| Objective:The tumour is a kind of disease that threatens mankind's healthy for a long time . There is no effective means used for early diagnosis and therapy at present. In the past 20 years, with the development of basic immunology, especially the appearance of the new investigation approach, people could effectively screen the tumour antigen,which offered a new train of thought for early diagnosis of the tumour and immunological therapy. But most of the antigen was the kind of self-antigen with the defects of poor-immunogenicity and easily-caused immunotolerance. By the theory of biologic evolution ,different organisms exist syngenes,when the nature evolves from the lower grade to the higer grade .And the father is the relation between the two organisms,the stronger the immunogennicity is .we used Zheng's plant protein(ZPP)-a kind of 50 Kda glycoprotein extracted from a lower plant,between species to elevate immunogenicity and overcome self-immune tolerance to tumor cells in the body.Methods:1. when lymphocyte is stimulated by antigen ,cell's DNA will synthesize and need some precursors of NDA. 3H-TdR as a specific precusor of DNA labeled with 3H, can be used to detect cell's proliferation by isotopic method . We cultured human peripheral blood lymphocytes (PBL)with the vegetable protein and 3H-TdR for 72 hours . The celles were harvested by the aid of cell harvester and the radioactivity of lymphocyte was measured and stimulation index(SI) was calculated. By the SI,we analysed whether the vegetable prtein could specificly stimulate the lymphocyte of tumor patients to proliferation and verified its tumor antigenicity. 2.Cytotoxic T lymphocyte (CTL) is the antitumorous effect cell .It can directly injure the tumour cell which consists of the causing-allergy antigen peptide - MHC molecule . Most CTL are CD3+CD4-CD8+ cells, their ability of discerning and wounding to aim cell is restricted by MHC - I molecule .The minority are cells of CD3+CD4+CD8-, whose capacity to discern and wound to the tumour cell is restricted by MHC -II molecule . So, the expression level of CD4+ and CD8+ in the peripheral blood lymphcyte cell may has something to do with the discernment of T cell and the effective function. According to this principle, we culitivated the PBL of cancer patients with ZPP and used FTTC-CD3+,FTTC-CD4+and FTTC-CD8+monoclonlal antibody dye the PBL before culitivation and after culitivation. Flowcytometer was used to examine the expression-rate of CD3+ ,CD4+,CD8+on the surface of T cells, with the purpose of verifying the tumour antigen activity of ZPP. 3.The hyperplasia experiment of tumour cell:By cultivating the original era tumour cell of the esophagus cancer, ZPP and 3H-TdR outside the body, determining ZPP's promoting hyperplasia function to the tumour cell ,we can verify its tumour activation of antigen on the other hand.Results:1.The vegetable protein significantly stimulated esophageal(cardic) cancer patients' lymphocytes to proliferation. The SI of atypical hyperplsia patients(3.36±0.71) and of early esophageal (cardic) (3.89±0.97) and of intermediate or late cancer patients(2.64±0.53) were higer than that of normal persons(1.15±0.34)(P<0.01).2.After the situation that the esophagus cancer group -PBL were stimulated by ZPP,the percentage of　CD3+,CD8+ cell had obviously raised compared to before；CD4+'s cell's percentage did not have obvious change；The percentage of CD 3+,CD4+,CD8+ which PHA stimulated didn't have obvious change.3.ZPP made great acceleration to the proliferation of the original tumour cell of the esophagus cancer, but it didn't have stimulative effect on the normal organization -cell.ConA made no stimulative effect neither on normal organization -cell nor on tumour cell.Conclusions: The vegetable protein has characteristics of tumor common antigen and can elicit specific cellular immune response. This vegetable protein can be used clinical for tumor early diagnosis and specific active immuno-therapy. |
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