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Detection of guinea pig macrophages by a new CD68 monoclonal antibody, PM-1K
Tomohiro Horikawa1, Yoshihiro Komohara, Emi Kiyota
1Department of Cell Pathology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
Abstract:
A new monoclonal antibody, PM-1K, was raised against 24-h cultured human peritoneal macrophages. In immunohistochemical assays, PM-1K recognized freshly isolated blood monocytes and most tissue macrophages as well as myeloid dendritic cells such as Langerhans cells and interdigitating cells. The molecular size of the antigen recognized by PM-1K was determined to be 110 kD by means of immunoaffinity purification. Because this affinity-purified antigen recognized by PM-1K was also recognized by anti-CD68 antibodies, it is believed to be one of the heterogeneous molecules of the CD68 antigen. Analysis showed interspecies reactivity of PM-1K with macrophages from guinea pigs, pigs, bovine species, and monkeys. Among these macrophages, those of the guinea pig reacted strongly with PM-1K. Patterns of PM-1K immunostaining in guinea pig tissues were similar to those found in human tissues. Studies with the immunoelectron microscope revealed reaction products of PM-1K in the cytoplasm, especially around endosomes. Since only a few antibodies are available to label guinea pig macrophages, PM-1K is considered to be one of the most suitable antibodies to examine macrophages in experimental guinea pig models.
Insights
A novel monoclonal antibody, PM-1K, targets the CD68 antigen on human and animal macrophages and myeloid dendritic cells. This antibody is valuable for studying macrophages in experimental models, particularly in guinea pigs.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophages and myeloid dendritic cells are crucial immune cells.
- Specific antibodies are needed to study these cells in various species.
- The CD68 antigen is a marker found on macrophages.
Purpose of the Study:
- To characterize a new monoclonal antibody, PM-1K.
- To determine the reactivity and specificity of PM-1K.
- To evaluate PM-1K's utility in studying macrophages, especially in guinea pigs.
Main Methods:
- Monoclonal antibody production against human peritoneal macrophages.
- Immunohistochemical and immunoelectron microscopic analyses.
- Immunoaffinity purification and molecular size determination.
- Interspecies reactivity testing.
Main Results:
- PM-1K recognizes human monocytes, tissue macrophages, and myeloid dendritic cells.
- The antigen recognized by PM-1K is an 110 kD molecule, identified as part of the CD68 antigen.
- PM-1K shows reactivity with macrophages from guinea pigs, pigs, cattle, and monkeys, with strong reactivity in guinea pigs.
- Immunostaining patterns in guinea pig tissues mirror human patterns.
Conclusions:
- PM-1K is a specific antibody for the CD68 antigen.
- PM-1K is a valuable tool for studying macrophages and myeloid dendritic cells across species.
- PM-1K is particularly useful for research involving guinea pig macrophages due to limited available antibodies.
