| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1986: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1985: ¥800,000 (Direct Cost: ¥800,000)
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| Research Abstract |
The brain may be an immunologically privileged site, in that it lacks lymphatic drainage and is shielded from the immune system by the blood-brain barrier. However, intense immune reactions, e.g., against viruses, can take place locally in the central nervous system (CNS). These seemingly contradictory facts are evidence for adaptive immunologic elements in the CNS. Recently, it has been shown that murine brain cells express Ia antigens, and that several established human glioma cell lines express HLA-DR antigens. Unlike the widely distributed classical transplantation antigens mapping in the K and D regions, surface Ia antigens have limited tissue distribution and are found predominantly on cells involved in immune responses. Ia-positive cells from both lymphoid and nonlymphoid tissues are associated with immune function, and the presence of Ia antigens might be a marker for immune competency. Therefore, we reasoned that the identification of Ia-bearing cells in the brain may lead to
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the elucidation of immune mechanisms within the brain.
Bone marrow suspensions were obtained by flushing femoral shafts of young B6C3 <F_1> mice (H-2b/k) with Hepes-buffered Eagle's (HE) medium. 1 x 108 bone marrow cells were reacted with appropriately diluted anti-thy 1.2 antibody for 30 minutes at <4^o> C. And then these cells were incubated with rabbit's complement for 45 minutes at room temperature, in order to remove donor's T cells. These cells were washed and <10^7> cells were injected intravenously into retire (6-10 month of age) C57BL/6 mice that had received 950 rads of whole body irradiation 1-2 hr prior to intravenous injection. After about 6 months, the recipients' spleens were removed and sigle suspensions were prepared from these spleens. These spleen cells were reacted with mouse monoclonal anti- <Ia^k> antibody and H- <2^(b/k)> phenotype of spleen cells were analyzed with a fluorescence-activated cell sorter (FACS IV). About 6 months after transplantation of the bone marrow, one-third (30 mice) of the recipients were alive and they were all chimera ( <F_1> into parent) mice. Then, all brains of chimera mice were placed in 4% paraform. The brains were sectioned and stained with anti-Iak antibody. However, <Ia^k> phenotype-bearing cells were unable to detect by immunofluorescence staining. Less
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