bluuebunny/crossref_metadata_2025 huggingface dataset:
Chromosomal numerical aberrations are frequent in oesophageal and gastric adenocarcinomas: a study using in‐situ hybridization
Authors: Beuzen F , Dubois S , Fléjou J‐F
Published: March 2003

Aims 

(i) To investigate and compare the numerical aberrations of chromosomes 7, 8, 11, 17 and Y in a series of 60 adenocarcinomas of the oesophagus, gastric cardia and gastric antrum; and (ii) to specify the sequence of chromosomal aberrations occurring during the neoplastic progression of Barrett's oesophagus.

Methods and results: 

Chromosomal in‐situ hybridization was performed on deparaffinized tissue sections from 20 Barrett's adenocarcinomas, 20 adenocarcinomas of the cardia and 20 adenocarcinomas of the antrum, with centromeric alpha satellite DNA probes specific for chromosomes 7, 8, 11, 17 and Y, labelled with digoxigenin. Signals were detected by immunoperoxidase staining. The copy number for each chromosome was counted in 200 tumour cells nuclei and 100 lymphocytes as controls. In parallel, the DNA content of the nuclear suspensions was measured by flow cytometry. Numerical abnormalities of the five chromosomes (loss of the Y chromosome, monosomy, trisomy, and tetrasomy) were frequently observed in the three groups of adenocarcinomas (from 40% to 65% of the cases). Sixty per cent to 75% of oesophagus and gastric adenocarcinomas were DNA‐aneuploid. Chromosomal aberrations progressively increased with advancing degrees of dysplasia in Barrett's mucosa, with an increasing frequency of trisomy and loss of the Y chromosome from non‐dysplastic Barrett's mucosa to invasive adenocarcinoma, and with monosomy and tetrasomy present only in invasive cancers.

Conclusions: 

Our study confirms the high frequency of chromosomal numerical aberrations in oesophageal and gastric adenocarcinomas, without differences between adenocarcinomas of the gastric cardia and antrum. We have shown that these alterations occur early during the neoplastic transformation of Barrett's mucosa.
Renal oncocytoma with prominent intracytoplasmic vacuoles of mitochondrial origin
Authors: Koller A , Kain R , Haitel A , Mazal P R , Asboth F , Susani M
Published: March 2003

Aims: 

We observed two oncocytomas with prominent intracytoplasmatic vacuoles. To investigate if this previously undescribed finding is a diagnostic feature and compatible with the diagnosis of oncocytoma, we characterized these vacuoles by electron microscopy and immunohistochemistry.

Methods and results: 

The tumours were analysed by transmission electron microscopy. Immunohistochemistry was performed with antimitochondrial antibody, anti‐Golgi‐zone antibody, anti‐lysozyme antibody and anti‐human‐trans‐Golgi‐network antibody. By electron microscopy, the vacuoles were found to be double‐membrane‐bounded, and some contained fragmented christae. Immunohistochemistry showed a positive reaction of the vacuoles with anti‐mitochondrial antibody. Staining with anti‐Golgi‐zone antibody, anti‐lysozyme antibody and anti‐human‐trans‐Golgi‐network antibody was negative.

Conclusion: 

Both tumours are benign oncocytomas. The phenomena of cells with prominent intracytoplasmatic vacuoles is not inconsistent with the diagnosis of oncocytoma. The vacuoles are of mitochondrial origin and may develop, by balloon degeneration, as a mechanism of mitochondrial involution and elimination.
Central neurocytomas are genetically distinct from oligodendrogliomas and neuroblastomas
Authors: Tong C Y K , Ng H‐K , Pang J C S , Hu J , Hui A B Y , Poon W‐S
Published: March 2003

Aims

Central neurocytoma is a rare central nervous system tumour typically found in the lateral ventricles and at the septum pellucidum. Histologically, it resembles oligodendrogliomas and yet ultrastructurally, it shows neuronal differentiation. Its molecular oncogenesis is not known. The aim of this study was to examine whether major genetic events found in oligodendrogliomas and neuronal tumours, namely allelic deletions of chromosomes 1p and 19q and N‐myc amplification, can be found in central neurocytomas. As there was one report describing gain of chromosome 7 in central neurocytomas, we also examined epidermal growth factor receptor (EGFR) amplification, as the EGFR gene is located at chromosome 7p.

Methods and results

Nine central neurocytomas and matched blood samples were examined for loss of heterozygosity (LOH) of 1p and 19q13.2–13.4 with 23 finely mapped microsatellite markers. N‐myc amplification was studied by fluorescence in‐situ hybridization using paraffin‐embedded sections. EGFR amplification was tested for by differential PCR. Six of nine (67%) tumours showed LOH at one or more loci at 1p and 5/9 (56%) of cases showed LOH at 19q. However, common regions of deletion cannot be identified. The majority of informative markers are retained at 1p (84%) and 19q (86%). Only one tumour showed amplification of N‐myc and none of the cases showed amplification of EGFR.

Conclusion

Central neurocytomas are genetically distinct from oligodendrogliomas, and chromosomes 1p and 19q probably do not play an important role in their pathogenesis. N‐myc and EGFR amplification are rare.
Lymphoepithelioma‐like carcinoma of the breast: lack of evidence of Epstein–Barr virus infection
Authors: Dadmanesh F , Peterse J L , Sapino A , Fonelli A , Eusebi V
Published: March 2003

Six cases of lymphoepithelioma‐like carcinomas of the breast were studied in order to better define the morphology of this type of breast tumour and to see if these neoplastic lesions are related to Epstein–Barr virus (EBV) infection by analogy with histologically similar tumours located in other organs.

The cases were studied with immunohistochemistry using several antibodies and with in‐situ hybridization method employing different types of nucleotides in four cases. The cases presented the distinctive morphology of the lymphoepithelioma‐like carcinomas as seen in other organs. Specifically Rigaud and Schminke's patterns of growth were observed. In addition, at the edge of the neoplastic lesions, a lymphoepithelial lobulitis was seen. In‐situ hybridization failed to reveal any positivity for EBV genome.

Lymphoepithelioma‐like carcinoma is a specific type of breast tumour, but its relation to EBV has yet to be proven. The differential diagnosis with medullary carcinoma is discussed.
Proliferating trichilemmal tumour: p53 immunoreactivity in association with p27Kip1 over‐expression indicates a low‐grade carcinoma profile
Authors: Fernández‐Figueras M T , Casalots A , Puig L , Llatjós R , Ferrándiz C , Ariza A
Published: March 2003

Proliferating trichilemmal tumour: p53 immunoreactivity in association with p27Kip1 over‐expression indicates a low‐grade carcinoma profile

Aims: Alterations of cell‐cycle regulatory molecules in tumorigenesis may predict the biological behaviour of neoplasms and greatly contribute to their proper classification. Since the behaviour of proliferating trichilemmal tumour (PTT) is controversial, we decided to explore the possible significance of altered p53 and p27Kip1 immunohistochemical expression patterns in PTT.

Methods and results: We evaluated the percentage and distribution of positive tumour cells and compared the results with those obtained from usual trichilemmal cysts (TC) and squamous cell carcinomas with trichilemmal differentiation (SCCT). PTT showed p53 immunoreactivity (50.4 ± 29.6, mean ± standard deviation) that was not statistically different from that seen in SCCT (75.2 ± 36.3). On the other hand, p53 immunostaining was virtually absent in TC cases (positivity for p53 was observed in only one instance in < 1% of cells). As for p27Kip1, the mean percentage of positive cells in PTT (82.7 ± 9.9) was slightly lower than in TC (90.6 ± 4.6) but significantly higher than in SCCT (53.4 ± 30).

Conclusions: The similar p53 immunoreactivity in both PTT and SCCT favour the interpretation of the former as carcinoma, albeit one whose behaviour would be tempered by the well‐known regulatory effect exerted by p27Kip1 on the cell cycle.
Clinical implications of telomerase detection
Authors: Matthews P , Jones C J
Published: March 2003

Clinical implications of telomerase detection

In 1994 a sensitive method for the detection of telomerase was described. This assay, which was based on the polymerase chain reaction, suggested that telomerase activity was associated with immortal and cancer cells. Since then more than a thousand studies have documented the expression and activity of the enzyme in diseased tissues, primarily tumours. This review gives an overview of the biological significance of telomerase expression and methods for detecting its activity. This is followed by an organ system‐based discussion of expression in normal tissues and disease states. We finish with speculation as to the future role of telomerase detection in diagnostic histopathology.
Pleomorphic lobular carcinoma of the breast: its cell kinetics, expression of oncogenes and tumour suppressor genes compared with invasive ductal carcinomas and classical infiltrating lobular carcinomas
Authors: Frolik D , Caduff R , Varga Z
Published: March 2003

Pleomorphic lobular carcinoma of the breast: its cell kinetics, expression of oncogenes and tumour suppressor genes compared with invasive ductal carcinomas and classical infiltrating lobular carcinomas

Aims: The study addresses whether pleomorphic lobular breast carcinomas represent a distinct entity with respect to proliferation and apoptosis as well the expression of the p53, bcl‐2 and Her2 protein.

Methods and results: The study included 30 cases of pleomorphic lobular carcinoma (PLC; G2 n=15, G3 n=15). Poorly differentiated invasive ductal carcinomas (IDC; n=15) and well‐differentiated infiltrating lobular carcinomas (ILC; n=15) were used as controls. Lymph node metastases were present equally in all groups. MIB‐1 labelling was counted as: PLC (G2) 8.36%; PLC (G3) 11.3%; IDC 44.26%; ILC 2.19% (P=0.0001, P=0.004, P=0.001). Apoptotic index was: PLC (G2) 0.82%; PLC (G3) 1.2%; IDC 2.09%; ILC 0.6% (P=0.009, P=0.001). Over‐expression of Her2 protein was detected in 53% of PLC (G3) tumours and was present only in scattered cases in the other groups. PLCs and ILCs were strongly positive for bcl‐2 and for hormone receptors, while p53+ cells were rare. IDCs exhibited a heterogeneous staining pattern for bcl‐2 and for hormone receptors, while p53+ cells occurred considerably more frequently. Stage could not be linked directly to proliferation or apoptosis.

Conclusion: Our data suggest that more frequent over‐expression of Her2 among PLCs (G3) as well as the generally low apoptosis can contribute to their aggressive behaviour.
CD99/MIC‐2 surface protein expression in breast carcinomas
Authors: Milanezi F , Pereira E M , Ferreira F V , Leitão D , Schmitt F C
Published: March 2003

CD99/MIC‐2 surface protein expression in breast carcinomas

Aim: To evaluate the expression of CD99/MIC‐2 surface protein in invasive breast carcinomas and demonstrate whether or not there is a relationship with tumour phenotype.

Methods and results: Thirty‐five invasive breast carcinomas, including five metaplastic carcinomas, were stained with CD99 primary antibodies using standard protocols based on streptavidin–biotin–peroxidase method. Four out of five metaplastic carcinomas expressed CD99/MIC‐2 protein, three of them were matrix‐producing carcinomas. From the other 30 cases, only an invasive apocrine carcinoma was positive. There was no statistical correlation between CD99 expression and the parameters analysed (histological typing and grading, proliferative index and nodal status).

Conclusions: CD99/MIC‐2 is expressed in breast carcinomas, especially in the matrix‐producing variant of metaplastic carcinomas, which impairs its use as a marker to differentiate metaplastic carcinomas from primary and metastatic sarcomas of the breast. It seems to have no prognostic implications. However, phenotype similarities with other chondromyxoid tumours that also express the protein, like mesenchymal chondrosarcomas, suggest a relationship between MIC‐2 reactivity and morphological differentiation.
Lipoblastoma‐like tumour of the vulva: report of three cases of a distinctive mesenchymal neoplasm of adipocytic differentiation
Authors: Lae M E , Pereira P F , Keeney G L , Nascimento A G
Published: March 2003

Lipoblastoma‐like tumour of the vulva: report of three cases of a distinctive mesenchymal neoplasm of adipocytic differentiation

Aims: We sought to delineate and describe three cases of a distinctive mesenchymal neoplasm of the vulva showing adipocytic differentiation and affecting young patients.

Methods and results: In three patients between 13 and 38 years of age, the vulvar tumours had well‐circumscribed borders and ranged in size from 35 to 100 mm. Histologically, they were well circumscribed and lobulated. The lobules were separated by thin fibroconnective tissue septa and were composed of slender spindle cells showing slightly eosinophilic cytoplasm with indistinct boundaries, uniform nuclei with finely granular chromatin, and no nucleoli. The cells were embedded in a richly myxoid stroma. The background in all three tumours was a ‘chicken‐wire’, capillary vascular network resembling that seen in myxoid liposarcomas. Two tumours had scattered signet‐ring‐type lipoblasts and the third a large number of such lipoblasts. Clusters of mature adipocytes were entrapped in the tumours. None had mitotic figures, necrosis, or pleomorphism. The neoplastic cells stained positively for vimentin and were negative for other immunohistochemical markers. Treatment for all three tumours was enucleation alone. After follow‐up of 10 years, 7 years, and 1 year, all patients are well with no evidence of disease.

Conclusions: The benign behaviour of these neoplasms militates against the diagnosis of liposarcoma. We believe these are benign lesions of adipocytic differentiation akin to infantile lipoblastomas.
Tumour‐associated tissue eosinophilia as a prognostic factor in oral squamous cell carcinomas
Authors: Dorta R G , Landman G , Kowalski L P , Lauris J R P , Latorre M R D O , Oliveira D T
Published: March 2003

Tumour‐associated tissue eosinophilia as a prognostic factor in oral squamous cell carcinomas

Aims:  Tumour‐associated tissue eosinophilia has been described in many sites, including head and neck. The mechanism of eosinophil recruitment and its role in tumours has not yet been defined, and its presence has been related to a favourable as well as unfavourable prognosis. The aim of this study was to evaluate the influence of tumour‐associated tissue eosinophilia on the prognosis of 125 oral squamous cell carcinoma patients.

Methods and results:  The number of eosinophils was obtained by morphometric analysis and ranged from 0 to 392 per mm2. Tumour‐associated tissue eosinophilia was classified according to intensity as mild, moderate, or intense and correlated statistically to the intensity of the mononuclear inflammatory infiltrate as well as to the location of the eosinophilic inflammatory infiltrate. The multivariate analysis demonstrated that intense tumour‐associated tissue eosinophilia is an independent favourable prognostic factor for oral squamous cell carcinomas.

Conclusion:  These findings suggest an anti‐tumoral role of eosinophils not as yet well understood that should be better investigated.
Malignant neoplasms infiltrating pseudoangiomatous’ stromal hyperplasia of the breast: an unrecognized pathway of tumour spread
Authors: Damiani S , Peterse J L , Eusebi V
Published: March 2003

Aims:  To describe five malignant tumours in the breast (two invasive carcinomas and three non‐Hodgkin's lymphomas) showing an unusual pattern of spread within anastomosing spaces consistent with those described in pseudoangiomatous stromal hyperplasia.

Methods and results:  The histological and immunohistochemical features of the cases are presented. In all cases, the neoplasms consisted of non‐cohesive E‐cadherin‐negative cells located within anastomosing vascular‐like channels. These channels showed morphological and immunohistochemical features consistent with those seen in pseudoangiomatous stromal hyperplasia: they were lined by flattened cells that expressed CD34, bcl2 and smooth muscle actin, while CD31 was consistently negative.

Conclusions:  The open channels of pseudoangiomatous stromal hyperplasia have been regarded as spaces due to loss of cohesiveness between stromal fibroblasts if not artefacts. The fact that neoplastic cells spread through these spaces suggests that the spaces are true open channels that may represent part of a complex network of stromal channels (pre‐lymphatics) connected to the main lymphatic system of the breast. This finding opens new insights into the routes of tumour spread.
Pagetoid variant of actinic keratosis with or without squamous cell carcinoma of sun‐exposed skin: a lesion simulating extramammary Paget's disease
Authors: Mai K T , Alhalouly T , Landry D , Stinson W A , Perkins D G , Yazdi H M
Published: March 2003

Background: Extramammary Paget's disease usually occurs in anogenital skin. We present five cases of squamous cell carcinoma in situ of sun‐exposed skin and non‐squamous cell carcinoma in situ actinic keratosis that displayed atypical keratinocytes disposed in intraepithelial cell nests and immunohistochemical staining simulating extramammary Paget's disease.

Methods and results: Two pilot cases—one squamous cell carcinoma in situ and one non‐squamous cell carcinoma in situ actinic keratosis with formation of intra‐epidermal nests of atypical keratinocytes with a pagetoid spread pattern—were encountered at our institution. Fifty‐four consecutive cases of squamous cell carcinoma in situ including bowenoid actinic keratosis and 34 cases of non‐squamous cell carcinoma in situ actinic keratosis were reviewed to identify pagetoid spread of atypical cells. Representative sections of all cases with pagetoid spread of atypical keratinocytes were submitted for special stains for mucin, and immunostaining for cytokeratin 7 (CK7), cytokeratin 20 (CK20), cytokeratin CAM 5.2 (CAM 5.2), carcinoembryonic antigen (CEA), vimentin and S100 protein. In the group of squamous cell carcinoma in situ, 10 cases displayed pagetoid spread of atypical keratinocytes with cytoplasm ranging from clear to pale and atypical hyperchromatic nuclei. One review squamous cell carcinoma in situ was multicentric with three separate lesions. The atypical keratinocytes tended to form well to poorly defined cell groups extending from the basal cell layer to the corneal layer. No similar cases were identified in the group of non‐squamous cell carcinoma in situ actinic keratosis. Two pilot cases and three of 10 review cases with a total of seven separate lesions displayed a moderate to marked immunohistochemical reactivity for CK7 similar to extramammary Paget's disease. CEA immunoreactivity was also detected in two of these cases. In addition, two of 44 squamous cell carcinomas in situ without pagetoid spread of atypical keratinocytes showed a moderate reactivity for CK7 in very occasional atypical keratinocytes. The remaining seven squamous cell carcinomas in situ with pagetoid spread of atypical keratinocytes were not immunoreactive for CEA and CK7. Immunostaining for CK20, vimentin, S100 protein was negative in all atypical cells in all study cases.

Conclusions: Actinic keratosis, particularly squamous cell carcinoma in situ of sun‐exposed skin, may have histopathological and immunohistochemical features similar to extramammary Paget's disease and probably represents a variant of actinic keratosis. Awareness of the pagetoid variant of actinic keratosis arising in sun‐exposed skin is helpful to avoid the over‐diagnosis of extramammary Paget's disease.
Inherent properties of somatic hypermutation as revealed by human non‐productive VH6 immunoglobulin rearrangements
Authors: VARADE  , CARNAHAN  , KINGSLEY  , INSEL
Published: March 2003

To study inherent properties of somatic hypermutation of human immunoglobulin genes in the absence of antigen selection, mutations of human non‐productive VH6 rearrangements enriched by subtractive hybridization were characterized. Ten unique clones arising from nine non‐productive rearrangements were isolated. The frequency of mutation was 3·0%. Analysis of these mutations showed intrinsic bias for transitions and cytosine (C) to guanine (G) and G to C transversions. Bias for the strand of DNA targeted by mutation was not evident. Replacement mutations in the complementarity‐determining region (CDR) occurred more frequently than expected based on the primary DNA sequence. This targeting of replacement mutations to the CDR may explain the conservation of the VH6 sequence in primates.
Cross‐talk between Vβ8+ and γδ+ T lymphocytes in contact sensitivity
Authors: DIELI  , PTAK  , SIRECI  , ROMANO  , POTESTIO  , SALERNO  , ASHERSON
Published: March 2003

We have previously reported that T lymphocytes proliferating in vitro to the hapten trinitrochlorobenzene (TNCB) exhibit a very restricted Vβ gene usage and response to TNCB is limited to T‐cell receptors (TCR) composed of Vβ8.2 in combination with Vα3.2, Vα8 and Vα10. This paper investigates the role played by T lymphocytes expressing the Vβ8.2 gene segment in the contact sensitivity (CS) reaction to TNCB in the intact mouse and in its passive transfer into naive recipient mice. Mice injected with monoclonal antibodies to Vβ8 are unable to develop CS upon immunization with TNCB and 4‐day TNCB‐immune lymph node cells from mice that had been depleted in vivo or in vitro of Vβ8+ T lymphocytes fail to transfer CS. However, when separated Vβ8+ and Vβ8− cells were used for passive transfer, it was found that Vβ8+ T lymphocytes failed to transfer CS when given alone to recipient mice and a Vβ8− population was absolutely required. Further analysis revealed that within the Vβ8− population, T lymphocytes expressing the γδ TCR were fundamental to allow transfer of the CS reaction. These γδ cells were found to be antigen non‐specific, genetically unrestricted and to rearrange the Vγ3 gene segment. This indicates that transfer of the CS reaction requires cross‐talk between Vβ8+ and γδ+ T lymphocytes, thus confirming our previous results obtained using TNCB‐specific T‐cell lines. Time–course experiments showed that Vβ8+ lymphocytes taken 4–24 days after immunization with TNCB were able to proliferate and produce interleukin‐2 (IL‐2) in response to the specific antigen in vitro. Similar time–course experiments were then undertaken using the passive transfer of the CS reaction system. The results obtained confirm that TNCB‐specific Vβ8+ T lymphocytes are present in the lymph nodes of immunized mice from day 4 to day 24, and reveal that γδ+ T lymphocytes are active for a very short period of time, i.e. days 4 and 5 after immunization. In fact, TNCB‐specific Vβ8+ cells are able to transfer CS when taken 4–24 days after immunization, providing the accompanying Vβ8− or γδ+ T lymphocytes are obtained 4 days after immunization. In contrast, injection of Vβ8+ T lymphocytes together with Vβ8− or γδ+ T lymphocytes that had been taken 2 or 6 days after immunization, failed to transfer significant CS into recipient mice. Taken together, our results confirm that cross‐talk between Vβ8+ and γδ+ T lymphocytes is necessary for full development of the CS reaction and may explain why the CS reaction in the intact mouse lasts up to 21 days after immunization while the ability of immune lymph node cells to transfer CS is limited to days 4 and 5 after immunization.
Maintenance of granulocyte numbers during acute peritonitis is defective in galectin‐3‐null mutant mice
Authors: COLNOT C. , RIPOCHE M.‐A. , MILON G. , MONTAGUTELLI X. , CROCKER P. R. , POIRIER F.
Published: March 2003

Galectin‐3, also known as the macrophage marker Mac‐2, is a member of a family of structurally related animal lectins that exhibit specificity for β‐galactosides. In order to investigate the role of galectin‐3 in acute inflammation, we have compared the number of leucocytes present in the peritoneal cavity of wild type and galectin‐3 null mutant mice after intraperitoneal (i.p.) injection of thioglycolate broth. At day 1 after injection, we found no difference in the recruitment of mononuclear phagocytes and granulocytes to the peritoneal cavity. However, 4 days after thioglycolate injection, galectin‐3 mutant mice exhibited a significantly reduced number of recoverable granulocytes compared to wild‐type animals. As mutant granulocytes did not exhibit an accelerated rate of apoptosis and their uptake by macrophages appeared to be unaffected by the mutation, the phenotype described here suggests that galectin‐3 participates in an additional level of control during the resolution of acute inflammation.
Dose‐dependent mechanisms relate to nasal tolerance induction and protection against experimental autoimmune encephalomyelitis in Lewis rats
Authors: LI H.‐L. , LIU J.‐Q. , BAI X.‐F. , VAN DER MEIDE P. H. , LINK H.
Published: March 2003

Nasal administration of soluble antigens is an exciting means of specifically down‐regulating pathogenic T‐cell reactivities in autoimmune diseases. The mechanisms by which nasal administration of soluble antigens suppresses autoimmunity are poorly understood. To define further the principles of nasal tolerance induction, we studied the effects of nasal administration of myelin basic protein (MBP) on experimental autoimmune encephalomyelitis (EAE) in the Lewis rat. EAE is a CD4+ T‐cell‐mediated animal model for human multiple sclerosis. Nasal administration of guinea‐pig (gp)‐MBP at a dose as low as 30 μg/rat can completely prevent gp‐MBP‐induced EAE, whereas nasal administration of bovine (b)‐MBP is not effective even at a much higher dosage. Cellular immune responses, as reflected by T‐cell proliferation and interferon‐γ (IFN‐γ)‐ELISPOT, were suppressed in rats receiving the two different doses (30 and 600 μg/rat) of gp‐MBP, but not after administration of b‐MBP. Rats tolerized with both doses of gp‐MBP had also abrogated MBP‐induced IFN‐γ mRNA expression in popliteal and inguinal lymph node mononuclear cells compared with rats receiving phosphate‐buffered saline nasally. However, adoptive transfer revealed that only spleen mononuclear cells from rats pretreated with a low dose, but not from those pretreated with a high dose, of gp‐MBP transferred protection to actively induced EAE. Low‐dose (30 μg/rat) gp‐MBP‐tolerized rats also had high numbers of interleukin‐4 (IL‐4) mRNA‐expressing lymph node cells, while high‐dose (600 μg/rat) gp‐MBP‐tolerized rats had low numbers of IL‐4 mRNA‐expressing lymph node cells. Our data suggest an exquisite specificity of nasal tolerance. Dose‐dependent mechanisms also relate to nasal tolerance induction and protection against EAE in the Lewis rat.
FTY720, a novel immunosuppressant, induces sequestration of circulating mature lymphocytes by acceleration of lymphocyte homing in rats, III. Increase in frequency of CD62L‐positive T cells in Peyer’s patches by FTY720‐induced lymphocyte homing
Authors: YANAGAWA  , MASUBUCHI  , CHIBA
Published: March 2003

FTY720, a novel immunosuppressant, sequesters circulating mature lymphocytes, especially T cells, within lymph nodes and Peyer’s patches by accelerating lymphocyte homing, and thereby causes lymphocyte depletion in the blood. The FTY720‐induced acceleration of lymphocyte homing appears to be mediated by lymphocyte homing receptors including CD62L, CD49d/β7, and CD11a/CD18. In this study, expressions of CD62L, CD49d and CD11a on T cells in the peripheral blood, lymph nodes and Peyer’s patches were analysed by flow cytometry in rats given FTY720 (1 mg/kg) orally. FTY720 markedly decreased the number of peripheral blood T cells, while not affecting CD62L, CD49d and CD11a expressions at 1–3 hr after administration. In contrast, both the frequency of CD62L‐positive T cells and intensity of CD62L expression on T cells were increased in Peyer’s patches but not lymph nodes at 3 hr after administration of FTY720. CD49d and CD11a expressions on T cells were unaffected by FTY720 in both Peyer’s patches and lymph nodes at the same point in time. On the other hand, analysis of lymphocyte homing with calcein‐labelled lymphocytes and anti‐CD62L monoclonal antibody (mAb) confirmed that FTY720 predominantly increased CD62L‐dependent lymphocyte homing to Peyer’s patches. These findings indicate that FTY720 increases the frequency of CD62L‐positive T cells by accelerating CD62L‐predominant homing in Peyer’s patches.
Human B cells secrete migration inhibition factor (MIF) and present a naturally processed MIF peptide on HLA‐DRB1*0405 by a FXXL motif
Authors: WYMANN  , BLÜGGEL  , KALBACHER  , BLESKEN  , AKDIS  , MEYER  , BLASER
Published: March 2003

A better knowledge of peptide structures interacting with major histocompatibility complex (MHC) molecules is of great interest for better understanding of the molecular basis of immune recognition. We have isolated naturally processed peptides from a continuously growing antigen‐presenting Epstein–Barr virus‐transformed human B‐cell line. HLA‐DR complexes were purified by specific affinity chromatography and complexed peptides were released by acid treatment. The isolated peptides were separated by reversed phase chromatography and fractions were analysed by Edman degradation at picomolar ranges. From 30 fractions that were examined seven peptides bound to the HLA‐DRB1*0405 and two peptides from the human leucocyte antigen (HLA) class II associated invariant chain bound to HLA‐DRB1*1302. In addition, a N‐terminal β‐chain peptide of the 0405 allele was identified. Evaluation of amino acid sequences revealed a refined FXXL motif for the 0405 allele, in which F (phenylalanine) stands for any aromatic amino acid and L (leucine) can be exchanged by either I (isoleucine) or V (valine). In total, three fractions contained a peptide derived from the human migration inhibition factor (MIF), a pro‐inflammatory cytokine that is normally produced by activated T lymphocytes and monocytes/macrophages. Indeed, cytokine analysis revealed high amounts of MIF secreted by the B‐cell line, confirming that MHC class II expressing cells can present any intrinsic peptide that contains the distinct motif for HLA‐binding. For MIF, the amino acid sequence Y36IAV39 represents the required binding motif for HLA‐DRB1*0405. Nevertheless, it is the first time that cytokine fragments were found to bind to HLA molecules on human B cells.
Tumour necrosis factor and interferon‐γ are required in host resistance against virulentRhodococcus equiinfection in mice: cytokine production depends on the virulence levels ofR. equi
Authors: KASUGA‐AOKI†  , TAKAI  , SASAKI  , TSUBAKI  , MADARAME  , NAKANE
Published: March 2003

Rhodococcus equiis a facultative intracellular bacterial pathogen that causes pneumonia in foals and immunosuppressed humans. There are at least three virulence levels ofR. equiand these pathogenicities are associated, in mice, with the presence of virulence plasmids. This study focused on cytokine secretion, in mice, in the course of a primary infection with sublethal doses ofR. equistrains of different virulence levels (virulent, intermediately virulent and avirulent). Tumour necrosis factor (TNF) and interferon‐γ (IFN‐γ), but not interleukin‐4 (IL‐4) and interleukin‐10 (IL‐10), were induced endogenously in mice in relation to the multiplication and clearance of virulent and intermediately virulent strains ofR. equi. These cytokines were not detected in mice infected with avirulentR. equi. Deaths occurred among mice treated with monoclonal antibodies (mAbs) against either TNF or IFN‐γ prior to sublethal dose infection with virulent and intermediately virulent strains ofR. equi, but not with avirulentR. equi. These results suggested that cytokine production depended largely on the virulence levels ofR. equi: TNF and IFN‐γ were required early during infection with virulentR. equito limit replication and clearance of bacteria within the organs, but they were not necessary for limiting infection with avirulentR. equi.
T‐cell populations in the pig intestinal lamina propria: memory cells with unusual phenotypic characteristics
Authors: HAVERSON  , BAILEY  , STOKES
Published: March 2003

We have previously presented evidence of a highly organized and compartmentalized structure of the small intestinal lamina propria of the pig. In this study, we conducted a detailed analysis of the T‐cell populations found at this site, and compared these T cells with cell populations found in other tissue sites and the periphery. We showed that the CD4+ and CD8+ T‐cell populations found in the pig gut are of memory phenotype, defined by CD45 isoform expression, but show few signs of recent activation. They show a high degree of phenotypic and therefore presumably functional homogeneity. Both CD4‐ and CD8‐positive cells show strong parallels in the patterns of surface molecule expression, suggesting similar pressures on differentiation. The unique combination of surface molecules found on lamina propria T cells is found only infrequently on cells in other lymphoid sites.
Identification of a gastritogenic epitope of the H/K ATPase β‐subunit
Authors: DE SILVA  , GLEESON  , TOH  , VAN DRIEL  , CARBONE
Published: March 2003

We have previously shown that autoimmune gastritis can be elicited in mice by immunization with the gastric parietal cell H/K ATPase αβ heterodimer and that tolerance specifically induced to the H/K ATPase β‐subunit protects mice from the development of gastritis. Here we have identified the immunodominant gastritogenic epitope of the H/K ATPase β‐subunit (H/Kβ). Epitope mapping was carried out with a panel of 21 overlapping peptides that spanned the entire sequence of the gastric H/K ATPase β‐subunit. T cells from gastric H/K ATPase‐immunized mice responded to only one of the overlapping peptides, namely H/Kβ253–277. Furthermore, a single subcutaneous immunization of 6‐week‐old BALB/c mice with the ATPase β‐subunit peptides resulted in a T‐cell response to only H/Kβ253–277. Multiple immunization with the overlapping H/K ATPase peptides demonstrated that H/Kβ253–277 was capable of inducing a mononuclear infiltrate specifically within the gastric mucosa. We conclude that H/Kβ253–277 is the dominant gastritogenic epitope of the gastric H/K ATPase.
Progressor but not regressor skin tumours inhibit Langerhans’ cell migration from epidermis to local lymph nodes
Authors: LUCAS  , HALLIDAY
Published: March 2003

Langerhans’ cells (LC) are found in high numbers infiltrating skin tumours, the functional significance of which remains unknown. To study the mechanism by which tumours increase the number of LC we developed a procedure whereby supernatant from cultured T7 tumour cells applied topically increases the number of LC. Tumour factors increased the number of resident epidermal LC and did not attract LC precursors into parental murine skin grafted onto F1 hybrids. There was no evidence for increased LC division in response to the tumour‐derived factors. LC migration from the epidermis to local lymph nodes, induced by topical fluorescein isothiocyanate (FITC), was inhibited by the tumour supernatant. To examine the functional significance of this, FITC‐induced migration of LC from the epidermis overlying progressor tumours, which evade immunological destruction, and regressor tumours, which are immunologically destroyed, was examined. The progressor tumour T7 growing subcutaneously in syngeneic mice inhibited FITC‐induced migration of LC from overlying epidermis. Furthermore, two progressor, but not two regressor murine skin tumour lines growing in BALB/c nu/nu mice inhibited LC migration from the epidermis. Our results demonstrate that progressor skin tumours produce factor(s) which inhibit LC migration from the epidermis to lymph nodes, leading to LC accumulation. Inhibition of LC migration by tumour‐derived factors may enable tumours to evade the activation of protective immunity as regressor tumours do not interfere with the normal trafficking of LC.
Administration of interleukin‐12 exerts a therapeutic instead of a long‐term preventive effect on mite Der p I allergen‐induced animal model of airway inflammation
Authors: Lee  , Fu  , Chiang
Published: March 2003

Interleukin‐12 (IL‐12) is a key cytokine, which promotes T helper type 1 (Th1) cell‐mediated immunity and inhibits Th2‐type responses. It has been previously shown that IL‐12 administration during active immunization following a single allergen exposure can prevent antigen‐induced increases in immunoglobulin E (IgE) formation, Th2 cytokine production and bronchoalveolar lavage (BAL) eosinophils in a murine model of allergic airway inflammation. Thus, these studies have now been extended and two IL‐12 treatment protocols on this murine model were evaluated. Administration of IL‐12 during the active immunization strikingly increased Der p I‐specific serum IgG2a and transiently decreased the levels of IgG1 and IgE antibodies following multiple allergen challenges. Such early treatment of IL‐12 down‐regulated IL‐5 production and modestly up‐regulated interferon‐γ production but did not effect BAL eosinophilia. These results suggest that repeated exposure to antigen and IL‐12 is necessary to maintain a persistent Th1‐recall response. Furthermore, administration of IL‐12 to actively immunized mice, in which Th2‐associated responses were established, had a significant effect on IgG2a synthesis and a modest effect on IgE levels, also down‐regulation of IL‐5 production, and markedly increased interferon‐γ production and abolished recruitment of eosinophils. Therefore, these data indicate that IL‐12 can inhibit antigen‐induced eosinophil infiltration into airways, despite the existence of a Th2‐associated response. Taken together, these studies suggest that IL‐12 may be useful as an immunotherapeutic agent in the treatment of such pulmonary allergic disorders as bronchial asthma.
Intravenous tolerization with type II collagen induces interleukin‐4‐and interleukin‐10‐producing CD4+ T cells
Authors: Gumanovskaya  , Myers  , Rosloniec  , Stuart  , Kang
Published: March 2003

Intravenous (i.v.) administration of type II collagen (CII) is an effective way to induce tolerance and suppress disease in the collagen‐induced arthritis (CIA) model. In this study, we demonstrated that a single i.v. dose of CII (as low as 0·1 mg/mouse) completely prevented the development of CIA. This suppression was accompanied by decreases in levels of antibody specific for the immunogen, bovine CII and autoantigen, mouse CII. Splenocytes obtained from CII‐tolerized mice and stimulated with CII in vitro produced predominantly the T helper 2 (Th2)‐type cytokines interleukin‐4 (IL‐4) and interleukin‐10 (IL‐10). In contrast, cells obtained from mice immunized with CII produced predominantly interferon‐γ (IFN‐γ). Two‐colour flow cytometric analysis of cytokine expression and T‐cell phenotype demonstrated that CD4+ cells and not CD8+ or γδ+ cells were the predominant regulatory cells producing IL‐4 and IL‐10. Transgenic mice bearing a T‐cell receptor (TCR) specific for CII had a greater increase in the number of IL‐4‐secreting CD4+ cells, as well as a marked increase of IL‐4 in culture supernatants. This cytokine was produced by transgene‐bearing T cells. Elucidation of mechanisms for the induction of tolerance in mature T cells is an important line of study in autoimmune models because of the potential application for treating organ‐specific autoimmune disease.
A recombinant BCG vaccine generates a Th1‐like response and inhibits IgE synthesis in BALB/c mice
Authors: Kumar  , Behera  , Matsuse  , Lockey  , Mohapatra
Published: March 2003

The tubercle vaccine, bacille Calmette–Guérin (BCG), is a strong inducer of T‐helper type 1 (Th1) responsiveness, and it has been suggested that recombinant BCG (rBCG), which produces and secretes antigens, may be used to prevent allergic diseases. The effects of rBCG vaccination on allergic responses in a murine model were examined in this study. A BCG–Escherichia coli shuttle vector was developed with the promoter and signal sequence of the α‐antigen of Mycobacterium bovis, and the vector was tested using E. coliβ‐galactosidase as the model antigen and allergen. This vector enabled the expression of the E. coliβ‐galactosidase gene in BCG, which was detected in its protein extract by immunoblotting analysis. Vaccination of mice with a single dose of 106 recombinant BCG generated a β‐galactosidase‐specific antibody response. The splenocytes of vaccinated mice compared with controls produced significantly higher amounts of interferon‐γ (IFN‐γ) (P<0·01) and interleukin‐2 (IL‐2) (P<0·05) and lower amounts of IL‐5 (P<0·01). Mice vaccinated with rBCG had significantly less (P<0·01) serum IgE compared with controls. These results together demonstrate that rBCG secreting antigens or allergens may be utilized for the induction of a Th1‐like response and the down‐regulation of IgE antibody response.
Spontaneously occurring neutralizing antibodies against granulocyte–macrophage colony‐stimulating factor in patients with autoimmune disease
Authors: Meager  , Wadhwa  , Bird  , Dilger  , Thorpe  , Newsom‐Davis  , Willcox
Published: March 2003

There is increasing evidence that spontaneous anticytokine autoantibodies are associated with chronic infections and autoimmune diseases. We report the sporadic occurrence in autoimmune diseases of such autoantibodies to granulocyte–macrophage colony‐stimulating factor (GM‐CSF), a cytokine involved in inflammation and the regulation of proliferation, differentiation and function of granulocytic and monocytic cell lineages. In 41 of 425 patients tested, we found low to moderate levels of autoantibodies binding to GM‐CSF in serum or plasma. These were most prevalent in patients with myasthenia gravis (MG). However, neutralizing autoantibodies against GM‐CSF were very rare, being found in only three patients. Two had autoimmune MG, one with thymoma (Patient A) and the other (Patient B) with ‘seronegative’ MG, i.e. without the antiacetylcholine receptor autoantibodies characteristic of most MG patients, and a third (Patient D) had multiple sclerosis. Only very limited amounts of Patient A and Patient D serum/plasma were available for analysis and therefore further studies were carried out on the more plentiful samples from Patient B. The anti‐GM‐CSF autoantibodies of Patient B were predominantly polyclonal immunoglobulin G and strongly neutralized recombinant human (rh) GM‐CSF derived from different expression systems. They had similar immunological and immunochemical characteristics to anti‐GM‐CSF antibodies that developed in immunocompetent colorectal carcinoma patients following (rh)GM‐CSF therapy. In serial samples from Patient B, the anti‐GM‐CSF autoantibodies were undetectable from diagnosis at age 8 years until at least age 13, but then developed spontaneously during (temporary) withdrawal of immunosuppressive treatment. Their neutralizing activity has persisted since their first detection at age 15 years 1 month, and was at its highest level recently at age 17 years 7 months. There was no obvious association with other autoimmune phenomena, nor were any haematological deficiencies overtly manifested, suggesting that any loss of GM‐CSF function may have been compensated for by other cytokines.
Characterization of the memory/activated T cells that mediate the long‐lived host response against tuberculosis after bacillus Calmette–Guérin or DNA vaccination
Authors: Silva  , Bonato  , Lima  , Faccioli  , Leão
Published: March 2003

The memory/activated T cells, which mediate the long‐lived host response against tuberculosis, in mice immunized with either bacillus Calmette–Guérin (BCG) or mycobacterium heat‐shock protein 65 (hsp 65) antigen expressed from plasmid DNA (DNA‐hsp 65), were characterized. Protection against Mycobacterium tuberculosis challenge by DNA‐hsp 65 vaccination was associated with the presence of lymph node T‐cell populations in which CD8+/CD44hi interferon‐γ (IFN‐γ)‐producing/cytotoxic cells were prominent even after 8 or 15 months of plasmid DNA‐mediated immunizations, whereas after BCG vaccination the majority were CD4+/CD44lo IFN‐γ‐producing T cells. When the cells were separated into CD4+CD8− and CD8+CD4− and then into CD44hi and CD44lo types, CD44lo cells were essentially unable to transfer protection in adoptive transfer experiments, the most protective CD44hi cells were CD8+CD4− and those from DNA‐vaccinated mice were much more protective than those from BCG‐immunized mice. The frequency of protective T cells and the level of protection were increased up to 8 months and decreased after 15 months following DNA or BCG immunizations.
Immune responses and protection against vaginal infection after nasal or vaginal immunization with attenuated herpes simplex virus type‐2
Authors: Parr E. L. , Parr M. B.
Published: March 2003

We compared nasal and vaginal immunizations using attenuated herpes simplex virus type‐2 (HSV‐2) for protection against vaginal infection with wild‐type HSV‐2. Mice were immunized once intranasally, intravaginally after progestin (DP) treatment, or intravaginally with scarification after oestradiol treatment. Compared with vaginal immunizations, nasal immunization did not increase immunoglobulin A (IgA) plasma cell numbers in the vagina or elicit a higher antiviral IgA titre in vaginal secretions. Both types of vaginal immunizations increased the number of immunoglobulin G (IgG) plasma cells in the vagina and the secretion/serum titre ratio of IgG antiviral antibody, indicating local production of virus‐specific IgG in these groups. Cell‐mediated immunity in the vagina, as indicated by memory T‐cell secretion of interferon‐γ (IFN‐γ) in situ 20 hr after HSV‐2 challenge, was essentially equivalent in the vaginally immunized groups but significantly lower in the nasal group, while lymphocyte recruitment to the vagina was similar in all three groups. All three immunizations protected all mice from neurological disease after challenge, but vaginal DP immunization induced the greatest immunity against reinfection of the vaginal epithelium.
Effects of α‐galactosylceramide (KRN7000), interleukin‐12 and interleukin‐7 on phenotype and cytokine profile of human Vα24+ Vβ11+ T cells
Authors: Van Der Vliet H. J. J. , Nishi N. , Koezuka Y. , Peyrat M. A. , Von Blomberg B. M. E. , Van Den Eertwegh A. J. M. , Pinedo H. M. , Giaccone G. , Scheper R. J.
Published: March 2003

The α‐galactosylceramide KRN7000 was reported to be presented by CD1d to natural killer (NK) T cells, cells that are thought to play an important role in the rejection of malignant tumours and in the regulation of several autoimmune diseases. Here we analysed human peripheral blood (PB) NK T cells (Vα24+ Vβ11+ T cells) before and after a short‐term culture in the presence of KRN7000. KRN7000 strongly activated PB Vα24+ Vβ11+ T cells and, when stimulated, the vast majority of these cells expressed interferon‐γ (IFN‐γ). Exposure of these KRN7000‐cultured Vα24+ Vβ11+ T cells to interleukin‐12 (IL‐12), but not to IL‐7, resulted in a relative increase in IFN‐γ‐expressing Vα24+ Vβ11+ T cells, compared with IL‐4‐expressing Vα24+ Vβ11+ T cells, indicating a shift towards a T‐helper type 1 (Th1) phenotype. KRN7000 strongly up‐regulated the expression of the cytotoxic molecule granzyme B (GrB) in Vα24+ Vβ11+ T cells. Although IL‐7 resulted in a decrease in GrB levels in KRN7000‐cultured Vα24+ Vβ11+ T cells, IL‐12 increased GrB levels in both Vα24+ Vβ11+ T cells and in Vα24+ Vβ11+ T‐cell clones and increased cytotoxicity against hCD1d‐transfected HeLa cells. Our data provide further insight into the characteristics of human Vα24+ Vβ11+ T cells and indicate that KRN7000 is a potent activator of Vα24+ Vβ11+ T cells. Combined with the established anti‐tumour effects of KRN7000 in mouse models, these results may support the use of KRN7000 as an anti‐tumour agent in man.