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Immunohistochemistry of lymphocytes and macrophages in human celloidin-embedded temporal bone sections with acute
T Ganbo1, I Sando, C D Balaban
1Elizabeth McCullough Knowles Otopathology Laboratory, Department of Otolaryngology, University of Pittsburgh School of Medicine, PA 15213, USA.
Insights
This study examined immune cells in human temporal bone sections from acute suppurative otitis media. Lymphocytes infiltrated the mucosa, while macrophages were found only in the effusion, suggesting a role in otitis media pathogenesis.
Area of Science:
- Immunology
- Otolaryngology
- Pathology
Background:
- Acute suppurative otitis media is a common infection.
- Understanding immune cell distribution is crucial for otitis media pathogenesis.
Purpose of the Study:
- To investigate the distribution of lymphocytes and macrophages in human temporal bone sections from acute suppurative otitis media.
- To evaluate the utility of archival temporal bone sections for otitis media research.
Main Methods:
- Immunohistochemistry using antibodies for CD3, CD43 (T-lymphocytes), CD20 (B-lymphocytes), CD45 (leukocyte common antigen), and CD68 (macrophages).
- Antigen retrieval techniques including sodium hydroxide-methanol solution and microwave treatment.
- Avidin-biotin-peroxidase complex technique.
Main Results:
- T-lymphocytes (CD3, CD43) and B-lymphocytes (CD20) were detected in the mucosa of the eustachian tube, tympanic cavity, and mastoid air cells.
- Macrophages (CD68) were primarily found within the effusion in the mastoid air cells.
- Immune cells, particularly T-lymphocytes, showed diffuse infiltration of the lamina propria.
Conclusions:
- Archival human temporal bone sections can be effectively used for immunohistochemical studies.
- The distribution of immune cells suggests specific roles in the inflammatory process of acute suppurative otitis media.
- This approach offers a promising avenue for investigating otitis media pathogenesis.
Abstract:
Immunohistochemical analyses were used to investigate the distribution of lymphocytes and macrophages in routine human temporal bone sections obtained from a subject with acute suppurative otitis media. Primary antibodies specific for human CD3 and CD43 (T-lymphocytes), CD20 (B-lymphocytes), CD45 (leukocyte common antigen), and CD68 (macrophages) were used. As a pretreatment, the sections were soaked in antigen retrieval solution (saturated sodium hydroxide-methanol solution in methanol at a ratio of 1:3). A second antigen retrieval procedure (microwave treatment in 1% zinc sulfate) was also employed for identifying CD3-positive cells. Then the avidin-biotin-peroxidase complex technique was performed. Positive reactions to all antibodies but anti-CD68 were observed in the mucosa of the eustachian tube, tympanic cavity, and mastoid air cells. Particularly, cells positive to anti-CD3 or anti-CD43 were making a diffuse invasion upon the lamina propria. CD68-positive cells were scattered only in the effusion of mastoid air cells. These results suggest that the retrospective immunohistochemical study of archival temporal bone sections is a promising approach to investigate the pathogenesis of otitis media.
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