A Study For Immunological Mechanisms Of Human Anaplastic Large Cell Lymphoma On BALB/C Mice Tumor Model

Objective: To build animal tumor model of anaplastic large cell lymphoma andinvestigate the pathogenesis of humoral immunology of tumor on BALB/c mice withimmune ability.Methods:1. Human anaplastic large cell lymphoma cell line Karpas-299were cultivated.2. The BALB/c mice were randomly divided into control group, cyclophosphamide groupand transplantation group. The cyclophosphamide group and transplantation groupBALB/c mice were pretreated by injecting with cyclophosphamide (250mg/kg/only).3. Toconstruct ALCL mouse tumor model, the cultivated tumor cell were implanted into theBALB/c mice with the subcutaneous injection after being pretreated3days later.According to tumor formation in body, the transplanted tumor were divided into tumorgroup and unpaired tumor group.4. After implantation24days, Routine HE staining andimmunohistochemical staining of tumor tissue pathological slices of each group wereperformed, Peripheral blood was collected to detect IL-2、IL-4、IgM、TNF-β、TGF-β、sCD30by ELISA.Results:1. After pretreatment with cyclophosphamide, the survival rate of mice were100%, and the the overall tumor rate was80%of the transplanted mouse.2.By HEstaining,tumor tissues showed characterization of malignant lymphoma: morphologicaldiversification, larger, abundant cytoplasm, prominent nucleoli light staining, eccentric,horseshoe-shaped, curled irregular nuclear mitotic visible. Necrosis of part tumor tissue.The IHC staining results for anaplastic large cell lymphoma: CD30, ALK is positive, CD20is negative.3. ELISA test results:(1).Compared with the control group: incyclophosphamide group, only TNF-beta were decreased (P<0.05). In mice without tumorgroup, IL-2, TNF-beta, sCD30, IgM content increased (P<0.05), IL-4, TGF-beta showedno difference (P>0.05). In tumor group, only cytokines IL-2were decreased (P<0.05);(2).Compared with CTX24group: In mice without tumor group, cytokines IL-2and TNF-betacontent increased (P<0.05), IL-4, IgM, TGF-beta, sCD30content showed no evidentdifferences(P>0.05); In tumor group, cytokines IL-2were decreased (P<0.05), otherindicators significantly increased (P<0.05);(3). Compared with transplantation groupwithout tumor: in tumor group cytokines IL-2showed decreased (P<0.05), other indicators significantly increased (P<0.05).Conclusion:1.We successfully construct the anaplastic large cell lymphoma BALB/cmice tumor models in mice with normal immune function.2. After pretreatment BALB/cmice, the cyclophosphamide play an important role in inducing immune tolerance, which isone of the foundational condition of tumor development.3. Through analyzing the changeof cytokines level in each mice animal model group, it is likely that, in the process ofbuilding a tumor, the ALCL tumor cells may improve the expression of immune inhibitingcytokines TNF-beta, sCD30, IL-4, TGF-beta, and reduced the level of immune cytokineIL-2in body, inhibiting the body’s anti-tumor effect, evading immune attack, in order topromote tumor growth and invasion.