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Monoclonal antibodies reactive with chicken peritoneal macrophages: identification of macrophage heterogeneity
Abstract:
Mouse monoclonal antibodies (MCAs) were prepared against chicken inflammatory macrophages for the purpose of analyzing macrophage heterogeneity. Macrophage-rich adherent peritoneal exudate cells harvested from Cornell K-strain chickens 42 hr after stimulation with Sephadex were used as immunogens in the production of the monoclonal antibodies. Eight hybridoma clones producing antibodies reactive with chicken peritoneal macrophages were subjected to characterization. While six of the monoclonal antibodies cross-reacted with various hematopoietic cell types, two MCAs (CMTD-1 and -2) were restricted in reactivity to macrophages. CMTD-1 was found to react with activated peritoneal macrophages generated by several irritants. In contrast, CMTD-2 identified a subpopulation of macrophages elicited by specific carbohydrate-based stimulants. This presumably was based on the cross-reaction of this MCA with specific carbohydrate linkages. Analysis using flow cytometry revealed the time-dependent appearance of CMTD-2-positive peritoneal macrophages between 24 and 52 hr after ip Sephadex injection. This subpopulation of peritoneal macrophages was found to be heterogeneous for the ability to undergo in vitro phagocytosis of sheep erythrocytes. CMTD-2-positive cells were also detected in the thyroids of 2-week-old Obese strain chickens with spontaneous autoimmune thyroiditis and at a low incidence in the spleens of normal chickens.
Insights
Researchers developed mouse monoclonal antibodies (MCAs) to study chicken macrophage heterogeneity. Two MCAs, CMTD-1 and CMTD-2, specifically identified macrophage subsets, revealing distinct cellular responses and presence in autoimmune conditions.
Area of Science:
- Immunology
- Cell Biology
- Veterinary Science
Background:
- Macrophage heterogeneity plays a crucial role in immune responses.
- Characterizing specific macrophage subsets in avian species is essential for understanding immune mechanisms.
- Existing tools for analyzing chicken macrophages were limited.
Purpose of the Study:
- To develop novel mouse monoclonal antibodies (MCAs) for identifying chicken macrophage subsets.
- To analyze macrophage heterogeneity in chicken peritoneal exudate cells.
- To investigate the distribution and characteristics of distinct macrophage populations.
Main Methods:
- Production of hybridoma clones secreting MCAs against chicken inflammatory macrophages.
- Characterization of MCA reactivity using flow cytometry and immunofluorescence.
- Analysis of macrophage populations elicited by various stimulants (Sephadex, other irritants).
Main Results:
- Two MCAs, CMTD-1 and CMTD-2, demonstrated specific reactivity to chicken macrophages.
- CMTD-1 recognized activated peritoneal macrophages induced by diverse irritants.
- CMTD-2 identified a distinct macrophage subpopulation elicited by carbohydrate-based stimulants, potentially via specific carbohydrate linkages.
- CMTD-2-positive macrophages showed heterogeneous phagocytic activity and were found in thyroids of chickens with autoimmune thyroiditis.
Conclusions:
- CMTD-1 and CMTD-2 are valuable tools for dissecting chicken macrophage heterogeneity.
- CMTD-2 identifies a specific macrophage subset involved in immune responses to certain stimulants and potentially linked to autoimmune conditions.
- Further research can utilize these MCAs to explore macrophage functions in health and disease in chickens.