Gene expression analysis of Wnt+ and Wnt- effector CD8 T cells. Mus musculus

Wnt signal transduction during an immune response is involved in the establishment of functional CD8 T cell memory P14 ConductinLacZ CD8 T cells (CD45.2) were transferred in CD45.1+.2+ recipient mice. The day after, recipients were infected with 200pfu LCMV WE. At day 8 after infection, cells from spleen and lymph nodes were harvested, magnetically enriched for CD8 T cells, loaded with the beta-galactosidase substrate (FDG) and sorted as 7AAD-negative CD45.- negative, beta-galactosidase positive versus negative cells. Overall design: P14 CD8 T cells were sorted at day 8 after LCMV infection according to LacZ activity for RNA extraction and hybridization on Affymetrix microarrays.

免疫应答过程中的Wnt信号转导（Wnt signal transduction）参与功能性CD8 T细胞记忆的建立。将携带ConductinLacZ标记的P14 CD8 T细胞（CD45.2）转输至CD45.1+.2+受体小鼠。造模次日，受体小鼠经200噬斑形成单位（pfu）的淋巴细胞脉络丛脑膜炎病毒WE株（LCMV WE）感染。感染后第8天，收获脾脏与淋巴结细胞，通过磁性分选富集CD8 T细胞，以β-半乳糖苷酶底物FDG进行标记，随后分选出7AAD阴性、CD45阴性的β-半乳糖苷酶阳性与阴性细胞。整体实验设计：在LCMV感染后第8天，依据β-半乳糖苷酶（LacZ）活性分选P14 CD8 T细胞，用于RNA提取，并在Affymetrix微阵列芯片上完成杂交实验。