Interstitial Mononuclear Cell Research Articles

Screening of vesicoureteral reflux in pediatric patients with kidney transplantation showing non‐specific interstitial fibrosis and tubular atrophy with interstitial Tamm–Horsfall protein deposits in protocol allograft biopsy

Interstitial fibrosis and tubular atrophy (IF/TA) in kidney allografts are induced by multiple factors, and although much effort has been devoted on the classification of IF/TA, clarification of the causes of non-specific IF/TA is equally important for appropriate therapy. Tamm-Horsfall protein (THP) in renal tissue can be a useful marker for the histological expression of urine backflow and suspected vesicoureteral reflux (VUR). Here, we examined the presence of VUR in pediatric recipients with interstitial THP deposits in kidney allografts to clarify the cause of non-specific IF/TA. Ten pediatric patients showing interstitial THP deposits with non-specific IF/TA were enrolled and voiding cystourethrography was performed. Major histological findings of these patients were interstitial mononuclear cell infiltration and fibrosis associated with interstitial THP deposits. The semiquantitative grading scores of IF/TA were as follows: (i) mild IF/TA (n = 3); (ii) moderate IF/TA (n = 3); and (iii) severe IF/TA (n = 4). In the severe grade, diffuse interstitial mononuclear cell infiltrates were prominent with the appearance of thyroidization classically observed in chronic pyelonephritis. Eight of ten patients (80%) had VUR into the graft. Although symptomatic pyelonephritis was not observed in any of the patients, asymptomatic bacteriuria was detected in 40.0% of the patients. There was no significant correlation between VUR grade and IF/TA histological score. The patients without VUR also showed mild or severe IF/TA. Therefore, VUR and urinary flow stasis accompanied by asymptomatic urinary tract infection appear to be the causes of interstitial mononuclear cell infiltration and fibrosis associated with interstitial THP deposits in kidney allografts.

Up-regulated renal expression of TNF-α signalling adapter proteins in lupus glomerulonephritis

The role of tumour necrosis factor (TNF-alpha) signalling adapters in lupus nephritis (LN) is poorly understood. This study investigated renal expression of TNF-alpha and TNF signalling adapter proteins, including TNF receptor-associated death domain protein (TRADD), receptor-interacting protein (RIP) and TNF receptor-associated factor-2 (TRAF-2) in patients with LN. The renal expression of proliferating cell nuclear antigen (PCNA) and CD68 was also measured. The study showed that glomerular and tubular expression of TNF-alpha, TRADD, RIP and TRAF-2 was significantly up-regulated in class III and IV LN in which the intense staining was observed on the crescents, proximal and distal tubules and interstitial mononuclear cells. The number of PCNA-positive cells and CD68-positive cells (macrophages) was increased obviously in class III and IV LN. There was a correlation between the expression levels of TNF-alpha, TRADD, RIP, TRAF-2 and the number of PCNA-positive or CD68-positive cells and active index of renal pathology. These findings suggest that TNF-alpha and TNF-alpha adapters in patients with LN play a role in immunopathogenic injury via transmitting abnormal cell proliferating and proinflammatory signals. The findings have provided further insights into the role of TNF-alpha and its adapter proteins in the pathogenesis of LN and have important therapeutic implications.

Hemin therapy attenuates kidney injury in deoxycorticosterone acetate-salt hypertensive rats

Upregulating the heme oxygenase (HO) system removes the prooxidant heme, and thus is cytoprotective. Additionally, the products from the HO pathway including, carbon monoxide, bilirubin, and biliverdin, scavenge reactive oxygen species, inhibit lipid peroxidation, and suppress tissue inflammation, while the iron formed enhances the synthesis of the antioxidant ferritin. Deoxycorticosterone acetate (DOCA)-salt hypertension, a model of human primary aldosteronism, causes oxidative stress and impairs renal function by stimulating inflammatory/oxidative transcription factors such as NF-kappaB and activating protein (AP-1). The effect of the HO system in end-organ damage in mineralocorticoid-induced hypertension has not been fully characterized. In this study, the administration of the HO inducer hemin lowered blood pressure (191 vs. 135 mmHg; n = 22, P < 0.01), increased creatinine clearance, and reduced kidney hypertrophy proteinuria, albuminuria, and histopathological lesions, including glomerular hypertrophy, glomerulosclerosis, tubular dilation, tubular cast formation, and interstitial mononuclear cell infiltration in nephrectomy/DOCA-high-salt-hypertension. The renoprotection was accompanied by reduced levels of NF-kappaB, AP-1, fibronectin, transforming growth factor (TGF)-beta, and 8-isoprostane, a marker of oxidative stress. Correspondingly, a robust increase in total antioxidant capacity, HO activity, cGMP, and an antioxidant like ferritin was observed in hemin-treated animals. Our findings suggest that suppression of oxidative/inflammatory insults alongside the corresponding decline of fibronectin and TGF-beta, an activator of extracellular matrix proteins, may account for the attenuation of renal histopathological lesions and the antihypertrophic effects of hemin. The multifaceted interaction among the HO system, TGF-beta, fibronectin, AP-1, and NF-kappaB may be explored to design new drugs against end-stage-organ damage.

Clinical Variables Associated With the Presence of Inflammatory Infiltrates in Patients With Dilated Cardiomyopathy Undergoing Heart Transplantation

Introduction Idiopathic dilated cardiomyopathy (DCM) is, together with ischemic heart disease, the major cause of end-stage heart failure leading to heart transplantation. However, an unknown percentage of patients with this diagnosis has inflammatory foci found in the histopathological study of the explanted heart. This fact suggests an undetected process of acute myocarditis as the cause of cardiac dysfunction. Objective The objective of this study was to identify clinical and echocardiographic variables related to the presence of myocardial infiltrates, as a potential guide to determine which patients should undergo endomyocardial biopsy in DCM. Materials and Methods We retrospectively analyzed 161 patients who underwent heart transplantation with a diagnosis of DCM between 1987 and 2007. The presence of inflammatory infiltrates was considered significant when the histopathological study of tissue blocks from the left ventricle showed 1 or more foci per cm 2 of perivascular or interstitial mononuclear or polymorphonuclear cells, whether or not in the presence of cytolysis. Results Seventeen patients (11%) had these inflammatory histological findings; of them, 6 (35%) showed preponderance of eosinophils and 7 (41%) showed areas of cytolysis. The DCM group with inflammatory infiltrates showed significant differences in terms of younger age (45 ± 15 vs 50 ± 11 years; P < .01) and smaller ventricular diameters ( P < .05). Male gender was more frequent in this group, and the patients had a poorer clinical status and greater dependence on inotropic drugs. Conclusions Inflammatory infiltrates are frequently present in DCM explanted hearts. Although there are no relevant clinical variables to identify subclinical myocarditis, these patients are younger and have smaller ventricular diameters and poorer functional status at the time of transplantation.

Vascular Endothelial Growth Factor-A and Platelet-Derived Growth Factor-B Combination Gene Therapy Prolongs Angiogenic Effects via Recruitment of Interstitial Mononuclear Cells and Paracrine Effects Rather Than Improved Pericyte Coverage of Angiogenic Vessels

Vessel stabilization and the inhibition of side effects such as tissue edema are essential in angiogenic gene therapy. Thus, combination gene transfers stimulating both endothelial cell and pericyte proliferation have become of interest. However, there is currently little data to support combination gene transfer in large animal models. In this study, we evaluated the potential advantages of such a strategy by combining the transfer of adenoviral (Ad) vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF)-B into rabbit hindlimb skeletal muscle. AdLacZ alone or in combination with AdVEGF-A were used as controls. Contrast-enhanced ultrasound, modified Miles assay, and immunohistology were used to quantify perfusion, vascular permeability, and capillary size, respectively. Confocal microscopy was used in the assessment of pericyte-coverage. The transfer of AdPDGF-B alone and in combination with AdVEGF-A induced prominent proliferation of alpha-smooth muscle actin-, CD31-, RAM11-, HAM56-, and VEGF- positive cells. Although, pericyte recruitment to angiogenic vessels was not improved, combination gene transfer induced a longer-lasting increase in perfusion in both intact and ischemic muscles than AdVEGF-A gene transfer alone. In conclusion, intramuscular delivery of AdVEGF-A and AdPDGF-B, combined, resulted in a prolonged angiogenic response. However, the effects were most likely mediated via paracrine mechanisms rather than an increase in vascular pericyte coverage.

Pathology of hypersensitivity pneumonitis

Hypersensitity pneumonitis, caused by inhalation of various antigens, is characterized by interstitial mononuclear cell infiltration, nonnecrotizing granulomas, cellular bronchiolitis, and fibrosis. The pathological picture of chronic hypersensitivity pneumonitis is, however, complicated; it is sometimes difficult to differentiate chronic hypersensitivity pneumonitis from idiopathic pulmonary fibrosis/usual interstitial pneumonia, nonspecific interstitial pneumonia, and connective-tissue-related lung disease. The clinical, radiological, and pathological features of chronic hypersensitivity pneumonitis have recently been described. This study reviews the previously reported information and provides new insights into the pathological features of chronic hypersensitivity pneumonitis. The pathological features of chronic hypersensitivity pneumonitis comprise overlapping usual interstitial pneumonia-like pattern with subpleural patchy fibrosis, alternating normal alveoli and fibroblastic foci, a nonspecific interstitial pneumonia-like pattern, and centrilobular fibrosis. In contrast to pathological features of acute and subacute hypersensitivity pneumonitis, epithelioid cell granulomas are sparse or absent, but giant cells are seen in the interstitium. Bridging fibrosis between peribronchiolar area and perilobular areas is an outstanding feature of chronic hypersensitivity pneumonitis. Autopsy cases of chronic hypersensitivity pneumonitis have demonstrated not only upper lobe contraction but also lower lobe contraction, mimicking usual interstitial pneumonia pattern and diffuse alveolar damage. The present review focuses on the pathological features of chronic hypersensitivity pneumonitis and presents that centrilobular fibrosis and bridging fibrosis are the important hallmarks of chronic hypersensitivity pneumonitis, even with a usual interstitial pneumonia-like pattern.

Cyclosporin A (CsA) modulates the glomerular production of inflammatory mediators and proteoglycans in experimental nephrosis.

Nephrosis is characterized by glomerular epithelial cell injury and a decrease in the glomerular basement membrane (GBM) proteoglycan content. Although CsA is a useful treatment for a group of patients with this disease, its mechanism of action is unclear. We have previously shown that in experimental nephrosis there is an increase in the glomerular production of tumour necrosis factor-alpha (TNF-alpha) and platelet-activating factor (PAF). Here we have studied the effect of CsA on kidney generation of TNF-alpha and PAF in puromycin aminonucleoside (PAN) nephrosis as well as on the synthesis of proteoglycans by cultured glomerular epithelial cells. Rats receiving CsA had, on day 8 of PAN injection, a significant reduction in proteinuria, blood cholesterol levels and in interstitial mononuclear cells. A diminution in glomerular production and urinary excretion of TNF-alpha and PAF was also noted. In in vitro studies, at 24 h of incubation PAF and TNF-alpha induced in glomerular epithelial cells a significant decrease in proteoglycan synthesis. Neither PAF nor TNF-alpha had any significant effect on glomerular epithelial cell proliferation. CsA alone induced a dose-response increase in proteoglycan synthesis and a slight decrease in cell proliferation. CsA also reversed the inhibitory effect of PAF and TNF-alpha on proteoglycan synthesis. However, CsA did not alter the pattern of proteoglycan production, remaining around 50% chondroitinase ABC-, 15% heparitinase-sensitive. Our results indicate that PAF and TNF-alpha could be implicated in the pathogenesis of nephrosis through the inhibition of proteoglycan synthesis by glomerular epithelial cells. The beneficial effect of CsA in nephrosis may be due to the recovery of the GBM charge selectivity caused by the normalization of glomerular PAF and TNF-alpha synthesis and the increase in proteoglycan synthesis by glomerular epithelial cells.

The effect of cyclosporin A on the interstitial mononuclear cell infiltration and the induction of Heymann's nephritis

Heymann's nephritis was induced with brush-border (BB) antigen. Interstitial mononuclear cell infiltration was studied with cytological examinations of fine-needle aspiration biopsies (FNAB), and with immunoperoxidase stains of frozen sections with monoclonal antisera. The effect of cyclosporin A (CyA), 20 mg/kg when administered intraperitoneally for 8 days in association with both initial immunization, and with the booster 4 weeks later, on the interstitial leukocyte infiltration and on the development of membranous glomerulonephritis (MGN) and proteinuria were investigated. Another group of rats was immunized, but not given CyA. Experimental animals were killed in groups 3, 6 and 20 weeks after initial immunization. CyA inhibited significantly the initial interstitial lymphocyte and blast cell response at 3 weeks (FNAB), but did not inhibit the secondary response after the booster. The anti-BB titre reacted in a similar fashion. Immunoperoxidase stains indicated a clearly suppressed T suppressor/cytolytic (T s/c) cell response. Glomerular basement membrane (GBM) deposits of IgG developed more slowly and were more scarce in the CyA-treated rats, when compared with the untreated group. Only one out of 15 CyA treated rats developed C3 deposits in the GBM during the course of the study, and none developed proteinuria, when most untreated rats (10/17) had C3 deposits and were nephrotic at 20 weeks. Thus, CyA depressed the initial interstitial cellular response after immunization with BB antigen, and also inhibited the development of antibody response, C3 deposits and proteinuria of Heymann nephritis. These effects of CyA may be contributed to an inhibited amplification of the autoimmune response associated with interstitial damage and continuous release of autoantigen.

Functional and phenotypic characteristics of testicular macrophages in experimental autoimmune orchitis

Testicular inflammation with compromised fertility can occur despite the fact that the testis is considered an immunoprivileged organ. Testicular macrophages have been described as cells with an immunosuppressor profile, thus contributing to the immunoprivilege of the testis. Experimental autoimmune orchitis (EAO) is a model of organ-specific autoimmunity and testicular inflammation. EAO is characterized by an interstitial inflammatory mononuclear cell infiltration, damage of the seminiferous tubules and germ cell apoptosis. Here we studied the phenotype and functions of testicular macrophages during the development of EAO. By stereological analysis, we detected an increased number of resident (ED2+) and non-resident (ED1+) macrophages in the testicular interstitium of rats with orchitis. We showed that this increase was mainly due to monocyte recruitment. The in vivo administration of liposomes containing clodronate in rats undergoing EAO led to a reduction in the number of testicular macrophages, which correlated with a decreased incidence and severity of the testicular damage and suggests a pathogenic role of macrophages in EAO. By immunohistochemistry and flow cytometry we detected an increased number of testicular macrophages expressing MHC class II, CD80 and CD86 costimulatory molecules in rats with orchitis. Also, testicular macrophages from rats with EAO showed a higher production of IFNgamma (ELISA). We conclude that testicular macrophages participate in EAO development, and the ED1+ macrophage subset is the main pathogenic subpopulation. They stimulate the immune response through the production of pro-inflammatory cytokines and antigen presentation and thus activation of T cells in the target organ.

The extent of parasite-associated necrosis in the placenta and foetal tissues of cattle following Neospora caninum infection in early and late gestation correlates with foetal death

The protozoan parasite Neospora caninum is the most frequently diagnosed abortifacient in the UK and a leading cause of abortion worldwide but the mechanisms leading to abortion are not fully understood. The distribution of parasites and the histopathological changes in the placenta and foetus were compared in 12 cows following experimental infection of cattle with N. caninum in early ( n = 6) and late ( n = 6) gestation, by PCR, immunohistology, light microscopy and transmission electron microscopy. Twelve uninfected pregnant cattle were used as controls. Infection in early gestation led to foetal death. In the placentae of cattle immediately following foetal death, N. caninum DNA was detected and there was evidence of widespread parasite dissemination. This was associated with extensive focal epithelial necrosis, serum leakage and moderate maternal interstitial mononuclear cell infiltration. In the foetuses, parasites were evident in all tissues examined and were associated with necrosis. In the placenta of cattle infected in late gestation, N. caninum DNA was detected sporadically but parasites were not evident immunohistologically. Small foci of necrosis were seen associated with mild interstitial mononuclear cell infiltration. Detection of N. caninum DNA in the foetuses was sporadic and parasites were demonstrated immunohistologically in brain and spinal cord only, with an associated mononuclear cell infiltration. This data is consistent with uncontrolled parasite spread in an immunologically immature foetus and could, via multiparenchymal necrosis of foetal tissues or the widespread necrosis and inflammation observed in the placenta, be the cause of Neospora-associated abortions.

Protease-Activated Receptor-2 Augments Experimental Crescentic Glomerulonephritis

Protease-activated receptor-2 (PAR-2) is a cellular receptor expressed prominently on epithelial, mesangial, and endothelial cells in the kidney and on macrophages. PAR-2 is activated by serine proteases such as trypsin, tryptase, and coagulation factors VIIa and Xa. It induces pleiotropic effects including vasodilatation, increasing plasminogen activator inhibitor (PAI-1) expression, mesangial cell proliferation, and cytokine production by macrophages. The role of PAR-2 in renal inflammation was studied in antiglomerular basement membrane antibody-induced crescentic glomerulonephritis (CGN) using PAR-2-deficient (PAR-2(-/-)) mice and wild-type littermate controls. PAR-2(-/-) mice had reduced crescent formation, proteinuria, and serum creatinine compared with wild-type mice 21 days after initiation of CGN. Glomerular accumulation of CD4(+) T cells and macrophages and the number of proliferating cells in glomeruli were similar in both groups. Glomerular fibrin deposition was significantly reduced in PAR-2(-/-) mice, and this was associated with reduced renal plasminogen activator inhibitor expression and increased renal matrix-metalloprotinase-9 activity. These results demonstrate a proinflammatory role for PAR-2 in CGN that is independent of effects on glomerular leukocyte recruitment and mesangial cell proliferation. PAR-2-mediated augmentation of renal plasminogen activator inhibitor expression and inhibition of matrix-metalloprotinase-9 activity may contribute to increased glomerular fibrin accumulation and glomerular injury in CGN.

A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, enhanced nitric oxide production in obstructed rat kidney

Background The persistent renal hemodynamic maladjustment caused by imbalances between vasoactivators predisposes the kidney to tubulointerstitial injury and ultimate interstitial fibrosis. The decoction (A&A) of a combination of roots of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has shown antifibrotic effects in rats with chronic kidney diseases and improvement of renal blood flow in rats with acute ischemic renal injury. In the present study, we investigated the effects and possible mechanisms of A&A on vasoactivators in the process of renal interstitial fibrosis. Methods Male Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After oral administration of A&A (14 g/kg/d) for 3, 7 and 10 days, morphological changes were evaluated by HE, Masson and Sirius red staining technique. The levels of Ang-II, ET-1, and the activities of different nitric oxide synthases (NOSs) in renal homogenate were measured by radioimmunoassay. The nitrite concentration as nitric oxide (NO) production was measured using the Griess reagent. Western blot analysis and immunohistochemical staining were performed to determine the expressions of eNOS, nNOS, and iNOS in the kidney. The ability of scavenging reactive oxygen species (ROS) was evaluated by spectrophotometry. Results Morphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which reduced by A&A administration ( p < 0.05, UAA vs. UUO group). The levels of Ang-II and ET-I were increased in obstructed kidneys, but not significantly changed after A&A administration. NO production did not change in obstructed kidney at day 3 but increased in day 7 and day 10. Administering A&A progressively increased NO production by 2.2, 1.2, and 1.2 fold at days 3, 7 and 10, respectively. The activities of constitutive NOS and iNOS were comparable between UUO group and sham group. In contrast, the activity of constitutive NOS was much higher in UAA than that of UUO rats, which increased 78%, 68% and 78% at days 3, 7 and 10 respectively, although the protein expression of eNOS, nNOS and iNOS in renal tissue had no change in UAA rats. The activities of scavenging ROS in UUO group were not significantly different from the sham group at days 3 and 7, but increased at day 10 (24.1 ± 15.0 vs. 10.1 ± 0.8 U/min/mg protein, p < 0.05). After A&A administration, the activities of scavenging ROS were significantly increased at days 3 and 7 (51.5 ± 17.9 vs. 11.7 ± 7.4 U/min/mg protein, p < 0.05; and 16.1 ± 5.6 vs. 7.7 ± 1.4 U/min/mg protein, p < 0.05) respectively, comparing with the UUO group. Conclusion The anti-fibrosis effects of A&A might be associated with enhancing NO production via eNOS activation and scavenging ROS, and in turn might improve ischemic microvasculature and attenuate interstitial fibrosis.

A CIAS1 mutation in a Japanese girl with familial cold autoinflammatory syndrome

Abbreviations ACU acquired cold urticaria CIAS1 cold-induced autoinflammatory syndrome 1 CINCA chronic infantile neurologic cutaneous and articular FCAS familial cold autoinflammatory syndrome MWS Muckle-Wells syndrome NOMID neonatal-onset multisystem inflammatory disease Here we describe a Japanese girl with familial cold autoinflammatory syndrome (FCAS) and identify the mutation in the CIAS1 gene, encoding a cryopyrin protein expressed mainly in phagocytic cells. A 14-year-old girl was referred to our hospital because of a rash, fever, and arthralgia. From 4 months of age, she had suffered from intermittent episodes of pruritic rash. At the age of 13 years, she had developed severe pain in the right hip joint and a gait disturbance and had been admitted to a local hospital. The patient then experienced intermittent right arthralgia and lumbago and was treated with nonsteroidal anti-inflammatory drugs. Physical examination revealed discrete urticarial plaques 2–10 mm in diameter surrounded by macular erythema distributed across the cheeks and lower and upper extremities (Fig. 1a). After exposure to cold air, the urticarial rash worsened and conjunctivitis appeared. The ice cube test result was negative. The patient’s father and paternal grandmother seemed to also have a similar urticarial rash and conjunctivitis after cold exposure. Laboratory tests showed an elevated white blood cell count of 13,500/μl with 79% neutrophils, an elevated erythrocyte sedimentation rate of 44 mm/h, and an elevated C-reactive protein concentration of 4.4 mg/dl. Skin biopsy revealed moderate superficial and minimal deep perivascular inflammation with interstitial neutrophils and mononuclear cells (Fig. 1b). Patients have long been diagnosed as having acquired cold urticaria (ACU) because FCAS has not been reported in Japan. However, in this case, an ice cube test was negative, and the patient was suspected Eur J Pediatr (2008) 167:245–247 DOI 10.1007/s00431-007-0451-x

Increased Apoptosis and Proliferative Capacity are Early Events in Cyst Formation in Autosomal-Dominant, Polycystic Kidney Disease

Previous studies have highlighted epithelial proliferation and apoptosis in the cyst lining as common features in animal models of cystic disease. In this study, we sought to evaluate the timing and extent of these changes in renal tissue obtained from patients with autosomal-dominant, polycystic kidney disease (ADPKD) subjected for nephrectomy for a variety of clinical indications. Cell proliferation was assessed using an antibody to proliferating cell nuclear antigen (PCNA), and apoptosis was evaluated by the use of terminal deoxynucleotidyl transferase (TdT) digoxigenin-deoxyuridine (dUTP) nick end-labeling technique (ApopTag). The origin of cystic structures was evaluated using antibodies to epithelial membrane antigen (EMA). The lineage of interstitial mononuclear cells was assessed by anti CD 45 and CD 68 monoclonal antibodies. We found an increased rate of proliferation within the epithelium, not only of cystic, but also of noncystic, tubules that was significantly higher than the corresponding values from normal kidney (p ≤ 0.0001). Apoptotic index values were significantly increased within the epithelium lining noncystic and cystic structures (p < 0.001). In the interstitium, increased proliferation and apoptosis rates were also noted. Interstitial infiltrates were dense and consisted mainly of CD 68–positive macrophages and CD 45–positive lymphocytes. The present study demonstrated that changes in cell turnover are early events in cyst formation. The observation of mild proportionate elevation of both proliferation and apoptosis values of the epithelium lining cysts explains the lack of increase risk of renal cell carcinoma in ADPKD. The development of heavy interstitial inflammation could contribute to progressive tubulointerstitial scarring, leading to progressive renal failure.

Role of Ellagic Acid against Cisplatin‐Induced Nephrotoxicity and Oxidative Stress in Rats

The aim of this study was to investigate the possible protective role of antioxidant treatment with ellagic acid on cisplatin-induced nephrotoxicity using biochemical and histopatological approaches. Adult male Sprague-Dawley rats were randomly divided into four groups. The control group received 0.9% saline; animals in the ellagic acid group received only ellagic acid (10 mg/kg); animals in the cisplatin group received only cisplatin (7 mg/kg); animals in the cisplatin + ellagic acid group received ellagic acid for 10 days after cisplatin. The effects of ellagic acid on cisplatin-induced nephrotoxicity were evaluated by plasma creatinine, urea, sodium and calcium concentrations; kidney tissue malondialdehyde, reduced glutathione (GSH), glutathione peroxidase (GSH peroxidase) and catalase activities and histopatological examinations. Administration of cisplatin to rats induced a marked renal failure, characterized by significant increases in plasma creatinine, urea and calcium concentrations. Cisplatin also induced oxidative stress, as indicated by increased kidney tissue concentrations of malondialdehyde, and reduced activities of GSH peroxidase and catalase. Furthermore, treatment with cisplatin caused a marked tubular necrosis, degeneration and desquamation, luminal cast formation, karyomegaly, tubular dilatation, interstitial mononuclear cell infiltration and inter-tubular haemorrhagia. Ellagic acid markedly reduced elevated plasma creatinine, urea and calcium levels and counteracted the deleterious effects of cisplatin on oxidative stress markers. In the same way, ellagic acid ameliorated cisplatin-induced pathological changes including tubular necrosis, degeneration, karyomegaly, tubular dilatation when compared to the cisplatin alone group. These results indicate that the antioxidant ellagic acid might have a protective effect against cisplatin-induced nephrotoxicity and oxidative stress in rat, but not enough to inhibit cisplatin-induced renal dysfunction.

Morphological studies in a model for dengue-2 virus infection in mice

One of the main difficulties in studying dengue virus infection in humans and in developing a vaccine is the absence of a suitable animal model which develops the full spectrum of dengue fever, dengue haemorrhagic fever, and dengue shock syndrome. It is our proposal to present morphological aspects of an animal model which shows many similarities with the dengue infection in humans. BALB/c mice were intraperitoneally infected with non-neuroadapted dengue virus serotype 2 (DENV-2). Histopathological and morphometrical analyses of liver tissue revealed focal alterations along the infection, reaching wide-ranging portal and centrolobular veins congestion and sinusoidal cell death. Additional ultrastructural observations demonstrated multifocal endothelial injury, platelet recruitment, and alterated hepatocytes. Dengue virus antigen was detected in hepatocytes and in the capillar endothelium of the central lobular vein area. Liver function tests showed high levels of aspartate transaminase and alanine transaminase enzyme activity. Lung tissue showed interstitial pneumonia and mononuclear cells, interseptal oedema, hyperplasia, and hypertrophy of the bronchiolar epithelial cells. DENV-2 led to a transient inflammatory process, but caused focal alterations of the blood-exchange barrier. Viremia was observed from 2nd to 11th day p.i. by isolation of DENV-2 in C6/36 mosquito cell line inoculated with the supernatant of macerated liver, lung, kidney, and cerebellum tissues of the infected mice.

Histopathology of discordant xenotransplantation

Purpose of review The combination of improved regimens of immunosuppression and supportive therapies with sustained depletion of native xenoreactive antibodies that bind to donor antigens, or with genetic deletion of donor antigens, has resulted in a significant extension of graft survival in large animal models of discordant xenotransplantation. This review focuses on organ transplantation in the pig-to-nonhuman primate and summarizes a shifting pattern of histopathologic findings associated with xenograft rejection. Recent findings Hyperacute rejection is preventable in renal and cardiac xenografts but still problematic in pulmonary xenografts. Interstitial mononuclear cell infiltrates of acute cellular rejection are no longer prominent in xenografts with prolonged survival. Acute humoral xenograft rejection has been modulated in xenograft kidneys and hearts and now involves thrombotic microangiopathy as a major feature. Graft vasculopathy typical of chronic rejection has only recently been identified. Summary For most discordant xenografts, thrombotic microangiopathy challenges our understanding of acute humoral xenograft rejection, the main cause of xenograft failure. Chronic rejection may be the next hurdle in xenotransplantation. As xenograft survival is prolonged, an ongoing scrutiny of the histologic features of these grafts will be crucial to our ability to characterize the pathophysiologic mechanisms of xenograft rejection or tolerance.

Post-transplantation nephrotic syndrome

CASE PRESENTATION A 60 year-old Caucasian female who developed chronic renal failure from focal segmental glomerulosclerosis (FSGS) received a living-related renal allograft from her 28year-old son. The patient presented initially at age 48 with signs of nephrotic syndrome (peripheral edema, hypoalbuminemia, and hypercholesterolemia) and a serum creatinine of 1.5 mg/dl (132mmol/l) (normal range 0.6–1.2 mg/dl (52.8–105.6mmol/l)). She was treated with prednisone for six months, followed by cyclosporine A, with only partial remission of proteinuria. Renal function slowly declined over the ensuing decade. At the time of initial presentation, serologic test results were negative or within normal limits for antinuclear antibody, C3 and C4 complement levels, hepatitis B surface antigen, hepatitis C antibody, and serum and urine protein electrophoreses. Her past medical history was significant for hypertension for 25 years, and hypothyroidism. She had no history of childhood urinary tract infections or urinary reflux and she denied regular intake of over-the-counter medications. The patient’s post-operative course was unremarkable and she was discharged seven days post-transplantation with a serum creatinine of 1 mg/dl. Immunosuppressive medications included FK506, mycophenolate mofetil, and prednisone. Four weeks post-transplant, the patient noted worsening lower extremity edema. A 24-hour urine collection contained 4.5 g protein (normal range 0–150 mg/day). Serum albumin was 3.6 g/dl (36 g/l) (normal range 3.5–5.0 g/dl (30–50 g/l)) and serum creatinine was 1.0 mg/ dl (88.4mmol). FK506 levels were within therapeutic range. A renal ultrasound showed no signs of urinary tract outflow obstruction and normal venous and arterial blood flow. A renal biopsy was performed. RENAL BIOPSY FINDINGS Twenty-four glomeruli were sampled for light microscopy, none of which showed segmental or global glomerulosclerosis (Figure 1). Peripheral capillary lumina were fully patent and glomerular basement membranes were uniformly normal in thickness, contour and texture. There was a minimal patchy interstitial mononuclear inflammatory cell infiltrate, involving o5% of the cortical area, with rare foci of mild tubulitis (o5 mononuclear inflammatory cells/ tubular cross-section). There was no significant tubular atrophy or interstitial fibrosis. Features of isometric tubular vacuolization or viral inclusions were not identified. Arterial vessels were unremarkable. Immunofluorescence microscopy showed no staining of peritubular capillaries for C4d and no glomerular staining for IgG, IgM, IgA, C3, C1, fibrinogen, albumin, kappa, or lambda. Electron microscopic examination of two glomeruli revealed diffuse (100%) podocyte foot process effacement and no other significant ultrastructural abnormalities (Figure 2).

Correlation of histopathological findings and magnetic resonance imaging in the spine of patients with ankylosing spondylitis

Ankylosing spondylitis (AS) is a chronic inflammatory disease which affects primarily the sacroiliac joints and the spine. In patients with active disease, magnetic resonance imaging (MRI) of the spine shows areas of bone marrow edema, the histopathological equivalent of which is unknown. In this study we correlate inflammation in the spine of patients with AS as revealed by histological examination with bone marrow edema as detected by MRI. We have compared the histopathological findings of zygapophyseal joints from 8 patients with AS (age: 30 to 64, disease duration 7 to 33 years) undergoing spinal surgery with findings in MRI. For histopathological analysis, we quantified infiltrates of CD3+, CD4+ and CD8+ T cells as well as CD20+ B cells immunohistochemically. Bone marrow edema was evaluated in hematoxylin and eosin stained sections and quantified as the percentage of the bone marrow area involved. All patients with AS showed interstitial mononuclear cell infiltrates and various degrees of bone marrow edema (range from 10% to 60%) in histopathological analysis. However, in only three of eight patients histopathological inflammation and edema in the zygapophyseal joints correlated with bone marrow edema in zygapophyseal joints of the lumbar spine as detected by MRI. Interestingly, two of these patients showed the highest histological score for bone marrow edema (60%). This first study correlating histopathological changes in the spine of patients with AS with findings in MRI scans suggests that a substantial degree of bone marrow inflammation and edema is necessary to be detected by MRI.

Involvement of CD44 in leukocyte recruitment to the rat testis in experimental autoimmune orchitis

Experimental autoimmune orchitis (EAO) is characterized by an interstitial mononuclear cell infiltrate and a severe lesion of the seminiferous tubules with germ cells that undergo apoptosis and sloughing. The aim of this study was to determine the role of CD44 in testicular leukocyte recruitment in EAO. The biological functions of CD44 have been attributed to the generation of a functionally active hyaluronan-binding phenotype. Orchitis was induced in Sprague-Dawley adult rats by active immunization with an emulsion of testicular homogenate and complete Freund's adjuvant using Bordetella pertussis as co-adjuvant. Control rats (C) injected with saline and adjuvants and normal (N) untreated rats were also studied. CD44 expression was analyzed by flow cytometry in peripheral blood mononuclear cells (PBMC) and lymph node cells isolated from rats at different times after the first immunization. We observed an increase in the mean fluorescence intensity of both samples in the C and experimental (E) groups only after the immunization period. A significant decrease in percentage of CD44+PBMC and in mean fluorescence intensity was observed in rats with orchitis compared with the C group. By in vitro hyaluronic acid-binding assay we demonstrated that the percentage of PBMC adhesion was higher in the E group compared with the C and N groups. By immunohistochemistry, we observed a significant increase in the number of CD44+cells in the testicular interstitium of rats with severe orchitis compared with the N and C groups. These results suggested that the CD44 molecule is involved in the homing of lymphomonocytes into the testes of rats with autoimmune orchitis.