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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

Tracking of the gene expression shift in memory-mECs in RA1 between PB and the synovium.Memory-mECs in the RA1 PB and RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1). cDNAs of RA1 memory-mECs in PB and the synovium obtained by the single-cell analysis were mixed (2 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. The log-transformed ratio (gene expression value in the synovium / gene expression value in PB) was shown.
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f6: Tracking of the gene expression shift in memory-mECs in RA1 between PB and the synovium.Memory-mECs in the RA1 PB and RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1). cDNAs of RA1 memory-mECs in PB and the synovium obtained by the single-cell analysis were mixed (2 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. The log-transformed ratio (gene expression value in the synovium / gene expression value in PB) was shown.

Mentions: The combined use of single-cell TCR repertoire analysis and single-cell transcriptome analysis enabled us to track the gene expression shift within the same clones by localization. Since memory-mECs in the RA1 PB and the RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1), we examined the phenotypic changes of memory-mECs caused by infiltration into the synovium (Fig. 6). Chemokine receptors, which are important for infiltration into the synovium (CXCR4 and CCR5), and effector molecules (GZMB) were up-regulated in the synovium. On the other hand, CD5, a marker of autoreactivity41, was stably expressed.


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)

Tracking of the gene expression shift in memory-mECs in RA1 between PB and the synovium.Memory-mECs in the RA1 PB and RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1). cDNAs of RA1 memory-mECs in PB and the synovium obtained by the single-cell analysis were mixed (2 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. The log-transformed ratio (gene expression value in the synovium / gene expression value in PB) was shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: Tracking of the gene expression shift in memory-mECs in RA1 between PB and the synovium.Memory-mECs in the RA1 PB and RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1). cDNAs of RA1 memory-mECs in PB and the synovium obtained by the single-cell analysis were mixed (2 pooled samples in total), and qPCR was performed. ACTB was used as an internal control gene. The log-transformed ratio (gene expression value in the synovium / gene expression value in PB) was shown.
Mentions: The combined use of single-cell TCR repertoire analysis and single-cell transcriptome analysis enabled us to track the gene expression shift within the same clones by localization. Since memory-mECs in the RA1 PB and the RA1 synovium were identical clones (corresponding to C1.1 in Fig. 1), we examined the phenotypic changes of memory-mECs caused by infiltration into the synovium (Fig. 6). Chemokine receptors, which are important for infiltration into the synovium (CXCR4 and CCR5), and effector molecules (GZMB) were up-regulated in the synovium. On the other hand, CD5, a marker of autoreactivity41, was stably expressed.

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