Expression data from effector CD4 T cells isolated from MRL/Faslpr mice.. Mus musculus

MRL/Faslpr mice is a lupus prone strain that exhibits lupus disease features at 12-16 weeks of age, including high-titer circulating anti-DNA antibodies, splenomegaly, lymphadnopathy, skin lesions, and IgG deposits in the kidney. At 16-24 weeks of age, CD4+ B220- CD44+ T cells were sorted into three populations based on the expression of two cell surface molecules, CD62L and PSGL1. CD62Lhi PSGL1hi, CD62Llo PSGL1hi, and CD62Llo PSGL1lo CD4+ T cells were isolated directly ex vivo. There was no treatment given to the animals. Naive (CD62Lhi CD44lo) CD4+ B220- T cells were isolated from young 6-8 week old female mice for comparison. We used a microarray to identify unique features of the CD62Llo PSGL1lo population in comparison to naïve CD4+ T cells and other activated CD4+ T cells. Overall design: Cells were isolated from the spleens of aged (16-24 weeks) female MRL/Faslpr mice directy ex vivo, and immediatley sorted into 3 populations; CD62Lhi PSGL1hi, CD62Llo PSGL1hi, and CD62Llo PSGL1lo. All 3 populations of cells were previously gated on TCRb+, CD4+, B220-, CD44+. Naive CD4 T cells were isolated directly ex vivo from the spleens of young (6-8 weeks) female MRL/Faslpr mice, and immediately sorted by gating on TCRb+, CD4+, B220-, CD62Lhi, CD44lo. Three indepedent sorts were performed. RNA was isolated using Qiagen's RNAeasy kit and total RNA was submitted to the W.M. Keck Foundation Biotechnology Resource Laboratory at Yale for amplification and hybridization to Affymetrix Mouse Genome 430 2.0 GeneChips.

MRL/Faslpr小鼠是一种狼疮易感品系，于12~16周龄时即可表现出狼疮疾病特征，包括高滴度循环抗DNA抗体、脾脏肿大、淋巴结病、皮肤病变以及肾脏IgG沉积。在16~24周龄时，研究人员基于两种细胞表面分子CD62L与PSGL1的表达水平，将CD4+ B220- CD44+ T细胞分为三个亚群：CD62Lhi PSGL1hi、CD62Llo PSGL1hi以及CD62Llo PSGL1lo CD4+ T细胞，所有细胞均直接从活体分离，实验动物未接受任何干预。为进行对照，研究人员从6~8周龄的年轻雌性小鼠体内分离获得初始型（CD62Lhi CD44lo）CD4+ B220- T细胞。本研究采用微阵列（microarray）技术，对比初始型CD4+ T细胞与其他活化型CD4+ T细胞，以鉴定CD62Llo PSGL1lo亚群的独特分子特征。

实验整体设计如下：细胞直接从16~24周龄雌性MRL/Faslpr小鼠的脾脏中分离，并即刻分选得到上述三个细胞亚群；所有分选前的细胞均先通过TCRb+、CD4+、B220-、CD44+设门。初始型CD4+ T细胞则直接从6~8周龄雌性MRL/Faslpr小鼠的脾脏中分离，通过TCRb+、CD4+、B220-、CD62Lhi、CD44lo设门进行分选。实验共独立重复分选3次。总RNA采用Qiagen的RNAeasy试剂盒提取，随后将总RNA样品送至耶鲁大学W.M. Keck基金会生物技术资源实验室进行扩增，并与Affymetrix小鼠基因组430 2.0基因芯片进行杂交。