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Distribution of dendritic cells expressing dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN, CD209):
Akihiro Masuda1, Toshio Nishikawa2
1Department of Surgical Pathology, Tokyo Women's Medical University, Tokyo, Japan. masuda@research.twmu.ac.jp.
Insights
Dendritic cells (DCs) expressing DC-specific ICAM-3-grabbing non-integrin (DC-SIGN) interact with various immune cells, including B cells, in lymphoid tissues. This study introduces a novel multiple immunohistochemistry method for detailed analysis of these cellular interactions.
Area of Science:
- Immunology
- Cell Biology
- Histology
Background:
- Dendritic cells (DCs) play a crucial role in initiating immune responses.
- DC-specific ICAM-3-grabbing non-integrin (DC-SIGN) is a key molecule expressed on certain dendritic cells.
- Understanding the interactions of DC-SIGN⁺ DCs with other immune cells is vital for deciphering immune regulation.
Purpose of the Study:
- To investigate the distribution and morphological interactions of DC-SIGN⁺ DCs in human tonsillar and lymphoid tissues.
- To evaluate a novel multiple immunohistochemistry (IHC) technique for analyzing complex cellular interactions in formalin-fixed paraffin-embedded sections.
Main Methods:
- Multiple immunohistochemistry (IHC) was employed on formalin-fixed paraffin-embedded tissue sections.
- The graphics editing program Photoshop was utilized to enable multi-staining with four or more antibodies.
- Conventional IHC with diaminobenzidine color development was repeated and images were processed for multi-staining.
Main Results:
- DC-SIGN⁺ DCs were predominantly found around lymphoid follicles, forming distinct DC-SIGN⁺ DC-rich areas.
- These DCs exhibited direct contact with T cells, fascin⁺ DCs, blood vessels, and multiple B cell subsets, including naive and memory B cells.
- The multiple IHC method proved effective for detailed analysis of complex tissue structures.
Conclusions:
- DC-SIGN⁺ DCs are strategically positioned to interact with a broad range of immune cells, including B cells, suggesting a significant role in regulating both T cell and B cell-mediated immune responses.
- The developed multiple IHC method is a simple, versatile, and valuable tool for analyzing intricate cellular interactions in various archival specimens.
Abstract:
The distribution of dendritic cells (DCs) expressing DC-specific ICAM-3-grabbing non-integrin (DC-SIGN, CD209) and the morphological interaction of DC-SIGN⁺ DCs with other cells, especially B cells, in tonsillar and other lymphoid tissues were investigated by multiple immunohistochemistry (IHC) using the graphics editing program Photoshop, which enabled staining with 4 or more antibodies in formalin-fixed paraffin sections. Images obtained by repetition of conventional IHC using diaminobenzidine color development in a tissue section were processed on Photoshop for multiple staining. DC-SIGN⁺ DCs were present in the area around the lymphoid follicles and formed a DC-SIGN⁺ DC-rich area, and these cells contacted not only T cells, fascin⁺ DCs, and blood vessels but also several subsets of B cells simultaneously, including naïve and memory B cells. DC-SIGN⁺ DCs may play an important role in the regulation of the immune response mediated by not only T cells but also B cells. The multiple IHC method introduced in the present study is a simple and useful method for analyzing details of complex structures. Because this method can be applied to routinely processed paraffin sections with conventional IHC with diaminobenzidine, it can be applied to a wide variety of archival specimens.
