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Establishment of Secreting Idiotype Immunogloblin BALB/c mouse B Cell Lymphoma Model
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Abstract:
Objective: To establish the secreting idiotype immunogloblin (Id) BALB/c mouse B cell lymphoma animal model for further study of Id-dendritic cells (DC) vaccines. Methods: BALB/c mouse B cell lymphoma cell lines were obtained by fusing SP2/0 cells with BALB/c mouse spleen cells after immunization with pristane and incomplete Freund s adjuvant (IFA) respectively. Enzyme-linked immunosorbent assay (ELISA) and flow cytometry (FCM) were used to screen the secreting Id clones and analysis the phenotypes. Both mice ascites induction and serum-free culture medium X-VIVO 20 were used to produce the Id of SB4 and SB5. By using euglobulin precipitation and gel filtration on Suphecray-200 to purify the Id. To assess the ability of tumorigenicity, （4～5）×10 5 SB4/5 cells were injected subcutaneously in the right axillary or into abdominal cavity of 6~8 week BALB/c mice. DC were induced from bone marrow progenitor cells of B cell lymphoma bearing mice with recombinant murine GM-CSF and IL-4. Results: Two secreting none auto-reactive Id malignant B cell lymphoma cell lines, named as SB4 and SB5, were screened. Chromosomes study, DNA index, cell cycle proliferative indexes, cell generation time and mitotic index analyses all demonstrated SB4 and SB5 possessing the characteristics of malignant tumor cell lines. Both Id proteins belonged to IgM/κ. SB4 and SB5 cells grew progressively in syngeneic mice. Tumors could be touched in all mice and the average life-span was 30± 4 days and typical DC could be induced. Conclusion: BALB/c mouse B cell lymphoma SB4 and SB5 cell lines created by cell fusion techniques could secrete non-auto-reactive Id and had the characteristics of B cell lymphoma cells.
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