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Development and characterization of macrophage hybridomas derived from murine peritoneal exudate cells
T Ashikaga1, Z Wang, M Yamamoto
1Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Abstract:
Mouse macrophage (Mphi) hybridoma clones were generated by somatic cell hybridization of myeloma X63 cells (H-2d) with C57BL/6 (H-2b) peritoneal exudate cells elicited with a streptococcal preparation, OK432, or thioglycollate medium. Although they hardly adhered to plastic dishes and could not be morphologically distinguished from parental X63 tumor cells, the clones retained Mphi characteristics. These included phagocytosis and production of lysozyme and nonspecific esterase, suggesting that they were hybridomas derived from Mphi. Some of them expressed various levels of Ia antigen and Fc receptor. Because they induced proliferation of T cells from Balb/c mice but not those from C57BL/6 mice, the Ia antigen of Mphi hybridoma was assumed to be derived from peritoneal Mphi. The level of proliferation induction was correlated to the level of Ia antigen expression. Several clones produced a factor that cytostatically inhibited growth of murine mammary carcinoma and was serologically identified with arginine deiminase.
Insights
Researchers created mouse macrophage (Mphi) hybridomas that retain Mphi functions and express Ia antigen. These hybridomas produced a factor inhibiting tumor growth, identified as arginine deiminase.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Hybridoma technology is crucial for producing monoclonal antibodies and studying cell function.
- Macrophage (Mphi) hybridomas offer a unique model for investigating Mphi biology and interactions.
- Understanding Mphi characteristics is vital for immunology and cancer research.
Purpose of the Study:
- To generate and characterize novel mouse macrophage (Mphi) hybridoma clones.
- To investigate the functional properties and antigen expression of these Mphi hybridomas.
- To explore the potential of Mphi hybridomas in producing anti-tumor factors.
Main Methods:
- Somatic cell hybridization of myeloma X63 cells with C57BL/6 peritoneal exudate cells.
- Characterization of hybridoma clones for Mphi-specific functions (phagocytosis, enzyme production).
- Analysis of Ia antigen and Fc receptor expression, and T cell proliferation assays.
Main Results:
- Successfully generated Mphi hybridoma clones retaining Mphi characteristics like phagocytosis and enzyme production.
- Observed expression of Ia antigen and Fc receptors on some hybridoma clones.
- Demonstrated that Ia antigen expression correlated with T cell proliferation induction.
- Identified a tumor growth-inhibiting factor produced by several clones, identified as arginine deiminase.
Conclusions:
- Mouse Mphi hybridomas can be successfully generated and retain key Mphi functions.
- Ia antigen expression on Mphi hybridomas is functional and influences T cell responses.
- These Mphi hybridomas represent a valuable tool for immunological studies and may yield novel anti-cancer agents.