Abstract: Objective To investigate the anti-tumor effect of sequential injection of heterogeneic lymphocyte (HL) and autogeneic lymphocyte (AL). Methods Inactivated heterogeneic lymphocyte (IHL) was isolated from spleen of C57BL/6 mice and inactivated with mitomycin. Hepa1-6 cells were inoculated into liver tissue of CB6F1 mice. Fibrin glue (FG) was prepared with cryoprecipitate collected from mouse plasma using freeze-thaw method. The experimental treatment consisted of two stages. At first stage (13 days), tumor-bearing mice were injected with FG-IHL in liver as experiment group, and injected with FG-phosphate buffer saline (FG-PBS) in liver as control group. Immunological indices such as tumor cell killing rate of the spleen lymphocytes and number of lymphocytes, CD8⁺T and NK in the two groups were detected, respectively. At the later stage (8 days), FG-AL was prepared from randomly selected tumor-bearing mice in experiment and control group, which was injected into the liver of rest tumor-bearing mice in the same group, respectively. After 8-day treatment, tumor size and inhibition rate were compared between the two groups. Results The tumor cell killing rate of AL in experiment group was significantly higher than that in control group (25.7±4.81% vs 5.9±1.64,P<0.01). Numbers of spleen lymphocyte, CD8⁺T and NK cell in experiment group were significantly higher than those in control group, respectively (all P<0.05). After 2-stage treatment, mean volume of tumor in experiment group was significantly smaller than that in control group (1.15±0.25 cm³ vs. 1.91±0.37 cm³, P<0.01). Tumor inhibiting rate in experiment group was 39.8%. Conclusion The treatment of sequential injections with FG-IHL followed by the FG-AL in situ could suppress the growth of transplanted tumor in mice, which might become a promising biotherapy to treat cancers.

Key words: Lymphocyte, Injection, Hepatocarcinoma, Fibrin glue