| Cataract is an important ocular disease that induces blindness. To date, extracapsular cataract extraction (ECCE) or phacoemulsification combined with artificial intraocular lens (IOL) implantation is considered to be the best treatment. However, posterior capsular opacification (PCO), the most common postoperative complication, may decrease the visual acuity again.Some clinical and experimental studies suggest that PCO is mainly due to the proliferation and migration of postoperative remnants of lens epithelial cells (LECs) on the posterior capsule. The etiology of PCO is incompletely known. The surgical trauma might activate LECs in a wound-healing process. Several experimental studies showed that some agents have been effective in inhibiting LECs proliferation and differentiation, and also in development of PCO, such as anti-metabolites, nonsteroid antiphlogistic drugs, cyclosporin A. But their clinical use is limited by intolerable toxic side effects to other ocular tissues. Thus, it is imperative to produce a monoclonal antibody specific for LECs as carrier of the target-directing agents designated to identify and selectively kill LECs and to prevent them from subsequent proliferation.We produced monoclonal antibody (McAb) specific for human lens epithelial membrane with hybridoma technique. The subclass of the McAb was determined by indirect enzyme-linked immunosorbent assay (ELISA) method and the specificity was confirmed both by immunohistochemical analysis and by indirect immunofluorescence techniques. We prepared large quantities of the McAb by the hybridoma cells inoculated to the peritoneal cavity of BALB/c mouse. The McAb was purified by hydroxyapatite chromatography. Cell enzyme-linkedimmunosorbent assay (CELISA) and SDS-polyacrylamide electrophoresis were used to identify the purified antibody. We evaluated its effect on LECs proliferation by MTT colorimetry and observed the relevant morphological changes of LECs with light microscopy and transmission electron microscopy. The main results are the following:1. LECs from human fetus, adult, senile cataract patients and rabbit were successfully cultured using modified explant technique in vitro and established LECs culture system. The technique is simple, easy to perform and to popularize.2. Monoclonal antibodies, HILE4 and HILE6 specific for human lens epithelial membrane were developed successfully after two intrasplenic injections of cultured human LECs. The subclass of the monoclonal antibodies is IgM. The intrasplenic immunization procedure could facilitate monoclonal antibody production in some instances in which small quantities of immunogen or limited time is available.3. These antibodies reacted specifically with the cell membrane of LECs and cross-reacted with rabbit lens epithelial membrane. No other tissue in human eyes, such as the cornea, iris, ciliary body, choroid, retina, sclera were associated.4. We prepared large quantities of HILE6 by the hybridoma cells inoculated to the peritoneal cavity of BALB/c mouse. The titer is 1:8000. The mouse ascitic fluid was purified by hydroxyapatite chromatography twice and the purity was up to mass fraction 90%. Hydroxyapatite chromatography is a simple, rapid and efficient procedure for the purification of monoclonal IgM antibody.5. HILE6 at appropriate doses possessed prohibitive effects on the proliferation of LECs and induced the morphological changes of lens epithelium, even to death.We successfully prepared monoclonal antibody against human lens epithelial membrane, which can serve as a carrier of the target-directing agents designated to actively identify and selectively kill LECs and to prevent them from subsequent proliferation. The agents maybe more efficient in prevention and treatment of PCO.
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