The importance of eosinophil cells in kidney allograft rejection

The composition of the intragraft cellular infiltrate was studied in 83 renal allograft recipients with the technique of fine-needle aspiration cytology in the first four weeks following kidney transplantation. We found a significantly (P less than 0.05) higher mean tissue eosinophil percentage in patients who had irreversible rejections with transplant loss than in those who had reversible rejections (12.54 +/- 2.31 versus 3.79 +/- 1.14, mean +/- SEM). Patients who had serious, dialysis-requiring rejections also showed a significantly (P less than 0.05) higher mean tissue eosinophil percentage than those who had reversible rejections (21.40 +/- 5.98 versus 3.79 +/- 1.14, mean +/- SEM). The frequency of the HLA B8 antigen was 46.2% in patients who had excessive tissue eosinophilia, whereas its frequency in all the studied patients was 18.3%. Based on our observations, the presence of more than 4% eosinophils in the tissue inflammatory exudate is a specific (91%) and fairly sensitive (78%) indicator of irreversible and severe acute rejections.     

Significance of interstitial lesions as the early indicator for acute vascular rejection in human renal allografts

To evaluate the relevance between interstitial lesions and acute vascular rejection of renal allograft, we examined 129 renal allograft biopsy samples obtained from 60 transplant patients within 90 d post-transplantation. Plasma cells, eosinophils and polymorphonuclear cells (PMNs) in the cortical interstitium were counted, respectively, and the number of each cells per 10 fields at a magnification of 200x was calculated. Then, the existence of interstitial edema was judged. We categorized 129 biopsy specimens into specimens before (Pre), on (R) and after acute rejection (Post). An increased number of eosinophils (mean +/- SE; 15.4 +/- 9.2 per 10 fields) was observed in R rather than in Pre (0.6 +/- 0.3, p < 0.05). The increasing number of plasma cells depends on a time course following the onset of acute rejection. The number is significantly larger in acute vascular rejection (4.3 +/- 1.1) than in non-vascular rejection (1.3 +/- 0.4, p < 0.01). In living donor transplants, the number of PMNs in patients before vascular rejection (median, 16.7 per 10 fields; range, 5.0-20.0 per 10 fields) was significantly larger than in normal or borderline patients (2.2; 0.0-36.7, p < 0.01). The incidence of interstitial edema in patients before vascular rejection (55.6%) was significantly higher than in normal or borderline patients (8.0%, p < 0.01). This study suggested that acute vascular rejection is relevant to interstitial lesions, such as for a plasma cell, eosinophil and PMN infiltrate, and edema. Also, these lesions seem to be the indicator of acute vascular rejection. These findings may contribute to the refinement of histological diagnosis of acute vascular rejection.     

Risk factors for chronic allograft nephropathy after renal transplantation: a protocol biopsy study

BACKGROUND: Chronic allograft nephropathy (CAN) leads to chronic allograft dysfunction and loss. Regular renal transplant biopsies may be useful to find risk factors for CAN. METHODS: We carried out 688 protocol biopsies in 258 patients at 6, 12, and 26 weeks after renal transplantation. Patients with signs of CAN in the biopsy 3 (N= 70, CAN group), and those without (N= 120, non-CAN group), were compared. RESULTS: Chronic tubulointerstitial changes increased from biopsy 1 to 3 (5% vs. 37%, P < 0.0001). Fifty-six of 190 patients had acute rejection within 6 months (30%), 33 of which were found in protocol biopsies (17%). On univariate analysis, the CAN group had CAN more often at biopsy 2 than the non-CAN group (23% vs. 4%, P < 0.0001), had a lower calculated creatinine clearance at biopsy 1 and 2 (49.4 +/- 25.8 vs. 57 +/- 20.2 mL/min, P= 0.01; 47.3 +/- 21.2 vs. 57.9 +/- 19.5 mL/min, P= 0.001, respectively), had a living donor less often than a brain dead donor (7% vs. 18%, P= 0.045), had a longer cold ischemia time (17.4 +/- 7 vs. 14.9 +/- 8.1 hours, P= 0.04), and had arterionephrosclerosis more often (24% vs. 12%, P= 0.02). On multivariate analysis, the differences in CAN at biopsy 2 (P= 0.001) and lower GFR at biopsy 2 (P= 0.002) were confirmed; in addition, nephrocalcinosis (P= 0.006) and acute rejection (P= 0.046) were found to occur more often. CONCLUSION: Chronic tubulointerstitial changes develop early after renal transplantation and are associated with reduced kidney function. Risk factors for CAN are arterionephrosclerosis (donor-related), nephrocalcinosis (related to preexisting hyperparathyroidism), a long cold-ischemia time (ischemia-perfusion-related), and acute rejection. Renal functional decline precedes morphologic changes of CAN, expressed as tubular atrophy and interstitial fibrosis.     

The effects of OKT3 therapy on infiltrating lymphocytes in rejecting renal allografts

OKT3 antibody therapy is effective in the treatment of renal allograft rejection. However its exact mode of action is unknown. Following OKT3 administration, peripheral blood lymphocytes fail to express the CD3 antigen, although other membrane antigens are relatively preserved. In this way the lymphocytes are unable to respond to foreign antigens. It is not known whether this modulation of CD3 on lymphocytes occurs within the rejecting allograft. Ten patients who received OKT3 therapy for steroid-resistant renal allograft rejection were studied. The aim of the study was to examine whether OKT3 antibody therapy altered the degree or the relative composition of the inflammatory infiltrate and to assess whether OKT3 treatment resulted in modulation of CD3 on intragraft lymphocytes. The study involved immunoperoxidase examination of biopsy material and flow cytometric analysis of peripheral blood lymphocytes obtained before and during treatment with OKT3 antibody. The results show that the total number of infiltrating leukocytes decreased after 5 days of treatment (3215 +/- 700 cells/l. 0 mm2 of tissue before vs. 1730 +/- 635 at day 5; P less than 0.001). The relative proportions of macrophages, total lymphocytes, CD4 +ve and CD8 +ve cells did not alter during therapy. Despite marked modulation of peripheral blood lymphocytes (CD3/CD2 ratio 0.89 +/- 0.13 before vs. 0.10 +/- 0.11 during treatment, P less than 0.001), there was no evidence of modulation of intragraft lymphocytes (CD3/CD2 ratio 0.98 +/- 0.14 before vs. 0.90 +/- 0.21 during treatment, P = NS). Although OKT3 antibody therapy is effective clinically at improving renal allograft rejection, this study demonstrates that it does not appear to cause modulation of the CD3 antigen on intragraft lymphocytes.     

Increased serum beta 2 microglobulin during rejection, cyclosporine-induced nephrotoxicity, and cytomegalovirus infection in renal transplant recipients

In 83 renal transplant recipients, serum beta 2 microglobulin (beta 2m) levels were significantly elevated during pretransplant uremia, rejection, cyclosporine-induced nephrotoxicity, and infections. In patients with normal serum creatinine, 74% had elevated serum beta 2m levels. None of the cyclosporine-treated patients had normal levels of beta 2m. Patients with stable renal allograft function receiving cyclosporine showed significantly higher serum beta 2m (P less than 0.001) and serum creatinine (P less than 0.01) levels than azathioprine treated patients. Patients with an irreversible rejection showed significantly higher serum concentrations of beta 2m than patients experiencing a reversible rejection (P less than 0.001). During cytomegalovirus (CMV) infection the serum beta 2m levels were elevated compared with other infections (P less than 0.001), while the serum creatinine was not. However, infected patients had higher serum levels of beta 2m and creatinine than patients with stable renal allograft function (P less than 0.001). Serum beta 2m may therefore be useful in the early diagnosis of CMV infection. To conclude, serum beta 2m levels cannot distinguish between rejection, cyclosporine nephrotoxicity, or infection.     

A Monocyte chemoattractant protein-1 (MCP-1) polymorphism and outcome after renal transplantation

Among the factors modulating transplant rejection and cardiovascular disease, chemokines and their respective receptors deserve special attention. In this respect, increased expression of MCP-1 and the corresponding receptor CCR2 have been demonstrated in renal transplant rejection and coronary artery disease. The impact of the MCP-1-2518G and CCR2-64I genotypes on renal allograft function was investigated in 232 patients who underwent transplantation over an 11-yr period. Genomic DNA was genotyped using PCR with sequence-specific primers followed by restriction fragment length polymorphism analysis. Eighteen (7.8%) patients were homozygous for the MCP-1-2518G mutation. The G/G allele of MCP-1 -2518 behaved as a determinant for long-term allograft survival and resulted in reduction of the mean graft survival, as compared with the heterozygous (A/G) or wild-type (A/A) allele (67 +/- 14 versus 95 +/- 4 mo; Log rank P = 0.0052). The 64I mutation of CCR2 had no effect on kidney graft failure (93 +/- 6 and 91 +/- 5 mo, respectively; P = 0.81). None of the investigated polymorphisms showed a significant shift in gene frequency in acute rejection and rejection-free groups. In conjunction with these findings, peripheral blood mononuclear cells from kidney transplant recipients carrying the G-allele were characterized by a 2.5-fold higher MCP-1 secretion (P < 0.05). In conclusion, recipients of renal transplants homozygous for the -2518 G mutation of the MCP-1 gene are at risk for premature kidney graft failure. This variant of MCP-1 may be a future predictor for long-term kidney graft failure.     

Applications and Limitations of Blood Eosinophilia for the Diagnosis of Acute Cellular Rejection in Liver Transplantation

This study evaluates the predictive value of the blood eosinophil count in the diagnosis of acute cellular rejection, its value as a marker of response to treatment, the diagnostic use in a subgroup of patients with normal transaminases and compares blood eosinophilia in patients with and without hepatitis C virus infection. A consecutive cohort of 101 liver transplant patients, 275 liver biopsies, and blood eosinophils recorded on the day or one day before biopsy were analyzed. An elevated eosinophil count has a positive predictive value for acute cellular rejection of 82%. A normal eosinophil count excludes moderate/severe rejection with a predictive value of 86%. The eosinophil count decreases in 69% of patients following treatment of acute cellular rejection with corticosteroids irrespective of treatment outcome. Acute cellular rejection in the presence of an elevated eosinophil count occurs significantly less often (p = 0.007) in patients with hepatitis C virus. An elevated eosinophil count is a valuable marker of acute cellular rejection. However, blood eosinophil levels should not be used to predict acute cellular rejection following treatment with corticosteroids. Blood eosinophilia, seen less often in patients with hepatitis C virus, may reflect an over-diagnosis of acute cellular rejection in these patients.     

Elevated numbers of circulating endothelial cells in renal transplant recipients

BACKGROUND: Damage of microvascular endothelial cells is a salient feature of acute vascular rejection and chronic allograft nephropathy, yet specific blood markers of ongoing endothelial injury are currently unavailable. Circulating endothelial cells have recently been established as a novel marker of endothelial damage in a variety of vascular disorders. METHODS: We studied 129 renal transplant recipients who underwent percutaneous graft biopsy. Circulating endothelial cells were isolated with immunomagnetic anti-CD146-coated Dynabeads. Cells were stained with acridine and counted. To verify their endothelial origin, staining for Ulex europaeus lectin 1 (UEA-1) was performed in parallel. Twenty-one healthy controls were also studied. RESULTS: On biopsy, seven patients had acute vascular rejection, 15 patients had acute tubulointerstitial rejection, 14 patients had borderline rejection, and 93 patients had no rejection. Patients with acute vascular rejection had the highest cell numbers (72+/-39 cells/mL) when compared with all other patients (P<0.02). Regardless of their biopsy findings, however, all other renal transplant recipients had significantly higher numbers of circulating endothelial cells (25+/-20 cells/mL) than healthy controls (7+/-5 cells/mL, P<0.001). Finally, there was a significant correlation of cell numbers and serum cyclosporine A trough levels. By contrast, there was no correlation with serum creatinine, age, or the number of immunosuppressive drugs. CONCLUSIONS: The number of circulating endothelial cells is a novel marker of endothelial damage in renal transplant recipients. Further studies must now evaluate the origin of these cells, corroborate the clinical significance of our findings, and delineate the influence of calcineurin inhibitors.     

Factors contributing to acute rejection in renal transplantation: the role of noncompliance

Early episodes of acute rejection after renal transplantation reflect inadequate immunosuppression at a time of heightened immune challenge. Late acute rejection episodes, however, are less likely related to inadequacy of immunosuppression and may be due to patient noncompliance or overzealous weaning of immunosuppression. We evaluated 443 consecutive renal transplant recipients to determine the incidence and etiology of acute rejection. All episodes were confirmed by ultrasound-guided biopsy. The cause of each acute rejection was determined by chart review. Medication compliance was determined by history at the time of admission for biopsy. Over a follow-up period of 42 +/- 22 months, 87 patients (20%) suffered acute rejection. There was a trend toward fewer episodes of acute rejection with thymoglobulin induction and tacrolimus-based immunosuppression. Younger recipients had an increased risk of acute rejection (odds ratio 0.47, range 0.24-0.91, P = .027). Patient noncompliance with immunosuppression was associated with late acute rejection (P = .0002). Acute rejection increased the risk of allograft failure (P < .0001). Modifiable factors, including the choice of immunosuppression, reduce the risk of acute rejection. More importantly, the transplant recipient plays a substantial role in the maintenance of their allograft health through compliance with immunosuppressive drug therapy. Future strategies to improve compliance, including increased vigilance in high-risk patient groups, frequent medication review, and laboratory testing, should be encouraged.     

Eosinophilic infiltrates in a pulmonary allograft: a case and review of the literature.

An unusual case of peribronchial eosinophilic infiltrates associated with peripheral blood eosinophilia in a lung transplant patient is described. The role that eosinophils play in lung allograft rejection is reviewed. Tissue eosinophils have been associated with acute pulmonary allograft rejection. Although, eosinophils in bronchoalveolar lavage fluid (BAL) have been observed in allograft rejection, this relationship is less well defined. The role of eosinophils in the pathophysiology of allograft rejection is unclear.     

Cohort study of the prognostic significance of acute transplant glomerulitis in acutely rejecting renal allografts.

BACKGROUND: Acute transplant glomerulitis is a unique lesion in renal allografts, the prognostic significance of which is controversial. We conducted this retrospective cohort study to examine the independent prognostic significance of moderate-to-severe transplant glomerulitis in acute rejection. METHODS: Renal allograft survival for patients with acute rejection were studied, comparing one group with significant glomerulitis (G, n=28) with those with no glomerulitis (NG, n=35). Clinical, biopsy, and demographic data and renal graft survival were compared, and the association of G with graft failure was examined. RESULTS: In the G versus NG group, a greater percentage of patients were highly sensitized (peak panel reactive antibody value >80%; P=0.009), had had a previous renal transplant (40% vs. 11%; P=0.02), or had suffered from delayed graft function (P=0.03). The G group had a trend toward earlier rejection episodes (P=0.07), a significantly higher serum creatinine at the time of index biopsy (P=0.01), a higher prevalence of vascular rejection (P=0.02), and less improvement in mean reciprocal serum creatinine at 1-2 weeks after biopsy (P=0.02). Although there was a trend toward shorter allograft survival in the G group (P=0.09), the level of significance of which increased with adjustment for transplantation time period and the duration of the transplant-biopsy interval (P=0.06), the relative risk for graft loss was no longer significant when additionally adjusted for index biopsy Banff score (relative risk, 0.97; P=0.97). CONCLUSION: In this study, G was significantly more common in highly sensitized patients and was strongly associated with vascular rejection biopsies but was not an independent predictor of graft survival.     

Eosinophil granule major basic protein in acute renal allograft rejection

Conventional staining techniques to determine the presence of tissue eosinophils underestimate their number and do not usually detect eosinophil degranulation. We have studied the involvement of eosinophils in acute renal allograft rejection by immunofluorescence localization of eosinophil granule major basic protein (MBP) in the kidney and by measurement of MBP in the plasma and urine by radioimmunoassay. Tissue eosinophilia and extracellular deposition of MBP indicative of eosinophil degranulation were observed in 94% and 87%, respectively, of patients with acute rejection as compared with 17% and 17%, respectively, of patients with cyclosporine nephrotoxicity. The urine levels of MBP were significantly elevated in acute rejection but not in cyclosporine nephrotoxicity. Plasma MBP concentrations were within the normal range in both acute rejection and cyclosporine nephrotoxicity. The presence of marked tissue eosinophilia and eosinophil degranulation did not always indicate irreversible rejection. Interleukin-2 and IL-2 receptors were also elevated in the urine during acute rejection. These results support a role for the eosinophil as an effector of tissue damage during rejection and suggest the potential usefulness of urine MBP determinations for the immunologic monitoring of transplanted patients.     

Heightened peripheral blood lymphocyte CD69 expression is neither sensitive nor specific as a noninvasive diagnostic test for renal allograft rejection

It has been reported that acute allograft rejection is associated with heightened expression of the peripheral blood lymphocyte (PBL) early activation marker CD69 and that this may serve as a potential biomarker of rejection. This study sought to determine whether PBL CD69 expression correlates with both acute clinical and subclinical renal allograft rejection as well as clinically inapparent cytomegalovirus (CMV) infection. Flow cytometric determination of PBL CD69 expression was performed at the time of clinical and protocol biopsies (n = 131) in 45 renal transplant recipients. Nineteen patients also underwent weekly monitoring of PBL CD69 expression for the initial 15 wk after transplantation. Simultaneous screening for CMV viremia was performed with a semiquantitative PCR assay. No differences were seen in either CD4+ or CD8+ lymphocyte CD69 expression between the biopsy diagnoses. CMV viremia however, independent of rejection, was associated with greater CD69 expression on CD8+ lymphocytes (17.8 +/- 10.4% versus 9.6 +/- 4.8%; P < 0.0001) but not CD4+ lymphocytes. No individuals experienced clinical CMV disease. Weekly monitoring of PBL CD69 expression did not change coincident with the diagnosis of rejection; however, CMV viremia coincided with a substantial rise in the proportion of CD8+69+ lymphocytes in a number of individuals. Thus, PBL CD69 expression is neither sensitive nor specific for the noninvasive diagnosis of renal allograft rejection. Furthermore, clinically inapparent CMV viremia is associated with heightened expression of this activation marker on CD8+ lymphocytes. This latter finding suggests that clinically inapparent CMV viremia may be a potential confounder for biomarkers of rejection that examine peripheral blood lymphocytes.     

Prediction of subclinical renal allograft rejection by vascular endothelial growth factor in serum and urine

BACKGROUND: Until now subclinical renal allograft rejection has only been recognized through a protocol biopsy. The aim of this study was to assess whether measurement of vascular endothelial growth factor (VEGF) in serum and urine could be adopted as a new noninvasive tool to predict subclinical rejection. METHODS: Concentration of VEGF in serum and urine was determined by ELISA in 132 recipients of a renal allograft with stable renal transplant function who were to undergo protocol biopsy and 80 healthy controls. A conventional receiver operating characteristic (ROC) curve was used to determine the sensitivities and specificities for patients with subclinical rejection. RESULTS: Levels of VEGF in serum (126.96 +/- 20.13 pg/mL; 95% confidence interval [95% CI], 83.10-170.83) and urine (16.14 +/- 4.09 ng/mmol creatinine; 95% CI, 7.21-25.06) of 13 patients with subclinical rejection significantly differed from those of 119 patients with no allograft rejection (No-AR) and health controls. The areas under the ROC curve were 0.771 (95% CI, 0.0.64-0.901) and 0.819 (95% CI, 0.662-0.976), respectively. Levels of VEGF in serum and urine after antirejection therapy (50.45 +/- 6.58 pg/mL and 2.60 +/- 0.83 ng/mmol creatinine, respectively) were lower than those at the time of protocol biopsy. No difference in urinary and serum VEGF expression was observed between cyclosporine and tacrolimus treatment. CONCLUSION: It is first reported that the monitoring of VEGF in serum and urine might be a new noninvasive approach to supplement a protocol biopsy for detection of subclinical rejection.     

The variable nature of chronic declines in renal allograft function

Despite having important implications for the design of therapeutic trials, the clinical setting, time of onset, and rate of progression for chronic declines in renal allograft function have not been well characterized. In the present investigation, monthly estimates of glomerular filtration rate (E-GFR) were made using creatinine clearance and interim serum creatinine levels. There were 200 patients transplanted from 1978 to 1982 (precyclosporine) who survived at least 12 months with a functioning allograft. Of these, 25 had irreversible declines in E-GFR (greater than 30%) attributable to acute rejection, 50 had gradual, chronic declines in E-GFR, and 125 maintained stable function. Patients with chronic declines in E-GFR more often returned to dialysis (56%, P less than 0.001) than those with irreversible, acute reductions (24%), or stable function (2%). Chronic declines in allograft function were modeled by one or two least-squares-fitted regression lines. In most cases, the onset was early, but in 26% chronic declines in E-GFR began 2.2 +/- 1.2 (mean +/- SD) years after transplantation. Among those with chronic declines in E-GFR, 20/50 (40%) had spontaneous improvements in the rate of progression after 2.7 +/- 1.1 years and survived 8.4 +/- 2.6 years with functioning grafts, while 30/50 (60%) continued to have progressive declines in E-GFR and survived 6.1 +/- 2.5 years (P less than 0.01). Although chronic declines in E-GFR were evident 3.2 +/- 1.7 years before graft failure, routinely measured clinical and laboratory parameters from the early posttransplant period failed to predict patients who developed chronic declines in E-GFR. Altogether these data suggest that chronic declines in allograft function have an unpredictable onset and variable clinical course.     

The enhancing and sensitizing effects of donor blood components, including dendritic cells, in a rat renal allograft model

In the DA-to-Lewis renal allograft model, donor whole blood enhanced renal allograft survival (14.5 +/- 7.6 days versus 6.9 +/- 0.6 days in controls [P less than 0.01]). The effect of individual cell components given in numbers equivalent to those present in the enhancing volumes of donor whole blood was studied. Immunization with donor red cells alone produced greater enhancement than that produced by whole blood (36.14 +/- 19.5 days [P less than 0.01]). B lymphocytes also enhanced allograft survival (16.0 +/- 3.9 days [P less than 0.01]). Although slight enhancement was observed with platelets (8.5 +/- 0.6 days) and 10(5) dendritic cells (8.4 +/- 0.5 days), in terms of allograft function dendritic cell immunization produced evidence of dose-dependent accelerated rejection. A similar finding was obtained with donor T cell immunization. Donor plasma had no effect. We conclude that, although donor blood has an overall enhancing effect on renal allograft survival in this model, the sensitizing and enhancing effects can be ascribed to individual types of cells.     

The role of percutaneous transluminal dilatation in the treatment of transplant renal artery stenosis

The assessment of percutaneous transluminal dilatation (PTD) as a new therapeutic approach for transplant renal artery stenosis (TRAS) was prompted by allograft loss following surgical treatment. Seventeen (7.0%) of 243 allograft patients had TRAS greater than 80% diagnosed by angiography. The outcome of PTD in 4 TRAS patients was compared to antihypertensive drugs alone in 5 and attempted surgical repair in 8. Patients receiving antihypertensive drugs alone required 3.2 drugs at optimal doses to maintain diastolic BP less than or equal to 90 mm Hg. Malignant hypertension developed in 1 non-compliant patient. Three of the 8 surgical repairs resulted in loss of functioning grafts due to postoperative renal artery thrombosis; the remaining 5 had substantial reduction in diastolic BP (pre 112 +/- 8 mm Hg, post 93 +/- 8 mm Hg, P less than .05). Four patients with main renal artery stenosis had successful, uncomplicated PTD with no graft loss, a significant reduction in diastolic BP (pre 108 +/- 10 mm Hg, post 88 +/- 5 mm Hg, P less than .05) and less antihypertensive drug requirement (pre 3.0 +/- .8 drugs, post 1.0 +/- .8 drugs, P less than .05). Restenosis occurred in 1 patient during the 6 to 10 month follow-up period. PTD compared favourably with conventional management of TRAS as a new therapeutic approach.     

Differentiation of cytomegalovirus infection from acute rejection using renal allograft fine needle aspirates.

Cytomegalovirus (CMV) infection is an important cause of renal allograft dysfunction and may be difficult to distinguish from acute transplant rejection both clinically and histologically. To establish the early diagnosis of CMV infection, we used immunohistochemical staining with antibodies against CMV early nuclear protein (CMV-A) and histocompatibility leukocyte class II antigen (DR) in renal transplant fine needle aspirates. Fifty-eight aspirates from 27 patients were assessed, 53 for CMV-A and 53 for DR. Positive staining was defined as greater than or equal to 35% stained tubular cells for CMV-A and greater than or equal to 30% stained tubular cells for DR. Clinical diagnoses were made retrospectively without using the information obtained from aspirate diagnoses. CMV-A staining was negative in 44 aspirates, none at the time of CMV infection. CMV-A was positive in nine aspirates, seven during CMV infection (78%, P less than 0.00001 versus CMV-A negative). DR staining was never present in the absence of acute rejection. All aspirates performed during acute rejection had positive DR staining (P less than 0.00001 versus DR negative). Aspirates with acute rejection comprised 80% of all DR-positive aspirates, whereas those with CMV infection included only 13%. The percent CMV-A staining increased with CMV disease progression; DR staining decreased after successful treatment of acute rejection. These data demonstrate that CMV-A staining is associated with CMV infection whereas DR staining is not. DR staining is specifically related to acute rejection. CMV-A and DR staining of fine needle aspirates is a potentially valuable diagnostic tool to distinguish rapidly between CMV infection and acute transplant rejection as the etiology of renal allograft dysfunction.