We have previously shown that the frequency of chronic nasal carriage of is higher in GPA patients compared to HC [30]. 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 Metixene hydrochloride hydrate analysis. CD4+ TEM 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 TEM1, TEM2, TEM17, and TEM17.1 cells, respectively. Results The percentage of CD4+ TEM cells was significantly increased in GPA patients in remission compared to HCs. Chemokine receptor co-expression analysis within the CD4+ TEM cell population demonstrated a significant increase in the proportion of TEM17 cells with a concomitant significant decrease in the TEM1 cells in GPA patients compared to HC. The percentage of TEM17 cells correlated negatively with TEM1 cells in GPA patients. Moreover, the circulating proportion of TEM17 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 TEM1 and TEM17 cells is modulated in CMV- seropositive GPA patients. Conclusions Our data demonstrates the identification of different CD4+ TEM cell subsets in peripheral blood of GPA patients based on chemokine receptor co-expression analysis. The aberrant balance between TEM1 and TEM17 cells in remission GPA patients, showed to be associated with disease pathogenesis in relation to organ involvement, and tendency to relapse. Electronic supplementary material The online version of this article (doi:10.1186/s13075-017-1343-8) contains supplementary material, which is available to authorized users. (% male)63 (% 44)42 (% 40)Age, mean (range)62.3 (26.8C85.2)57.2 (21.5C86.8)PR3-ANCAa, (% positive)39 (% 62)PR3-ANCA titer, median (range)1:40 (0C1:640)Creatinine umol/L, median (range)86 (52C224)CRP mg/L, median (range)2.7 (0.3C99)eGFR ml/min*1.73 m2, median (range)64 (21C109)CMV seropositive, (% positive) (N.D.)33 (% 54) (2)21 (% 58) (6) (% positive) (N.D.)27 (% 44) (1)BVAS, mean0Disease duration in years, Metixene hydrochloride hydrate median (range)9.6 (1.9C42.7)No. of total relapses, median (range)1 (0C7)Relapserb, (%)43 (% 68)Disease type, (% generalized)52 (% 83)Treatment at time of sampling, (%)?Azathioprine3 (% 5)?Azathioprine + prednisolone12 (% 19)?Prednisolone6 (% 10)?Mycophenolate mofetil + prednisolone7 (% 11)?Methotrexate1 (% 2)?No immunosupressive treatment34 (% 54)Co-trimoxazole, high dose/low dose/no dose17/15/31No. of organs involved, median (range)3 (1C7)Clinical manifestations, (%)?Renal35 (% 56)?ENT45 (% 71)?Joints36 (% 57)?Pulmonary40 (% 63)?Nervous system20 (% 32)?Eyes24 (% 38)?Cutaneous13 (% 21)?Other7 (% 11) Open in a separate window Characteristics at sampling time point Birmingham Vasculitis Activity Score, cytomegalovirus, C-reactive protein, estimated glomerular filtration rate, ear, nose and throat, granulomatosis with polyangiitis, healthy control, antineutrophil cytoplasmic antibodies targeting proteinase 3, GPA patient in remission, nasal carriers were determined as described previously [27]. Briefly, nasal isolates were sampled by rotating a sterile cotton swab in each anterior nary. Swabs were inoculated on 5% sheep-blood and salt mannitol agar for 72?h at 35?C. was identified by coagulase and DNase positivity. Metixene hydrochloride hydrate Patients were considered to be chronic nasal carriers when 50% of their nasal cultures grew test was used for data with Gaussian distribution and the Mann-Whitney test for data without Gaussian distribution. For intra-individual comparison of values at multiple time points during follow-up, repeated measures analysis of variance was used if data were normally distributed and a Friedman test was used if data had a non-Gaussian distribution. The association between clinical parameters and CD4+ TEM cell subsets in inclusion samples of r-GPA patients was investigated using the Spearmans rank correlation coefficient. In order to account for interactions of CMV Metixene hydrochloride hydrate and age on the percentage Metixene hydrochloride hydrate of CD4+T cells subsets and CD4+TEM cell subsets we used a linear (Enter) regression analysis. Non-normally distributed data were log-transformed. Differences were considered statistically significant at two-sided values equal to or less than 0.05. Results Higher frequency of CD4+ TEM cells in peripheral Rabbit Polyclonal to ABCA6 blood of GPA patients in remission We have previously reported that r-GPA patients have an increased percentage of circulating CD4+ TEM cells compared to HC [16]. Here, we confirm that within the CD4+ T cell population in.