bluuebunny/crossref_metadata_2025 huggingface dataset:
Glioblastoma multiforme in a mature ovarian teratoma with recurring brain tumours
Authors: Yadav A , Lellouch‐Tubiana A , Fournet J C , Quazza J E , Kalifa C , Sainte‐Rose C , Jaubert F
Published: March 2003

Aims

We report a case study to highlight the occurrence of glioblastoma multiforme in an ovarian teratoma.

Methods and results

A 10‐year‐old girl presented with a left frontal lobe primitive neuroectodermal tumour which was successfully treated. After 6 uneventful years, she developed glioblastoma multiforme located posterior to the site of the initial tumour. Six years later, she presented with a mature cystic teratoma containing glioblastoma multiforme.

Conclusions

Glioblastoma in an ovarian teratoma is an exceptional event, which might have an initial clinical presentation as a metastatic brain tumour. Alternatively, recurring glial tumours may occur in a genetically predisposed person; the role of radiation and chemotherapy in this context remains to be elucidated.
Collagenous spherulosis mimicking keratinizing squamous metaplasia in a borderline endometrioid tumour of the ovary
Authors: Treilleux  , Godeneche  , Duvillard  , Clement‐Chassagne  , Suignard  , Bailly
Published: March 2003

Aims

Collagenous spherulosis (CS) is a rare lesion which is an incidental finding in breast and salivary glands. It is characterized by fibrillar spherules exhibiting an intrinsic radiating or concentric pattern which are surrounded by myoepithelial cells. This entity can be misdiagnosed as adenoid cystic carcinoma and in‐situ ductal carcinoma.

Methods and results

We report here the first case of CS arising in a borderline endometrioid tumour of the ovary where it merged with squamous metaplasia.

Conclusion

This observation illustrates another pitfall of CS which can be misidentified as keratin pearls. The pathogenesis remains unclear but it has been claimed that the accumulation of basement membrane material may be due to the proliferation of pre‐existing myoepithelial cells that secrete matrix components. Since ovarian tumours do not contain myoepithelial cells, one should assume that the epithelial cells differentiate towards myoepithelial cells as it has been shown in vitro and ex vivo.
Hyperexpression of ICAM‐1 and CD36 in placentas infected with Plasmodium falciparum: a possible role of these molecules in sequestration of infected red blood cells in placentas
Authors: Sartelet  , Garraud  , Rogier  , Milko‐Sartelet  , Kaboret  , Michel  , Roussilhon  , Huerre  , Gaillard
Published: March 2003

Aims

During pregnancy, Plasmodium falciparum malaria is frequent and associated with maternofetal complications. This could be the consequence of sequestration by several adhesion molecules of parasite‐infected red blood cells in syncytiotrophoblast. To investigate the expression of ICAM‐1 and CD36, two of the adhesion molecules for Plasmodium falciparum, an immunohistochemical study was carried out in malaria‐infected placentas.

Methods and results

Thirty‐five infected and 35 noninfected samples were chosen randomly. According to the histological classification of Bulmer, the infected placentas were separated in three groups: active, active chronic and past‐chronic infection. CD36 was localized in the cytoplasm of stromal cells of terminal villi of infected or noninfected placentas, but not in syncytiotrophoblast. ICAM‐1 was detected in the cytoplasm of stromal and endothelial villous cells in both infected and noninfected placentas and in syncytiotrophoblast of eight infected placentas showing more frequently active than active chronic or past‐chronic infection (P < 0.001). The percentage of cells immunostained for CD36 or ICAM‐1 was evaluated in the terminal villi. The proportion of villous cells, with ICAM‐1 and CD36 immunostaining, was significantly higher in infected vs. noninfected placentas (P < 0.0001) and CD36 was detected more in acute inflammatory vs. past‐chronic inflammatory placentas (P < 0.05).

Conclusions

The higher expression of ICAM‐1 in infected placentas and its localization in syncytiotrophoblast particularly during acute infection, suggest ICAM‐1 can act directly in the sequestration of parasite‐infected red blood cells (IRBCs). On the other hand, the expression of CD36 is influenced by the presence of IRBCs without being directly implicated in sequestration of IRBCs. The hyperexpression of these two molecules could explain the high frequency of malaria during pregnancy.
Lack of cyclin D1 overexpression in gastric carcinogenesis
Authors: Blok  , Craanen  , Van Diest  , Dekker  , J Tytgat
Published: March 2003

Aims

Cyclin D1 overexpression was examined in early gastric carcinomas and precursor lesions with the following aims; (1) to assess the chronology of cyclin D1 overexpression in various stages of gastric carcinogenesis, (2) to correlate cyclin D1 overexpression with the Lauren type, the grade of differentiation and the type of growth pattern of the tumours and (3) to correlate cyclin D1 overexpression with clinical parameters, in particular lymph node metastasis and overall prognosis.

Methods and results

Forty‐five paraffin‐embedded gastrectomy specimens from early carcinomas were examined for the presence of various precursor lesions. The Lauren type, the grade of differentiation and the type of growth pattern were reassessed for all early carcinomas. Cyclin D1 overexpression was examined using the monoclonal antibody DCS‐6. Cyclin D1 overexpression was absent from all precursor lesions. Ten early carcinomas (22%) were cyclin D1 positive without significant differences when stratified according to Lauren type, grade of differentiation, type of growth pattern or lymph node status. Univariate analysis failed to show a significant difference in 5‐year surival rate between cyclin D1 positive and negative early carcinomas (90% vs. 94%).

Conclusions

Cyclin D1 protein overexpression does not play a role in the progression from normal to neoplastic gastric mucosa and does not discriminate between intestinal and diffuse type early gastric carcinomas of Caucasian origin. Moreover, mechanisms other than cyclin D1 protein overexpression underlie the reported difference in biological behaviour of early gastric carcinomas with different types of growth pattern. Finally, although it appears that cyclin D1 does not have prognostic significance, studies on larger numbers, including advanced carcinomas, are warranted.
Diversion colitis in children: an iatrogenic appendix vermiformis?
Authors: Vujanić  , Dojčinov
Published: March 2003

Aims

Diversion colitis (DC) is a localized, relatively benign, iatrogenic condition which occurs in almost 100% of diverted colonic segments in patients who undergo ileostomy/colostomy for various reasons. The aim of this study was to establish histological features of DC in children.

Methods and results

Twenty‐three cases of DC following colostomy for Hirschsprung's disease in young children were analysed. The distinguishing features included prominent follicular lymphoid hyperplasia (100%), chronic mucosal inflammation (100%), accompanied by a variable degree of acute inflammation (78%) and Paneth cell metaplasia (26%). Less frequent histological findings were as follows: mild goblet cell depletion (22%), foci of cryptitis (13%), crypt abscesses (13%) and mild architectural distortion (22%). A previously unrecognized feature was the presence of mucosal aggregates of eosinophils, found in 43% of cases. A striking similarity between the normal appearance of the vermiform appendix and pathological features in DC was noted and the possible relationship between the two is discussed.

Conclusions

Histological features of DC in children are very similar to those described in adults. They should help to distinguish it from ulcerative colitis and Hirschsprung's‐associated enterocolitis in order to prevent inappropriate therapy and follow‐up. There are many similarities between DC and the normal appendix vermiformis.
Validation of immunolocalization of the urokinase receptor expression in ductal carcinoma in situ of the breast: comparison with detection by non‐isotopic in‐situ hybridization
Authors: Hildenbrand R , Leitz M , Magdolen V , Luther T , Albrecht S , Graeff H , Stutte H‐J , Bleyl U , Schmitt M
Published: March 2003

Aims: Ductal carcinoma in situ (DCIS) is a pre‐invasive form of mammary carcinoma with no microscopic evidence of cancer cell invasion through the basement membrane. However, for initiation of invasion, tumour cells have to acquire and focus proteolytic activity on to the cell surface in order to infiltrate the surrounding extracellular matrix. The receptor (uPA‐R or CD87) for the serine protease urokinase‐type plasminogen activator (uPA) plays a central role in invasion and metastasis. This study was performed to determine and localize m‐RNA and protein of uPA‐R in ductal carcinoma in situ of the breast.

Methods and results

We analysed uPA‐R mRNA and protein expression by in‐situ hybridization and immunohistochemistry, respectively, in 50 formalin‐fixed, paraffin‐embedded specimens of DCIS. Three different antibodies were used to stain cell‐associated uPA‐R; chicken polyclonal antibody (pAb) HU277 and monoclonal antibodies (mAb) IID7 and 3936. In all cases, myoepithelial and stromal cells reacted with either antibody. Especially, reaction of macrophage‐like cells with mAb 3936 resulted in a well‐marked and bright staining. Applying mAb IID7, in 46 of the 50 breast specimens tumour cells showed a positive immunoreaction. Likewise pAb HU277 stained tumour cells in 40 of the 50 cases, whereas mAb 3936 reacted with only 24 of the 50 tissue sections. Endothelial cells were marked by both mAb IID7 and pAb HU277 (46/50 and 35/50, respectively); mAb 3936 did not label at all. All of the cell types stained by mAb IID7 and pAb HU277 also displayed reactivity with uPA‐R mRNA‐specific antisense oligonucleotides in in‐situ hybridization.

Conclusions

Our results reveal the presence of the tumour invasion‐related receptor for the protease uPA not only in invasive ductal breast carcinoma but also in different types of DCIS.
When can complete regression of low‐grade gastric lymphoma of mucosa‐associated lymphoid tissue be predicted after Helicobacter pylori eradication?
Authors: Yamashita H , Watanabe H , Ajioka Y , Nishikura K , Maruta K , Fujino M A
Published: March 2003

Aims: 

Recent studies suggest that primary low‐grade gastric lymphomas of mucosa‐associated lymphoid tissue (MALT) are cured in many cases between 1 and 18 months after H. pylori eradication. The aim of this study is to elucidate when complete regression (CR) of MALT lymphoma can be histologically predicted after H. pylori eradication.

Methods and results: 

Twenty‐one patients with low‐grade gastric MALT lymphoma were treated with triple therapy (amoxicillin, clarythromycin and proton pump inhibitor) for 14 days. Subsequently, they were followed up by sequential endoscopy and biopsy (number of biopsy specimens for each endoscopy is 3–8, with an average of 4) from 91 to 657 days (average: 309 ± 165 days). Eradication of H. pylori infection was achieved in all patients. Nine patients were free of lymphoma at 1 to 2 months after eradication and remained in CR at 163–657 days. Twelve patients showed residual lymphoma at 1 to 2 months after eradication. Five out of 12 patients revealed only one or two small foci of lymphoma‐cell aggregation and showed a high incidence (80%) of CR at the latest biopsy (135–434 days, average 276 ± 115 days after eradication), while seven patients showed diffuse remains of lymphoma cells and indicated CR in only one case (14%) at 362 days, partial regression in five cases at 130–431 days (average 227 ± 114 days), and no change in one case at 91 days after eradication.

Conclusions: 

These results suggest that CR of low‐grade MALT lymphoma can be predicted at 1 to 2 months after eradication therapy by checking histological changes of MALT lymphoma cells.
Congenital gastrointestinal stromal tumour is morphologically indistinguishable from the adult form, but does not express CD117 and carries a favourable prognosis
Authors: Bates A W , Feakins R M , Scheimberg I
Published: March 2003

Aims: The histological and immunohistochemical features of a congenital stromal tumour of the jejunum are compared with those of adult gastrointestinal stromal tumours (GIST). The literature concerning the diagnosis and prognosis of congenital small intestinal stromal tumours is reviewed.

Methods and results

A term female infant presented with intestinal obstruction, from birth. Histology of a 15‐mm jejunal nodule showed a predominantly spindle‐cell tumour with epithelioid areas. There was a low mitotic count and mild nuclear pleomorphism, extensive necrosis and haemorrhage, and focal calcification. Immunohistochemically, tumour cells stained for muscle specific actin and vimentin. Staining for CD117 (c‐kit), S100, desmin and CD34 was negative. The features were compared to those of seven adult cases: no morphological feature was specific to the congenital tumour, which was smaller than the adult cases. There were no ultrastructural features specific for a particular line of differentiation. Immunohistochemical staining patterns were similar, except for CD117, which was strongly positive in all adult tumours, but negative in the congenital tumour.

Conclusions

This congenital jejunal stromal tumour morphologically resembled adult GIST, but lack of c‐kit expression suggests that it is nosologically distinct. Despite the presence of histological features which would cause the tumour to be categorized as malignant in an adult, it is apparent from previous reports of congenital small intestinal stromal tumours that the prognosis is favourable.
Effects of long‐term treatment with the anti‐androgen bicalutamide on human testis: an ultrastructural and morphometric study
Authors: Morgante E , Gradini R , Realacci M , Sale P , D'eramo G , Perrone G A , Cardillo M R , Petrangeli E , Russo Ma , Di Silverio F
Published: March 2003

To assess the effects of more than 4 years' treatment with the anti‐androgen bicalutamide on human testis by clinical, ultrastructural and morphometric analysis.

Two patients (aged 74 and 69 years) with prostate cancer were treated for more than 4 years with bicalutamide 50 mg daily. Clinical characterization and follow‐up included luteinizing hormone (LH), follicle‐stimulating hormone (FSH), testosterone and prostate‐specific antigen (PSA) measurements and clinical response of the tumours. Due to progression of the disease, patients underwent surgical orchidectomy as a further androgen withdrawal therapy. Testis biopsies were studied by light and electron microscopy and analysed by morphometry. Control samples were obtained from the normal testis of two patients with testicular cancer who underwent orchidectomy. Clinical follow‐up showed a good response in the control of tumour growth and serum PSA decreased to < 4 ng/mL; concentrations of serum LH, FSH and testosterone were within the normal range. Testicular morphology of treated patients was unexpectedly well preserved; the organization of seminiferous tubules was normal with all the germ line elements and mature spermatozoa present. In some areas, a net increase of peritubular connective tissue was evident which may be a consequence of the age of the patients.

Long‐term bicalutamide (50 mg) treatment appears to have very little impact on testis ultrastructure and sperm maturation, while it is effective in the control of androgen‐dependent prostatic tumours.
Myofibroblastic differentiation in malignant fibrous histiocytoma (pleomorphic myofibrosarcoma): a clinicopathological study
Authors: Montgomery E , Fisher C
Published: March 2003

Myofibroblastic differentiation in malignant fibrous histiocytoma (pleomorphic myofibrosarcoma): a clinicopathological study

Aims: We compared the clinical and pathological features of pleomorphic malignant fibrous histiocytoma (MFH)‐like soft tissue sarcomas with and without myofibroblastic differentiation on electron microscopy.

Methods and Results: Fifty‐three soft tissue tumours designated as MFH by light and electron microscopy were reassessed. Eighteen were specifically diagnosed and excluded, and follow‐up (FU) information obtained for 24 of the other 35 cases. Myofibroblastic ultrastructure was seen in 7/24 (29%). Seventeen of 24 (71%) lacked myofibroblasts on electron microscopy, which showed fibroblastic or undifferentiated cells. Histologically, all tumours but one had storiform‐pleomorphic areas; one myofibroblastic neoplasm was fascicular and myxoid. No other morphological differences were seen. In seven myofibroblastic cases, smooth muscle in four cases and muscle‐specific actin in two cases, desmin in three cases and S100 in one case were present. In 15 other tumours, smooth muscle in five cases and muscle‐specific actin in one case, and desmin in one case were present; none of these cases expressed S100. CD34 was found in the myxoid areas of one myofibrosarcoma and 3/15 other tumours. Positivity for bcl‐2 was seen only in non‐myofibroblastic sarcomas (4/14). On follow‐up (median 41 months), 2/7 (29%) myofibroblastic tumours recurred, 5/7 (71%) metastasized, and 3/7 (43%) patients died of disease. Among the non‐myofibroblastic sarcomas, with a median follow‐up of 47 months, 6/17 cases (35%) recurred, 10/17 (59%) metastasized, and 7/17 patients (41%) died of disease.

Conclusions: Pleomorphic sarcomas with and without myofibroblastic differentiation on electron microscopy are clinically and histologically similar. The former display myoid immunohistochemical markers more frequently.
Malignant epithelioid mesothelioma: anti‐mesothelial marker expression correlates with histological pattern
Authors: Attanoos R L , Webb R , Dojcinov S D , Gibbs A R
Published: March 2003

Malignant epithelioid mesothelioma: anti‐mesothelial marker expression correlates with histological pattern

Aims: Malignant epithelioid mesothelioma shows marked cytoarchitectural diversity. The aim of the study was to evaluate how immunoreactivity with mesothelial markers related to histological pattern.

Methods and results: Ninety‐two cases of malignant epithelioid mesothelioma (60 pleural, 32 peritoneal) were examined and classified as exhibiting tubulopapillary, adenomatoid, solid, small cell or pleomorphic patterns. All cases were immunohistochemically stained with thrombomodulin, calretinin, CD44H, and cytokeratin 5/6. Cases of malignant mesothelioma exhibited a number of different histological forms. Immunohistochemical expression of each mesothelial marker tested was not homogeneous across different histological patterns of malignant epithelioid mesothelioma, even within the same tumour section. Calretinin (with nuclear expression) was identified to show the highest overall sensitivity and lowest range variation in staining (67% sensitivity in small cell areas to 100% expression in pleomorphic areas). Cytokeratin 5/6 and thrombomodulin yielded similar overall sensitivity. Thrombomodulin appeared to demonstrate higher sensitivity for small cell variant tumour (83% sensitivity). A notable advantage with cytokeratin 5/6 was that expression was more diffuse in nature rather than the focal membranous elaboration seen in thrombomodulin. The widest range of staining was seen in small cell mesothelioma (83% sensitivity with thrombomodulin to 17% sensitivity with cytokeratin 5/6) and in tubulopapillary areas (90% sensitivity with calretinin to 38% sensitivity with CD44H).

Conclusions: Calretinin appears most useful and shows the highest overall sensitivity for malignant epithelioid mesothelioma, with good expression in areas displaying a tubulopapillary, adenomatoid, solid and pleomorphic pattern. For small cell mesothelioma, thrombomodulin appears to confer higher sensitivity and is advocated, in this setting, as the first line mesothelial marker. Cytokeratin 5/6 is a useful and easily interpretable mesothelial marker. CD44H is not of particular use in the diagnosis of malignant epithelioid mesothelioma. Accurate interpretation of immunohistochemistry in mesothelioma requires an awareness of the immunophenotypic heterogeneity identified in different histological forms of the tumour, and this is of particular importance in small biopsies.
Genomic imbalances detected by comparative genomic hybridization are prognostic markers in invasive ductal breast carcinomas
Authors: Zudaire I , Odero M D , Caballero C , Valenti C , Martínez‐Penuela J M , Isola J , Calasanz M J
Published: March 2003

Genomic imbalances detected by comparative genomic hybridization are prognostic markers in invasive ductal breast carcinomas

Aims: The aim of this work is the study of the prognostic significance of the chromosomal aberrations described in a series of invasive ductal breast carcinomas.

Methods and results: We analysed by comparative genomic hybridization a group of 70 formalin‐fixed paraffin‐embedded invasive ductal breast carcinomas. Aberrations showed a frequency similar to previous studies using frozen tumours. Interestingly, we identified gains involving 6q16‐q24 more frequently than in other series. We analysed the association among the chromosomal imbalances, 11 histopathological factors, relapse rate and overall survival of patients. Associations showed 16q losses as a potential marker of good prognosis, as they were more frequent in node‐negative (P=0.025) and in oestrogen‐positive tumours (P < 0.001). Furthermore, 100% of bcl‐2+ tumours presented this aberration compared with 29.3% in bcl‐2– (P=0.014). 1q, 11q, 17q and 20q gains were associated with poor prognosis: 95% of cases with 1q gains were bigger than 20 mm (P=0.041). Tumours with 1q and 11q gains showed a higher relapse rate (P=0.063; P=0.066). Within the good prognosis group of lymph node‐negative patients, 17q and 20q gains identify a subgroup with increased relapse rate (P=0.039).

Conclusions: Chromosomal imbalances, together with histopathological factors, may help to predict outcome in breast cancer patients.
Immunohistochemical detection of 8‐hydroxydeoxyguanosine, a marker of oxidative DNA damage, in human chronic cholecystitis
Authors: Seki S , Kitada T , Yamada T , Sakaguchi H , Nakatani K , Onoda N , Satake K
Published: March 2003

Immunohistochemical detection of 8‐hydroxydeoxyguanosine, a marker of oxidative DNA damage, in human chronic cholecystitis

Aims: Recent studies suggest that oxidative DNA damage induced during chronic inflammation may play a role in carcinogenesis in some organs. Although gallbladder carcinomas are frequently observed with a background of chronic cholecystitis, little is known about oxidative DNA damage in chronic cholecystitis. The aims of this study were to investigate the expression of 8‐hydroxydeoxyguanosine (8‐OHdG), a biomarker of oxidative DNA damage, in normal and chronically inflamed human gallbladder mucosa and compare its expression with clinicopathological findings.

Methods and results: 8‐OHdG expression was immunohistochemically examined using a monoclonal antibody against 8‐OHdG in human gallbladder specimens. In normal gallbladder (n=5), no 8‐OHdG expression was observed. In contrast, nuclear expression of 8‐OHdG was detected in 28 of 31cases (90.3%) in gallbladder epithelial cells with chronic cholecystitis. The positive cells were predominantly observed in the areas of active inflammation with prominent cell infiltration. Quantitative analysis revealed that the number of 8‐OHdG+ cells (labelling index) significantly (rs=0.671, P < 0.05) correlated with the degree of the activity of mucosal inflammation, while gender, age, and the presence of gallstones did not influence the index.

Conclusions: Oxidative DNA damage is common in chronic cholecystitis, suggesting a possible link between chronic inflammation and gallbladder carcinogenesis.
Imaging bioluminescent indicators shows Ca2+ and ATP permeability thresholds in live cells attacked by complement
Authors: SALA‐NEWBY  , TAYLOR†  , BADMINTON  , REMBOLD  , CAMPBELL
Published: March 2003

A series of permeability thresholds to Ca2+ metabolites and macromolecules, occurring at different times when cells are attacked by complement, has been established by imaging HeLa cells transiently expressing a recombinant cytosolic fusion protein of firefly luciferase and aequorin (luciferase–aequorin) to measure changes in ATP and cytosolic free Ca2+. Nuclear fluorescence of propidium was used as a measure of permeability to small molecules, and luciferase activity imaged to assess lysis. The rise in cytosolic free Ca2+ observed after C9 attack preceded by at least 60 s both the increase in propidium fluorescence, measured in single cells, and the decrease in ATP monitored by luciferase light emission. These effects were dependent on the concentration of C9. At concentrations of C9 up to 4 μg/ml no loss of luciferase–aequorin protein was detected at the end of the experiment. Thus the membrane integrity of the cells remained intact, even though the cells were permeable to propidium. These results confirmed our earlier observations that propidium permeability in cells attacked by complement was not a reliable measure of cell death. They also show that it is vital to take account of cellular heterogeneity if the mechanisms by which cells respond to membrane pore former attack are to be correctly interpreted.
Effect of T‐helper cytokine environment on specificity of T‐cell responses to mycobacterial 65 000 MW heat‐shock protein
Authors: SIEW  , BEECH  , THOMPSON  , ELSON
Published: March 2003

The purpose of this work was to determine if the fine specificity of T cells differed between mice immunized with an antigen in a T helper 1 (Th1) cytokine‐dominated environment as compared with a T helper 2 (Th2) cytokine‐dominated environment. It was found that splenic T cells from mice immunized with mycobacterial heat‐shock protein (hsp 65) and interleukin‐12 (IL‐12) produced less interleukin‐4 (IL‐4) and more interferon‐γ (IFN‐γ) in response to stimulation with hsp 65 in vitro than did T cells from mice immunized with hsp 65 alone. The T‐cell proliferative response to hsp 65 did not differ between the two groups of mice, although the responses were higher than those of T cells from non‐immunized mice. Strikingly, T cells from mice given hsp 65 and IL‐12 gave significantly higher responses to six peptides (corresponding to the sequence of hsp 65) to which T cells from mice immunized with hsp 65 alone did not respond. It is considered that different epitopes are presented to T cells (possibly owing to changes in antigen processing) if the environment is shifted, by IL‐12, from Th2 towards Th1 cytokines.
Characterization of the gene for the membrane and secretory form of the IgM heavy‐chain constant region gene (Cμ) of the cow (Bos taurus)
Authors: MOUSAVI  , RABBANI  , PILSTRÖM  , HAMMARSTRÖM
Published: March 2003

Our present understanding of the evolution of immunoglobulins is derived from a few vertebrate species. In order to obtain additional information on the development of the humoral immune system, we cloned and determined the nucleotide sequence of the bovine cDNA and genomic IgM heavy‐chain constant region gene (Cμ). The gene contains four constant region domain‐encoding exons (CH1 to CH4) and two exons encoding the transmembrane domain (TM1, TM2), expressed in the membrane‐bound receptor form of the IgM. The sequence of a cDNA clone encoding the 3′ portion of the membrane form of the μ‐chain revealed that the TM1 exon is spliced to the CH4 exon, as occurs in other mammals. Comparison of deduced amino acid sequence data from different vertebrates revealed a high similarity to sheep Cμ (88%) and a lower degree of similarity to pig (62%), rat (62%), rabbit (58%) human (56%), hamster (55%), mouse (54%), chicken (28%) and horned shark (22%) Cμ.
Recognition of a common mycobacterial T‐cell epitope in MPB59 of Mycobacterium bovis
Authors: LIGHTBODY  , GIRVIN  , POLLOCK  , MACKIE  , NEILL  , POLLOCK
Published: March 2003

Bovine tuberculosis, which persists as a residual level of infection in many European countries, has implications not only for the economy of farming communities but also for human health. The aim of this study was to identify a common mycobacterial antigen which was recognized in bovine tuberculosis and to characterize the response to this antigen at the epitope level. A T‐cell clone, phenotype CD4+, raised from an animal experimentally infected with Mycobacterium bovis was shown to proliferate in response to a panel of sonicates derived from different mycobacterial species indicating recognition of an antigen with broad specificity. This antigen was subsequently shown to be MPB59. Recognition of MPB59 at the epitope level was determined in experimental and field cases of bovine tuberculosis using a panel of synthetic peptides (20‐mers with 10‐residue overlaps) incorporating the signal sequence and mature protein. The results showed that in vitro interferon‐γ was predominantly produced in response to adjacent peptides numbers 10 and 11, suggesting that the dominant epitope was contained in the overlap, correlating to residues 101–110 (YYQSGLSIVM). This epitope was recognized by 54% of tuberculous cattle of mixed breeds, which suggests that it may be genetically permissive in terms of major histocompatibility complex presentation. Sequence analysis confirmed that there were only minor differences in the amino acid composition within this region for various mycobacterial species, which could explain the common T‐cell recognition described in this study. Common recognition of this epitope indicates that it would have limited potential for use as a diagnostic reagent per se but may have potential for inclusion in a subunit vaccine.
Gene transfer of costimulatory molecules into a human colorectal cancer cell line: requirement of CD54, CD80 and class II MHC expression for enhanced immunogenicity
Authors: LINDAUER  , RUDY  , GÜCKEL  , KNEBEL DOEBERITZ  , MEUER  , MOEBIUS
Published: March 2003

Colorectal cancer is considered a non‐immunogenic malignany. One strategy to augment the immunogenicity of such tumours is represented by the expression of costimulatory molecules by gene transfer. Using transfected variants of the human colorectal cancer cell line SW480 we tested various costimulatory molecules (CD80, CD86, CD54) and a class II major histocompatibility complex (MHC) allele (HLA‐DR3) alone or in combination on their ability to support primary T‐lymphocyte activation in vitro. Expression of CD80 or CD86 similarly as the combination of both was not sufficient to induce proliferation of human allogeneic T cells. Expression of CD54 together with CD80 strongly augmented the costimulatory function of CD80, as observed in the presence of a CD3 monoclonal antibody (mAb), but did not lead directly to a T‐cell response against modified tumour cells. Importantly, SW480 cells coexpressing CD54, CD80 and the HLA‐DR3 allele effectively promoted T‐lymphocyte proliferation. Moreover, the use of such CD54+/CD80+/HLA‐DR3+ SW480 variants for repetitive stimulations resulted in the generation of T‐cell lines predominantly composed of CD8+ T cells exhibiting class I MHC restricted cytolytic activity towards untransfected SW480 tumour cells. This demonstrates that the generation of immunogenic tumour cell variants, i.e. for the use as cellular vaccines, requires multiple genetic alterations in the case of non‐immunogenic human tumours cells, such as colorectal cancer cells.
Active suppression induced by cutaneous exposure to bacterial superantigen is prevented by interleukin‐12 treatment in vivo
Authors: SALOGA  , ENK  , BECKER  , BELLINGHAUSEN  , KÜHN  , KNOP
Published: March 2003

Exposure to the bacterial superantigen staphylococcal enterotoxin B (SEB) leads to inhibition of several immune responses and the induction of regulatory cells. The aim of this study was to characterize these regulatory cells further and to investigate the effect of interleukin‐12 (IL‐12) on superantigen‐induced suppression. For this purpose BALB/c mice were injected subcutaneously with low doses of SEB that did not deplete the SEB‐reactive Vβ T cells. Intravenous transfer of unseparated local‐draining lymph node cells from these SEB‐treated animals suppressed the proliferative response of mononuclear spleen cells of naive syngeneic recipients for at least 3 weeks. The regulatory cells did not produce the type 2 cytokines, interleukin‐4 (IL‐4) or interleukin‐10 (IL‐10), or increased amounts of transforming growth factor‐β (TGF‐β). Depletion of CD8+ or SEB‐reactive Vβ7+ and Vβ8+ T cells, prior to transfer, abrogated the suppressive effect. Intraperitoneal injections of IL‐12 into donors, prior to SEB treatment, prevented the induction of functional regulatory cells, and treatment of recipients with IL‐12, prior to receipt of cells from SEB‐treated donors, prevented the suppressive effect of regulatory cells that were already induced. The data indicate that exposure to minute amounts of superantigens directly induces superantigen‐reactive and CD8+ regulatory T cells and that superantigen‐induced suppression can be prevented and reversed by IL‐12 treatment in vivo.
Imm_unization of mice with phosphatidylcholine drastically reduces the parasitaemia of subsequent Plasmodium chabaudichabaudi blood‐stage infections
Authors: BORDMANN  , RUDIN  , FAVRE
Published: March 2003

It has been suggested that phospholipids and antibodies directed against phospholipids are important in the pathology of malaria. We have investigated the influence of imm_unizations with phospholipids on the course of subsequent blood‐stage Plasmodium chabaudi chabaudi infections in ICR inbred mice. We observed a significant reduction in the parasitaemia following imm_unization with phosphatidylcholine (PC), but not with phosphatidylethanolamine (PE) imm_unization. At the peak of the infection, PC‐imm_unized mice displayed a T‐helper 2 (Th2)‐type cytokine production pattern, whereas PE‐imm_unized or non‐treated controls displayed a cytokine production pattern of the T‐helper 1 (Th1) type. Serum imm_unoglobulin transfer from PC‐imm_unized mice protected naive mice in a similar fashion to PC‐imm_unization, demonstrating that the observed reduction of parasitaemia was caused by the presence of PC‐specific antibodies.
Vβ11+ T‐lymphocyte expansion by toxic shock syndrome toxin‐1 differs in mice bearing H‐2q versus H‐2b haplotypes
Authors: ZHAO  , BRUNSBERG  , HOLMDAHL  , TARKOWSKI
Published: March 2003

We have recently demonstrated that toxic shock syndrome toxin‐1 (TSST‐1) expanded Vβ11+ T lymphocytes contribute to Staphylococcus aureus arthritis and sepsis‐induced mortality. Interestingly, Vβ11+ T‐cell mediated joint pathology varies in different mouse strains. In this study, we characterized the in vitro pattern of Vβ11+ T‐cell expansion by TSST‐1 in mice with various genetic backgrounds. Mice expressing major histocompatibility complex (MHC) class II I‐E molecules did not expand Vβ11+ T cells upon stimulation with TSST‐1. Using B10 congeneic I‐E negative mouse strains, we found that the TSST‐1‐expanded Vβ11+ T cells in B10Q (H‐2q) and B10M (H‐2f) mice but not in B10B (H‐2b) mice. Antigen‐presenting cells (APC) from B10Q mice, L cells and lymphoma cell line transfected with a q gene did not restore the deficient Vβ11+ T‐cell expansion by TSST‐1 in purified T cells from B10B mice. In contrast, I‐Ab APC were able to stimulate Vβ11+ T cells from H‐2q mice. Furthermore, Vβ11+ T cells in H‐2b mice did expand when exposed to staphylococcal enterotoxin A (SEA). These findings suggest that the T‐cell repertoire, skewed by clonal deletion and inactivation of self‐reactive T cells, accounts for the different magnitude of Vβ11+ T‐cell expansion among the different mouse strains.
Lysosomal routing of FcγRI from early endosomes requires recruitment of tyrosine kinases
Authors: NORMAN  , HARRISON  , DAVIS  , FLOTO  , ALLEN
Published: March 2003

The high‐affinity receptor for imm_unoglobulin G (FcγRI) plays a central role in the clearance of imm_une complexes by mediating their internalization and delivery to lysosomes. In monocytic U937 cells, receptor internalization is independent of tyrosine kinase activity. However, the tyrosine kinase inhibitor, genistein, prevents further progress of the receptor to lysosomes and traps it in a sub‐plasma membrane early endosome. Similarly, FcγRI expressed in COS cells is able to internalize imm_une complexes but is unable to translocate to lysosomes. This suggests that FcγRI, whose cytoplasmic tail is devoid of known signalling motifs, must recruit tyrosine kinases via its γ‐chain to achieve lysosomal delivery. We show that a chimera of the extracellular domain of FcγRI and the cytoplasmic tail of the γ‐chain is both internalized and efficiently trafficked to lysosomes. Our study suggests that a key function of the γ‐chain is recruitment of tyrosine kinases to initiate the intracellular signalling pathways required to target FcγRI following imm_une complex aggregation to lysosomes and not to initiate endocytosis per se.
Modulation of virus‐induced delayed‐type hypersensitivity by plasmid DNA encoding the cytokine interleukin‐10
Authors: Manickan E. , Daheshia M. , Kuklin N. , Chun S. , Rouse B. T.
Published: March 2003

This report evaluates the efficacy of eukaryotic expression plasmids encoding cytokines at modulating the induction and expression of cutaneous delayed‐type hypersensitivity (DTH) responses to virus infections. Mice given a single intramuscular administration of cytokine DNA were subsequently infected with either herpes simplex virus (HSV) or vaccinia virus, then tested for DTH. Responses in animals given interleukin‐10 DNA were markedly suppressed for at least 5 weeks after pretreatment. Animals also expressed diminished T‐cell proliferative responses and modest changes in the balance of T helper type 1 and 2 T‐cell reactions. Treatment of animals already sensitized to express DTH, also showed inhibited responses, these taking 6–7 days after treatment to become apparent. Our results show the potency and convenience of plasmid DNA encoding cytokines to modulate inflammatory reactions. Advantages and risks of the cytokine DNA approach are briefly discussed.
Role of mRNA stability in the different patterns of cytokine production by CD4+ cells from young and old mice
Authors: PIOLI C. , PUCCI S. , BARILE S. , FRASCA D. , DORIA G.
Published: March 2003

CD4+ cells from young (3 months) and old (19 months) mice were stimulated by plate‐bound anti‐CD3 monoclonal antibody (mAb) alone or also by soluble anti‐CD28 mAb. Supernatants were analysed by enzyme‐linked immunosorbent assay (ELISA) to determine cytokine concentrations. Total RNA was extracted from cells, reverse transcribed and the cDNA amplified by polymerase chain reaction (PCR) to evaluate the amount of specific mRNA. The results indicate that anti‐CD3 alone is not sufficient to induce interleukin‐2 (IL‐2) production in CD4+ cells from both young and old mice. However, anti‐CD28, together with anti‐CD3 mAb, induces a much higher production of IL‐2 in CD4+ cells from young as compared with old mice. Conversely, interferon‐γ (IFN‐γ) production is also induced by anti‐CD3 alone and is higher in CD4+ cells from old as compared with young mice. Upon addition of anti‐CD28 mAb, IFN‐γ production increases in both groups, but it remains much higher in old than in young mice. Also the production of IL‐4 and IL‐10 is induced by anti‐CD3 mAb but it is increased by the addition of anti‐CD28 mAb. CD4+ cells from old mice produce more IL‐4 and IL‐10 as compared with cells from young mice. The amounts of cytokine specific mRNA in CD4+ cells from young and old mice parallel the cytokine levels in culture supernatants. Results on the mRNA turnover indicate that when CD4+ cells are stimulated by anti‐CD3 or costimulated also by anti‐CD28 mAb, the IFN‐γ, IL‐4 and IL‐10 specific mRNAs are more stable in old than in young mice, suggesting that mRNA stability has a relevant role in the different patterns of cytokine production.
Binding of HIV‐2 envelope glycoprotein to CD8 molecules and related chemokine production
Authors: AKIMOTO  , KANEKO  , SEKIGAWA  , HASHIMOTO  , KANEKO  , YAMAMOTO
Published: March 2003

We recently found that human immunodeficiency virus (HIV)‐2 envelope glycoprotein, but not that of HIV‐1, could bind to CD4 and CD8 molecules on T cells, and that the binding site of HIV‐2 envelope glycoprotein was located on the α‐chain (but not the β‐chain) of CD8. This study showed that the binding of HIV‐2 envelope glycoprotein could induce phosphorylation of protein tyrosine kinase p56lck in CD8+ T cells. We also found that production of β‐chemokines in response to HIV‐2 envelope glycoprotein was significantly higher than that in response to HIV‐1 envelope glycoprotein, and that CD8+ T cells were the main source of β‐chemokines production among the T‐cell population. These findings indicate the possibility that the binding of envelope glycoprotein to CD8 molecules are related to signal transduction into CD8+ T cells and the resultant β‐chemokine production in HIV‐2 infection. Our results may help to explain the differences in disease manifestations between HIV‐1 and HIV‐2, including the lower virulence of HIV‐2 and the longer survival of HIV‐2‐infected individuals.
Autoreactive CD4− CD8−αβ T cells to vaccinate adjuvant arthritis
Authors: HAQUE  , KIMOTO  , INADA  , TOKUNAGA  , KOHASHI
Published: March 2003

Studies suggested that experimental autoimmune diseases can effectively be prevented and treated by application of normal autoreactive T cells or autoreactive T cells in an attenuated form. In this study, several autoreactive CD4− CD8− T‐cell clones (A2, A6, and A13 cells) were isolated for the first time from the draining lymph nodes of Lewis rats with adjuvant arthritis (AA). Surprisingly, intraperitoneal inoculation with A13 cells, but not A2 or A6 cells protected rats from AA both clinically and histologically. It was demonstrated that A13 cells were CD4− CD8−αβ T cells, and showed proliferative responses to irradiated syngeneic spleen cells (antigen‐presenting cells; APC). Interestingly, A13 cells proliferated against concanavalin A (Con A) and staphylococcal enterotoxin B (SEB), but did not show any proliferation to Mycobacterium tuberculosis (Mt), or its 65 000 MW heat‐shock protein (HSP). Rats protected from AA by inoculation with A13 cells showed a specific anti‐idiotypic delayed‐type hypersensitivity reaction compared with other autoreactive T cells (A2 or A6 cells). These findings demonstrate that AA can be suppressed by autoreactive CD4− CD8−αβ T cells, and these cells may be used as therapeutic agents in experimental autoimmunity.
Rat NKR‐P1+ CD3+ T cells: selective proliferation in interleukin‐2, diverse T‐cell‐receptor‐Vβ repertoire and polarized interferon‐γ expression
Authors: BADOVINAC  , BOGGIANO  , TRAJKOVIĆ  , FREY  , VUJANOVIĆ  , GOLD  , MOSTARICA‐STOJKOVIĆ  , VUKMANOVIĆ
Published: March 2003

Cells expressing markers of both natural killer and T lymphocytes (NK T cells) in humans and mice express a restricted T‐cell receptor (TCR) repertoire, are of CD4− CD8− or CD4+ CD8− phenotype, and upon anti‐CD3 stimulation secrete large amounts of interleukin‐4 (IL‐4) and interferon‐γ (IFN‐γ). NK T cells may be the primary source of IL‐4‐promoting T helper type 2 (Th2) responses and/or they might be involved in regulating the balance between Th1‐ and Th2‐type immune responses, and may consequently affect susceptibility to autoimmune diseases associated with a skewed Th phenotype. We show that rat NK T cells selectively proliferate to IL‐2, and use this fact to analyse cytokine production by NK T cells in two rat strains differentially susceptible to Th1‐ or Th2‐type autoimmune diseases. Analysis by reverse transcription–polymerase chain reaction revealed that, in contrast to mouse, rat NK T cells secrete exclusively IFN‐γ and not IL‐4 after anti‐CD3 stimulation, and use a wider TCR‐Vβ repertoire, suggesting that rat NK T cells are not essential for the development of Th2‐type CD4+ T‐cell responses.
Vitamin D differentially regulates B7.1 and B7.2 expression on human peripheral blood monocytes
Authors: CLAVREUL  , D’HELLENCOURT  , MONTERO‐MENEI  , POTRON  , COUEZ
Published: March 2003

The hormonal active form of vitamin D3, 1,25‐dihydroxyvitamin D3 (1,25(OH)2D3), inhibits (through an unknown mechanism) the ability of monocytes/macrophages to induce T‐cell activation. For T cells to be optimally activated, recognition of antigen/major histocompatibility complexes (MHC) by the T‐cell receptor (TCR) must be accompanied by a second costimulatory signal. Considerable experimental data now suggest that this costimulatory signal is predominantly generated by B7.1 and/or B7.2 molecules, expressed on antigen‐presenting cells (APC), when engaged to their counter‐receptor, CD28, present on T cells. To determine whether the inhibitory effect of 1,25(OH)2D3 on monocytes/macrophages might involve modulation of the expression of B7.1 and B7.2 molecules, we analysed (by flow cytometry) the influence of 1,25(OH)2D3 and an analogue, KH 1060, on the expression of these two molecules at the surface of resting human peripheral blood monocytes. In parallel, we tested the effect of these two agents on human monocyte expression of cell‐surface markers (CD14 and CD4) and antigen‐presenting molecules (MHC class I and MHC class II). Our results showed that both 1,25(OH)2D3 and KH 1060 inhibited the basal expression of B7.2 in a dose‐ and time‐dependent manner, without affecting B7.1. Moreover, these two compounds increased CD14 and reduced MHC class II and CD4 expression. Furthermore, the effect of 1,25(OH)2D3 on B7 molecule expression in combination with lipopolysaccharide (LPS) or cytokines, including interleukin‐10 (IL‐10), interferon‐γ (IFN‐γ) and tumour necrosis factor‐α (TNF‐α), was studied. The 1,25(OH)2D3‐induced B7.2 down‐regulation was still detectable when monocytes were activated by IL‐10, IFN‐γ and TNF‐α but not with LPS. Moreover, the induction of B7.1 by TNF‐α was inhibited by addition of 1,25(OH)2D3. We conclude that the ability of 1,25(OH)2D3 to decrease B7.2 expression on human monocytes might contribute to its inhibitory effect on APC‐dependent T‐cell activation and to its immunosuppressive properties observed in autoimmune diseases and organ transplantation.
Effects of type I/type II interferons and transforming growth factor‐β on B‐cell differentiation and proliferation. Definition of costimulation and cytokine requirements for immunoglobulin synthesis and expression
Authors: ESTES  , TUO  , BROWN  , GOIN
Published: March 2003

In this report, we sought to determine the role of selected type I interferons [interferon‐α (IFN‐α) and interferon‐τ (IFN‐τ)], IFN‐γ and transforming growth factor‐β (TGF‐β) in the regulation of bovine antibody responses. B cells were stimulated via CD40 in the presence or absence of B‐cell receptor (BCR) cross‐linking. IFN‐α enhanced IgM, IgG2 and IgA responses but did not enhance IgG1 responses. BCR signalling alone was more effective at inducing IgG2 responses with IFN‐α than dual cross‐linking with CD40. Recombinant ovine IFN‐τ was less effective at inducing IgG2 responses when compared with IFN‐α, though IgA responses were similar in magnitude following BCR cross‐linking. At higher concentrations, IFN‐τ enhanced IgA responses greater than twofold over the levels observed with IFN‐α. Previous studies have shown that addition of IFN‐γ to BCR or pokeweed mitogen‐activated bovine B cells stimulates IgG2 production. However, following CD40 stimulation alone, IFN‐γ was relatively ineffective at stimulating high‐rate synthesis of any non‐IgM isotype. Dual cross‐linking via CD40 and the BCR resulted in decreased synthesis of IgM with a concomitant increase in IgA and similar levels of IgG2 production to those obtained via the BCR alone. We also assessed the effects of endogenous and exogenous TGF‐β on immunoglobulin synthesis by bovine B cells. Exogenous TGF‐β stimulates both IgG2 and IgA production following CD40 and BCR cross‐linking in the presence of IL‐2. Blocking endogenous TGF‐β did not inhibit the up‐regulation of IgG2 or IgA by interferons.