Browsing by Autor "Hideyuki Nagasawa"

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A Comparison of the Phenotype of Dendritic Cells Derived from Discrete Peyer's Patch Macrophages of Non-Infected and Toxoplasma Gondii Infected Mice
(Japanese Society of Veterinary Science, 2003) Levi Makala; Julio C. S. Reyes; Yoshifumi Nishikawa; Yoshinori Tsushima; Xuenan Xuan; Xiaohong Huang; Hideyuki Nagasawa
A comparison of the expression of surface membrane antigens between dendritic cells (DC) derived from Peyer's patch macrophages (DPP-DC) of non-infected and Toxoplasma gondii (T. gondii) infected mice was performed. C57BL/6J mice aged 6-8 weeks of both sexes were infected orally with a 0.5 ml suspension containing 2 x 10(4) bradyzoites of the Beverley strain of T. gondii, sacrificed on day 8 and DC generated using discrete Peyer's patch macrophages (DPP-Mø) as progenitor cells. When a comparison of the expression of surface membrane antigens between the antigen presenting cells (APC) obtained from discrete Peyer's patches of non-infected and T. gondii infected mice was carried out, no significant differences were observed in the macrophage progenitor and DC populations expression of F4/80, DEC-205, CD11c, CD80 (B7-1) and CD34. However, a significant decrease in MHC class II antigen levels and a down regulation of the co-stimulatory molecule CD86 (B7-2) were noted. B7-1 appeared to be the dominant co-stimulatory ligand, whereas B7-2, which was down regulated during T. gondii infection, had a weak expression. Taken together, these results may help clarify the role of DC in the complex network regulating surface membrane antigens, as well as, their capacity for antigen uptake, processing and presentation during toxoplasmosis.
Phenotype and Function of Murine Discrete Peyer's Patch Macrophage Derived-Dendritic Cells
(Japanese Society of Veterinary Science, 2003) Levi Makala; Julio C. S. Reyes; Yoshifumi Nishikawa; Yoshinori Tsushima; Xuenan Xuan; Xiaohong Huang; Badgar Battsetseg; Tomohide Matsuo; Hideyuki Nagasawa
The phenotype and function of peritoneal cavity macrophage-derived dendritic cells (PEC-DC) was previously reported. In this study we have gone further in using our established culture system to generated discrete Peyer's patch dendritic cells (DPP-DC) from murine discrete Peyer's patch macrophages (DPP-Mø), following stimulation with granulocyte macrophage colony stimulating factor (GM-CSF) plus interleukin 4 (IL-4) for 7 days. DPP-Mø from murine small intestines were obtained by mechanical disruption of discrete Peyer's patches (DPP), followed by metrizamide density gradient centrifugation to remove Peyer's patch resident DC and debri, after which an overnight adherent step in tissue culture medium was carried out for macrophage enrichment. Characterization of the generated DPP-DC was carried out using well-established criteria of morphology, expression of membrane antigens and capacity for antigen presentation. Dendritic cells expressed DEC-205, F4/80 and CD34 at high levels, but exhibited very low CD11c levels. They were shown to present soluble protein antigen to CD3(+) spleen T cells. A comparison of the surface antigen expression in the progenitor DPP-Mø population and the generated DPP-DC showed a significant decrease in MHC class II levels and a marked down regulation of the co-stimulatory molecule CD86 (B7-2). High expression of the haemopoietic progenitor marker CD34 indicates that the generated DC, possess a haemopoietic rather than myeloid origin. Taken together, these results may provide a better understanding of the complex network regulating mucosal immune responses.