The function analysis of the apoptosis of pancreatic β-cells in autoimmune (Type I)diabetes using microdissection methods
| Project/Area Number |
10670439
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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 |
内科学一般
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| Research Institution | Fukuoka University |
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Principal Investigator |
ANZAI Keizo Fukuoka Univ., School of Medicine. Assist. Prof., 医学部, 助手 (60258556)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUMIYA Junji Fukuoka Univ., Hospital, Instructor, 病院・講師 (70206556)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Type I diabetes / NOD mice / Apoptosis |
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| Research Abstract |
We produced frozen sections from the pancreas of non-obese diabetic (NOD) mice. which used as model animal of autoimmune diabetes (type I). The apoptotic cells were detected within islets of non-diabetic NOD mice by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) methods (Topics of the internal medicine advance, l998).
The micodissection methods using micromanupulator was adopted in order to analyze mechanism of pathogenesis of the apoptosis, and either pancreatic β cells or infiltrating lymphocytes around and within islets from pancreas section were cut off and were collected. and mRNA was extracted.
In addition, We analyzed gene expression in either islets or lymphocytes, for example IL-4. IFN-γ. FasL, Fas. perforin, granzyme. TNF-receptor family, iNOS and Bcl-2 family associated with apoptosis. by RT-PCR methods. By arranging result at present the contribution preparation is being carried out.
We measured the serum soluble Fas and FasL in the human NK leukemia patient by ELISA methods (British J Haematology. l998). The NOD mice is also being examined the serum soluble Fas and FasL used the similar measuring method.
In order to clarify the difference in population of infiltrated lymphocytes and in the ratio of CD40. CD40L, CD28, CD86 positive cells between diabetic NOD mice and non-diabetic NOD mice (40 wk-old and over), islets with insulitis were immunohistochemically examined for subsets of infiltrated lymphocytes. We found no difference in between the two groups.
In addition, in order to clarify the difference in cytokine and TCR repertoire between B cell deficient NOD mice or B cell deficient streptozocin induced diabetic AKR/J mice and control littermates with B cells. we studied the cytokine and TCR repertoireby infiltrated T cells into islets in both groups by RT-PCR from islets (International Immunology, in press).
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Report
(3 results)
Research Products
(12 results)
