Histopathology

Expression of CD44 in premalignant and malignant Barretts oesophagus.

Year 1998
Lagorce-Pages C. Paraf F. Dubois S. Belghiti J. Flejou JF.
Department of Pathology, hopital Beaujon, Clichy, France.
AIMS: To establish the prevalence of CD44 protein expression in a large surgical series of Barrett's adenocarcinoma and associated preneoplastic lesions and to correlate this expression with clinicopathological data and prognosis. METHODS AND RESULTS: CD44H and variants (V4/V5, V6) expression was detected by immunohistochemistry in formalin-fixed, paraffin wax tissue samples of Barrett's mucosa (50) and Barrett's adenocarcinoma (73) obtained from surgical resections from 73 patients. This expression was correlated with pathological features of the tumour and prognosis. CD44H and V6 expression was found in 62% and 55% of Barrett's specialized mucosa negative for dysplasia and in 70% and 63% of Barrett's adenocarcinoma, respectively. CD44H and V6 expression was restricted to the lower part of the crypts in Barrett's specialized mucosa negative for dysplasia and reached the upper part of the crypts in high-grade dysplasia. A significant relation was found between CD44V6 expression and depth of tumour invasion in the oesophageal wall (P = 0.05), neoplastic vascular invasion (P = 0.0001), neoplastic perineural invasion (P = 0.0004) and stage in Rosenberg's classification (P = 0.02). Cancers with CD44V6 expression had a significantly poorer prognosis (5-year survival: 17%) than those without (5-year survival: 44%) (P = 0.02) in univariate analysis. However, multivariate analysis showed that CD44V6 expression had no independent prognostic value when tumour invasion and lymph node involvement were taken as explanatory variables. CONCLUSION: CD44H and V6 are frequently expressed in Barrett's oesophagus. The pattern of expression that we observed from mucosa negative for dysplasia to adenocarcinoma suggests that CD44H and V6 may be involved in the carcinogenesis of Barrett's mucosa. CD44V6 expression in adenocarcinoma is correlated to aggressive pathological features.

Multiple polypoid lesions of primary mucosa-associated lymphoid-tissue lymphoma of colon.

Year 1998
Yatabe Y. Nakamura S. Nakamura T. Seto M. Ogura M. Kimura M. Kuhara H. Kobayashi T. Taniwaki M. Morishima Y. Koshikawa T. Suchi T.
Department of Pathology and Clinical Laboratories, Aichi Cancer Center Hospital, Japan.
AIMS: This study was focused on unusual cases of mucosa-associated lymphoid tissue (MALT) lymphoma presenting with multiple polypoid lesions of the colon and rectum with a special reference to the differential diagnosis of mantle cell lymphoma. METHODS AND RESULTS: The lesions of these five cases grossly showed a segmental distribution of nodular protrusions in three patients and of innumerable small polyps in two. These patterns of involvement simulated those of multiple lymphomatous polyposis, known as the gastrointestinal presentation of mantle cell lymphoma (MCL), and caused a differential diagnostic problem between MALT lymphoma and MCL, which have different prognostic and therapeutic implications. Their histological features are almost indistinguishable from each other, especially in the small biopsy specimens via endoscope. The most important procedure for their differentiation is cyclin D1 immunohistochemistry and its negative reaction provides strong indication of MALT lymphoma. Of interest, one case showed a rare karyotypic abnormality of t(11;18)(q21;q21), which has been reported specifically in MALT lymphoma. CONCLUSIONS: This study has indicated that the multiple polypoid lesions of the colon occur not only in MCL, but also in MALT lymphoma, making differential diagnosis between the two entities necessary, and cyclin D1 immunohistochemistry is indispensable for distinguishing between them.

Up-regulation of lysozyme production in colonic adenomas and adenocarcinomas.

Year 1998
Yuen ST. Wong MP. Chung LP. Chan SY. Cheung N. Ho J. Leung SY.
Department of Pathology, University of Hong Kong, Queen Mary Hospital, Hong Kong.
AIMS: The presence of lysozyme protein in some gastric adenomas and adenocarcinomas has been well documented. There have been relatively few studies investigating the presence of lysozyme in tumours of the large intestine and they show contrasting results. We aim to investigate the cellular source and expression of lysozyme in colonic adenomas and adenocarcinomas. METHODS AND RESULTS: We randomly selected 29 and 27 colonic adenomas and adenocarcinomas, respectively. Using in-situ hybridization (ISH) and immunohistochemistry (IHC), we found an up-regulation of lysozyme in the dysplastic epithelium of all the adenomas studied, with more than 80% of cases expressing moderate to strong signals. Although the up-regulation of lysozyme was also observed in adenocarcinomas, only 30% of the cases showed moderate to strong signals, mostly with an uneven distribution. Down-regulation of lysozyme in the severely dysplastic and invasive foci were noted in some cases of adenoma with malignant transformation. Normal colonic glands were consistently negative for lysozyme at both the mRNA and the protein level, but inflamed and immature regenerative colonic epithelium at the crypt base showed positive signals in a similar pattern to those observed in the dysplastic epithelium of the adenomas. CONCLUSIONS: Our results confirm that colonic epithelium can produce lysozyme and its expression is up-regulated in the dysplastic epithelium in adenomas and in invasive cancer cells. It is interesting that regenerative colonic epithelium showed a similar pattern of lysozyme expression as in adenomas. The loss of lysozyme secreting phenotype in most of the invasive tumours suggests that lysozyme may not confer an advantage to tumour progression.

Expression of vascular endothelial growth factor in digestive neuroendocrine tumours.

Year 1998
Terris B. Scoazec JY. Rubbia L. Bregeaud L. Pepper MS. Ruszniewski P. Belghiti J. Flejou J. Degott C.
Service d'Anatomie Pathologique, Hopital Beaujon, Clichy, France.
AIMS: Angiogenesis is a complex multistep process essential for tumour growth. Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen and vascular permeability-inducing agent. Recent studies have shown that VEGF expression is correlated to microvessel density and tumour progression. The aim of this study was to analyse VEGF expression in a series of gastrointestinal neuroendocrine tumours. METHODS AND RESULTS: Surgical specimens from 28 gastrointestinal carcinoids and 20 pancreatic endocrine tumours were examined for VEGF expression by immunohistochemistry. Intense cytoplasmic staining for VEGF was observed in several cells of the islets of Langerhans and in neuroendocrine cells of normal digestive mucosa. All midgut carcinoids showed strong VEGF expression in tumoral cells. Positive VEGF immunostaining was observed in 16 of 20 neuroendocrine pancreatic tumours but it was usually much lower than in midgut carcinoids. Western blotting analysis in eight cases identified a major band at 30-32 kDa. No correlation between VEGF expression and tumour stage was found. CONCLUSIONS: This study demonstrates that neuroendocrine cells are a major source of VEGF, particularly in midgut carcinoids. This finding suggests that the presence of VEGF may be required to maintain the differentiated state of capillary vessels in these hypervascular tumours. Such secretion, in conjunction with the other growth factors synthesized by these neuroendocrine tumours, may have an important role in tumour growth.

Spontaneous regression of hepatocellular carcinoma.

Year 1998
Kaczynski J. Hansson G. Remotti H. Wallerstedt S.
Department of Medicine, Goteborg University, Sweden.
AIMS: To determine the clinical and histopathological features of a case of a spontaneous regression of hepatocellular carcinoma (HCC). CASE DETAILS: HCC was found incidentally in a 73-year-old man during a laparotomy for evaluation of gastric retention. Despite no treatment being given, he improved gradually with no sign of tumour as evidenced by coeliac angiography 15 months later, as well as by explorative laparotomy after another 2 years. The patient died 15 years after the primary diagnosis of HCC, without known evidence of tumour recurrence. The patient's clinical records were reviewed, and paraffin-embedded liver tissue was re-evaluated. Both histological and immunohistochemical features were compatible with the diagnosis of a well differentiated HCC. Conceivable causes of the spontaneous regression of this and other reported cases are discussed, but the phenomenon remains enigmatic. CONCLUSIONS: This case with a histologically proven HCC and a very long follow-up time confirms that spontaneous regression does occur. Since the case was found incidentally our report also implies that this may not be as rare as reported.

Immunoperoxidase staining for cytokeratins 8 and 18 is very sensitive for detection of occult node metastasis of colorectal cancer: a comparison with genetic analysis of K-ras.

Year 1998
Sasaki M. Watanabe H. Jass JR. Ajioka Y. Kobayashi M. Hatakeyama K.
First Department of Pathology, Niigata University, School of Medicine, Japan.
AIMS: Recent reports suggest that genetic examination of K-ras or p53 mutation is more sensitive for the detection of occult lymph node metastasis in colorectal carcinomas than conventional examination by haematoxylin and eosin (H & E) staining or immunohistochemistry for gene products. The aim of this study was, first, to define the microscopic characteristics of metastatic cancer cells in lymph nodes stained by the anti-cytokeratin antibody CAM5.2 for cytokeratins 8 and 18, and, second, to compare the detection rate of occult lymph node metastasis for immunohistochemical vs genetic methods. METHODS AND RESULTS: K-ras mutations were first examined in primary tumours of seven cases which showed distant metastasis or local recurrence within 5 years of the initial surgery in spite of the original reporting of no lymph node metastasis by routine H & E staining. K-ras mutations were positive in three cases in primary tumours and lymph nodes, and the remaining four primary tumours were negative for p53 mutation as well as K-ras mutation. Therefore, genetic analysis of occult lymph node metastasis was uninformative, but occult metastasis was detected by cytokeratin staining in two of these four cases. Comparative study of cytokeratin-positive cells was performed on each of the 43 lymph nodes from three cases with K-ras mutations. Cancer cells were detected in 28 of the 43 lymph nodes (65.1%) by cytokeratin staining and in 10 of the 43 corresponding lymph nodes (23.3%) by genetic analysis. Artefactual contamination by cancer cells was present in eight of the 28 cytokeratin positive lymph nodes, and three of the eight nodes were genetically positive. CONCLUSIONS: This study suggests that cytokeratin immunohistochemistry is more sensitive and specific for the detection of occult lymph node metastasis than genetic diagnosis by K-ras mutation in cases with genetic alterations as well as in cases without them.

Primary intestinal gamma-delta T-cell lymphoma with evidence of Epstein-Barr virus.

Year 1998
Lavergne A. Brocheriou I. Delfau MH. Copie-Bergman C. Houdart R. Gaulard PH.
Service Central d'Anatomie et Cytologie Pathologiques, Hopital Lariboisiere, Paris, France.
AIMS: Primary intestinal T-cell lymphomas account for about 5% of all primary gastrointestinal lymphomas and are mostly associated with coeliac disease. They usually express the CD3-associated T-cell receptor alpha/beta heterodimer and HML1, and some are related with Epstein-Barr virus (EBV). As far as we know, the present report describes the first case of primary gamma-delta (gamma delta) EBV-associated intestinal T-cell lymphoma without enteropathy. Only hepatosplenic, nasal and cutaneous gamma delta T-cell lymphomas have previously been described. METHODS AND RESULTS: Our case concerned a 43-year-old man with no history of coeliac disease, who presented with multifocal small bowel involvement showing high grade T-cell lymphoma with medium sized and large pleomorphic cells and a small pleomorphic T-cell component. Angioinvasion and angiocentricity were occasionally present. Immunohistochemical studies of lymphoma cells showed a T-cell gamma delta phenotype (CD3+, CD2+, TCR delta 1+, V delta 2+ and beta F1-) without expression of CD4, CD8, CD5, or HML1. Most tumour cells were positive for the cytotoxic granular proteins TiA1 and granzyme B. Rearrangement of the TCR gamma chain gene was demonstrated by polymerase chain reaction and in-situ hybridization with EBER probes revealed strong nuclear positivity in virtually all neoplastic cells. CONCLUSION: We described the first case of primary intestinal gamma delta T-cell lymphoma without enteropathy in which EBV might fulfil a pathogenic role.

Autopsy findings in 27 children with haemophagocytic lymphohistiocytosis.

Year 1998
Ost A. Nilsson-Ardnor S. Henter JI.
Department of Pathology and Cytology, Karolinska Hospital, Karolinska Institute, Stockholm, Sweden.
AIMS: Primary haemophagocytic lymphohistiocytosis (HLH) is a fatal childhood disorder. The diagnosis is difficult to establish, clinically as well as histopathologically, and it is markedly underdiagnosed. Because of these difficulties, we wanted to elucidate the histopathological findings in population-based patient material. METHODS AND RESULTS: The post-mortem findings in 27 children with primary HLH diagnosed in Sweden between 1971 and 1986 was reviewed. Twelve of these patients had an affected sibling and three additional children had parental consanguinity. Some of the children showed generalized disease, whereas in others only one or a few organs were affected. The major histological alteration was an accumulation of primarily lymphocytes, but also of histiocytes, some of which exhibited evidence of haemophagocytosis. The haemophagocytic activity may be difficult to detect if there are pronounced post-mortem changes, particularly in the spleen, and it is therefore preferable to perform the autopsy as soon as possible after death in order to minimize autolysis. Haemophagocytosis was most commonly observed in the spleen (17/24), the lymph nodes (17/23) and the bone marrow (9/23), indicating that a negative bone marrow examination does not rule out this diagnosis. Three additional patients had discrete signs of haemophagocytosis in the bone marrow. In the spleen, the lymph nodes and the bone marrow, lymphocytic depletion, pronounced in some cases, could be observed, even without prior treatment with steroids or cytostatics. In the liver, most of the patients demonstrated an infiltration of lymphocytes into the portal tracts similar to that seen in chronic persistent hepatitis (22/27), a finding which is uncommon in infancy and therefore suggestive of the diagnosis HLH. Other organs involved included the thymus, lungs intestine, pancreas, kidney, heart and striated muscle. CONCLUSIONS: The diagnosis of HLH must be based on clinical, histological and additional laboratory findings. A negative bone marrow examination is common. Previous treatment with steroids and/or cytostatic drugs may attenuate or even eliminate the typical histological findings. Liver findings similar to those in chronic persistent hepatitis are common.

CD44 variant expression in inflammatory colonic mucosa is not disease specific but associated with increased crypt cell proliferation.

Year 1998
Fromont Hankard G. Cezard JP. Aigrain Y. Navarro J. Peuchmaur M.
Department of Pathology, Robert Debre Hospital, Paris, France.
AIMS: A recent study reported increased epithelial expression of CD44 variants in ulcerative colitis (UC) compared with Crohn's disease (CD). However, the use of CD44 expression for diagnostic purposes in inflammatory bowel disease has been controversial, and the meaning of the appearance of CD44 variants in epithelial colonic cells remains unknown. We investigated the relationship between CD44 isoform expression and crypt cell proliferation in paediatric colitis. METHODS AND RESULTS: The expression of CD44v3, CD44v6, and MIB1 (proliferation marker) was studied by immunohistochemistry on surgical colonic samples of UC (n = 13), CD (n = 10), colostomy resections with non-specific mucosal inflammation (NSMI) (n = 3), and normal controls (N = 5). The proliferation index (% of MIB1 positive cells) was assessed in both v6 positive and v6 negative crypts. Epithelial expression of CD44v3 and v6, negative in normal controls, was variable and focal in colitis. No preferential expression of CD44 variants was found in UC. The proliferation index was dramatically increased in v6 positive crypts compared with v6 negative crypts in UC, CD and NSMI. CONCLUSIONS: These data suggest that CD44 variant staining is not a useful marker for distinguishing paediatric UC from CD, and that CD44v6 expression in the inflamed colonic mucosa is not disease specific but is related to crypt cell proliferation.

Detection of P-glycoprotein in frozen and paraffin-embedded gastric adenocarcinoma tissues using a panel of monoclonal antibodies.

Year 1998
Lacueva FJ. Teruel A. Calpena R. Medrano J. Mayol MJ. Perez-Vazquez MT. Rufete C. Camarasa MV. Ferragut JA.
Department of Pathology, Alicante University, Spain.
AIMS: Most chemotherapeutic regimens used against gastric carcinoma include anthracyclines whose effectiveness can be impaired by the presence of P-glycoprotein. In order to obtain a reliable pattern of P-glycoprotein expression in these tumours an immunohistochemical study using a panel of anti-P-glycoprotein antibodies was performed in frozen and paraffinized tissues. METHODS AND RESULTS: Frozen and paraffinized samples from 25 gastric carcinomas were immunohistochemically analysed using a panel of four anti-P-glycoprotein monoclonal antibodies including C219, MRK16, JSB-1 and C494. Semiquantitative analysis indicated that moderate or high P-glycoprotein levels were detected in 40% to 76% of gastric adenocarcinomas, depending on the anti-P-glycoprotein antibody used. The antibody C494 was the most sensitive in detecting P-glycoprotein in both frozen and paraffinized gastric carcinoma samples. Moreover, C494 showed a pattern of staining exclusively associated with the plasma membrane, in contrast to the cytoplasmic with reinforcement of plasma membrane pattern displayed by the other three antibodies. Significant differences in P-glycoprotein levels were obtained when C494 and MRK16 were used in frozen tissues. Finally, detection of P-glycoprotein in frozen samples did not improve when compared to paraffinized ones. CONCLUSIONS: It appears that P-glycoprotein is frequently expressed in gastric adenocarcinomas, and the use of C494 complemented by JSB-1 is recommended for reliable detection of P-glycoprotein in this neoplasm.

No evidence for c-erbB-2 overexpression in gastric carcinogenesis.

Year 1998
Blok P. Craanen ME. Dekker W. Offerhaus GJ. Tytgat GN.
Department of Gastroenterology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.
AIMS: Conflicting data on c-erbB-2 overexpression in gastric carcinomas can be found in the literature with regard to overall prevalence, prognostic significance and the histological type according to Lauren. The majority of these studies have focused on advanced gastric carcinomas whereas data on c-erbB-2 overexpression in early gastric carcinomas, especially of Caucasian origin, are relatively sparse. We therefore examined a series of Caucasian early gastric carcinomas to assess overall c-erbB-2 overexpression and to correlate c-erbB-2 overexpression, if any, with the type of growth pattern and the Lauren type. METHODS AND RESULTS: Forty-five paraffin-embedded gastrectomy specimens from early carcinomas were examined for the presence of chronic active gastritis, chronic atrophic gastritis, subtypes of intestinal metaplasia and dysplasia. The Lauren type and the type of growth pattern were reassessed for all early carcinomas. c-erbB-2 overexpression was assessed with monoclonal antibody 3B5 and polyclonal antibody A485. Complete absence of c-erbB-2 overexpression was observed in chronic active gastritis, chronic atrophic gastritis, subtypes of intestinal metaplasia, and dysplasia. Moreover, c-erbB-2 overexpression was found absent in both intestinal-type (n = 20) and diffuse-type early gastric carcinomas (n = 25), irrespective of growth type. CONCLUSIONS: c-erbB-2 overexpression does not play a role in the progression from normal to neoplastic gastric mucosa and should be considered as a late event in gastric carcinogenesis. Moreover, c-erbB-2 overexpression does not discriminate between intestinal and diffuse type early gastric carcinomas of Caucasian origin. Finally, it appears that mechanisms other than c-erbB-2 overexpression underlie the reported differences in biological behaviour of early gastric carcinomas with different types of growth pattern.

Metallothionein and apoptosis in primary human hepatocellular carcinoma and metastatic adenocarcinoma.

Year 1998
Deng DX. Chakrabarti S. Waalkes MP. Cherian MG.
Department of Pathology, University of Western Ontario, London, Canada.
AIMS: Differences in expression of metallothionein (MT) have been reported in various human tumours. MT is mainly expressed in proliferating epithelial tumour cells but in human hepatocellular carcinoma (HCC) there is only a minimal expression of MT. Since MT is a zinc binding protein and certain inducers of MT including zinc play a role in apoptosis, studies were undertaken to compare the expression of MT and the presence of apoptotic cells (APPC) in both primary HCC and metastatic adenocarcinoma. METHODS AND RESULTS: Histological sections of 13 cases of primary HCC and eight cases of metastatic adenocarcinoma were obtained from archival samples. They were stained for MT using a polyclonal antibody which crossreacts readily with human MT and for APPC by the TUNEL technique. Normal human liver had consistent MT staining with no detectable APPC. The primary HCC showed moderate MT staining with a small number of APPC while metastatic adenocarcinoma showed no MT staining with a large number of APPC. CONCLUSIONS: These results suggest a relationship between absence of MT and appearance of APPC in human liver tumours, especially in metastatic adenocarcinomas.

Localized amyloidosis and gastrointestinal lymphoma: a rare association.

Year 1998
Goteri G. Ranaldi R. Pileri SA. Bearzi I.
Institute of Pathological Anatomy and Histopathology, University of Ancona, Italy.
AIMS: Five cases of primary gastrointestinal (GI) lymphoma (three in the stomach, one in the ileum (IPSID) and one in the colon) associated with localized AL amyloidosis were studied to identify morphological or immunohistochemical features which could explain the amyloid deposition. METHODS AND RESULTS: All the cases were low-grade marginal zone B-cell lymphomas; one case of gastric lymphoma and the IPSID also had a high-grade component. The lymphomas had a monoclonal plasma cell population, with different light and heavy-chain type expression in the five cases. Plasma cell differentiation was closely associated with the amyloid deposits. The latter were an incidental microscopic finding in one case, but produced tumoral masses in the other. CONCLUSIONS: The presence of amyloid in primary GI lymphoma is rare, but can have diagnostic value. In the present study, neither particular features of the lymphomatous proliferation nor specific agents are identified. Therefore, the factors predisposing to amyloid deposition require elucidation.