Division of Rheumatology, David Geffen School of Medicine at UCLA, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Genes Immun. 2010 Jun;11(4):294-309. doi: 10.1038/gene.2010.6. Epub 2010 Mar 4.
Tolerizing mice polygenically predisposed to lupus-like disease (NZB/NZW F1 females) with a peptide mimicking anti-DNA IgG sequences containing MHC class I and class II T cell determinants (pConsensus, pCons) results in protection from full-blown disease attributable in part to the induction of CD4(+)CD25(+)Foxp3+ and CD8(+)Foxp3+ regulatory T cells. We compared 45 000 murine genes in total white blood cells (WBC), CD4(+) T cells, and CD8(+) T cells from splenocytes of (NZBxNZW) F1 lupus-prone mice tolerized with pCons vs untreated naïve mice and found two-fold or greater differential expression for 448 WBC, 174 CD4, and 60 CD8 genes. We identified differentially expressed genes that played roles in the immune response and apoptosis. Using real-time PCR, we validated differential expression of selected genes (IFI202B, Bcl2, Foxp3, Trp-53, CCR7 and IFNar1) in the CD8(+)T cell microarray and determined expression of selected highly upregulated genes in different immune cell subsets. We also determined Smads expression in different immune cell subsets, including CD4(+) T cells and CD8(+) T cells, to detect the effects of TGF-beta, known to be the major cytokine that accounts for the suppressive capacity of CD8(+) Treg in this system. Silencing of anti-apoptotic gene Bcl2 or interferon genes (IFI202b and IFNar1 in combination) in CD8(+) T cells from tolerized mice did not affect the expression of the other selected genes. However, silencing of Foxp3 reduced expression of Foxp3, Ifi202b and PD1-all of which are involved in the suppressive capacity of CD8(+) Treg in this model.
用模拟抗 DNA IgG 序列的肽对多基因易患狼疮样疾病的小鼠（NZB/NZW F1 雌性）进行耐受处理（pConsensus，pCons）可导致部分免受完全发作疾病的保护，这归因于诱导 CD4(+)CD25(+)Foxp3+和 CD8(+)Foxp3+调节性 T 细胞。我们比较了来自（NZBxNZW）F1 狼疮易感小鼠脾细胞中 pCons 处理的与未处理的幼稚小鼠的总白细胞（WBC）、CD4(+)T 细胞和 CD8(+)T 细胞中的 45000 个鼠基因，发现 448 个 WBC、174 个 CD4 和 60 个 CD8 基因的表达有两倍或更多的差异。我们确定了在免疫反应和凋亡中起作用的差异表达基因。使用实时 PCR，我们验证了 CD8(+)T 细胞微阵列中选定基因（IFI202B、Bcl2、Foxp3、Trp-53、CCR7 和 IFNar1）的差异表达，并确定了不同免疫细胞亚群中选定高度上调基因的表达。我们还检测了 TGF-beta 已知是该系统中 CD8(+)Treg 抑制能力的主要细胞因子，在不同免疫细胞亚群中 Smads 的表达，包括 CD4(+)T 细胞和 CD8(+)T 细胞。在耐受小鼠的 CD8(+)T 细胞中沉默抗凋亡基因 Bcl2 或干扰素基因（IFI202b 和 IFNar1 联合）不会影响其他选定基因的表达。然而，Foxp3 的沉默降低了 Foxp3、Ifi202b 和 PD1 的表达-所有这些都参与了该模型中 CD8(+)Treg 的抑制能力。
