| Project/Area Number |
03454184
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Jichi Medical School |
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Principal Investigator |
NAKANO Masayasu JICHI MEDICAL SCHOOL, DEPARTMENT OF MICROBIOLOGY, PROFESSOR, 医学部, 教授 (70048958)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Shinji JICHI MEDICAL SCHOOL, DEPARTMENT OF MICROBIOLOGY, ASSISTANT, 医学部, 助手 (50195989)
MATSUURA Motohiro JICHI MEDICAL SCHOOL, DEPARTMENT OF MICROBIOLOGY, ASSOCIATE PROFESSOR, 医学部, 助教授 (20150089)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1991: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | Bacterial Infection / Bacterial Lipopolysaccharide (LPS) / Macrophages / Protein Phosphorylation / Plastin / Legionella / Salmonella / Monokine / 腫瘍壊死因子(TNF) / 緑膿菌 / マクロファ-ジ / Lーアルギニン |
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| Research Abstract |
(1) The most dominantly phosphorylated cytosolic protein, 65kDa protein (pp65), in the murine (C3H/HeN) peritoneal macrophages after stimulation with bacterial lipopolysaccharide (LPS) was isolated and purified. It was a substrate of serine kinase and possessed the amino acid sequences similar to human L-plastin. This protein was also found in the bone marrow cells, liver cells (presumably Kupffer cells) and spleen cells, while it could not be seen in the red blood cells and thymocytes. Identical protein was detected in the peritoneal macrophages of LPS-low responder C3H/HeJ mice. However, the protein in C3H/HeJ macrophages was unable to be phosphorylated by the stimulation with LPS. The posphorylation of pp65 was affected by inhibitors (IH) of protein kinase C (PKC) or calmodulin-dependent kinase, though it was not affected by IH of tyrosine kinase. PKC IH affected the expression of TNF-alpha and IL-1beta mRNA and the production their active molecules. However, calmodulin-dependent IH inhibited mRNA expression and production of IL-1, but did not inhibit those of TNF. Furthermore, the previous treatment of macrophages with nonpyrogenic lipid A analogues induced refractory states on pp65 phosporylation to LPS stimulation.
(2) Murine peritoneal macrophages that had been infected with virulent strain of Legionella pneumophila or Salmonella typhimurium exhibited the protein phosphorylation of pp76(L. p.) or pp85 and 72 (S. t.) which was specific to the infection of these organisms. Live organisms themselves or Heat-killed organisms of these strains did not cause the phosphorylation.
(3) It was demonstrated that cytokines such as TNF-alpha, TNF-beta, IL-1beta and IL-6 enhanced the killing of the organisms of Salmonella or Pseudomonas aeruginosa by the peritoneal macrophages of mice.
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