Utility of Major Leukocyte Subpopulations for Monitoring Secondary Cytomegalovirus Infections in Renal-Allograft Recipients by PCR

DOI: 10.1128/JCM.36.4.1008-1014.1998

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Fig. 1.
Correlation of pp65 antigen-positive cells with CMV DNA levels in mixed PBL (○), PMNL (x) and PBMC (•) of patients with active CMV infection determined by Spearman regression analysis. (A) Forty-three samples from 23 patients were analyzed before onset of ganciclovir therapy. Significant correlation was observed for mixed PBL (r _S = 0.979; P < 0.001) and PMNL (r _S = 0.973; P < 0.001) but not for PBMC (r _S = −0.150;P = 0.20). (B) Thirty samples were obtained from 18 patients during antiviral treatment. Again, CMV DNA load in mixed PBL (r _S = 0.964; P < 0.001) and PMNL (r _S = 0.958; P < 0.001), but not in PBMC (r _S = 0.068;P = 0.25), correlated significantly with pp65 antigen-positive cell counts.

Table 1.

Frequencies of positive CMV PCR results in leukocyte subsets and plasma

Group	No. of patients	Total no. of samples	No. of samples positive^a (%)
Group	No. of patients	Total no. of samples	Mixed PBL	PBMC	PMNL	Plasma^b
Active infection
Before therapy, pp65 positive	23	43	43 (100)	43 (100)	43 (100)	39 (90.7)
Therapy
pp65 positive	18	30	30 (100)	30 (100)	30 (100)	19 (63.3)
pp65 negative^c	18	55	47 (85.5)	45 (81.8)	4 (7.3)	6 (10.9)
Latent infection^d	17	60	35 (58.3)	31 (51.7)	3 (5.0)	5 (8.0)

↵a For leukocytes, ≥5 copies/10⁵ total PBL; for plasma, ≥5 CMV DNA copies/10 μl.
↵b pp65-positive samples before versus during therapy, P < 0.05 (χ² test); pp65-positive samples before therapy versus pp65-negative samples after therapy, P < 0.001; pp65-positive samples before therapy versus latent infection, P < 0.001.
↵c Mixed PBL versus PBMC, no significant difference (McNemar’s test); mixed PBL versus PMNL, P< 0.001; PBMC versus PMNL, P < 0.001.
↵d Mixed PBL versus PBMC, no significant difference (McNemar’s test); mixed PBL versus PMNL, P< 0.001; PBMC versus PMNL, P < 0.001.

Table 2.

CMV DNA load in leukocyte subsets and plasma

Group	No. of patients	No. of samples	Median no. of pp65-positive cells^f (range)	Median no. of CMV DNA copies^a (range)
Group	No. of patients	No. of samples	Median no. of pp65-positive cells^f (range)	Mixed PBL^b	PBMC^c	PMNL^d	Plasma^e
Active infection
Before therapy, pp65 positive	23	43	152 (4–563)	950 (10–2,580)	35 (10–60)	920 (5–2,520)	45 (10–105)
Therapy
pp65 positive	18	30	26 (5–122)	190 (10–870)	40 (10–55)	170 (5–805)	25 (5–45)
pp65 negative	18	55	NA^g	35 (5–60)	30 (10–55)	7.5 (5–15)	10 (5–20)
Latent infection	17	60	NA	30 (10–65)	35 (5–60)	10 (5–20)	10 (5–15)

↵a Per 10⁵ total PBL (leukocytes) or per 10 μl (plasma). Differences in CMV DNA levels between groups were analyzed by the Mann-Whitney U test.
↵b pp65-positive samples before versus after onset of therapy, P < 0.001; pp65-positive samples versus pp65-negative samples after onset of therapy, P< 0.001; pp65-negative samples after onset of therapy versus latent infection, no significant difference.
↵c No significant differences.
↵d Before versus during therapy,P < 0.001. Only four samples after antiviral treatment and three samples in patients without active infection were PCR positive (Table 1).
↵e Before versus during therapy,P < 0.001. Only six samples after antiviral treatment and five samples in patients without active infection were PCR positive (Table 1).
↵f pp 65 antigen-positive cell counts: before versus after therapy, P < 0.001. All samples were pp65 antigen negative when ganciclovir therapy was discontinued.
↵g NA, not applicable.