BKV Clearance Time Correlates With Exhaustion State and T-Cell Receptor Repertoire Shape of BKV-Specific T-Cells in Renal Transplant Patients
This study was approved by our local ethical review committee in compliance with the declaration of Helsinki. Informed and written consent was obtained from all patients (Ethic Committee Charité University Medicine, Berlin, Germany, EA2/028/13). The study cohort consisted of 7 kidney transplant recipients with sustained BKV reactivation (Table 1). The HLA typing for each patient and donor is summarized in Figure 1.
TABLE 1
Table 1. Characteristics of patients with BKV reactivation.
FIGURE 1
Figure 1. Recipient and donor HLA type. HLA type of the patients and their kidney donors. Black square indicate presence of the HLA type, white indicate absence.
BKV Viremia
BKV-DNA copy numbers were determined in serum as previously described (16). Briefly, DNA was isolated from serum using a QIAamp DNA Mini Kit (Qiagen) according to the manufacturer's instructions. Primers and probes were designed to amplify the VP1 region of BKV. A plasmid standard containing the VP1 coding region was used to determine the copy number per ml. Samples exceeding the detection level (>1,000 copies/ml) were considered positive.
BKV Stimulation
PBMC were isolated from 40 ml freshly drawn heparin-treated peripheral blood using the Ficoll-Hypaque (PAA Laboratories) gradient method and collected in RPMI-1640 medium (supplemented with L-glutamine (2 mmol/l), fetal calf serum (10%), penicillin (100 IU/ml), and streptomycin (0.1 mg/ml), all from Biochrom) and s...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
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