| Three nasopharyngeal carcinoma(NPC) transplantable tumors and two cell lines were established and characterized to provide in vivo and in vitro system for the study of EBV specific cellular immunity and it's application in treatment of nasopharyngeal carcinoma. Part IBALB/C nude mice and scid mice were used as transplantation host. Tumor tissues were obtained by forceps from nasopharyngeal tumors of patients and transplanted to subcutaneous in axilla or back of mices with punture trocar. Twenty six cases of transplantation were performed, of which 14 in nude mices(16 mices. 21 sites) and 12 in scid mices(13 mices, 14 sites). The cell lines were derived from the established transplantable tumors. The tissues were cultured in 199 medium with 10% calf serum in 50ml culture bottle after being minced into small cluster less than 1mm in diameter.Characteristics including the transplanting efficiency, growth rate, gross appearance, morphosis under optic micrcope and electron microscope and karyotypes chromosome were investigated. Detection of LMP-1 and LMP-2 were performed for the transplantable tumors and cell lines with immunohistochemical and immunofluorescence method.Results: Three transplantable tumor named as CSNET-1, CSNET-2, CSNET-3 were successfully established from 26 specimens of human nasopharyngeal carcinoma. Both of CSNET-2 and CSNET-3 were generated and passes in scid mice. The CSNET-1 was primarily generated in nude mice and than translated to scid mice after 3 passages. To date the 4 CSNET-1, CSNET-2, CSNET-3 have been passed to 11th, 14th and 9th generation, respectively. The overall success rate of transplantation were 91%(39/43), 97%(29/30) and 94%(34/36); The medium time of latent growth were 23, 38 and 20 days; and longest persistence of tumor in vivo were 93, 38 and 44 days respectively for the above tumors. Two cell lines derived from CSNET-1 (9th generation) and CSNET-2(4th generation) named as CSNE-1 and CSNE-2 have been successfully passed to 26th and 40th generations in vitro for 12 and 30 months. The karyotypes of all transplantable tumors displayed as typical human origin with hyperdiploid chromosomal and multiple structure aberration. Histologically, the tumors and cell lines posses the characteristics of poor differentiated squamous carcinoma under either microscope or electron microscope. Some particles mimic mature EBV were found in CSNET-1 under electron microscope.LMP-1 was detectable in CSNET-1, CSNET-2, CSNET-3, CSNE-1 and CSNE-2 by immunohistochemical(envisfon method) test. LMP-2 assay showed positive results in CSNET-1, CSNET-2 and CSNET-3 with immunohistochemical method (ABC), and in CSNE-2 with immunofluorescence method. Part IIT cell subgroups of peripheral blood were tested and compared for 25 healthy donors and 58 patients of NPC. Indirect erythrocyte wreath method was used for the test.Responses of T cell to LMP-2 peptides were studied with ELISPOT for 18 healthy donors, 17 patients of NPC and 2 patients with other cancers. The peptides used were CLG, LLW, FLY, LLS and LTA.Results: The quantity of CD3+cell was statistically less in NPC patients than in healthy donors (51.8±4.3 vs 58.3±3.0, t=6.81,p=0.00). In addition, the ratio of CD4+ cell to CD8+ cell was significant less in NPC patients than in healthy donors (1.13±0.24 vs 1.41±0.18,t=5.28,p=0.00).In the study of ELISPOT, 16 cases (13 of healthy donors and 3 of NPC patients)showed positive response to all peptides, 14 cases (1 of healthy donors, 11 of NPC patients and 2 of other cancer patients) showed negative response to all peptides, 7 cases showed positive response to partial peptides. The positive response rate was significantly higher in healthy donors than in NPC patients ( x 2=14.7, P<0.01) Conclusions:The above results demonstrate that the transplantable tumors CSNET-1, CSNET-2, CSNET-3 and the cell line CSNE-1, CSNE-2 of NPC are human origin and are able to pass steadily...
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