| Project/Area Number |
05670230
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | NAGASAKI UNIVERSITY |
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Principal Investigator |
AOSAI Fumie Nagasaki University School of Medicine, Assistant Professor, 医学部, 講師 (80150316)
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| Co-Investigator(Kenkyū-buntansha) |
YANO Akihiko Nagasaki University School of Medicine, Professor, 医学部, 教授 (20135122)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Antigenic peptide of Toxoplasma16FA02 : T cell epitope / T.gondii-infected cell specificCTL / MHC binding peptides / HLA-A2 / HLA-A2 |
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| Research Abstract |
Naturally processed peptides derived from Toxoplasma gondii (T.gondii) -infected cells were acid extracted and detected by cytotoxic T lymphocytes (C) induced from peripheral blood lymphocytes of a patient with chronic toxoplasmosis. The CTL lines were obtained by weekly in vitro stimulation with a T.gondii-infected human B lymphoma line, ARH,which shares HLA-A2 and Cw4 determinants with the patient. The lytic activity of these CTL lines against T.gondii-infected ARH and ARH pulsed with fraction 29 of a reversed-phase high-performance liquid chromatography (RP-HPLC) extract from T.gondii-infected ARH was inhibited by an anti-HLA-A,B,C monoclonal antibody (mAb) and an anti-HLA-A2 mAb. Anti-HLA-DR mAb failed to block the lytic activity. Thus, the presentation of peptides by T.gondii-infected cells for CTL is mediated by HLA-A2 molecules. The peptides bound to HLA-A2 molecules were purified with an anti-HLA-A2 mAb-coupled immunoadsorbent column from T.gondii-infected ARH cell lysate. The peptides dissociated from the complex of HLA-A2 molecules and fractionated by RP-HPLC in fraction 29 were submitted for sequential NH_2-terminal Edman degradation. The amino acid sequence analysis of fraction 29 revealedthat a dominant leucine was found at position 2, isoleucine was found at position 3, phenylalanine was found at position 8, and methionine and phenylalanine were dominant at position 9. Thus, the amino acid sequence of the HLA-A2-bound peptide in fraction 29 was in part consistent with the predictive algorithm of HLA-A2-binding peptide motifs.
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