BACKGROUND: Stage 5 chronic kidney disease (CKD5) is linked to complex yet not fully understood disturbances in immune system. This study aimed to investigate these disturbances by exploring the detailed composition of peripheral blood immune cell compartments in CKD5 patients and to provide integrative, multivariable dissection of how common inflammatory risk factors shape the immune landscape. METHODS: This cross-sectional study included 107 patients with chronic kidney disease stage 5 (CKD5) and 29 healthy blood donors as controls. Peripheral blood B cells, T cells, and dendritic cells were measured using a standardized and validated flow cytometry panel. The impact of selected clinical factors on immune cell composition was initially evaluated using a robust multivariate method (PERMANOVA). Variables that significantly affected immune cell composition were then included in a subsequent series of Poisson regression models, assessing predictors influence on the counts of individual immune cell subpopulations. RESULTS: Compared to healthy controls, CKD5 patients presented with B cell lymphopenia across all measured subsets except for plasmablasts, T cell lymphopenia with an immunosenescent phenotype predominantly in the CD4+ compartment, and significantly higher counts of LIN-HLA-DR+ antigen-presenting cells, mainly due to an increase in myeloid dendritic cell subpopulations. PERMANOVA identified smoking, CMV seropositivity, age, dialysis treatment, and atherosclerotic cardiovascular disease as factors significantly influencing peripheral blood immune composition. Subsequent Poisson regression models revealed that smoking was associated mainly with an increase in switched memory B cells, CMV seropositivity with an increase in CD4+ and CD8+ TEMRA cells, age with a decrease in naive CD8+ T cells, and dialysis treatment with an increase in marginal-zone B cells. CONCLUSIONS: Patients with CKD5 exhibit distinct composition of peripheral blood immune cells, further modified by other factors associated with systemic inflammatory response. These factors should be considered in immunomonitoring protocols and may enhance prediction of clinical outcomes such as vaccine responses.

• Klíčová slova
• B cells, T cells, chronic kidney disease, dendritic cells, flow cytometry, inflammation, peripheral blood immune cell composition,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The power to predict kidney allograft outcomes based on non-invasive assays is limited. Assessment of operational tolerance (OT) patients allows us to identify transcriptomic signatures of true non-responders for construction of predictive models. METHODS: In this observational retrospective study, RNA sequencing of peripheral blood was used in a derivation cohort to identify a protective set of transcripts by comparing 15 OT patients (40% females), from the TOMOGRAM Study (NCT05124444), 14 chronic active antibody-mediated rejection (CABMR) and 23 stable graft function patients ≥15 years (STA). The selected differentially expressed transcripts between OT and CABMR were used in a validation cohort (n = 396) to predict 3-year kidney allograft loss at 3 time-points using RT-qPCR. FINDINGS: Archetypal analysis and classifier performance of RNA sequencing data showed that OT is clearly distinguishable from CABMR, but similar to STA. Based on significant transcripts from the validation cohort in univariable analysis, 2 multivariable Cox models were created. A 3-transcript (ADGRG3, ATG2A, and GNLY) model from POD 7 predicted graft loss with C-statistics (C) 0.727 (95% CI, 0.638-0.820). Another 3-transcript (IGHM, CD5, GNLY) model from M3 predicted graft loss with C 0.786 (95% CI, 0.785-0.865). Combining 3-transcripts models with eGFR at POD 7 and M3 improved C-statistics to 0.860 (95% CI, 0.778-0.944) and 0.868 (95% CI, 0.790-0.944), respectively. INTERPRETATION: Identification of transcripts distinguishing OT from CABMR allowed us to construct models predicting premature graft loss. Identified transcripts reflect mechanisms of injury/repair and alloimmune response when assessed at day 7 or with a loss of protective phenotype when assessed at month 3. FUNDING: Supported by the Ministry of Health of the Czech Republic under grant NV19-06-00031.

• Klíčová slova
• Chronic antibody-mediated rejection, Kidney graft failure, Operational tolerance, Peripheral blood transcripts, RNA sequencing,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Kidney transplant recipients (KTRs) with "operational tolerance" (OT) maintain a functioning graft without immunosuppressive (IS) drugs, thus avoiding treatment complications. Nevertheless, IS drugs can influence gene-expression signatures aiming to identify OT among treated KTRs. METHODS: We compared five published signatures of OT in peripheral blood samples from 18 tolerant, 183 stable, and 34 chronic rejector KTRs, using gene-expression levels with and without adjustment for IS drugs and regularised logistic regression. FINDINGS: IS drugs explained up to 50% of the variability in gene-expression and 20-30% of the variability in the probability of OT predicted by signatures without drug adjustment. We present a parsimonious consensus gene-set to identify OT, derived from joint analysis of IS-drug-adjusted expression of five published signature gene-sets. This signature, including CD40, CTLA4, HSD11B1, IGKV4-1, MZB1, NR3C2, and RAB40C genes, showed an area under the curve 0⋅92 (95% confidence interval 0⋅88-0⋅94) in cross-validation and 0⋅97 (0⋅93-1⋅00) in six months follow-up samples. INTERPRETATION: We advocate including adjustment for IS drug therapy in the development stage of gene-expression signatures of OT to reduce the risk of capturing features of treatment, which could be lost following IS drug minimisation or withdrawal. Our signature, however, would require further validation in an independent dataset and a biomarker-led trial. FUNDING: FP7-HEALTH-2012-INNOVATION-1 [305147:BIO-DrIM] (SC,IR-M,PM,DSt); MRC [G0801537/ID:88245] (MPH-F); MRC [MR/J006742/1] (IR-M); Guy's&StThomas' Charity [R080530]&[R090782]; CONICYT-Bicentennial-Becas-Chile (EN-L); EU:FP7/2007-2013 [HEALTH-F5-2010-260687: The ONE Study] (MPH-F); Czech Ministry of Health [NV19-06-00031] (OV); NIHR-BRC Guy's&StThomas' NHS Foundation Trust and KCL (SC); UK Clinical Research Networks [portfolio:7521].

• Klíčová slova
• Biomarkers, Immunosuppressive Drugs, Kidney, Operational Tolerance, RT-qPCR, Transplantation,
• MeSH
• dospělí MeSH
• genové regulační sítě * účinky léků   MeSH
• imunosupresiva farmakologie   terapeutické užití   MeSH
• konsensus MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• logistické modely MeSH
• rejekce štěpu genetika   prevence a kontrola   MeSH
• senioři MeSH
• stanovení celkové genové exprese metody   MeSH
• studie případů a kontrol MeSH
• transplantace ledvin škodlivé účinky   MeSH
• transplantační tolerance * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• imunosupresiva MeSH