1989 Fiscal Year Final Research Report Summary
Gene cloning for vaccine in Pneumocystis carinii pneumonia
| Project/Area Number |
63480208
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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 |
Respiratory organ internal medicine
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| Research Institution | Kagoshima University (1989)
The University of Tokyo (1988) |
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Principal Investigator |
TANABE Kiyokatsu The Faculty of Medicine, Kagoshima University. Associate prof., 医学部, 助教授 (80134625)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Yoshikazu The Institute of Medical Science, University of Tokyo. Associate prof., 医科学研究所, 助教授 (40114590)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Pneumocystis Carinii Pneumonia / Gene Cloning / Recombinant DNA / Vaccine / Monoclonal Antibody / Opportunistic Infection / Amino-acid Sequence |
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| Research Abstract |
Pneumocystis carinii is a pathogen which causes fatal pneumonia in patients with acquired immune deficiency syndrome. To facilitate the basic study of P. carinn, we analyzed the major surface proteins by immunochemical and bioclinical methods. The major protein components of both cysts (resting form) and trophozoites (vegetative form) are part of a group of proteins called P115 with apparent masses of 105 to 120 kilodiltons.
They represent an unusually large portion of the total proteins of this organism. The purified proteins exhibited six isoelectric variants when analyzed by two-dimensional gel electrophoresis. A monoclonal antibody raised against cysts recognized all six variants and reacted with epitopes that were located in the cell wall, thereby indicating that P115 is an immunoreactive surface component. Data are presented that the isoelectric variants contain identical or closely related protein components and that they are mamiose-ricn glycoproteins.
Deglycosylated P115 migrates primarily as a single more acidic protein in two-dimensional gels, suggesting that the isoelectric variants may be due primarily to differences in glycosylation. The majority of sera tested from humans with diagnosed pneuinocystosis reacted strongly with the P115 proteins.
The cytoplasmic 5S ribosomal RNA (5S rRNA) sequence from Ptietimocystis carinn was determined. A sequence comparison matrix of 382 eukaryote 5S rRNA sequences and an evolutionary tree were constructed to establish the phylogenetic position of Ptieumocystis. The data suggest that Pneumocystis is associated with the Rhizopoda/ Myxomycota/ Zygomycota group (= 'Protista fungi') but not with common fungi, such as Ascomycota or Basidiomycota, nor with other protozoa.
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[Publications] Tanabe, K., Takasaki, S., Watanabe, J., Kobata, A., Egawa, K., & Nakamura, Y.: "Glycoproteins composed of major surface immuno determinants of Pneumocystis carinii." Inf. Immu., 57:1363-1368, 1989.
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