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Quantitative and qualitative characterization of expanded CD4+ T cell clones in rheumatoid arthritis patients.

Ishigaki K, Shoda H, Kochi Y, Yasui T, Kadono Y, Tanaka S, Fujio K, Yamamoto K - Sci Rep (2015)

Bottom Line: Our in-depth characterization of ECs in RA successfully demonstrated the presence of the specific immunological selection pressure, which determines the phenotype of ECs.Moreover, transcriptome tracking added novel aspects to the underlying sequential immune processes.Our approach may provide new insights into the pathophysiology of RA.

View Article: PubMed Central - PubMed

Affiliation: Department of Allergy and Rheumatology, Graduate School of Medicine, the University of Tokyo, Tokyo, Japan.

ABSTRACT
Rheumatoid arthritis (RA) is an autoimmune destructive arthritis associated with CD4(+) T cell-mediated immunity. Although expanded CD4(+) T cell clones (ECs) has already been confirmed, the detailed characteristics of ECs have not been elucidated in RA. Using combination of a single-cell analysis and next-generation sequencing (NGS) in TCR repertoire analysis, we here revealed the detailed nature of ECs by examining peripheral blood (PB) from 5 RA patients and synovium from 1 RA patient. When we intensively investigated the single-cell transcriptome of the most expanded clones in memory CD4(+) T cells (memory-mECs) in RA-PB, senescence-related transcripts were up-regulated, indicating circulating ECs were constantly stimulated. Tracking of the transcriptome shift within the same memory-mECs between PB and the synovium revealed the augmentations in senescence-related gene expression and the up-regulation of synovium-homing chemokine receptors in the synovium. Our in-depth characterization of ECs in RA successfully demonstrated the presence of the specific immunological selection pressure, which determines the phenotype of ECs. Moreover, transcriptome tracking added novel aspects to the underlying sequential immune processes. Our approach may provide new insights into the pathophysiology of RA.

No MeSH data available.


Related in: MedlinePlus

Gene expression profiles of PB memory-mECs.cDNAs of PB memory-mECs (corresponding to C1.1 for RA1 and C2.1 for RA2 as shown in Fig. 1) and memory-NECs obtained by the single-cell analysis were mixed (4 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. *p < 0.05, **p < 0.005.
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f4: Gene expression profiles of PB memory-mECs.cDNAs of PB memory-mECs (corresponding to C1.1 for RA1 and C2.1 for RA2 as shown in Fig. 1) and memory-NECs obtained by the single-cell analysis were mixed (4 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. *p < 0.05, **p < 0.005.

Mentions: In order to further characterize the memory ECs in RA, we next performed single-cell transcriptome analysis of the most expanded CD4+ T cell clones (mECs) and non-expanded CD4+ T cell clones (NECs) in the peripheral blood of 2 patients, RA1 and RA2 (Fig. 4). We defined mECs and NECs as the CD4+ T cell clones detected in the single-cell analysis of each sample with the highest frequency and only once, respectively. We focused on “the most expanded” clones for this purpose because multiple single cells are required for the reliable characterization of the gene expression profile of each clone. We also confined our target to memory-phenotype CD4+ T cell clones (memory-mEC and memory-NECs). By comparing memory-mECs with memory-NECs from the same patient, we could extract the “clonal expansion” molecular signatures, without being influenced by inter-individual variability.


Quantitative and qualitative characterization of expanded CD4+ T cell clones in rheumatoid arthritis patients.

Ishigaki K, Shoda H, Kochi Y, Yasui T, Kadono Y, Tanaka S, Fujio K, Yamamoto K - Sci Rep (2015)

Gene expression profiles of PB memory-mECs.cDNAs of PB memory-mECs (corresponding to C1.1 for RA1 and C2.1 for RA2 as shown in Fig. 1) and memory-NECs obtained by the single-cell analysis were mixed (4 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. *p < 0.05, **p < 0.005.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4542667&req=5

f4: Gene expression profiles of PB memory-mECs.cDNAs of PB memory-mECs (corresponding to C1.1 for RA1 and C2.1 for RA2 as shown in Fig. 1) and memory-NECs obtained by the single-cell analysis were mixed (4 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. *p < 0.05, **p < 0.005.
Mentions: In order to further characterize the memory ECs in RA, we next performed single-cell transcriptome analysis of the most expanded CD4+ T cell clones (mECs) and non-expanded CD4+ T cell clones (NECs) in the peripheral blood of 2 patients, RA1 and RA2 (Fig. 4). We defined mECs and NECs as the CD4+ T cell clones detected in the single-cell analysis of each sample with the highest frequency and only once, respectively. We focused on “the most expanded” clones for this purpose because multiple single cells are required for the reliable characterization of the gene expression profile of each clone. We also confined our target to memory-phenotype CD4+ T cell clones (memory-mEC and memory-NECs). By comparing memory-mECs with memory-NECs from the same patient, we could extract the “clonal expansion” molecular signatures, without being influenced by inter-individual variability.

Bottom Line: Our in-depth characterization of ECs in RA successfully demonstrated the presence of the specific immunological selection pressure, which determines the phenotype of ECs.Moreover, transcriptome tracking added novel aspects to the underlying sequential immune processes.Our approach may provide new insights into the pathophysiology of RA.

View Article: PubMed Central - PubMed

Affiliation: Department of Allergy and Rheumatology, Graduate School of Medicine, the University of Tokyo, Tokyo, Japan.

ABSTRACT
Rheumatoid arthritis (RA) is an autoimmune destructive arthritis associated with CD4(+) T cell-mediated immunity. Although expanded CD4(+) T cell clones (ECs) has already been confirmed, the detailed characteristics of ECs have not been elucidated in RA. Using combination of a single-cell analysis and next-generation sequencing (NGS) in TCR repertoire analysis, we here revealed the detailed nature of ECs by examining peripheral blood (PB) from 5 RA patients and synovium from 1 RA patient. When we intensively investigated the single-cell transcriptome of the most expanded clones in memory CD4(+) T cells (memory-mECs) in RA-PB, senescence-related transcripts were up-regulated, indicating circulating ECs were constantly stimulated. Tracking of the transcriptome shift within the same memory-mECs between PB and the synovium revealed the augmentations in senescence-related gene expression and the up-regulation of synovium-homing chemokine receptors in the synovium. Our in-depth characterization of ECs in RA successfully demonstrated the presence of the specific immunological selection pressure, which determines the phenotype of ECs. Moreover, transcriptome tracking added novel aspects to the underlying sequential immune processes. Our approach may provide new insights into the pathophysiology of RA.

No MeSH data available.


Related in: MedlinePlus