 123, almost all MHPCs were included in the dull population (Rhlow), but not in the bright one (Rhhigh). Morphologically, most Rhlow cells were round, blastic cells containing a large nucleus with nucleoli and narrow cytoplasm. Based on these results, we suggest that all of the MHPCs in canine BM show the Rhlow WGAhigh LD phenotype, and may contain hematopoietic stem cells, which are the primitive HPCs.  相似文献