Previous research into the molecular mechanisms that underlie Ag-specific CD8؉ T cell differentiation and function has largely focused on the role of proteins. However, it is now apparent that the mammalian genome expresses large numbers of long (>200 nt) nonprotein-coding RNAs (ncRNAs), and there is increasing evidence that these RNAs have important regulatory functions, particularly in the regulation of epigenetic processes underpinning cell differentiation. In this study, we show that CD8 ؉ T cells express hundreds of long ncRNAs, many of which are lymphoid-specific and/or change dynamically with lymphocyte differentiation or activation. Numerous ncRNAs surround or overlap immunologically important protein-coding genes and can be predicted to function via a range of regulatory mechanisms. The overlap of many long ncRNAs expressed in CD8 ؉ T cells with microRNAs and small interfering RNAs further suggests that long ncRNAs may be processed into and exert their effects via smaller functional species. Finally, we show that the majority of long ncRNAs expressed in CD8 ؉ T cells harbor signatures of evolutionary conservation, secondary structures, and/or regulated promoters, further supporting their functionality. Taken together, our findings represent the first systematic discovery of long ncRNAs expressed in CD8؉ T cells and suggest that many of these transcripts are likely to play a role in adaptive immunity. The Journal of Immunology, 2009, 182: 7738 -7748.
C ytotoxic CD8ϩ T cells represent a critical arm of the adaptive immune system. They originate from common lymphoid precursors in the bone marrow and undergo subsequent selection in the thymus. Following viral infection, naive CD8 ϩ T cells develop into effector cells that, upon activation, kill infected host cells and limit viral spread. Once an infection is cleared, most CD8 ϩ T cells undergo programmed cell death, leaving behind a minority of self-renewable memory CD8 ϩ T cells that are able to rapidly mount a protective response upon subsequent viral challenge. Intensive study over the past few decades has considerably advanced our knowledge of the functional and phenotypic changes that occur throughout the CD8 ϩ T cell life cycle. Nevertheless, our knowledge of the underlying regulatory basis for CD8 ϩ T cell differentiation and function remains incomplete. In the past decade, it has become increasingly evident that nonprotein-coding RNAs (ncRNAs) 4 fulfill critical regulatory roles in mammalian biology. Most attention has focused on short RNAs, including small interfering RNAs (siRNAs), microRNAs (miRNAs), and PIWI-interacting RNAs, which regulate gene expression at the transcriptional and/or posttranscriptional levels (reviewed in Refs. 1 and 2). Although less prominent, long ncRNAs (Ͼ200nt) have also been recognized as serving important biological functions (reviewed in Ref.3). Air and HOTAIR, for instance, coordinate and regulate gene expression within the Igf2r and HOX loci, respectively, whereas Xist does so across an entire chromosome (4 -6). In ...