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Published on: November 16, 2015
Distribution of lectin receptors in the human hyperplastic tonsil: histochemical and flow cytometric analyses
A B Sarker1, T Akagi, T Yoshino
1Second Department of Pathology, Okayama University Medical School, Japan.
Insights
This study maps lectin receptors on human tonsil cells using immunohistochemistry and flow cytometry. Lectin binding varied by cell type and fixative, with Lotus receptors found exclusively on B-lymphocytes.
Area of Science:
- Immunology
- Histology
- Cell Biology
Background:
- Lectin receptors play crucial roles in cellular recognition and function.
- Understanding lectin distribution in lymphoid tissues like the human tonsil is vital for immunological research.
Purpose of the Study:
- To investigate the distribution and expression patterns of various lectin receptors on human tonsil tissue components.
- To evaluate the impact of different tissue fixation methods on lectin binding affinity.
Main Methods:
- Utilized 16 biotinylated lectins for immunohistochemical analysis on frozen and paraffin-embedded human tonsil sections.
- Employed the avidin-biotin-peroxidase complex (ABC) method for staining.
- Performed dual-color flow cytometry on cell suspensions using fluorescein isothiocyanate-conjugated lectins and phycoerythrin-labeled antibodies.
Main Results:
- Lectin binding intensity was significantly dependent on the fixative used, with ethanol-fixed sections yielding optimal results.
- Con-A, RCA-1, LcH, WGA, MPA, PHA, PSA, PNA, SJA, and GSA-1 showed broad reactivity across tonsil components.
- Lotus and BPA binding was specific to germinal center lymphocytes, while SBA and DBA reacted with macrophages/histiocytes, vascular endothelium, and epithelial cells. Lotus receptors were exclusively on B-lymphocytes.
- Flow cytometry confirmed widespread lymphocyte surface staining for most lectins, with Lotus receptors identified on B-lymphocytes.
Conclusions:
- Fixation methods critically influence lectin receptor detection in human tonsil tissue.
- Specific lectin-receptor interactions provide markers for distinct cell populations within the human tonsil, particularly B-lymphocytes.
Abstract:
The distribution of lectin receptors in the human tonsil was studied using 16 biotinylated lectins. The avidin-biotin-peroxidase complex (ABC) method was used on frozen and paraffin-embedded tissue sections. Cell suspensions were also analysed by dual flow cytometry using respective fluorescein isothiocyanate-conjugated lectins and phycoerythrin-labeled anti-CD3 and anti-human immunoglobulin. Frozen sections fixed with acetone and paraffin-embedded materials fixed in three solutions were compared for lectin affinity; ethanol-fixed sections gave best results followed by frozen and buffered formalin-fixed ones, then nonbuffered formalin. Con-A, RCA-1, LcH, WGA, MPA, PHA, PSA, PNA, SJA and GSA-1 reacted with all tissue components of the tonsil in immunohistochemical studies, but binding intensity was fixative dependent. Binding of Lotus and BPA to lymphocytes was limited to germinal center lymphocytes. Other tissue components were also reactive but staining intensity was weaker in Lotus compared with BPA. SBA and DBA did not react with lymphocytes, but reacted with macrophages/histiocytes, vascular endothelia, and epithelial cells. LBA and LPA were constantly negative with all tissue components irrespective of fixatives. Flow cytometric analyses showed that all but three (DBA, LBA and LPA) partially or totally stained lymphocyte surfaces. Lotus receptors were expressed exclusively on B-lymphocytes.
