Histopathology of serial graft biopsies from liver transplant recipients

Serial graft biopsies (n = 78) from 12 liver transplant recipients (followed clinically up to 47 months) were studied with the use of histology, histochemistry, immunostaining, and electron microscopy. Planned-protocol needle biopsy specimens were taken from the graft before removal from the donor, 1 hour after transplantation, on the eighth day, and at yearly intervals. Nonprotocol biopsies were taken when deterioration of the clinical condition made a decision on changes in the regimen necessary. The protocol biopsies provided a baseline for graft condition and diagnostic histopathologic features. In these biopsies signs of hyperacute rejection, chronic rejection, or the recipient's previous liver disease were not observed. Mild acute rejection was regularly present on the eighth day, possibly due to a lag phase in the effect of immunosuppression. The syndromes in the nonprotocol biopsies included "pure" parenchymal cholestasis, reversible acute rejection resembling chronic active hepatitis, viral infection, and acute bacterial cholangitis. Each of these syndromes correlated with a separate histopathologic entity. Therefore, these entities proved to be of diagnostic value. It is concluded that a graft biopsy may substantially add to the pathogenetic interpretation, differential diagnosis, and management of major graft syndromes in orthotopic liver transplant recipients.     

Pathologic analysis of right-lobe graft failure in adult-to-adult live donor liver transplantation

Live donor adult liver transplantation (LDALT) utilizing right-lobe grafts is now acceptable as an alternative to cadaveric orthotopic liver transplantation (OLT). However, some LDALTs fail and require urgent OLT or result in recipient death. Our aim was to determine the basis of LDALT failure. Liver specimens from 49 LDALT recipients were evaluated and the findings correlated with clinical outcome. Ten patients (20.4%) had either early (< or = 1 month) or late (> 1 month) graft failure. Eight early failures, 7 of which occurred among our first 25 cases, were due to extensive liver parenchymal necrosis as a result of hepatic artery thrombosis (n=3), portal vein thrombosis (n=1), hyperperfusion syndrome (n=1), complete graft thrombosis (n=1) with Factor V Leiden on a regimen of therapeutic heparin (n=1), sepsis and concomitant graft dysfunction with venous outflow tract injury (n=1), and venous outflow tract thrombosis and parenchymal thermal injury with sepsis (n=1). Preoperative, intraoperative, or postoperative severe vessel wall injury was evident in 6/8 early failures. Two patients had late graft failure, 1 from recurrent hepatitis C and 1 with sepsis/multisystem organ failure. There were no significant differences in graft size, rejection episodes, or operative or ischemic times between patients with and without graft failure. In conclusion, LDALT failed in 10/49 (20%) of our patients, with 8/10 occurring within 1 month post-LDALT owing to vascular/thrombotic complications experienced during the early phase of our institutional experience. Perioperative vessel wall injury appeared to be a major factor in predicting early graft loss.     

Assessment of donor liver steatosis: pathologist or automated software?

Steatosis in donor liver biopsy specimens has been shown to correlate with graft dysfunction after orthotopic liver transplantation. This 2-part (laboratory pilot, clinical retrospective) study compared the traditional interpretation of steatosis by a pathologist with an automated measurement determined by an image analysis system. In our pilot study, Sprague-Dawley rats were studied prospectively by feeding them a choline-deficient diet for up to 7 days. In our clinical group, data from 49 consecutive recipients of cadaveric liver transplantation were reviewed retrospectively. In both studies, the percentages of microvesicular fat, macrovesicular fat, and total fat content within liver biopsy specimens were determined by an automated image analysis software program and a pathologist using the same set of slides. The association between fat content of the donor liver and patient survival and graft survival, along with levels of aspartate aminotransferase, alanine aminotransferase, prothrombin time, and total bilirubin after transplantation, were also examined in the clinical study. A direct correlation was observed between levels of macrovesicular fat determined by a pathologist and the automated software using livers from rats fed a choline-deficient diet and livers from deceased donors. A significant association was observed between macrovesicular fat content in the donor liver biopsy and graft survival by both techniques. We conclude that an image analysis system can be used to automate the determination of fat content in liver biopsy specimens, and that its findings correlate with both the visual interpretation by a pathologist and graft survival. Further study is needed to determine the role of an automated technique in the evaluation of donor livers for transplantation.     

Severe steatosis as the initial histologic manifestation of recurrent hepatitis C genotype 3

Steatosis is a common finding that is seen in patients with both chronic hepatitis C and alcoholic liver disease; however, the extent of involvement in the former is generally minimal to mild. We present 2 patients who underwent live donor liver transplantation for end-stage liver disease that was caused by chronic hepatitis C (genotype 3) and alcohol abuse. Both patients presented with liver allograft dysfunction, with liver biopsy findings of moderate to marked steatosis. Exclusion of a relapse of alcohol use required intense questioning of both the patients and their families. A definitive diagnosis of recurrent hepatitis C was established by viral markers with institution of the proper therapy and resolution of graft dysfunction. We conclude that recurrent hepatitis C, particularly genotype 3, may present with severe steatosis. Recognition of this phenomenon is important, and confirmation with viral markers is necessary to provide optimal patient care.     

Steatosis and fibrosis in patients with chronic hepatitis C

BACKGROUND: Steatosis is present on liver biopsy in approximately 50% of patients with hepatitis C; its association with stage of fibrosis has been reported, but its relation to other fibrosis associated factors is unknown. Aim: To study the relation between steatosis and other histological features in patients with hepatitis C, and changes in steatosis with time. METHODS: Cross sectional study: 233 routine liver biopsies from 219 patients with hepatitis C; hepatectomy specimens from 65 patients transplanted for hepatitis C cirrhosis. Longitudinal study: 41 patients with two biopsies and 10 patients with three biopsies performed over 2-8 years. Biopsies were scored by the Ishak scheme, and degree of steatosis assessed subjectively. Multivariate analysis was used to study the interaction of fibrosis associated factors. Changes in steatosis over time in individual patients were explored in the longitudinal study. RESULTS: Steatosis was present in 50% of biopsies. It correlated strongly with fibrosis in non-cirrhotic samples, but declined in cirrhosis, and was unusual in transplant hepatectomy specimens. On multivariate analysis of non-cirrhotic biopsies, steatosis was associated with increasing patient age and remained significantly associated with fibrosis independent of portal inflammation and interface hepatitis. In the longitudinal study, steatosis persisted and increased over time, except in patients developing cirrhosis. CONCLUSIONS: Steatosis is associated with fibrosis independently of necroinflammation, but declines in cirrhosis. It may represent a pathogenic pathway distinct from necroinflammatory activity in the generation of liver fibrosis, and should be included in the assessment of biopsies for clinical and research purposes.     

Reparative regeneration of the human liver grafted from a cadaver and a relative donor

The morphological features and dynamics of regeneration of the grafted human liver were studied by using data on 609 needle biopsies obtained for morphological monitoring in 38 recipients during transplantation of the liver from cadavers and relative donors. The biopsy specimens of donor organs taken prior to grafting served as a control. Irrespective of the type of grafting, regenerative processes in the grafted liver were shown to have common regularities and to run by using the mechanisms of both intracellular regeneration and proliferation. Their maximum rate was seen within a month after grafting of a cadaverous organ and within 3 months after grafting a part of the liver from a relative donor. In cadaverous transplantation, regenerative processes depended mainly on the degree of ischemic lesions; in relative donor organ transplantation, that depended on the fitness of graft mass (the index K being close to 0.5), proliferative processes and the time of higher hepatocytic ploidity increased. An adequate evaluation of the status of a cadaverous organ, estimation of the optimum mass of a grafted hepatic part from a relative donor, and effective immunosuppressive therapy ensure required reparation rates and a complete recovery of the structure of a graft.     

Keratinocyte dysplasia in hematopoietic stem cell transplantation recipients in the day-28-to-84 posttransplantation period

Severe keratinocyte dysplasia (SKD) has been reported as a common event in the early posttransplantation period of hematopoietic stem cell transplantation patients. The purpose of our study is to determine the possible causes of SKD during the intermediate posttransplantation period and to ascertain its prevalence in skin biopsies. Skin biopsy slides, obtained from hematopoietic stem cell transplantation recipients who were days 28 to 84 posttransplantation, were evaluated for SKD. Forty-four examples of SKD were identified in 467 slides, or 9%. Thirty-seven patients were evaluated as cases in a case-control design. SKD was strongly associated with a conditioning regimen containing busulfan with an odds ratio of 7.25 (P = .0002). In?a multivariate-adjusted analysis, SKD was not associated with cyclophosphamide, fludarabine, total-body irradiation, or a nonmyeloablative conditioning regimen. SKD was not associated with clinical acute graft-versus-host disease. SKD histology gradually resolved, reaching a normal histology after an average of 241 days. This study finds that severe keratinocyte dysplasia in the period 28 to 84 days post-HSCT is strongly associated with a busulfan-conditioning regimen.     

Utility and Safety of Liver Biopsy in Patients with Undetermined Liver Blood Test Anomalies after Allogeneic Hematopoietic Stem Cell Transplantation: A Monocentric Retrospective Cohort Study

Liver blood test anomalies are common after allogeneic hematopoietic stem cell transplantation (allo-HSCT), but their cause often remains difficult to identify. Our objective was to evaluate the safety and utility of liver biopsies in patients who underwent allo-HSCT. In a retrospective single-center cohort study, we reviewed all cases of patients who underwent liver biopsy between June 2005 and July 2017. During this period, 54 biopsies were performed in 45 patients, in which 38 patients underwent allo-HSCT for malignant and 7 for nonmalignant hematological disorders. Median time between allo-HSCT and liver biopsy was 213 days. Seven biopsies were percutaneous, and 47 were transjugular. No adverse event related to the biopsy procedure occurred; 94.5% biopsies (51 of 54) led to a histological diagnosis. Cholestatic graft-versus-host disease was histologically demonstrated in 16 biopsies (30%); hepatitis-like graft-versus-host disease in 9 biopsies (17%); nonalcoholic steatohepatitis in 6 biopsies (9%); regenerative nodular hyperplasia in 4 biopsies (5%); and drug-induced liver injury, sinusoidal obstruction syndrome, and viral hepatitis each in 3 biopsies (5%). Association between clinical, laboratory, imaging and pathological features was poor. Only 34% of physicians’ prebiopsy hypotheses were confirmed by pathological findings. Patient management was influenced by liver biopsy results in 65% of cases, allowing us to identify a new diagnosis (n?=?13), rule out a differential diagnosis (n?=?14), or confirm the main hypothesis (n?=?6). In conclusion, liver biopsy is a safe and useful technique to investigate liver blood test anomalies following allo-HSCT.     

Oil Red O-assessed macrosteatosis in liver transplant donor biopsies predicts ischemia–reperfusion injury and clinical outcome

Steatosis in donor livers is an accepted adverse prognostic factor after liver transplantation. While its semiquantitative assessment shows varying reproducibility, it is questioned as a standard method. Additionally, the influence of hepatic steatosis on ischemia/reperfusion injury (I/R injury) has not been evaluated in biopsies after reperfusion. We compared different staining and analyzing methods for the assessment of donor liver steatosis in order to predict I/R injury and clinical outcome after transplantation. To do this, 56 paired pre- and post-reperfusion liver biopsies were analyzed for macro- (MaS)/micro- (MiS) and total steatosis in cryo and permanent sections by special fat (Oil Red O or ORO) and standard stains. Computerized morphometrical analyses were compared to the semiquantitative assessment by a pathologist. I/R injury was determined histopathologically and by M30 immunohistochemistry. We found ORO to be more sensitive in detecting hepatic steatosis with higher reproducibility for MaS. Semiquantitative analyses were highly reproducible and not inferior to computerized morphometry. Categorized MaS as determined by ORO correlated with the extent of I/R injury, initial poor function, liver enzymes, and survival. Therefore fat stains like ORO are a reliable and easy method comprising significant advantages in the evaluation of hepatic steatosis and are thereby of prognostic value. Computerized analysis is a precise tool though not superior to semiquantitative analyses.     

Frozen section diagnosis in donor liver biopsies: observer variation of semiquantitative and quantitative steatosis assessment

Donor livers are not generally accepted for liver transplantation if intraoperative frozen section histology on wedge biopsies provides evidence for more severe steatosis. In this reliability study, assessment of steatosis in donor liver biopsies by different approaches (frozen sections vs. paraffin sections; macrovesicular steatosis vs. microvesicular steatosis), different observers, and different evaluation methods (conventional microscopy vs. point grid analysis on digital microphotographs) was compared. One hundred twenty consecutive donor liver biopsies were investigated. Intraoperative diagnosis was made on hematoxylin and eosin (H&E)-stained frozen sections. The residual portion of each biopsy was analyzed later on H&E-, diastase-resistant PAS-, and Elastica van Gieson-stained paraffin sections. Microvesicular steatosis and macrovesicular steatosis were classified semiquantitatively into 5?% steps. Additionally, point grid counting was applied on ten digital microphotographs per slide. The values for steatosis revealed a wide range of data between 0 and 70 or 85?% (mean values, 12.0-18.3?%), considering all types of specimens. The results of the two observers were highly correlated for macrovesicular steatosis (r?≥?0.925) and for microvesicular steatosis (r?≥?0.880). The values for macrovesicular and microvesicular steatosis, however, showed poor correlation (r?≤?0.581). The rate of agreement between the two observers ranged between 84.2 and 95.8?% (κ, 0.763-0.937), depending on the threshold setting. For point grid analysis, significantly lower mean values and ranges for both types of steatosis compared to conventional histopathology were found (p?相似文献   

The clinical and pathologic implications of plasmacytic infiltrates in percutaneous renal allograft biopsies

Plasmacytic infiltrates in renal allograft biopsies are uncommon and morphologically distinctive lesions that may represent variants of acute rejection. This study sought significant clinical and pathologic determinants that might have influenced development of these lesions and assessed their prognostic significance. Renal allograft biopsies (n = 19), from 19 patients, with tubulointerstitial inflammatory infiltrates containing abundant plasma cells, composing 32 +/- 8% of the infiltrating mononuclear cells, were classified using Banff '97 criteria. Clonality of the infiltrates was determined by immunoperoxidase staining for kappa and lambda light chains and polymerase chain reaction for immunoglobulin heavy-chain gene rearrangements, using V(H) gene framework 3 and JH consensus primers. In situ hybridization for Epstein-Barr virus encoded RNA (EBER) was performed in 17 cases. The clinical features, histology, and outcome of these cases were compared with kidney allograft biopsies (n = 17) matched for time posttransplantation and type of rejection by Banff '97 criteria, with few plasma cells (7 +/- 5%). Sixteen of 19 biopsies (84%) with plasmacytic infiltrates had EBER-negative (in 14 cases tested) polyclonal plasma cell infiltrates that were classifiable as acute rejection (types 1A [4], 1B [10], and 2A [2]). These biopsies were obtained between 10 and 112 months posttransplantation. Graft loss from acute and/or chronic rejection was 50% at 1 year and 63% at 3 years, and the median time to graft failure was 4.5 months after biopsy. There was no significant difference in overall survival or time to graft failure compared with the controls. Three of 19 biopsies (16%) had EBER-negative polyclonal plasmacytic hyperplasia, mixed monoclonal and polyclonal polymorphous B cell hyperplasia, and monoclonal plasmacytoma-like posttransplantation lymphoproliferative disease (PTLD) and were obtained at 17 months, 12 weeks, and 7 years after transplantation, respectively. Graft nephrectomies were performed at 1, 19, and 5 months after biopsy, respectively. Plasmacytic infiltrates in renal allografts comprise a spectrum of lesions from acute rejection to PTLD, with a generally poor prognosis for long-term graft survival.     

Early CD3 peripheral blood chimerism predicts the long-term engrafting unit following myeloablative double-cord blood transplantation

After double-cord blood transplantation, long-term hematopoietic dominance of a single-cord blood donor graft is established in the majority of patients; however, the mechanism behind this observation remains largely unknown. Beginning at day 7 posttransplantation, we prospectively measured weekly lineage-specific peripheral blood donor chimerisms in patients undergoing myeloablative double-cord blood transplantation to evaluate whether the degree of early donor contribution to specific lineage(s) would predict the long-term engrafting unit. Our results demonstrate that the donor unit with higher CD3 chimerism at day 7 became the dominant engrafting unit in 26 of 31 evaluable patients (P = .0002) and in 34 of 34 evaluable patients at day 14 (P < .0001). Similarly, higher donor unit CD33 chimerism was associated with dominant engraftment in 8 of 8 (day 7) and in 31 of 32 (day 14) evaluable patients. No statistically significant correlation between the dominant unit and order of infusion, infused total nucleated cells, CD34, or CD3 cell doses, unit viability, or HLA disparity was observed. The correlation of higher early posttransplantation donor CD3 peripheral blood chimerism with the dominant unit suggests a rapid immune-mediated response as a primary mechanism of action for long-term single-donor dominance. This finding may have clinical implications for early selection of the winning unit after double-cord blood transplantation and for novel cord blood manipulation strategies.     

乙型肝炎病毒感染者肾移植前肝穿刺活检与预后分析

背景：乙肝病毒感染者肾移植手术国内有较多报道,但乙肝病毒感染者肾移植前肝穿刺活检观察有限。 目的：对慢性肾功能衰竭合并不同程度慢性乙型病毒性肝炎患者进行肾移植前肝穿刺活检,移植后2年随访观察转归情况。 方法：对接受肾移植的21例乙型肝炎病毒感染的尿毒症患者进行肝穿刺活检。根据肝活检组织病理学改变,分为轻度(n=9)、中度(n=7)、重度(n=5)3组。肾移植后随访观察2年。3组中各有2例进行重复肝活检组织病理学检查。 结果与结论：轻度慢性乙型肝炎组在随访2年中各项观察指标均无明显变化。中度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性明显高于正常水平,随访至终点时,2例重复肝活检病理显示已处于重度病变。重度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性持续高于正常水平;18个月开始,血清白蛋白水平低于正常值,球蛋白水平高于正常值;随访至终点时,有4例呈肝硬化改变。提示不同程度的慢性乙型病毒性肝炎患者肾移植后预后不同,肝活检是评价肝脏病变程度的重要手段,具有指导肾移植选择的作用。     

Kidney transplant pathology

Biopsy of the transplanted kidney plays an important role in the care and treatment of patients after kidney transplantation. Today the renal biopsy is a standard procedure which is performed early after renal transplantation in the case of a primary non-functioning graft or a significant rise in serum creatinine. On the other hand, a kidney biopsy is performed if an acute or creeping rise in serum creatinine or acute onset of proteinuria or erythrocyturia is observed during follow-up. Furthermore, zero biopsies or intraoperative biopsies of the graft are important in order to obtain information about the initial quality of the graft. This is particularly important in view of the shortage of donor organs and the resulting necessity to accept increasingly marginal organs, such as for example in the ESP program. In addition, an increasing number of transplant centres perform protocol biopsies, i.e. biopsies that are not based on clinical indication, but are performed at a certain time point after transplantation to detect subclinical rejections as well as histological alterations pointing to chronic allograft damage. Additionally, there is much scientific interest in protocol biopsies.     

活体肝移植中轻度脂肪变性供肝的应用

背景：在活体肝移植中使用脂肪变性供肝不但影响供者的安全,同时也影响受者的生存。 目的：评价活体肝移植中使用轻度脂肪变性供肝时供者的安全性及受者预后情况。 方法：回顾性分析104例成人间右半肝活体肝移植的资料,根据移植过程中供肝活检病理标本的脂肪变性程度将所有病例分成4个组。比较各组移植供受者移植后2周的肝体积增生率,分析104例成人间活体右半肝肝移植受者移植后死亡情况及原因。 结果与结论：4组病例在供受者移植后肝功能的恢复和受者后移植预后无明显差别,没有肝功能延迟恢复和原发无功发生。轻度大泡性脂肪肝者只要残肝足够可以成为合适的活体肝移植供者。使用轻度大泡性脂肪肝并不增加受者病死率和移植物失功。     

Recurrent hepatitis B in liver allograft recipients. Differentiation between viral hepatitis B and rejection.

The histologic findings in the original liver obtained from 9 liver allograft patients with active B virus hepatitis were compared with 28 posttransplant pathology specimens. All specimens were studied with the use of light and immunohistochemical microscopy in conjunction with pertinent clinical data. Eight of the 9 patients had chronic active hepatitis B (HB) with cirrhosis, prior to transplant, one of which had coexistent hepatocellular carcinoma. The ninth patient had fulminant hepatic necrosis secondary to acute HB prior to transplantation. In all of the patients with chronic HB prior to transplantation who survived more than 2 months after transplantation recurrent infection of the graft developed despite perioperative HB immunoglobulin therapy. The patient with acute fulminant hepatitis B pretransplant has done well postoperatively and has evidence of HB virus immunity (positive anti-HBs) 15 months after transplantation. Examination of tissue specimens obtained during episodes of allograft dysfunction in these 9 patients indicate that pathologic alterations of active HB infection of the allograft are associated with a preferential lobular insult, whereas those occurring in rejection preferentially involve portal tract structures. Serologic data combined with biopsy histopathologic data are essential in distinguishing between the two quite different events.     

Transplantationspathologie der Leber

Histopathology plays an important role in the diagnosis of graft complications following liver transplantation (LTX) and includes the diagnosis of underlying liver disease, assessment of the donor liver before LTX and control biopsies after LTX. Within the early period after LTX (time zero and first month) preservation/reperfusion injury, as well as hyperacute and acute rejection may occur. Surgical vascular or biliary complications can cause parenchymal morphological changes. Within 12 months following LTX, histological signs of opportunistic infections and chronic rejection are frequent findings and disease recurrence is typical beyond the first year. Drug toxicity in liver allograft recipients can be induced by immunosuppressive therapy or other drugs. A high percentage of adult patients reveal histological features of idiopathic chronic hepatitis 6 months after LTX. Histopathological differential diagnosis of the combined underlying causes or complications is often difficult.     

Centrilobular histopathologic changes in liver transplant biopsies

We evaluated centrilobular histologic changes seen on post-orthotopic liver transplantation (OLT) biopsies to refine the pathologic diagnosis by systematic study of morphologic and clinical data with possible identification of prognostic criteria. A total of 110 biopsies with zone 3 pathology from 59 patients were reviewed and correlated with clinical findings. Within the first 6 months post-OLT (group I), 39 of 47 patients had combinations of centrilobular hepatocytic dropout, ballooning, and cholestasis on single or multiple biopsies attributed to perioperative ischemic/perfusion injury; 12 of 39 patients with all 3 features present had increased incidence of biliary complications and sepsis and decreased 1-year patient and graft survival; 17 of 39 patients with 2 of the 3 features had increased biliary complications but not decreased 1-year survival; and the remaining 8 of 47 patients had central venulitis associated with acute cellular rejection. After 6 months post-OLT (group II), 14 patients, including 2 from group I, had biopsies with centrilobular pathology; 8 of 14 had central venulitis related to rejection (acute, 4; chronic, 4), and fibrosis was seen in 8 (rejection, 6; cardiac problems, 2). In conclusion, combinations of centrilobular hepatocytic ballooning, dropout, and cholestasis are seen in association with reversible or irreversible ischemic/perfusion damage in the early post-OLT period. The presence of all 3 features is associated with a poor outcome. Central venulitis as a feature of acute/chronic rejection is seen at any time post-OLT and is not a predictor of poor graft/patient survival.