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Crucial Role of Dendritic Cells in the Neuro-immune Cross-talk Mechanism

Author: ZhuChunFang
Tutor: WuKaiYun
School: Suzhou University
Course: Human Anatomy and Embryology
Keywords: FR anterograde tracing Neural immune pathways Lymphoid organs Dendritic cells Rat
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 17
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Abstract


Objective To investigate the lymphoid organs of dendritic cells in the nerve - the role of the immune pathway. Experimental Methods 1, FR anterograde tracing experiment group: SD rats 3, weighing 260 g ± 20 g, male. With SD rats supine, hands touched SD rats sternum, above it along the midline of the neck cut about 3cm incision, exposing the right superior cervical ganglion, with micro syringe connected to the neck of a glass microtubules ganglion slow injection 1μl? 3% FR, leaving needle 15 minutes; suture wounds, survived nine days after perfusion drawn. ? The control group: SD rats 2, weighing 260 g ± 20 g, male. With SD rats supine, hands touched SD rats sternum, above it along the midline of the neck cut about 3cm incision, exposing the right superior cervical ganglia (SCG), removal of superior cervical ganglion in the neck ganglion resection at the drop of 1μl? 3% FR, stitching wounds, survived nine days after perfusion drawn. ? 2, FR labeled double-labeled with the cell membrane staining elected FR labeled lymph node biopsy rinsed with 0.01M PBS (5min × 3 times), a solution of DMSO diluted DiOC18 (3) (1:80), room temperature 30min, 0.01M PBS rinse (5min × 3 times); room temperature DAPI nuclear staining 30min, 0.01M PBS rinse (5min × 3 times); liquid paraffin were mounted in a confocal microscope. 3, FR labeled with a fluorescent double-labeling immunohistochemistry elected FR labeled lymph node biopsy rinsed with 0.01M PBS (5min × 3 times), serum for 30min, drying, were dropping 0.01M PBS appropriate dilution different an anti-CD3 (1:500), CD14 (1:500), CD22 (1:500), S100 (1:500), NPY (1:500) and VIP (1:500), 4 ° C overnight; Section two days rinsed with 0.01M PBS (5min × 3 times), and an anti-dropping corresponding biotinylated secondary anti-IgG (1:100), 37 ° C 30min, 0.01M PBS rinse (5min × 3 times); add 0.01M PBS diluted SABC-FITC (1:100), 37 ° C 30min, 0.01M PBS rinse (5min × 4 times); room temperature DAPI nuclear staining 30min, 0.01M PBS rinse (5 min × 3 times); liquid paraffin were mounted in a confocal microscope. Results a. Superior cervical ganglion FR FR anterograde marker injection into the neck ganglion, a confocal fluorescence microscope cervical lymph node biopsy, lymph node can be seen in the red fluorescent marker into the nerve terminal fibers, indicating that from the neck ganglion sympathetic nerve endings are FR tag, but marked to nerve fibers located only a few lymph node tissue surrounding the nucleus, which indicates that this is surrounded by sympathetic nerve fibers in the nerve cells in the immune pathway may play a key role. Two. FR labeled with the cell membrane results from the double-labeling results Anterograde tracing known sympathetic around only a few cells in lymph nodes distribution, in order to determine the sympathetic nerve endings and labeled FR around the positional relationship between cells, we used DiOC18 (3 ) marks the cell membrane, confocal microscopy to mark sympathetic nerve endings and labeled red fluorescent green fluorescent membrane overlap, the results indicate that the sympathetic nerve endings located in the cell membrane. Three. FR anterograde labeled with a fluorescent double-labeling immunohistochemistry results in order to determine FR labeled sympathetic nerve endings around the types of cell distribution in lymphoid organs, we were using anti-T cell (CD3), B cells (CD22), giant macrophages (CD14) and dendritic cells (S100) specific antibodies to double-labeled. FR markers found sympathetic nerve endings around the cells do not express CD3, CD22 and CD14 expression and S100, which results show that the sympathetic nerve endings FR marker does not dominate T cells, B cells and macrophages, dendritic cells only control. In addition, also found a sympathetic nerve endings FR labeled NPY and VIP coexistence. Conclusion 1 This is the first confirmed morphologically lymphoid organs sympathetic dominance only dendritic cells, and other cells (such as T, B, macrophages) has no direct relations of domination, this finding is neuroimmunomodulation pathway research foundation. 2 lymphoid organs dominated mainly sympathetic neuropeptides NPY and VIP two kinds of coexistence of these two neuropeptides in the nerve may play a key role in immune regulation. 3 lymphoid organs of dendritic cells (DC) is a key nerve cells in the immune pathway may be a nerve - immune dialogue is, for it is in the role of neural immune regulation remains to be further studied.

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