Mechanisms and rescue strategies of calcineurin inhibitor mediated tolerance abrogation induced by anti-CD4 mAb treatment.

To ensure safety tolerance induction protocols are accompanied by conventional immunosuppressive drugs (IS). But IS such as calcineurin inhibitors (CNI), for example, cyclosporin A (CsA), can interfere with tolerance induction. We investigated the effect of an additional transient CsA treatment on anti-CD4mAb-induced tolerance induction upon rat kidney transplantation. Additional CsA treatment induced deteriorated graft function, resulting in chronic rejection characterized by glomerulosclerosis, interstitial fibrosis, tubular atrophy and vascular changes. Microarray analysis revealed enhanced intragraft expression of the B cell attracting chemokine CXCL13 early during CsA treatment. Increase in CXCL13 expression is accompanied by enhanced B cell infiltration with local and systemic IgG production and C3d deposition as early as 5 days upon CsA withdrawal. Adding different CNIs to cultures of primary mesangial cells isolated from glomeruli resulted in a concentration-dependent increase in CXCL13 transcription. CsA in synergy with TNF-α can enhance the B cell attracting and activating potential of mesangial cells. Transient B cell depletion or transfer of splenocytes from tolerant recipients 3 weeks after transplantation could rescue tolerance induction and did inhibit intragraft B cell accumulation, alloantibody production and ameliorate chronic rejection.

Permanent CNI treatment for prevention of renal allograft rejection in sensitized hosts can be replaced by regulatory T cells.

Recent data suggest that donor-specific memory T cells (T(mem)) are an independent risk factor for rejection and poor graft function in patients and a major challenge for immunosuppression minimizing strategies. Many tolerance induction protocols successfully proven in small animal models e.g. costimulatory blockade, T cell depletion failed in patients. Consequently, there is a need for more predictive transplant models to evaluate novel promising strategies, such as adoptive transfer of regulatory T cells (Treg). We established a clinically more relevant, life-supporting rat kidney transplant model using a high responder (DA to LEW) recipients that received donor-specific CD4(+)/ 8(+) GFP(+) T(mem) before transplantation to achieve similar pre-transplant frequencies of donor-specific T(mem) as seen in many patients. T cell depletion alone induced long-term graft survival in naïve recipients but could not prevent acute rejection in T(mem)(+) rats, like in patients. Only if T cell depletion was combined with permanent CNI-treatment, the intragraft inflammation, and acute/chronic allograft rejection could be controlled long-term. Remarkably, combining 10 days CNI treatment and adoptive transfer of Tregs (day 3) but not Treg alone also induced long-term graft survival and an intragraft tolerance profile (e.g. high TOAG-1) in T(mem)(+) rats. Our model allows evaluation of novel therapies under clinically relevant conditions.

Putative biomarker genes for grading clear cell renal cell carcinoma.

OBJECTIVE: The initial objective of this renal cancer study was to identify gene sets in clear cell renal cell carcinoma (ccRCC) to support grading of ccRCC histopathology. MATERIALS AND METHODS: Preselected ccRCC tumor tissues of grade 1 (G1, n = 14) and grade 3 (G3, n = 15) as well es 14 normal kidney tissues thereof were subjected to microarray expression analysis using Human Genome U133 Plus 2.0 Array. Event ratio scoring, hierarchical clustering and principal component analysis were used to determine gene sets that distinguish expression profiles from normal kidney tissue, G1 and G3 tumor tissues. RESULTS: An initial set of 73 genes provided seven gene subclusters (SC01 to SC07) that distinguish RNA expression profiles from G1, G3 tumor and normal kidney tissues. A ranked list of 24 genes was determined that separated G1 from G3 tumors in high concordance with histopathological grading confirmed by immunohistochemical analysis of ceruloplasmin protein expression. CONCLUSION: A final set of 24 genes has been determined awaiting further validation on the RNA as well as on the protein level by studying an additional cohort of ccRCC patients. A reliable separation of G1 and G3 tumor grades will be instrumental to foster and direct the administration of upcoming targeted therapeutics of ccRCC tumors in a more predictive and reliable manner.

[Small intestine, pancreas and islet cell transplantation]. / Dünndarm-, Pankreas- und Inselzelltransplantation.

The past decade has seen substantial improvements in patient and graft survival after intestinal transplantation. This improvement has been achieved by advances in donor and recipient selection, patient management, immunosuppression and surgical techniques. Intestinal transplantation is therefore considered a therapeutic option in the treatment of short bowel syndrome. Mile stones include the development of the calcineurin inhibitor Tacrolimus for immunosuppression as well as induction therapy using immune modulating substances like interleukin-2 receptor antagonists and antilymphocyte preparations. In addition to improvements in immunosuppression, antimicrobial prophylaxis and diagnosis of rejection, advances in surgical techniques have been crucial to achieving increased graft survival. Pancreas transplantation, generally with simultaneous kidney transplantation, is now available as a treatment option for patients with labile diabetes mellitus (usually type 1). Allogeneic islet transplantation was developed in the 1990s as a minimally invasive alternative to pancreas transplantation. Pancreatic islets are isolated enzymatically from the donor pancreas, in most cases infused into the portal vein and thus engrafted into the liver. Currently, technical and medical problems as well as high costs prevent the application of islet transplantation as a therapeutic option for a larger number of patients with diabetes mellitus.

Immunohistochemical expression of retinoblastoma protein and p16 in renal cell carcinoma.

INTRODUCTION: The regulation proteins retinoblastoma protein (pRb) and p16 play an important role in the cell cycle as tumor suppressors. pRb is the main substrate for the function of cyclin-dependent kinases during the cell cycle in the transition from G(1) to S phase. In this study, the immunohistochemical expression of pRb and p16 in renal cell carcinoma (RCC) were examined. METHODS: Paraffin-embedded specimens from 94 patients with RCC were examined immunohistologically, using primary antibodies for p16 and pRb (Novocastra) and a biotin-conjugated anti-mouse IgG secondary antibody. Microscopically, the expression of p16 and pRb was evaluated by examination of the staining intensity of 100 cells of each specimen, and compared with epidemiological parameters (tumor size, TNM, nuclear grade and follow-up). Statistical analyses were conducted by SPSS, version 15.0 (SPSS® Inc., Chicago, Ill., USA), the χ(2) test (Fisher's exact test), the Kaplan-Meier method and Mantel's log rank test. RESULTS: All 94 tumors showed a positive reaction for pRb (weakly positive in 67.0%; strongly positive in 33.0%). p16 was expressed in only 52.1% (weakly positive in 48.9%; intermediately positive in 3.2%; no strongly positive expression). The expression of p16-positive tumors was significantly associated with the expression of pRb (p = 0.040). Tumor size, grading, lymph node and distant metastases did not correlate with p16/pRb expression. CONCLUSION: pRb and p16 control the cell cycle as tumor suppressors. Therefore, in many tumors they are dysregulated. There are distinct differences in expression in various individual RCC. However, in a limited number of cases there was no significant correlation with clinical parameters.

[Erdheim-Chester disease of the orbit with compressive optic neuropathy]. / Erdheim-Chester-Erkrankung der Orbita mit Kompressionsneuropathie.

A 60-year-old man presented with left exophthalmos and deterioration in visual acuity of slow evolution. Bilateral orbital Erdheim-Chester disease was diagnosed. Systemic evaluation revealed a retroperitoneal fibrosis. Treatment with interferon-alpha followed, but bilateral compressive optic neuropathy with visual acuity deterioration and visual field defects evolved. Bilateral orbital decompression was performed.

Up-regulation of ICAM-1 during cold ischemia triggers early neutrophil infiltration in human pancreas allograft reperfusion.

BACKGROUND: Graft pancreatitis is induced by ischemia/reperfusion injury in which neutrophil infiltration is believed to be a crucial early event. This observation suggests the presence of adhesion molecules already at the time of reperfusion. Therefore, this study was performed to evaluate the pattern of ICAM-1 and P-Selectin expression on human pancreas allografts following cold ischemia and reperfusion. PATIENTS AND METHODS: We performed an analysis of pancreas biopsy specimens taken from 13 patients undergoing pancreas transplantation compared with pancreas specimens from 10 patients following resection. Cryostat sections were stained with monoclonal antibodies against CD11b, a neutrophil marker, and the adhesion molecules ICAM-1 and P-Selectin. RESULTS: Extensive infiltration of CD11b-positive cells was detected in venules and capillaries of pancreas allografts after reperfusion (18.38 +/- 0.87) compared with controls (T1 4.22 +/- 0.55) or with tissue specimens at about 10 hours of cold ischemia (2.60 +/- 0.35; P < .001). Similarly, the pattern of P-Selectin showed a moderate expression before organ harvest (1.54 +/- 0.21) and in samples during cold ischemia (1.46 +/- 0.24) followed by a significantly greater number of P-Selectin-positive cells after reperfusion (2.54 +/- 0.18; P = .005). ICAM-1 was only weakly expressed on the surface of the venular endothelium in all controls (0.77 +/- 0.12). In contrast to P-Selectin, ICAM-1 showed prominent up-regulation during cold ischemia (2.23 +/- 0.23; P < .001) with no further increase after reperfusion (2.23 +/- 0.17). CONCLUSION: The data suggested that ICAM-1 was already up-regulated during cold ischemia, possibly representing the mechanism of early neutrophil infiltration observed in human pancreatic ischemia/reperfusion injury.

[Pathology and pathologists in fiction revisited]. / Pathologie und Pathologen in der Belletristik: Eine Nachlese.

Pathology and pathologists are rarely the subjects of works of fiction. In the existing sources, the kind of representation naturally depends on the occupations and attitudes of the respective authors. The surgeon and gynecologist Carl Ludwig Schleich recollected Rudolf Virchow's free and easy handling of an autopsy assistant and his simultaneous understanding for a mourning husband. The dermatologist Gottfried Benn processed his disturbing impressions of pathology as an expressionistic dialogue between professor and students, with a violent ending. The writer and dramatic adviser Günther Weisenborn recalled unpleasant details about the autopsy course in his earlier medical studies, which he linked with individual views about the life of a deceased young woman. Praise, so to speak, to the dissecting pathologist have been sensitively written by the lawyer Maxence van der Meersch and by the surgeon Peter Bamm. Finally, the bestselling novelist Arthur Hailey gives an excellent fictional portrayal of the microscopic pathologist in The Final Diagnosis.

Intracardiac erythropoietin injection reveals antiinflammatory potential and improved cardiac functions detected by Forced Swim Test.

Systemic administration of erythropoietin (Epo) protects the myocardium from an ischemic insult and promotes beneficial remodeling. We hypothesized that intracardiac injection of Epo may exhibit cardioprotective potential with reduced systemic toxicity. Following myocardial infarction (MI), Epo was injected directly into the border of the infarction. Six weeks after an MI, we evaluated infarction size, angiogenesis, and pathologic effects of the treatment. Myocardial performance was assessed with a Forced Swim Test adapted to the study. Anti-inflammatory and cellular proliferative effects of Epo were analyzed by measuring expression of integrin-beta and CdK4 by reverse transcriptase-polymerase chain reaction (RT-PCR). The findings indicated improved cardiac status with direct Epo administration. Exercise capacity detected by the Forced Swim Test was significantly increased. There was radical reduction of absolute infarction size, ventricular dilatation, and hypertrophy in the Epo group. Integrin-beta was down-regulated and CdK4 expression was increased significantly with Epo. In conclusion, the study demonstrated that intramyocardial Epo injection, following MI, reduced inflammation, enhanced angiogenesis and proliferation, improved myocardial functions, and did not lead to intramural thrombus formation.

Vaginal metastasis of lung cancer: a case report.

BACKGROUND: Lung cancer is the second most common malignant tumor, with increasing incidence in the female population. The most frequent metastatic sites are the regional lymph nodes and surrounding areas as well as liver, adrenal gland, bones and brain. Metastases in the vagina of primary lung cancer have not been previously reported. CASE REPORT: Lung cancer was diagnosed in a 67-year-old, postmenopausal woman. Two years following partial lung resection (right apical lobe, R0-resection, CR), the patient complained of increasing problems with urination. A suspect tumor was identified with palpation and confirmed sonographically. Histological and immunohistochemical examinations of a vaginal excisional biopsy revealed metastatic adenocarcinoma, with the staining reactivity as primary lung neoplasm. Anterior exenteration was performed. CONCLUSION: Some cases of vaginal metastases from extragenital tumors have been previously reported. This is the first report of vaginal metastases from primary lung cancer. We suggest that adenocarcinoma especially tend to form metastases in the female genital tract. The present case emphasizes that, in women with unclear symptoms and findings in the small pelvis (e.g. urination problems, suspect vaginal tumor), the formation of such metastases should be taken into account.

Tumour budding as prognostic factor in stage I/II colorectal carcinoma.

AIMS: The term tumour 'budding' has been coined for the detachment of tumour cells from the neoplastic glands of adenocarcinomas and is presumed to be an early step in the metastatic process. A limited number of studies have shown budding to be an adverse prognostic factor. METHODS AND RESULTS: All primary single, non-metachronous TNM stage I/II colorectal carcinomas without neoadjuvant treatment resected in the years 1994-1999 were included (n = 186). Tumour buds were counted in pan-cytokeratin immunostains in a 0.785-mm2 field of vision (250 x). During follow-up 21 patients had distant metastases and 12 patients died of their disease. Budding was determined at 14 and 20.46, median and mean, respectively (range 0-120). A cut-off of 25 was found to be sensitive (0.76) and specific (0.739). Kaplan-Meier survival analysis showed high budding to be a strong adverse prognosticator. By Cox regression, high budding together with venous angioinvasion were independent prognostic factors. CONCLUSIONS: This study confirms the prognostic value of budding in a contemporary series of colorectal carcinomas that by TNM were low risk. Technically easy, rapid and robust to determine, budding quantified in pan-cytokeratin stains significantly aids in the identification of high-risk patients and is recommended for more general use in surgical pathology.

[Paleopathological skeleton findings. Macroscopical and radiographical studies of 364 individuals from a medieval graveyard]. / Paläopathologische Skelettbefunde. Makroskopische und röntgenologische Untersuchungen an 364 Individuen eines mittelalterlichen Friedhofes.

Paleopathological examinations can give an idea of diseases and living conditions in ancient populations. An archaeological collection of 364 late medieval/early modern skeletons from the thirteenth to eighteenth centuries, excavated from a church cemetery in the Rostock town center, was examined palaeopathologically. The type and frequency of certain diseases within this north German urban population are described. The majority of the skeletons were from adults with a remarkably low percentage of children. Skeletal malformations (e.g. gap formations of the spinal column) were not abnormally represented. With the exception of single individuals, metabolic disorders or unusual infectious diseases could not be diagnosed. Degenerative diseases often found at the joints and the spinal column showed substantially lower prevalences in comparison with reference rural populations. Individual cases of benign and rare malignant neoplasms could be documented. Traumatic injuries as well as dental pathological changes were rare. In summary it can be concluded that the individuals buried here belonged to a better social class within the medieval population of Rostock.

[Pathology in Rostock]. / Pathologie in Rostock.

The name of Rostock was first mentioned in 1161 by the Danish historian Saxo Grammaticus. As the oldest university in Northern Europe, the Alma mater rostochiensis was inaugurated in 1419 and is proudely called Light of the North ("Leuchte des Nordens"). Its Medical Faculty belonged to the three founding faculties. As elsewhere, the roots of Rostock pathology hark back to anatomy. A Theatrum anatomicum existed since 1790. First lectures on pathology were read by Johann Wilhelm Josephi (1763-1845) who was Head of Anatomy in the so-called Dissection House ("Zergliederungshaus") situated at the Old Market of Rostock. In 1844, anatomy together with its pathology rooms moved into the Garden House ("Gartenhaus") on the university yard. From 1878 to 1930, the Pathology represented one section of the downtown Medical Studies Building. From 1930 up to now, the Pathology Institute is situated in the Strempel Street at the corner of the clinical center. The Rostock Pathology Chair was established in 1865. Since that time, the institute had ten directors. Inter alios, Ernst Schwalbe (1871-1920) was a famous teratologist at the beginning of the 20th century. Walther Fischer (1882-1969) was Head of Institute for 24 years and became well-known as oncopathologist. After World War II, Alexander Bienengräber (1911-1990) reconstructed the institute in all ist compartments to a modern standard. At present, about 40 persons, with eight pathologists among them, represent the staff of the institute. 150 medical students are taught in each semester. Scientific topics concern oral, colorectal and thyroid carcinoma, pancreatitis as well as renal and transplant pathology. Nearly 15,000 histology, 20,000 cytology, and 150 autopsy cases are presently examined per year.

[Pancreas transplant pathology: clinicopathologic and experimental findings]. / Pathologie der Pankreastransplantation: Klinisch-pathologische und experimentelle Befunde.

134 pancreas transplantations (113 simultaneous pancreas-kidney, 5 pancreas after kidney, 16 pancreas transplants alone) done in Rostock from VI/95 to III/04 were evaluated in respect to pancreas transplant lesions. Additionally, 36 pancreas specimen of Brown Norway rats experimentally transplanted into diabetic Lewis rats were examined. From 55 out of the 134 pancreas transplant patients, 122 partly repeated pancreas graft specimen examinations were carried out morphologically. The principal lesions in the human pancreas transplants were acute (enzymatic) necrotizing transplant pancreatitis (41 samples), acute (13) and chronic (14) transplant rejection specimen as well as primary or secondary graft thrombosis (12 probes). 23 probes were zero-hour biopsies and 2 showed normal tissue. From 69 out of the 118 pancreas transplant patients with an additional kidney graft, a total of 159 renal transplant probes were examined. They showed the following lesions: acute tubular damage or acute renal failure (23), acute (56) or chronic (22) kidney graft rejection, acute tubular cyclosporine or FK 506 toxicity (53), and histologically normal graft tissue (8 cases). As in other grafted organs, the changes occurring in the transplanted pancreas consist of varying lesions related and/or not related to pancreas transplant rejection. A concise classification and a reproduceable grading schedule are suggested for diagnostic, differential diagnostic, therapeutic, and prognostic purposes. Pancreatic rejection lesions can be classified according to a proposed Rostock '04 working classification of pancreas allograft rejection into three grades (I: mild, II: moderate, III: severe) both for acute and chronic pancreas rejection. There was no direct correlation of the findings in 21 patients with simultaneously studied pancreas and renal transplant biopsies. In contrast to renal grafts, pancreatic rejection signs were often superimposed by acute transplantation pancreatitis with or without secondary graft thrombosis, nonenzymatic necroses or infection. Experimental acute pancreas transplant rejection in rats showed quite similar findings to human grafts and was also graded into three different acute rejection stages.

[Glomerular diseases in renal biopsy. Correlation of clinical syndromes with histological types]. / Glomeruläre Krankheiten in der Nierenbiopsie. Korrelation klinischer Syndrome mit histologischen Typen.

Renal biopsies studied by light and electron microscopy as well as by immunohistochemistry seem to be continually necessary for an adequate typing and therapy of glomerulonephritis (GN) and other glomerular diseases. Basing on 653 renal biopsies examined in Rostock from 1990 to 1999, the morphological classification in a total of 585 cases with glomerular diseases is presented in comparison to their clinical syndromes according to the WHO classification. A nephrotic syndrome was most frequent and clinically reported in 258 of the 585 biopsy cases with glomerular diseases (44%). It was seen in 77 of the 87 cases with minimal change nephropathy (55%), in 46 of the 74 cases with focal segmental glomerulosclerosis (62%), and in 19 of the 24 diffuse membranous GN cases (79%). The majority of the varying histological subtypes of diffuse GN was not combined with a specific clinical syndrome except of diffuse crescentic GN presenting with a rapidly progressive nephritic syndrome in 14 of 16 cases (88%). IgA nephropathy was the most often diagnosed entity of glomerular diseases found in 122 of the 502 cases with primary or secondary GN (24%). It is obvious that a given morphological GN type can but must not be combined with a specific clinical GN syndrome, so that the clinical importance of renal biopsy is stressed.