Literature Interpretation: Mechanism of Lymphocyte Infiltration in Hashimoto's Thyroiditis Revealed by Multiplex Fluorescence IHC

1. Overview of Hashimoto's Thyroiditis (HT)

Hashimoto's Thyroiditis (HT) is a highly prevalent thyroid disease, first discovered by the Japanese scholar Hakaru Hashimoto in 1912, hence the name. Despite being termed "thyroiditis", it differs from common inflammatory conditions. HT is an organ-specific autoimmune disease that develops on the basis of genetic defects and susceptibility, triggered or exacerbated by various adverse lifestyle factors, infections, and other stress responses. Pathologically, the thyroid tissue of HT patients is characterized by lymphocytic infiltration, fibrosis, interstitial atrophy, and eosinophilic changes of acinar cells, so it is also known as chronic lymphocytic thyroiditis.

In recent years, the incidence of HT has increased rapidly, with a particular predilection for females. To date, there is no definitive cure for HT, and most patients eventually progress to hypothyroidism. To develop better therapeutic strategies, exploring the pathogenesis of HT has become a focus of scientific research.

2. Core Literature Research: Key Findings and Technical Support

In February 2022, Professor Huaidong Song's research team from the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine published a research paper entitled "Lymphocyte Infiltration and Thyrocyte Destruction is Driven by Stromal and Immune Cell Components in Hashimoto’s Thyroiditis" in Nature Communications, a sub-journal of Nature. This study revealed the mechanism by which lymphocytes ectopically infiltrate thyroid tissue and destroy thyroid follicular cells in HT patients, providing new strategies for the treatment of HT. Notably, the 4-Color Multiplex Fluorescence IHC Staining Kit (Catalog No.: abs50012) from the Absin product line of ANT BIO PTE. LTD. was a key technical tool employed in this research, laying a solid foundation for the accurate observation of cell localization and interaction.

3. Detailed Research Results

3.1 Clustering Analysis of Thyroid Tissue Cells in HT Patients

The research team performed clustering analysis on the total cells of thyroid tissue from HT patients using UMAP plots, identifying different cell clusters. The results showed that infiltrating immune cells accounted for 65-82% of all cells in the thyroid tissue of HT patients. Further analysis revealed that stromal cells are closely associated with immune cell infiltration in thyroid tissue.

3.2 Spatial Distribution and Role of ACKR1+ ECs and CCL21+ Fibroblasts

Using multiplex fluorescence immunohistochemistry technology (as shown in the figures below), the research team observed that ACKR1+ ECs (EC2) are distributed in the venules of thyroid tissue in HT patients. A subset of these cells is surrounded by myofibroblasts, showing double positivity for ACTA2 and CCL21, and localized on the intima of High Endothelial Venules (HEVs) or CCL21+ fibroblasts. In the thyroid tissue of HT patients, a large number of aggregates were found around lymphocytes.

In contrast, ACKR1+ ECs and CCL21+ fibroblasts or myofibroblasts were rarely detected in the thyroid tissue of non-HT control groups. This indicates that these cell subsets may play a key role in the pathological process of HT.

3.3 Critical Role of Inflammatory Macrophages and Dendritic Cells

Another important finding of the research team was the identification of inflammatory macrophages and dendritic cells that specifically exist in the thyroid tissue of HT patients. Both cell types highly express inflammatory chemokines and cytokines, such as IL-1β. IL-1β can stimulate thyroid cells to secrete IL-6, alter the integrity of epithelial cells, and inhibit the growth of thyroid cells. Previous studies have reported that IL-1β can also induce the expression of FAS on Thyroid Follicular Cells (TFCs) and mediate TFC apoptosis through the FAS-FASL signaling pathway.

These results suggest that thyroid-specific inflammatory dendritic cells and macrophages in the thyroid tissue of HT patients may play a crucial role in thyrocyte destruction via the IL-1β signaling pathway.

4. Hypothetical Model of Stromal Cell-Mediated Immune Infiltration in HT

Based on the above research results, Professor Huaidong Song's team established a hypothetical model to illustrate the role of stromal cells in immune infiltration in HT patients: In the thyroid tissue of HT patients, blood immune cells migrate under the action of CCL19/CCL21 chemokines secreted by CCL21+ myofibroblasts and CCL21+ fibroblasts. Meanwhile, ACKR1+ endothelial cells promote the migration of immune cells by transferring CCL2 and CCL5 secreted by CCL21+ myofibroblasts/fibroblasts and immune cells into the lumen of HEVs.

Subsequently, these infiltrating immune cells are attracted to the sites of scattered CCL21+ fibroblasts. In certain regions of the thyroid, Tertiary Lymphoid Organs (TLOs) with germinal centers are formed. CCL21+ fibroblasts are distributed in the T cell zones, potentially playing roles in structural organization and T cell signal transduction.

5. Research Images

Figure 1. Multiplex fluorescence immunohistochemical staining results of HT thyroid tissues. a-b: Merge images of VWF, ACKR1, CD45, and DAPI (Scale bar: 50μm); c: Merge image of ACTA2, CD45, ACKR1, and DAPI (Scale bar: 50μm).

Figure 2. Additional multiplex fluorescence staining results. Left: Merge image of ACKR1, MECA-79, and DAPI (Scale bar: 50μm); Right: Merge image of CD36, ACTA2, ACKR1, and DAPI (Scale bar: 25μm).

Figure 3. Co-localization and RNA expression analysis. Left: Merge image of MECA-79; Right: Merge image of CCL21/MECA-79/CD45 and CCL21 (RNA) expression with ACTA2 (Scale bar: 25μm).

Figure 4. Hypothetical model of immune cell infiltration in HT. Schematic diagram showing the interaction between HEVs, myofibroblasts, fibroblasts, immune cells, and thyroid follicular cells, as well as the role of chemokines (CCL19/CCL21, CCL2, CCL5) in immune cell migration and infiltration.

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8. Disclaimer

This article is AI-compiled and interpreted based on the original work in document 1225.docx (Absin Multicolor Literature Interpretation: Hashimoto’s Thyroiditis). All intellectual property rights (such as images, data) related to the original research belong to Nature Communications and Professor Huaidong Song's research team. If there is any infringement, please contact us immediately and we will take prompt action.

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