bluuebunny/crossref_metadata_2025 huggingface dataset:
A marginal zone pattern may be found in different varieties of non‐Hodgkin's lymphoma: the morphology and immunohistology of splenic involvement by B‐cell lymphomas simulating splenic marginal zone lymphoma
Authors: Piris M A , Mollejo M , Campo E , Menárguez J , Flores T , Isaacson P G
Published: March 2003

Aims: Splenic marginal zone lymphoma (SMZL) is characterized by a micronodular infiltrate of the splenic white pulp, centred on pre‐existing follicles, with a peripheral rim of ‘marginal’ zone B‐cells, always accompanied by a variable degree of red pulp infiltration. These histological features can be closely mimicked by a variety of other small B‐cell lymphomas when they involve the spleen, which makes recognition of SMZL difficult. We therefore have compared the histopathological and immunohistochemical features of other non‐Hodgkin's lymphoma (NHL) types with those of SMZL.

Methods and results

We selected cases of splenic involvement by different types of B‐cell lymphoma, including mantle cell lymphoma (MCL), follicular lymphoma (FL), immunocytoma (IM) and lymphocytic lymphoma (B‐CLL). A micronodular pattern and marginal zone differentiation were both found to be frequently present in FL and MCL, and with lesser frequency in IM and B‐CLL. The main morphological feature useful for differential diagnosis was the cytological composition of the white pulp tumoral nodules. SMZL is distinguished by characteristic dimorphic cytology, different from the monomorphic cytology of MCL, and the distinctive mixture of centroblasts and centrocytes which is the rule in FL. B‐CLL could also be identified on the basis of the polymorphic cytology including small lymphocytes and prolymphocytes, whereas cases diagnosed as IM show prominent plasmacytic differentiation, lacking the features of the other lymphoma types. Immunohistochemistry was particularly useful for the differential diagnosis. Thus the recognition of MCL was facilitated by the identification of cyclin D1 and CD43 reactivity, while FL could be recognized by the lack of IgD expression or the distinctive pattern of Ki67 staining found in SMZL. B‐CLL cells were CD23 + , CD43 +.

Conclusion

The results of this study provide morphological and immunohistological information useful in the recognition of the different varieties of NHLs when involving the spleen and the differential diagnosis of SMZL.
The investigation of sudden cardiac death
Authors: Davies
Published: March 2003

Pathologists are faced with an increasing complexity in the cardiac diseases that cause sudden natural death in the absence of coronary artery disease. A significant proportion of such natural sudden deaths are due to familial heart muscle disease (cardiomyopathy). The phenotypic characteristics of both hypertrophic cardiomyopathy and arrythmogenic right ventricular dysplasia are wider than previously thought and the hearts may be very close to normal on naked eye examination. Detailed histology of the myocardium is needed to identify such cases. Up to 200 sudden deaths a year in England occur in young, apparently fit individuals in whom toxicology and detailed examination of the heart for structural abnormalities is negative. Genetic defects in ion channels (long QT interval) are now known to be one cause of this phenomenon. In investigating a case of sudden death without cause, a study of the family − if they wish it − may be helpful in arriving at a cause.
High mitotic index associated with poor prognosis in gastrointestinal autonomic nerve tumour
Authors: Tornóczky T , Kálmán E , Hegedûs G , Horváth Ö P , Sápi Z , Antal L , Jáksó P , Pajor L
Published: March 2003

Aims

Three gastrointestinal autonomic nerve tumours (GANT) were characterized by immunohistochemistry and flow cytometry. Two of the three cases occurred in the small intestine, while the third was found in the stomach. Besides the immunohistochemical and ultrastructural description, the aim of this study was to examine the relation between the known and accepted predictive factors (ploidy data, the S‐phase fraction, the mitotic and MIB‐1 index and the size of the tumour) and the survival of the patients.

Methods and results

The immune profile showed that 3/3 cases were vimentin and NSE, 2/3 were synap‐tophysin and PGP 9.5 positive, while 1/3 also showed S100 positivity. Ultrastructurally, all the cases had dense core granules, one of them contained skenoid fibres. The flow cytometry revealed diploid DNA in all cases, however, significant differences could be seen in the proliferative activity of the individual neoplasms.

Conclusions

In spite of the published data of gastrointestinal stromal tumours (GIST) generally, neither the MIB‐1 index and the ploidy data nor the size of the primary tumour helped to predict the clinical progression of the examined GANTs. However, the high proliferative activity (57 mitoses/10 HPF) and the elevated S‐phase fraction (24%) was associated with advanced, metastatic and recurring disease in case 3. On the basis of these three cases, high mitotic activity is the most reliable factor in predicting aggressive clinical behaviour.
Low‐grade metaplastic carcinoma of the thymus
Authors: Yoneda S , Marx A , Heimann S , Shirakusa T , Kikuchi M , Müller‐Hermelink H K
Published: March 2003

Aims

Five cases of a characteristic low‐grade thymic epithelial tumour are described that we suggest calling metaplastic carcinoma of the thymus.

Methods and results

The patients' ages ranged from 44 to 71 (mean 56.2) years. Four of the patients were male. Three of five tumours showed invasion into mediastinal fat or pleura but, otherwise, all were well circumscribed. No metastases were present. Histologically, the tumours showed a biphasic pattern with solid carcinomatous areas merging gradually with a spindle cell component. Lymphocytes were rare. Cytological atypia and mitotic activity were variable in the solid areas, but slight or absent in the spindle cell component. On immunohistochemistry, the tumours showed expression of cytokeratin, vimentin and/or epithelial membrane antigen, both in the carcinomatous and spindle cell components. In two cases, actin expression was also present in both components. In one case, chromogranin, S100 protein, glial fibrillary acidic protein and neuron‐specific enolase were expressed in at least some cells of both components. None of the patients had myasthenia gravis. All patients are alive without evidence of recurrence or metastasis.

Conclusion

Metaplastic carcinoma of the thymus is a distinct clinicopathological entity that should be distinguished from the usually benign medullary thymomas and from the clinically aggressive carcinosarcomas and sarcomatoid carcinomas.
Malignant myoepithelioma of the vulva resembling a rhabdoid tumour
Authors: Hinze P , Feyler S , Berndt J , Knolle J , Katenkamp D
Published: March 2003

Aims

We report an example of malignant myoepithelioma of the vulva, which has not been hitherto described. We discuss the differential diagnosis and briefly review the literature.

Methods and results

The lesion was found in an 81‐year‐old woman as an indolent 40 mm tumour. The neoplastic cells showed a myoid, spindled, epithelioid and plasmacytoid phenotype. Hyalinization of extracellular material and myxoid changes were present. There was a partly solid and microcystic pattern and a tight cohesiveness of cells was lacking. The circumscribed multinodular tumour somewhat resembled an extrarenal rhabdoid tumour, having large tumour cells with prominent nucleoli and large amounts of acidophilic cytoplasm. Immunohistochemically, the tumour cells were immunoreactive for cytokeratin, vimentin, muscle‐specific actin, alpha‐smooth muscle actin, and S100 protein, but not for desmin, epithelial membrane antigen, factor VIII‐related antigen, CD34 and CD31.

Conclusions

The histological and cytomorphological appearance of the tumour well as the immunohistochemical findings suggest the diagnosis of malignant myoepithelioma, possibly derived from minor vestibulary glands or ectopic breast tissue. Differential diagnoses are, in particular, extrarenal rhabdoid tumour and ‘proximal type’ epithelioid sarcoma. Differentiation is important, because the tumours show a different behaviour and prognosis.
CD44v6 expression in inflammatory bowel disease is associated with activity detected by endoscopy and pathological features
Authors: Camacho F I , Muñoz C , Sánchez‐Verde L , Sáez A I , Alcántara M , Rodríguez R
Published: March 2003

Aims

Overexpression of CD44v6 in colon crypt epithelial cells has been suggested to have diagnostic potential in differentiating ulcerative colitis from other forms of colon inflammation, including Crohn's disease. Our aim was to determine the value of CD44v6 expression in inflammatory bowel disease (IBD) and to look for possible associations between CD44v6 expression and activity of this disease.

Methods and results

CD44v6 expression was studied using immunohistochemical techniques in 100 surgical and endoscopic colon samples of ulcerative colitis (n = 71) and Crohn's disease (n = 29), and in every case disease activity was studied by endoscopy and microscopic examination. Fifty‐five of 71 (77.5%) samples of ulcerative colitis showed monoclonal antibody 2F10 stained colon epithelium, as did 16 of 29 (55.2%) samples of Crohn's disease. CD44v6 was detected in 88.2% (15 of 17) of cases of IBD with severe disease activity and in 100% of eight cases of severe ulcerative colitis. Our study showed a strong association between CD44v6 expression and the activity of IBD (P = 0.007).

Conclusions

CD44v6 expression in IBD is significantly associated with activity detected by means of endoscopy and pathological features. Our data suggest that CD44v6 expression may have some usefulness in conjunction with other factors as a means of evaluating the disease activity. Moreover, CD44v6 expression was higher in ulcerative colitis than Crohn's disease (P = 0.02), although this does not confirm the utility of monoclonal antibody 2F10 in differential diagnosis between ulcerative colitis and Crohn's disease, as there was a notable percentage of positive samples of Crohn's disease.
Inflammatory aetiology of primary oesophageal achalasia: an immunohistochemical and ultrastructural study of Auerbach's plexus
Authors: Raymond  , Lach  , Shamji
Published: March 2003

Aim

Achalasia is a disease of the oesophagus characterized by increased lower oesophageal sphincter (LOS) tone, absence of LOS relaxation with swallowing and aperistalsis of the body of the oesophagus. The aetiology and pathogenesis of idiopathic achalasia is still controversial.

Methods and results

We examined 16 oesophageal biopsies and one low oesophagectomy specimen from patients with achalasia. The control group was composed of five autopsy cases with no history of oesophageal disorders, three cases of diffuse oesophageal spasm, one of gastro‐oesophageal reflux disease and one patient with oesophageal carcinoma. Sections were immunostained for neurofilaments NF70 and NF200, S100 protein and neurone‐specific enolase. Biopsies with inflammatory infiltrates, were in addition immunostained with antibodies against leucocyte common antigen as well as for CD20, CD43, CD68 and CD45RO. All biopsies were examined after plastic embedding, and electron microscopy (EM) was performed on samples containing autonomic plexus. An inflammatory infiltrate of varying intensity was present along the nerve fascicles and around ganglion cells in 90% of the cases of achalasia. T‐lymphocytes predominated in all these cases. The autonomic nerves showed loss of fibres and degenerative changes which were discernible only by EM. Although there was no convincing neuronal loss or signs of active neuronal degeneration in biopsied cases, the oesophagectomy specimen revealed total absence of neurones and significant loss of nerve fibres. The control group showed normal plexuses and no inflammation.

Conclusion

Degeneration and significant loss of nerve fibres associated with predominant T‐cell lymphocytic inflammatory infiltrate around the myenteric plexus support the concept for the inflammatory, probably autoimmune, aetiology of autonomic nervous system injury in primary achalasia.
CD101 expression by Langerhans cell histiocytosis cells
Authors: Bouloc  , Boulland  , Geissmann  , Fraitag  , Andry  , Teillac  , Bensussan  , Revuz  , Boumsell  , Wechsler  , Bagot
Published: March 2003

Aims

Our objective was to study the expression of a recently identified cell surface molecule, CD101 and in Langerhans cell histiocytosis (LCH) patients as CD101 has been shown to be present on dendritic cells. We wanted to determine if CD101 expression could be helpful for the diagnosis of LCH in conjunction with other markers (CD1a, S100 protein), and could be predictive of the evolution and dissemination of the disease.

Methods and results

The expression of CD101 was studied by immunohistochemical technique in 11 cases of Langerhans cell histiocytosis on frozen sections. The expression of CD101 was positive in nine cases, high in six cases and low in three cases. There was no expression in the other two cases. No correlation with the evolution, the localization or the dissemination of the disease could be evidenced.

Conclusions

CD101 is a new phenotypic marker that might be useful in combination with other markers for the diagnosis of LCH. However, as the anti‐CD101 antibody works only in frozen sections, its value is limited compared to anti‐CD1a antibody.
Lymphoproliferative disorders in children with primary immunodeficiencies: immunological status may be more predictive of the outcome than other criteria
Authors: Canioni D , Jabado N , MacIntyre E , Patey N , Emile J F , Brousse N
Published: March 2003

Aims:

Lymphoproliferative disorders (LPDs) are a severe complication in primary immunodeficiency and post‐transplant patients. In primary immunodeficiency patients, LPDs are not well‐known and, thus, we tried to evaluate their distinctive features and to determine prognostic factors predictive of clinical outcome by comparison with LPDs in post‐transplant children.

Methods and results:

Clinical records and histopathology of 18 LPDs occurring in primary immunodeficieny children were compared with those of 10 LPDs in post‐transplant children, together with results of in‐situ hybridization for the detection of Epstein–Barr virus (EBV)‐RNA and molecular biological techniques. LPDs were frequently extranodal, EBV‐associated, and were more commonly pleomorphic in primary immunodeficiency than in post‐transplant patients. A low T‐cell count and abnormal T‐cell function indicated bad prognosis in both groups. Polymorphic LPDs (PLPDs) were most frequent (n = 19), whereas lymphomas were rare (n = 7), and pseudo‐tumoral lymphoid hyperplasias (n = 2) were observed only in primary immunodeficiency. Comparative p53/bcl‐2 staining revealed a p53 overexpression in lymphomas compared with PLPDs; CD20/CD79a showed a similar staining in lymphomas, whereas PLPD expressed mainly CD20. TCR and IgH rearrangements did not help in distinguishing PLPDs from lymphomas, but detection of IgH clonality by Southern blot indicated poor prognosis, whereas oligoclonality by Southern blot regardless of PCR clonality and especially a polyclonal profile by Southern blot and PCR indicated a relatively good prognosis.

Conclusions:

This study documents the pleomorphism of LPDs in primary immunodeficiency compared to post‐transplant children, even if some LPDs are similar in both groups (PLPDs). No criteria are useful enough to ascertain the diagnosis of malignancy in this series. Some molecular biological criteria help to predict the clinical outcome which, nevertheless, seems to depend more on the degree of immunosuppression and on T‐lymphocyte presence and function.
Estimation of age of human cadavers by immunohistochemical assessment of advanced glycation end products in the hippocampus
Authors: Sato Y , Kondo T , Ohshima T
Published: March 2003

Advanced glycation end products (AGEs) are known to accumulate in long‐lived tissue proteins during normal ageing. In this study, we examined the expression of AGEs in human hippocampus using immunohistochemistry and determined its utility for estimating the age of cadavers of unknown age.

Hippocampus tissues were obtained at autopsy from 31 individuals, including 10 fire victims, aged 0–96 years within 3 days postmortem. Immunostaining using anti‐AGE antibody demonstrated that the perikarya of pyramidal neurones in the hippocampus was immunoreactive for the anti‐AGE antibody, and the immunoreactivity was increased with age. Quantitative analysis of the AGE‐immunoreactivity in the pyramidal neurones of the CA4 region revealed a significant correlation between the AGE‐immunoreactivity and the age in nonfire death cases with a correlation coefficient of 0.91 (P < 0.01). The significant correlation could be obtained even in fire death cases affected by the unusual environmental condition.

These results suggest that the immunohistochemical analysis of AGEs in human hippocampus may be useful for the age estimation of cadavers with unknown age.
High expression of MDR‐1 gene and P‐glycoprotein in initial and re‐biopsy specimens of relapsed B‐cell lymphoma
Authors: Liu Q , Ohshima K , Kikuchi M
Published: March 2003

Multidrug resistance (MDR) is a major obstacle in the treatment of lymphoma. The expression of MDR‐1 mRNA and P‐glycoprotein (MDR‐1/P‐gp) has been linked to MDR. We aimed to investigate the expression of MDR‐1/P‐gp in B‐cell lymphoma.

Samples at diagnosis and relapse from 10 patients with B‐cell lymphoma were obtained. We also obtained 14 unselected control cases of B‐cell lymphoma at diagnosis. The expression of mRNA and protein were determined semiquantitatively by RT‐PCR and immunohistochemistry. High MDR‐1 and P‐gp expressions were found in seven and seven of 10 samples obtained at diagnosis, eight and eight of 10 samples obtained at relapse, and three and four of 14 control cases at diagnosis, respectively. The results of RT‐PCR paralleled those of immunohistochemistry. Concordance of high MDR‐1/P‐gp expression was noted in 27 of 34 samples (r = 0.73, P = 0.001). There were no significant changes in MDR‐1/P‐gp expression in all cases at relapse and during the clinical course following chemotherapy. In the 14 control cases, the average survival time was 12.7 months in MDR‐1/P‐gp positive cases and 29.0 months in the MDR‐1/P‐gp negative cases (P = 0.20).

Our results showed that at least some B‐cell lymphomas express MDR‐1/P‐gp, which could be detected by different methods, and suggested that high MDR‐1/P‐gp expression in tumour cells may be associated with a high probability of relapse and poor prognosis.
Female adnexal tumours of probable Wolffian origin: an immunohistochemical study comparing tumours, mesonephric remnants and paramesonephric derivatives
Authors: Tiltman A J , Allard U
Published: March 2003

To establish an immunohistochemical profile of presumed female adnexal mesonephric tumours (FATWO) for diagnostic purposes and to compare the findings with those of mesonephric and paramesonephric derivatives in order to establish supportive evidence for a mesonephric origin.

Standard immunohistochemistry was performed on formalin‐fixed tissues. Tumours, mesonephric remnants and paramesonephric structures generally show positive staining for vimentin, CAM 5.2 and cytokeratins 7 and 19 but are negative for CK20 and 34βE12. EMA is positive in both mesonephric and paramesonephric derivatives but is negative in the tumours. Glutathione S‐transferase μ (GSTµ) is generally positive in both tumours and mesonephric derivatives but negative in paramesonephric structures.

Immunohistochemistry plays little part in the diagnosis of FATWO. The tumours are generally cytokeratin and vimentin‐positive and EMA‐negative. GSTµ, as a marker for the mesonephric duct, is a useful adjunct. Our findings of the study support but do not prove that FATWOs are of mesonephric origin.
Accuracy of frozen section diagnoses of breast lesions after introduction of a national programme in mammographic screening
Authors: Scheiden R , Sand J , Tanous A M , Knolle U , Capesius C , Wagnon M Ch , Faverly D
Published: March 2003

Accuracy of frozen section diagnoses of breast lesions after introduction of a national programme in mammographic screening

Aims: By introducing mammography screening programmes, the size of the detected breast lesions became smaller and the histopathological interpretation problems greater. The study’s aim was to analyse the risks and possible limitations of the frozen section method.

Methods and results: Frozen section consultations of breast lesions (n=559) 2 years before and 6 years after launching a national mammographic screening programme in 1992 were evaluated in regard of the benign/malignant ratio, tumour size, preoperative frozen section results and final permanent section diagnoses. The breast frozen section examinations of 1990 compared with those from 1998 declined from 70.7% (299/423) to 62.2% (260/418) (P < 0.01), the benign/malignant ratio from 1.09 to 0.54 (P < 0.0001), the rate of the conclusive, correct frozen section diagnoses from 96.3% to 91.9% (P < 0.03). The sensitivity dropped from 92.3% to 87.6%, the negative predictive value from 95.7% to 88.3%, whereas the negative likelihood ratio rose from 0.08 to 0.12. The ‘small’ (≤ 10 mm) invasive breast carcinomas increased from 14.2% to 22.3% (P < 0.01) and the ‘in situ’ carcinomas from 2.1% to 6.6% (P < 0.05).

Conclusions: The declining sizes of breast tumours (≤ 10 mm), especially from radiologically detected lesions and sometimes without a macroscopic correlate, create new limitations and changing indications in the histopathological interpretation. Considering the performance of new diagnostic methods (i.e. large core needle biopsies), frozen sections of surgical specimens should not be the primary diagnostic procedure for breast lesions and should be performed only after other preoperative methods have failed.
Pedunculated peritoneal surface polyps in pseudomyxoma peritonei syndrome
Authors: Sugarbaker P H , Yan H , Shmookler B
Published: March 2003

Pedunculated peritoneal surface polyps in pseudomyxoma peritonei syndrome

Aims: Pseudomyxoma peritonei syndrome is a rare disease that originates from an adenomatous lesion of the appendix that, from pressure, perforates to gain access to the free peritoneal cavity. The relative sparing of the small bowel surfaces allows for complete cytoreduction even though many kilograms of mucinous tumour exist at other sites within the abdomen and pelvis. The purpose of this study was to examine the mechanism whereby the small bowel remains free of gross tumour and peritoneal surface polyps form.

Methods and results: Peritoneal surface polyps were harvested and examined grossly and histologically. A hypothesis for their formation on small bowel and small bowel mesentery was proposed. Polyps are known to be associated with repeated motion of enteric contents moving past adenomatous tissue so that, over time, an elongated stalk is created. We have repeatedly observed pedunculated polyps on the peritoneal surface of the small bowel in patients with pseudomyxoma peritonei syndrome. No other site within the peritoneal cavity has had a pseudomyxoma polyp located upon its surface.

Conclusions: The peristaltic motion of the small bowel causes adherent adenomatous tissue to develop a stalk on the peritoneal surface. Motion not only creates polypoid lesions but also repeatedly clears mucinous tumour cells from the small bowel surface. With pseudomyxoma peritonei and with other types of cancerous dissemination, prevention of adherence by motion may interfere with the implantation of malignant cells.
CD9 immunohistochemical staining of breast carcinoma: unlikely to provide useful prognostic information for routine use*
Authors: Jamil F , Peston D , Shousha S
Published: March 2003

CD9 immunohistochemical staining of breast carcinoma: unlikely to provide useful prognostic information for routine use

Aims: CD9, a cell membrane glycoprotein, is found in a variety of tumour cells and is believed to regulate cell motility and possibly cell growth. It has been reported that the absence of CD9 is associated with increased aggressiveness of breast carcinoma, but no detailed studies of the distribution of CD9 in normal and abnormal breast tissue are available. This investigation was aimed at studying the distribution of CD9 in a wide variety of breast biopsies including normal, benign, and malignant cases, and assessing its usefulness as a prognostic marker in breast cancer.

Methods and results: Sections of 113 breast biopsies from female and male patients including 10 normal, 23 benign, and 80 malignant cases were examined. The monoclonal antibody CD9 (Novacastra Ltd, Newcastle‐upon‐Tyne, UK) was used with the avidin–biotin complex immunoperoxidase technique. The results were assessed semiquantitatively using a four scale system of 3+, 2+, 1+ and negative. All normal and benign epithelial cells were strongly stained (3+). In female breast carcinomas, 40% were 3+, 49% were 2+, and 11% were 1+. Both cases of male breast carcinomas scored 3+. For female breast cancers, the results were then correlated to tumour grade, the presence or absence of lymph node metastases, and oestrogen and progesterone receptor status. No significant statistical correlation was found with any of these parameters. We then examined 11 axillary lymph nodes with metastases from some of the above cases. Three of these cases had a CD9 score of 3+, seven were 2+, and one was 1+. The metastatic tumours in all 11 cases were strongly stained (3+).

Conclusions: Immunostaining for CD9 is unlikely to provide any useful additional prognostic information for clinical purposes.
Photoreceptor cell differentiation in retinoblastoma demonstrated by a new immunohistochemical marker mucin‐like glycoprotein associated with photoreceptor cells (MLGAPC)
Authors: Tsuji M , Goto M , Uehara F , Kaneko A , Sawai J , Yonezawa S , Ohba N
Published: March 2003

Photoreceptor cell differentiation in retinoblastoma demonstrated by a new immunohistochemical marker mucin‐like glycoprotein associated with photoreceptor cells (MLGAPC)

Aims: For further understanding of specific differentiation in retinoblastoma, we studied the expression of newly detected mucin‐like glycoprotein associated with photoreceptor cells (MLGAPC), which is specific for photoreceptor cells of retina and analogous to interphotoreceptor matrix proteoglycan‐1 (IMPG1).

Methods and results: Surgically enucleated retinoblastomas (n=21; undifferentiated type, n=15, differentiated type, n=6) were immunohistochemically studied with a polyclonal antibody against MLGAPC, and 17/21 cases (81%) showed positive staining of tumour cells. We classified various staining patterns and structures into four groups: type 1 showing a granular intracellular scattered staining pattern with round small cells; type 2 showing a reticular staining pattern between spindle‐shaped tumour cells; type 3 showing radiating staining from the centre of Homer–Wright rosettes; type 4 showing ring‐shaped, radiating and granular staining associated with Flexner–Wintersteiner rosettes. Eleven of 15 undifferentiated retinoblastomas (73%) showed type 1 or 2, and all the six differentiated cases showed type 3 or 4. Image analysis of immunostaining revealed an increase in MLGAPC‐positive area from 0.48% in undifferentiated cases to 1.60% in differentiated cases, and a negative correlation was shown between mitotic frequency and MLGAPC‐positive area.

Conclusions: This study proved MLGAPC as a valuable marker of retinoblastoma, and that photoreceptor differentiation takes place even in `undifferentiated' retinoblastoma.
Expression of human mucin genes in normal kidney and renal cell carcinoma
Authors: Leroy X , Copin M‐C , Devisme L , Buisine M‐P , Aubert J‐P , Gosselin B , Porchet N
Published: March 2003

Expression of human mucin genes in normal kidney and renal cell carcinoma

Aims: Human mucins are large O‐glycoproteins expressed by epithelial cells. Mucins are thought to be implicated in cell protection, cell adhesion and signalling. The aim of this study was to investigate the expression of the human mucin genes (MUC1–4, 5AC, 5B, 6–7) in normal kidney and renal cell carcinoma.

Methods and results: We analysed by in‐situ hybridization, immunohistochemistry and reverse transcriptase‐polymerase chain reaction (RT‐PCR) the expression of these genes in normal adult kidney (n=14) and renal cell carcinomas (n=29). MUC1, MUC3 and MUC6 were expressed both in normal kidney and in renal carcinomas. In normal kidney, MUC1 was expressed in the distal convoluted tubules and in collecting ducts, whereas MUC3 was restricted to the proximal tubules. MUC4 was strongly expressed in epithelial urothelial cells of pyelocalyceal cavities. MUC6 was only detected by RT‐PCR. In renal carcinoma, we showed a heterogeneous expression of MUC1 and MUC3 with an over‐expression of MUC3 in renal clear cell carcinoma. The level of MUC3 expression by in‐situ hybridization was associated with the nuclear grade in clear cell carcinoma.

Conclusions: This study is the first large series investigating human mucin gene expression in the kidney. MUC1, MUC3 and MUC6 are expressed in normal and tumour kidney. The over‐expression of MUC3 in renal cell carcinomas favours its implication in renal tumorigenesis.
Wide spectrum screening keratin as a marker of metaplastic spindle cell carcinoma of the breast: an immunohistochemical study of 24 patients
Authors: Adem C , Reynolds C , Adlakha H , Roche P C , Nascimento A G
Published: March 2003

Wide spectrum screening keratin as a marker of metaplastic spindle cell carcinoma of the breast: an immunohistochemical study of 24 patients

Aims: Metaplastic spindle cell carcinomas may be difficult to distinguish histologically from other spindle cell lesions in the breast. Variable staining with cytokeratin immunomarkers has been reported for metaplastic carcinomas. We evaluated the diagnostic utility of anti‐cytokeratin polyclonal antibody, wide spectrum screening keratin, to assess spindle cell breast lesions.

Methods and results: Twenty‐four patients with spindle cell breast carcinoma and 31 patients with benign or malignant spindle cell tumours were studied using a panel of antibodies directed against multiple cytokeratins (AE1/AE3, CAM5.2, wide spectrum screening keratin), epithelial membrane antigen (EMA), and vimentin. Sites of origin for the 31 controls included breast, bone, and soft tissue. All but one (95.8%) metaplastic carcinomas stained positively with wide spectrum screening keratin. Only rare or focal immunoreactivity was observed with AE1/AE3 in four cases; however, sensitivity of AE1/AE3 was improved in 13 cases using steam EDTA as an antigen retrieval technique. Three cases were immunoreactive with CAM5.2 and eight cases were immunoreactive with EMA. All control cases lacked immunoreactivity with the cytokeratin panel and EMA. The spindle cells in the metaplastic breast tumours (88%) and in the controls (97%) stained with vimentin.

Conclusions: Wide spectrum screening keratin may be the most useful and convenient antibody in differentiating metaplastic spindle cell carcinoma from other spindle cell lesions in the breast.
Extra‐abdominal desmoid fibromatosis: two familial cases with synchronous and metachronous multicentric hyalinizing nodules
Authors: Watanabe K , Ogura G , Tajino T , Suzuki T
Published: March 2003

Extra‐abdominal desmoid fibromatosis: two familial cases with synchronous and metachronous multicentric hyalinizing nodules

Aims:  Extra‐abdominal desmoid fibromatosis is an uncommon tumour. We present here two exceptional familial cases of extra‐abdominal desmoid fibromatosis, one of which was synchronous and metachronous.

Methods and results:  The first patient was a 37‐year‐old woman who had noted a tumour growing on the dorsum of her right foot when she was 12 years old. She underwent excision of the tumour but in the following year the tumour recurred locally and grew into multiple nodules. Subsequently, multicentric tumours appeared in her knee, distal and posterior aspects of her thigh, right back and right anterior shoulder. Polyostotic fibrous dysplasia of the femur and cranium was found on radiological examination. The second patient was a 74‐year‐old man, the uncle of the first patient. He underwent an excisional operation of a tumour on the internal malleolus surface of his fibula when he was 46 years old. The tumour recurred 7 years later and was excised. His post‐operative course has been uneventful. The histology of the primary and recurrent tumours was distinctive and consistently showed hyalinizing scar‐like features.

Conclusions:  Familial cases of extra‐abdominal desmoid fibromatosis with extensive multicentric lesions and distinctive hyalinizing scar‐like features are described. Recently, attenuated familial adenomatous polyposis with familial desmoid fibromatosis has been recognized, and familial desmoid fibromatosis without adenomatous polyposis may also be one of its variants. Although the present cases have no history of colon polyposis or carcinoma, monitoring of the intestinal tract would seem to be indicated.
Investigation of the role of delayed‐type‐hypersensitivity responses to myelin in the pathogenesis of Theiler’s virus‐induced demyelinating disease
Authors: BORROW  , WELSH  , TONKS  , DEAN  , BLAKEMORE  , NASH
Published: March 2003

The contribution of autoimmune responses to the pathogenesis of Theiler’s virus‐induced demyelinating disease was investigated. Delayed‐type hypersensitivity responses to myelin were examined in both symptomatic and asymptomatic mice at different times post‐infection, in order to determine whether autoreactivity correlates with the development of demyelination. The results indicate that although autoimmune responses probably do not play a major role in the initiation of demyelination at early times post‐infection, autoreactivity to myelin antigens dose eventually develop in symptomatic animals, perhaps through the mechanism of epitope spreading. Autoimmunity to myelin components is therefore an additional factor that may contribute to lesion progression in chronically diseased animals.
Regulation of interleukin‐6 and interleukin‐6R α (gp80) expression by murine immunoglobulin‐secreting B‐cell hybridomas
Authors: IWASAKI  , HAMANO  , FUJIMOTO  , KAKISHITA
Published: March 2003

We have examined the contribution of endogenous interleukin‐6 (IL‐6) to the differentiation of murine B‐cell hybridomas. AT73 was established by somatic hybridization between BALB/c mice B cells and 2.52M, a hypoxanthine‐aminopterine‐thymidine (HAT) medium‐sensitive B‐cell line mutant. It spontaneously secreted IgM, and addition of exogenous IL‐6 augmented IgM secretion. Triggering of CD40 led to an augmentation of IL‐6 expression and IgM secretion. Blocking the binding of IL‐6 to its cellular receptor through the use of inhibitory monoclonal antibodies inhibited CD40‐induced IgM secretion, suggesting a possible autocrine role of IL‐6 for the differentiation of a CD40‐activated B‐cell hybridoma. Co‐triggering with CD40 and B‐cell receptor or activation through CD40 and IL‐4 led to a synergistic augmentation of IL‐6 expression as well as additive IgM secretion; this was followed by a marked decrease in the expression of B‐cell surface markers on the cell membrane. Furthermore, under conditions where IL‐6 expression was augmented, gp80 expression was down‐regulated, suggesting a negative feedback mechanism in this B‐cell hybridoma. These findings provide a role by which T‐cell‐dependent activation through CD40 regulates an IL‐6 autocrine loop, controlling B‐cell differentiation.
Lymphotoxin‐α is an important autocrine factor for CD40+interleukin‐4‐mediated B‐cell activation in normal and atopic donors
Authors: WORM M. , EBERMAYER K. , HENZ B.
Published: March 2003

Stimulation of human B cells with anti‐CD40+interleukin‐4 (IL‐4) results not only in proliferation and immunoglobulin E (IgE)‐production, but also increased production of the cytokine lymphotoxin‐α (LT‐α) (formerly also known as tumour necrosis factor‐β (TNF‐β)). Here, we studied the role of LT‐α (TNF‐β) in B cells following stimulation with anti‐CD40+IL‐4 from normal versus atopic donors. Anti‐CD40+IL‐4 stimulation of peripheral blood mononuclear cells (PBMC) from atopic donors resulted in enhanced production of soluble LT‐α (TNF‐β) and increased membrane LT‐α (TNF‐β) expression on the B cells compared with normal donors. Functional evaluation of LT‐α (TNF‐β) in CD40+IL‐4‐stimulated B cells shows that recombinant LT‐α (TNF‐β) induces proliferation of B cells and enhances CD40+IL‐4‐mediated B‐cell proliferation and IgE synthesis in both normal and atopic donors in a dose‐dependent manner. These findings were supported by semiquantitative analysis of ε‐germline transcripts using reverse transcription–polymerase chain reaction (RT–PCR) showing increased ε‐germline transcription in the presence of LT‐α. Furthermore, addition of anti‐LT‐α (anti‐TNF‐β) to CD40+IL‐4‐stimulated B cells partially inhibited proliferation and IgE synthesis in a dose‐dependent manner indicating a role of endogenous LT‐α (TNF‐β) production by B cells during continued CD40+IL‐4 stimulation. These data suggest that LT‐α (TNF‐β) plays a potentially significant role during B‐cell proliferation and IgE synthesis. Moreover, LT‐α (TNF‐β) production seems to be differentially regulated in B cells from normal and atopic donors.
Immuno‐stimulatory effects of bacterial‐derived plasmids depend on the nature of the antigen in intramuscular DNA inoculations
Authors: LEE S. W. , SUNG Y. C.
Published: March 2003

The CpG motifs of bacterial‐derived plasmids augment antigen‐specific immune responses and steer those responses towards the T helper 1 (Th1) type. In this study, we have addressed the immuno‐stimulatory effect of intramuscular co‐administration of CpG motifs containing vector DNA on the modulation of immune responses to the haemagglutinin (HA) and the nucleoprotein (NP) proteins of influenza virus. The co‐administration of vector DNA with a HA‐encoding plasmid DNA showed a significant enhancement in the total IgG response, the generation of cytotoxic T lymphocyte (CTL), and the T‐cell proliferative response. In the case of NP‐encoding plasmid DNA inoculations, the co‐administration of vector DNA slightly decreased the total IgG response, although the IgG2a/IgG1 ratio and the CTL responses to NP were significantly increased. These observations suggest that the immuno‐stimulatory effects of bacterial‐derived plasmids depend upon the nature of the co‐administered antigen.
CD4+ T‐cell‐mediated cytotoxicity against staphylococcal enterotoxin B‐pulsed synovial cells
Authors: KAWAKAMI  , MATSUOKA  , TSUBOI  , URAYAMA  , NAKASHIMA  , KAWABE  , KOJI  , AOYAGI  , MAEDA  , EGUCHI
Published: March 2003

Apoptosis of synovial cells in rheumatoid arthritis (RA) synovium determined in vivo is suggested to counteract the overgrowth of synovium. Immunohistological examination has revealed the infiltration of activated CD4+ T cells, which express Fas ligand (FasL), in RA synovium. The presence of a putative antigen (Ag) of autoimmune disorders in a target organ may induce the activation of specific T cells in the inflammatory region such as RA synovium. We examined the possible role of CD4+ T cells activated by synovial cells in a staphylococcal enterotoxin B (SEB)‐dependent manner, inducing synovial cell apoptosis. Synovial cells were cultured with or without interferon‐γ (IFN‐γ) and further incubated with CD4+ T cells in the presence of SEB. After the cocultivation, both the cytotoxicity and FasL expression of CD4+ T cells were investigated. Constitutive Fas expression was detected on both unstimulated and IFN‐γ‐stimulated synovial cells. CD4+ T cells did not kill SEB‐pulsed unstimulated synovial cells efficiently. In contrast, when CD4+ T cells were incubated with IFN‐γ‐stimulated synovial cells with SEB whose human leucocyte antigen (HLA)‐DR and ‐DQ expression was markedly induced, significant cytotoxicity by these cells against synovial cells was detected. The addition of anti‐HLA‐DR and ‐DQ monoclonal antibodies (mAbs) or human Fas chimeric protein (hFas‐Fc) reduced this cytotoxicity. FasL expression of CD4+ T cells cocultured with IFN‐γ‐stimulated synovial cells with SEB was significantly induced. Furthermore, the addition of mAbs against CD54, CD58 and CD106 inhibited both the cytotoxicity and FasL expression of CD4+ T cells induced by IFN‐γ‐stimulated synovial cells in the presence of SEB, indicating the importance of costimulatory molecules on synovial cells in activating CD4+ T cells. Our results suggest that CD4+ T cells are activated by synovial cells by an SEB‐dependent manner and express FasL, inducing Fas‐mediated apoptosis of the latter cells. These phenomena may regulate the overgrowth of synovial cells in RA synovium.
NADPH diaphorase‐positive dendritic profiles in rat thymus are discrete from autofluorescent cells, immunoreactive for inducible nitric oxide synthase, and show strain‐specific abundance differences
Authors: DOWNING  , VIRAG  , JONES
Published: March 2003

Predisposition to autoimmune disorder in Lewis rats has been associated with abnormal hypothalamic regulation of circulating steroids, leading to inadequate suppression of T helper 1 (Th1) cell‐mediated inflammatory reactions. In addition, autoimmune syndromes can be triggered within formerly resistant animals, following damage to the negative selection process of the thymus. A contribution to the autoimmune‐susceptible phenotype may therefore derive from the status of thymic tolerance. One mechanism of intrathymic negative selection may involve nitric oxide. Because inducible nitric oxide synthase (iNOS) is known to be inhibitable by steroids, its expression might be different within strains having neuroendocrine disturbance. We report on a study to compare intrathymic iNOS expression in autoimmune‐prone Lewis rats with other resistant strains. Interdigitating cells (IDC), darkly stained for diaphorase, were confirmed as immunoreactive for iNOS. They were located towards the medullary side of an accumulation of unstained, but autofluorescent cells (presumed to be macrophages) that circumscribes the corticomedullary zone. The role of iNOS+ IDC in the apoptotic deletion of T cells has been suggested by other studies. Despite the blunted steroidal condition reported for Lewis, nitrergic cell abundance was shown, by quantitative analysis of histochemical stain, to be on average approximately twofold lower compared with resistant strains (Fischer and Sprague–Dawley). This trend was evident in males and females, and confirmed by independent observers. We hypothesize that an intrathymic, iNOS‐dependent mechanism may be important for the suppression of potentially autoreactive T‐cell clones.
Splenic B cells are required for tolerogenic antigen presentation in the induction of anterior chamber‐associated immune deviation (ACAID)
Authors: D’ORAZIO  , NIEDERKORN
Published: March 2003

Ocular immune privilege is the result of a number of protective mechanisms, including a specialized immune response to antigen encountered in the anterior chamber of the eye. Anterior chamber‐associated immune deviation, or ACAID, is characterized by the antigen‐specific, selective down‐regulation of systemic cell‐mediated and humoral immune responses. One current hypothesis of the initiation of ACAID predicts that ocular APC process antigen and then migrate out of the eye and to the spleen where various regulatory T‐cell populations are generated. A novel in vitro model of the ACAID spleen was developed to study the cells involved in the generation of suppressed T‐cell immunity. ACAID APC co‐cultured with whole splenocytes or splenic B and T cells induced efferent suppressors of delayed‐type hypersensitivity (DTH). However, ACAID APC co‐cultured with splenic T cells did not generate efferent suppressors of DTH. The requirement for B cells was confirmed with B‐cell knockout mice. ACAID APC co‐cultured with splenocytes from B‐cell knockout mice did not induce efferent suppressors of DTH. Moreover, ACAID could not be induced in B‐cell knockout mice in vivo. The reconstitution of B‐cell knockout mice with wild‐type B cells restored ACAID. In summary, these data confirm the role for B cells in the splenic phase of ACAID. A putative mechanism predicts that ACAID APC release antigenic peptides to B cells in the spleen. B cells then present antigen in a tolerogenic manner leading to the generation of regulatory T cells.
B‐1‐like cells exist in sheep. Characterization of their phenotype and behaviour
Authors: CHEVALLIER  , BERTHELEMY  , LAINÉ  , LE RHUN  , FÉMÉNIA  , POLACK  , NAESSENS  , LEVY  , SCHWARTZ‐CORNIL
Published: March 2003

Two populations of B lymphocytes, B‐1 (CD5+ and/or CD11b+) and B‐2 (CD5− and CD11b−) cells have been described. In mice, which is the species of reference for B‐1 and B‐2 cell studies, these two subsets present different developmental schemes, phenotypes, antibody repertoires, localization and behaviours. Interestingly, in sheep, B cells rearrange their immunoglobulin (Ig) loci around the neonatal period, similarly to murine B‐1 cells. However, the phenotype of the sheep B cells has not been characterized with regards to their developmental pathway. In this report, we show that two sheep B‐cell subsets can be distinguished on the basis of CD11b expression. Relative to CD11b− B cells, the CD11b+ B cells frequently co‐express CD5, CD11c, higher levels of surface IgM (sIgM), show larger cell size and higher cell‐cycling activity, and thus present a B‐1‐like phenotype. However, unlike murine B‐1 cells, sheep B‐1 like cells mainly localize in blood, display a higher propensity to spontaneous apoptosis relative to B‐2‐like cells, and proliferate after sIgM stimulation. Our data show that despite neonatal immunoglobulin loci rearrangements, sheep B cells do not all express a B‐1‐like phenotype. However, B‐1‐and B‐2‐like cells co‐exist and present phenotypic and behavioural specificities. Nevertheless, sheep B‐1‐and B‐2‐like cells differ from the murine B‐1 and B‐2 cells in their cell behaviour. These subsets can thus not be considered as true homologues among species.
High‐level production of alternatively spliced soluble interleukin‐6 receptor in serum of patients with adult T‐cell leukaemia/HTLV‐I‐associated myelopathy
Authors: HORIUCHI  , AMPOFO  , KOYANAGI  , YAMASHITA  , WAKI  , MATSUMOTO  , YAMAMOTO  , YAMAMOTO
Published: March 2003

We have previously shown, using human T‐cell lymphocytotrophic virus‐I (HTLV‐I)‐infected cell lines, that soluble interleukin‐6 receptor (sIL‐6R) is generated through an alternative splicing mechanism. In this study, we examined human sera for the presence of alternatively spliced soluble IL‐6R (AS‐sIL‐6R). We produced a monoclonal antibody (mAb) recognizing the unique sequence of AS‐sIL‐6R peptide, generated by an altered reading frame. We also made recombinant AS‐sIL‐6R protein in Spodoptera frugiperda‐9 (Sf‐9) cells carrying baculovirus, which encoded altered sIL‐6R or conventional IL‐6R cDNA. mAbs specifically recognized AS‐sIL‐6R, but not conventional IL‐6R, as demonstrated by Western blot analyses, fluorescence‐activated cell sorter, immunofluorescence analyses and enzyme‐linked immunosorbent assay (ELISA). We adapted an ELISA system and used it for detection of altered sIL‐6R in sera from 23 healthy persons, 12 patients with adult T‐cell leukaemia (ATL) and 13 patients with HTLV‐I‐associated myelopathy (HAM). Serum levels of AS‐sIL‐6R were 6·4 or 6·1 times greater in ATL (28·7±20·4 ng/ml, P<0·0001) and in HAM patients (27·5±12·1 ng/ml, P<0·0001) than in healthy individuals (4·5±2·1 ng/ml). High levels of AS‐sIL‐6R were also observed in plasma from rheumatoid arthritis patients and in persons with elevated levels of alanine aminotransferase (ALT), antinuclear antibody (ANA), or α‐fetoprotein (AFP). However, in human immunodeficiency virus‐1 (HIV‐1), hepatitis B virus (HBV) or hepatitis C virus (HCV)‐infected individuals, AS‐sIL‐6R levels were not elevated. In this study, we confirmed that AS‐sIL‐6R is indeed present in human sera. These observations suggest that alternative splicing of IL‐6R mRNA is of consequence in ATL, HAM and in some autoimmune diseases. The HTLV‐I‐infected T cells appeared to play an important role in AS‐sIL‐6R production.