| Project/Area Number |
12660285
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied veterinary science
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| Research Institution | YAMAGUCHI UNIVERSITY |
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Principal Investigator |
IWATA Hiroyuki Yamaguchi University, Faculty of Agriculture, Professor, 農学部, 教授 (40193750)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Ken Yamaguchi University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (90284273)
HAYASHI Toshiharu Yamaguchi University, Faculty of Agriculture, Professor, 農学部, 教授 (90111484)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | α1-acid glycoprotein / monoclonal antibody / L-selectin / glycan chain / bovine leukemia / E-selecin / sialyl lewis X antigen / シアソルルイスX抗原 |
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| Research Abstract |
In order to clarify the changes of glycan chain structure of acute phase glycoproteins in certain diseases, bovine α 1-acid glycoprotein ( α1AGP ) was studied on the preparation of monoclonal antybody ( Mab ), cDNA cloning of L-selectin and its expression, analysis of glycan chain, and functional analysis of lymphocytes.
1. Preparation of monoclonal antibody: α1AGP was purified from bovine sera and used for immunogen to mice. Three Mabs obtained ( IgM with κ light chain ) recognized bovine α1AGP on Western blot analysis. The Mabs was not reacted with α1AGP after cleavage with glycan chain on Western blot analysis.
2. cDNA cloning of bovine L-selectin and its protein expression: Bovine L-sel cDNA was cloned and its ORF was 1110 bp which encode 370 amino acids. In bacterial expression, the recombinat protein was insoluble. In baculovirus-insect cell expression the protein was expressed as membrane proteins, which reacted with the specific antibody.
3. Analysis of glycan chain: α1AGP was resolved into two fractions with high affinity to ConA in normal health, but 4 fractions with lower affinity in addition to the two fractions in bovine leukemia. As well as E-sel, the reactivity of antibody against sialyl Lewis X to bovine α1AGP was weak, but seemed to be stronger in bovine leukemia than in normal health.
4. Functional analysis of bovine lymphocytes: Cytokine mRNA was analyzed on bovine lymphocytes. IL-2 mRNA expression in lymphocytes vary in individuals, but IL-4 was decreased, and IFN-γ was increased with individual differences.
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