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First published online November 24, 2003
Journal of Experimental Biology 207, 55-65 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00710

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Characterization of a novel set of resident intrathyroidal bone marrow-derived hematopoietic cells: potential for immune-endocrine interactions in thyroid homeostasis

John R. Klein^* and Heuy-Ching Wang

Department of Basic Sciences, Dental Branch, University of Texas Health Science Center at Houston, Houston, TX 77030, USA

View larger version (80K): [in a new window]   Fig. 1. Expression of CD11b and CD11c in normal mouse thyroid. Thyroid tissues from normal healthy mice have significantly more CD11b+ than CD11c+ cells as determined by the reactivity of PE-anti-CD11b and PE-anti-CD11c mAbs. Reactivity of PE-labeled control antibody to mouse thyroid tissue sections is shown, and the location of some thyroid follicles are indicated (f). Note the high density of CD11b+ cells with typical dendritic cell morphology in four sections from the same thyroid, and the sparse number of CD11c+ cells in that tissue. Sections are from the thyroid from one BALB/c mouse, but were typical of four mice examined.

View larger version (123K): [in a new window]   Fig. 2. Expression of dendritic cell markers within the thyroid. Thyroid tissues from normal healthy mice have many widely dispersed CD11b+ cells in areas near thyroid epithelial cells and follicles, but lack cells expressing F4/80, CD40, CD80, CD19, CD3, CD8 and Ly-6G. Tissues are from the same animal, but are typical of the results from two mice.

View larger version (140K): [in a new window]   Fig. 3. Both (A) CD11b+ and (B) CD80+ cells are present in the spleen of 3A9 transgenic mice 48 h after i.p. injection with hen egg lysozyme (HEL) (GC, germinal centers). (C) CD80+ cells are not present in the thyroid of HEL-injected 3A9 mice despite the presence of (D) CD11b+ cells in that tissue. Both (E) CD11b+ and (F) CD80+ cells are present in the thyroid pericapsular lymph nodes of HEL-injected mice.

View larger version (103K): [in a new window]   Fig. 4. Presence of EGFP+ cells in the thyroid of EGFP+ EGFP– bone marrow radiation chimeras. Unstained thyroid tissue sections from EGFP+ EGFP– bone marrow radiation chimeras have intrathyroidal cells with endogenous green fluorescence. Note the presence of EGFP+ cells surrounding thyroid follicles from 1–20 weeks post-bone marrow transfer. Magnification 400x.

View larger version (99K): [in a new window]   Fig. 5. EGFP+ cells are abundant in the thyroid and the mesenteric lymph node, but not in the kidney or the liver, of EGFP+ EGFP– bone marrow radiation chimeras 2 weeks post-bone marrow transfer.

View larger version (114K): [in a new window]   Fig. 6. Presence of EGFP+ cells, CD11b+ cells and LCA+ cells in the thyroid of EGFP+ EGFP– bone marrow radiation chimeras (week 3). Thyroid tissue sections from EGFP+ EGFP– bone marrow radiation chimeras have EGFP+ cells (unstained section), and cells reactive with anti-CD11b and anti-CD45 LCA, suggesting that EGFP+ cells originate from bone marrow hematopoietic cells and not from bone marrow stromal cells. Magnification 400x.

View larger version (97K): [in a new window]   Fig. 7. Bone marrow-derived dendritic cells (DCs) generated in vitro migrate to the thyroid. Bone marrow cells from EGFP+ mice grown with 10 ng ml–1 GM-CSF express high levels of (A) CD11b, of which (B) 40% expressed CD11c. (C–F) Unstained thyroid tissue sections from non-irradiated EGFP– recipient mice 25 days post-DC transfer indicate the presence of EGFP+ intrathyroidal cells. Magnification 400x.

View larger version (100K): [in a new window]   Fig. 8. Double staining of thyroid tissue from normal BALB/c mice, demonstrating the co-expression of (A) CD11b and (B) TSHß; encircled area demarcates a region of CD11b+ cells not containing intracellular TSHß. 18 h post-i.p. injection of green fluorescent microspheres, there is an accumulation of microspheres in regions of high DC/m density in the (C) spleen, despite the absence of microspheres in the (D) thyroid of the same animals.

View larger version (36K): [in a new window]   Fig. 9. RT-PCR amplification of a 473 bp TSHß product from pituitary and thyroid tissues. The control lane consisted of PCR amplification using reagents and primers in the absence of added nucleic acids.