Persistent expansion of circulating CD4⁺ effector memory T cells (T_EM) in patients with granulomatosis with polyangiitis (GPA) suggests their fundamental role in disease pathogenesis. Recent studies have shown that distinct functional CD4⁺ T_EM cell subsets can be identified based on expression patterns of chemokine receptors. The current study aimed to determine different CD4⁺ T_EM cell subsets based on chemokine receptor expression in peripheral blood of GPA patients. Identification of particular circulating CD4⁺ T_EM cells subsets may reveal distinct contributions of specific CD4⁺ T_EM subsets to the disease pathogenesis in GPA.

Peripheral blood of 63 GPA patients in remission and 42 age- and sex-matched healthy controls was stained immediately after blood withdrawal with fluorochrome-conjugated antibodies for cell surface markers (CD3, CD4, CD45RO) and chemokine receptors (CCR4, CCR6, CCR7, CRTh2, CXCR3) followed by flow cytometry analysis. CD4⁺ T_EM memory cells (CD3⁺CD4⁺CD45RO⁺CCR7^-) were gated, and the expression patterns of chemokine receptors CXCR3⁺CCR4^-CCR6^-CRTh2^-, CXCR3^-CCR4⁺CCR6^-CRTh2⁺, CXCR3^-CCR4⁺CCR6⁺CRTh2^-, and CXCR3⁺CCR4^-CCR6⁺CRTh2^- were used to distinguish T_EM1, T_EM2, T_EM17, and T_EM17.1 cells, respectively.

The percentage of CD4⁺ T_EM cells was significantly increased in GPA patients in remission compared to HCs. Chemokine receptor co-expression analysis within the CD4⁺ T_EM cell population demonstrated a significant increase in the proportion of T_EM17 cells with a concomitant significant decrease in the T_EM1 cells in GPA patients compared to HC. The percentage of T_EM17 cells correlated negatively with T_EM1 cells in GPA patients. Moreover, the circulating proportion of T_EM17 cells showed a positive correlation with the number of organs involved and an association with the tendency to relapse in GPA patients. Interestingly, the aberrant distribution of T_EM1 and T_EM17 cells is modulated in CMV- seropositive GPA patients.

Our data demonstrates the identification of different CD4⁺ T_EM cell subsets in peripheral blood of GPA patients based on chemokine receptor co-expression analysis. The aberrant balance between T_EM1 and T_EM17 cells in remission GPA patients, showed to be associated with disease pathogenesis in relation to organ involvement, and tendency to relapse.