Marker for detection of IL-17-producing helper T-cell, and method for detection of IL-17-producing helper T-cell
A cell and labeling technology, applied in biochemical equipment and methods, microbial determination/inspection, biological testing, etc., can solve problems such as unclear reasons
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Embodiment 1
[0106] Example 1: Analysis of highly expressed genes in cultured Th17 cells derived from mice
[0107] 1. Isolation of Naive T Cells from Mouse Spleen
[0108] The spleen of the BALB / c mouse was removed to obtain a sample containing spleen cells. After the erythrocytes in the sample were hemolyzed with ammonium chloride, CD8, B cells, monocytes, macrophages, granulocytes, and immature cells were removed from the sample with magnetic beads (manufactured by POLYSCIENCE). Cellular components of red blood cells, crude CD4 positive (CD4 + ) T cells. CD4 + T cells were sorted by flow cytometry, and fractions of naive T cells (CD4 + / CD25neg / CD44low / CD62high). Likewise, naive T cells were purified from splenocytes of C57 / BL6 mice.
[0109] 2. Differentiate and culture Th1, Th2, Treg, Th17 cells from naive T cells
[0110]The naive T cells derived from BALB / c mice obtained in the above 1. were mixed with 0.5-2.0×10 6 Cells / 2ml / well were seeded onto 24-well plates containing an...
Embodiment 2
[0143] Example 2: Analysis of highly expressed genes in disease model mice
[0144] 1. Making disease model mice
[0145] 1) Making SKG arthritis model mice (hereinafter referred to as "arthritis model mice")
[0146] Arthritis model mice were produced by the following method
[0147] a) Preparation of bacteria components and administration to mice
[0148] Suspend the thermocurdlan (curdlan) (SIGMA company) produced by Alcaligenes faecalis in PBS (phosphate buffered saline) to obtain thermocurdlan preparation (50mg / ml) (hereinafter referred to as thalline Element). The bacterium component was administered to the abdominal cavity of 7-8 week-old spontaneous SKG arthritis mice (female) (Nature, vol 426, pp.454-460 (2003), purchased from clea-japan company) 200 μl per mouse . Four weeks later, 200 μl was intraperitoneally administered to each mouse.
[0149] b) Judging the severity of arthritis
[0150] 0 points: normal
[0151] 1 point: Mild joint inflammation
[0152]...
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