Table 1.
Clinical characteristics of patients at baseline.
Fig 1.
Characteristic of circulating Th17.1 cells in patients with rheumatoid arthritis (RA).
a. Flow cytometry plots explaining the gating strategy for the identification of peripheral blood Treg, Th1, Th2, Th1&2, Th17, and Th17.1 subpopulations. CD4+ T cell subsets in the peripheral blood of adult patients with RA were analyzed using flow cytometry. b. The MDR-1 activity of the indicated Th subset assessed using multicolor flow cytometry with rhodamine 123 (Rh-123). Total CD4+ T cells isolated from the peripheral blood were labeled with Rh-123. After a 1-h efflux period at 37 °C in the presence of vehicle (dimethyl sulfoxide) or MDR-1 inhibitors (cyclosporine A), cells were stained with antibodies to CCR6, CXCR3, and CD45RO, and the Rh-123 efflux of each Th subset was analyzed using flow cytometry. Data shown are flow cytometry plots representing three independent experiments performed on cells isolated from different donors with RA. c. The proportion of cells with CD28 expression within the indicated CD4+ T cell subpopulations of patients with RA. Data are representative of at least three independent experiments.
Fig 2.
Early change in the cell proliferation state of each Th subset by ABA treatment.
The peripheral blood mononuclear cells of patients with RA were obtained at baseline and 4 and 24 weeks of ABA treatment (0 and 4 weeks, n = 40; 24 weeks, n = 29). a. Sequential changes in the proportion of each T cell subset among CD4 T cells in the peripheral blood induced by ABA treatment. Data were analyzed using the Friedman rank sum test. b. Flow cytometry plot showing the frequency of Ki67 expression in the indicated CD4+ Th cells in patients with RA assessed using the intracellular staining of Ki67 antigen and analyzed using multicolor flow cytometry. c. Sequential changes in the proportion of Ki67 expression in each Th subset induced by ABA treatment (0 and 4 weeks, n = 40). Data were analyzed using the Wilcoxon signed rank test.
Fig 3.
Clinical significance of Th17.1 levels in abatacept treatment response.
ABA treatment was continued for 24 weeks. Subsequently, the correlation between the ABA response and proportion of Th subset at baseline was analyzed. The following indicators were used to evaluate response to ABA treatment: changes in the DAS28-CRP scores from baseline at 24 weeks after ABA treatment (ΔDAS28-CRP 24 weeks), disease activity evaluation after 24-week ABA treatment (remission and low, moderate, and high disease activities [LDA, MDA, and HDA, respectively]), and treatment response evaluation using the EULAR response criteria (good responder (GR), moderate responder (MR), and nonresponder [NR]). a. ABA treatment response after 24 weeks was evaluated as GR (n = 14), MR (n = 13), and NR (n = 13). The proportion of the indicated Th subsets among CD4+ lymphocytes at baseline in each group was plotted and displayed as box plot. b. Scatterplot shows the ratio or absolute number of Th17.1 cells at baseline and ΔDAS28-CRP 24 weeks. The regression line (red line) and its 95% CI (gray zone) are also shown in the plot. c. d. Patients were stratified into two groups (lower and higher) based on the median proportion of Th17.1 cells. The 100% stacked bar chart shows the EULAR response rate (c) and disease activity score (d) at 24 weeks after the initiation of ABA treatment in three groups. Data were analyzed using the Kruskal–Wallis and Mann–Whitney tests for between-group comparisons with Bonferroni correction (a), Spearman’s rank correlation coefficient (b), and Fisher’s exact test and Bonferroni correction for paired comparisons (c, d).
Table 2.
Clinical characteristics of the Th17.1-lower and Th17.1-higher patients at baseline.
Fig 4.
Prediction of therapeutic response to ABA based on the proportion of Th17.1 at baseline.
a. The difference in ABA therapeutic response between the Th17.1-lower (binary by median) and Th17.1-higher groups after the adjustment of patient background factors using inverse probability weighting (IPW). The size of the balloon plot indicates the weighting using the IPW method in each case. The red lines indicate the weighted mean (horizontal line) and SD (vertical line) after IPW adjustment. Moreover, the black lines indicate the non-weighted mean (horizontal line) and SD (vertical line). Data were analyzed using the weighted (red) or non-weighted (black) t-test. b. The adjusted odds ratio for the achievement of GR with baseline Th17.1-lower relative to Th17.1-higher. Logistic regression analysis was conducted using the IPW method to calculate the odds ratio adjusted for patient characteristics. Forest plot shows the unadjusted and adjusted odds ratios and 95% CI and P value. ABA, abatacept; IPW, inverse probability weighting; SD, standard deviation; OR, odds ratio; CI, confidence interval.