Figure 3.

PSC-associated polymorphism rs56258221 (BACH2/MIR4464) contributes to increased polarization of CD4⁺ T_N cells toward pro-inflammatory phenotypes

(A and B) Re-clustering of CITE-Seq data from CD4⁺ T_N (cluster C0 from Figure 1J) resulted in four subclusters of CD4⁺ T_N (SC0-3) (A), which were assigned to cellular states via analysis of differentially expressed genes (B). SC0 did not show differentially expressed genes.

(C) Expression levels of BACH2 among each cluster.

(D) Bar plot comparing the frequencies of SC0-3 between SNP carriers and non-carriers.

(E) Schematic depiction of the workflow for in vitro polarization assays.

(F) After 12 days of polarizing culture of CD4⁺ T_N from people with PSC (n = 18) and HD (n = 12), the frequency of CD4⁺IL-17A⁺ cells was determined.

(G) After 7 days of polarizing culture of CD4⁺ T_N from people with PSC (n = 21) and HD (n = 15), the frequency of T CD4⁺CXCR3⁺TBET⁺IFNg⁺TNFa⁺ cells was determined.

(H) After 7 days of polarizing culture of CD4⁺ T_N from people with PSC (n = 18) and HD (n = 14), the frequencies of CD4⁺CD25⁺CD152⁺FOXP3⁺ cells were determined.

(I–K) Data on people with PSC from T_H17 (F), T_H1 (G), and iT_REG (H) polarization were separated by genotype for polymorphism rs56258221. Characteristics of the clinical cohort are included in Tables S4–S6. Statistics: normality distribution was tested by Kolmogorov-Smirnov test; normal distribution: Welch’s t test; no normal distribution: Mann-Whitney U test. p < 0.05 was considered statistically significant. Data are presented as mean ± SD and deriving from n ≥ 2 repeats per experiment.