1991 Fiscal Year Final Research Report Summary
Study on preparation of an immunomissile against mammalian lens epithelial cells
| Project/Area Number |
01570984
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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 |
Ophthalmology
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| Research Institution | Tokai University, School of Medicine |
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Principal Investigator |
OZAKI Lieko Tokai Univ. Ophthalmol. Assistant Prof., 医学部, 講師 (90135187)
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| Project Period (FY) |
1989 – 1991
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| Keywords | Lens Epithelial Cell-Membrae / Immunomissile / Monoclonal antibody / Ricin-A Chain / cell-fusion / mouse myeloma cell / Bovine Lens Epithelial cell |
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| Research Abstract |
One of the main factors of visual disturbance in the society of increasing aged people is sinile cataract. One of the main complications of an intraocular lens implantation, which has got world-wide admission now, is the postoperative proliferation of lens epithelial cells(LECs)that causes after-cataract, tilt and shift of an inserted intraocular lens and uveitis by inflammatory secretions. from LECs after extracapsular cataract extraction. The complications from the proliferated LECs decreases the vision again and increases the astigmatism. As surgical removal of all of the LECs is impossible, it is imperative to prepare the immunomissile(IM)which has the possibility to hit all of the target cells without damaging non-specific cells.
From 1989 to 1990, mouse monoclonal anti-mammalian LEC immunoglobulin G_1(XC3 series of IgG_1 : XC3-IgG_1)was prepared : Immunized mouse spleen cells(BALB/c)by bovine LECs and mouse myeloma cells(BALB/c, P3Ul-4)were fused with polyethylene glycol, cloned b
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y limiting dilution, specified by immunomicroscopy on frozen sections of both eye- and other main tissues, and classified by immunodiffusion method. From 1991 to 1992, both of XC3-IgG_1 and ricin-A chain(VEC) were purified and conjugated together using SPDP(SIGMA). IM was injected into each one of the pair lens capsules of dog during endocapsular lens extraction under general anesthesia. Mouse ascites from parental myeloma cells(P3U1-4)was used as a negative control. Postoperative study was performed by slit-lamp and light microscopically until six months. The findings revealed that our IM supressed the postoperative proliferation of LECs effectively and safely without remarkable toxic change in eye tissues.
Gene cloning to propagate a specific antigenic segment of LEC-membrane proteins using our monoclonal antibodies, and production of mono- and/or polyclonal antibodies using the propagated antigenic segment(s), are planned for getting enough of IM for the first stage of clinical use of IM. Less
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