In vivo confocal microscopy qualitative investigation of the relationships between lattice corneal dystrophy deposition and corneal nerves.

BACKGROUND: To investigate the corneal neurotropic phenomenon in patients with lattice corneal dystrophy (LCD) with in vivo laser scanning confocal microscopy (IVCM). METHODS: IVCM was performed on a total of 15 patients (28 eyes) with LCD annually at a follow-up. A collection of the data was acquired to be analyzed. RESULTS: As indicated by the analysis, the LCD patients' normal corneal stromal nerves (Grade 0) presented a decline with the prolongation of the follow-ups, corresponding to a gradual increase in grade I and II involving amyloid-wrapped nerve fibers, which demonstrated that the growing amount of amyloid deposit due to the corneal nerve invasion increased slowly over time. CONCLUSIONS: The neurotropic phenomenon could increase with its severity in the corneal lesion of the patients with LCD, and also reflect the distribution of the corneal nerves, to some extent. IVCM provides a rapid, noninvasive way to observe the corneal nerves, which can be an efficient means of better understanding the development of LCD.

Transplanted corneal graft with metastatic cholangiocarcinoma to the donor eye.

OBJECTIVES: To report a case of corneal transplantation from a donor died of cholangiocarcinoma with metastasis to the eye. METHODS: A patient with limbal dermoid received corneal transplant from a donor died of cholangiocarcinoma. Pathologic examination of the remaining donor limbal tissue revealed metastasis of tumor to the limbal vessels. Prompt exchange of the graft was performed. Before the second corneal transplantation, the surrounding tissue of the recipient bed was excised and sent for histopathologic examination. RESULTS: No tumor transmission was noted surrounding the recipient bed. The second graft remained clear and the patient remained cancer free after regular examination for over a year. CONCLUSIONS: This is the first case to report that cholangiocarcinoma can metastasize to the corneal limbus. To avoid transmission of malignancies from the donor to the recipient, we suggest that donor tissue with history of malignancy should not be used for limbal allografting, and that frozen-section examination of donor limbal tissue is recommended before the transplantation.

[An experiment study of in vitro induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells to kill SO-RB(50)].

OBJECTIVE: To investigate induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells (DC) to kill retinoblastoma cells SO-RB(50). We hope to offer new approach for the treatment of patients with retinoblastoma. METHODS: DC was pulsed with RB tumor lysates in vitro and incubated with autologous lymphocytes to induce antigen specific CTL (cytotoxic T lymphocyte, CTL). SO-RB(50) cells were used as target cells and Raji cells were used as control target cells. Cytotoxicity of CTL was evaluated by MTT method (methyl thiazolyl letrazolium). The specific cytotoxicity of CTL to SO-RB(50) and Raji cells was compared. The cytotoxicity of CTL from RB and normal subjects was compared between these two groups. RESULTS: Antigen specific CTL showed greater cytotoxicity to SO-RB(50) than Raji cells, the difference was statistically significant, P < 0.01. The cytotoxicity was dose-dependent to the ratio of CTL/target cell. The nonspecific cytotoxicity to Raji cells was the same in CTL from RB patients and normal subjects, P > 0.05. The specific cytotoxicity of CTL from RB patients to SO-RB(50) was weaker than that from the healthy subjects, P < 0.01. CONCLUSION: DC pulsed with RB tumor lysate in vitro can induce antigen specific CTL which can kill the SO-RB(50) target cells specifically. This method may have potential value of therapy for the RB patients.

[Preparation of anti-human retinoblastoma monoclonal antibody by solid tumor cell immunization and study on characteristics of its antigen].

OBJECTIVE: To prepare anti-human retinoblastoma (Rb) monoclonal antibody (McAb) by immunization with solid tumor cells and preliminarily study the characteristics of its antigen. METHODS: Three Balb/C mice were immunized by intraperitoneal injection of Rb solid tumor cells acquired from enucleation. Spleen lymphocytes were separated from them and fused with myeloma cell line SP2/0. The anti-human Rb McAb was selected by Rb solid tumor cell, SO-Rb(50), SO-Rb(70), etc as antigens. The characteristics of its antigen were preliminarily studied by immunofluorescence, immunohistochemistry and Western blot. RESULTS: After repeatedly cloning them with micro-manipulation equipment, we successfully established 3 hybridoma cell lines secreting anti-human Rb McAb. Of all, 3C6 hybridoma cells could steadily secrete anti-human Rb McAb after they were frozen, resuscitated and passaged repeatedly for 2 years. Its subgroup was IgG(1). Both immunofluorescence and immunohistochemistry examinations demonstrated that the corresponding antigen of McAb 3C6 was specifically and highly expressed in Rb tumor cell membrane and cytoplasm. The other tumor tissues and human normal eye tissues were negative. Western blot analysis preliminarily demonstrated that McAb 3C6 could bind 25 000 protein band of Rb antigen. CONCLUSIONS: The anti-human Rb McAb 3C6 established by immunization of solid tumor cells is specifically and highly expressed in Rb tumor cells and has no cross-response with other tumor tissues and normal eyeballs. The antigen molecular weight bounded to this antibody is 25 000 or so, which further shows that it is a possibility for a new unidentified gene to be concerned with the tumor formation of Rb.