DNA cassette encoding the conserved epitope in CMV AD2 site I was cloned into the expression vector pGEX-5X (Amersham Bioscience [now GE Healthcare], Piscataway, NJ, USA). GST fusion proteins containing the gH epitopes from the AD169 and Towne strain were used to detect CMV gH type-specific antibodies
as previously reported . OD values specific to each antigen were obtained by subtracting the OD values for GST as described previously . An arbitrary cutoff for ELISA (OD = 0.25) was defined as the mean plus two standard deviations of OD values of a panel of healthy CMV seronegative volunteers . Detection of strain-specific gH-antibodies in the recipients’ serum samples, which matched those of their donors, was considered gH-m antibody positivity. The basic characteristics of the renal transplant recipients are summarized in Table 1. Fifty-two of the 77 recipients AZD0530 chemical structure had antibodies against gB. There were no differences between patients with
and without gB antibodies in other relevant variables, namely age, sex, number of HLA mismatches and immunosuppression protocols. The transplant recipients were followed up for 6 months after transplantation. Rejection was suspected when serum creatinine concentrations increased more than 25% above the basal level in the absence of urinary tract obstruction or renal Fenbendazole graft artery stenosis, as described previously . The first rejection episode
was confirmed histologically by biopsying the grafts. Everolimus molecular weight Preemptive therapy was employed when CMV infection and/or CMV end-organ disease were diagnosed, as described previously . Using StatView 5.0, Fisher’s exact test was used to evaluate the rate of acute rejection in different gB serostatus groups. Statistical significance was set at P < 0.05. The incidence of biopsy-proven acute rejection was calculated using the Kaplan–Mayer method, and comparisons were carried out by the log-rank test using SPSS. Subsequent to their entry into the study, 27/77 recipients (35%) in a D + /R+ setting experienced biopsy-proven rejection during the 6 months after transplantation. Among these 27 D + /R+ patients with rejection, 23 (85%) had antibodies against CMV gB. The incidences of acute rejection among recipients with (gB+) and without (gB−) antibodies against gB AD2 were 44% and 16%, respectively. The rate of acute rejection was significantly higher in gB+ recipients than in gB− recipients (Table 2). Figure 1 shows Kaplan–Meier curves for the cumulative probability of freedom from biopsy-proven acute rejection. There were significant differences between the gB+ group and the gB− group according to the log-rank test (P = 0.025).