bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytoskeletal and kinetic epithelial differences between NSAID gastropathy and Helicobacter pylori gastritis: an immunohistochemical determination
Authors: Lauwers G Y , Furman J , Michael L E , Balis U J , Kubilis P S
Published: March 2003

Cytoskeletal and kinetic epithelial differences between NSAID gastropathy and Helicobacter pylori gastritis: an immunohistochemical determination

Aims: Distinguishing histological features between non‐steroidal anti‐inflammatory drug (NSAID) gastropathy and Helicobacter pylori gastritis have been accepted. However, the molecular basis explaining these dissimilar histologies has not been elucidated. In an attempt to clarify this question we investigated the differences in the structural cytoskeleton and proliferative activity of these two gastropathies.

Methods and results: We assessed the distribution of five cytokeratins (CK) (CK7, 8, 18, 19 and 20) and Ki67 for the ability to distinguish NSAID from H. pylori gastropathies. In H. pylori gastritis, CK7, 8, 18 and 19 were expressed comparably to normal mucosa from the deep foveolae up to the tips of the glands. The detection of CK20, normally expressed in the upper foveolar region and surface, was decreased with only an epithelial surface reaction. In NSAID gastropathy, CK expression was increased in intensity, with normal distribution for CK8, 18 and 19. Modification of localization was noted for CK7 and 20, with labelling extending toward the deep foveolar region. Unlike H. pylori gastritis, no surface epithelial labelling with Ki67 was noted with NSAID gastropathy but downward elongation of the proliferative zone occurred instead.

Conclusions: Contrasting cytostructural alterations and distinct proliferative patterns distinguish NSAID gastropathy from H. pylori gastritis, possibly reflecting different injury pathways.
Primary squamous cell carcinoma of the thyroid gland: an entity with aggressive clinical behaviour and distinctive cytokeratin expression profiles
Authors: Lam K‐Y , Lo C‐Y , Liu M‐C
Published: March 2003

Primary squamous cell carcinoma of the thyroid gland: an entity with aggressive clinical behaviour and distinctive cytokeratin expression profiles

Aims: Primary squamous cell carcinoma of the thyroid gland is uncommon. This study aims to identify the clinicopathological features and the pattern of expression of cytokeratins and oncoproteins in this tumour.

Methods and results: Histological slides from Chinese patients with thyroid cancer treated in our institution from 1980 to 1999 were reviewed. Patients with primary squamous cell carcinoma of the thyroid were identified and their clinical records were analysed. The expression of cytokeratins (CKs), p53 and p21 in these cases were also studied by an immunohistochemical method. Four women (mean age 71 years) with squamous cell carcinoma of thyroid were found. The main presenting features were signs and symptoms of airway obstruction in three patients and neck swelling in one. The tumours were located at the right lobe (n=2), left lobe (n=1) or in both lobes of the thyroid (n=1). One patient died shortly after admission and the other three died within 4 months after thyroidectomy. The p53 protein was positive in 50% (2/4) of the tumours and p53+ tumours were poorly differentiated. The tumours were negative for p21. CK19 was expressed in all the tumours while CK7 expression was noted in 3/4 of the tumours. One carcinoma showed focal positivity to CK18. The tumours were negative for CKs 1, 4, 6, 10/13 and 20. The pattern of cytokeratin expression in squamous cell carcinoma of the thyroid gland was different from carcinoma showing thymus‐like differentiation (CASTLE) of the thyroid gland and oesophageal squamous cell carcinoma.

Conclusions: Squamous cell carcinoma of the thyroid has aggressive clinical behaviour and characteristic CK expression pattern. p53 over‐expression in these tumours was associated with tumour differentiation.
Endothelial nitric oxide synthase is strongly expressed in malignant mesothelioma but does not associate with vascular density or the expression of VEGF, FLK1 or FLT1
Authors: Soini Y , Puhakka A , Kahlos K , Säily M , Pääkkö P , Koistinen P , Kinnula V
Published: March 2003

Endothelial nitric oxide synthase is strongly expressed in malignant mesothelioma but does not associate with vascular density or the expression of VEGF, FLK1 or FLT1

Aims: To investigate endothelial nitric oxide synthase (eNOS) expression in malignant mesothelioma and its association with expression of vascular endothelial growth factor (VEGF), its receptors FLK1 and FLT1, and vascular density.

Methods and results: eNOS, VEGF, FLK1 and FLT1 were studied in 36 histological mesothelioma samples by immunohistochemistry. Two mesothelioma (M14K, M38K) and one non‐neoplastic mesothelial cell line (MET‐5A) were studied for eNOS mRNA expression by reverse transcriptase‐polymerase chain reaction (RT‐PCR). Vascular density was determined by staining the samples with an antibody to factor VIII. RT‐PCR showed that mesothelioma cells synthesize eNOS in vitro. eNOS immunoreactivity was found in 32/36 (89%) tumours. VEGF, FLK1 and FLT1 expression was found in 17 (45%), 24 (69%) and 25 (71%) cases, respectively. FLK1 or FLT1 immunoreactivity was more often seen in epithelioid and biphasic mesotheliomas than in sarcomatoid ones (P=0.007 and P=0.011, respectively). There was a significant association between FLK1 and FLT1 immunoreactivity (P=0.032). No significant association was found between FLK1, FLT1, VEGF and eNOS immunoreactivity and vascular density.

Conclusions: eNOS is strongly expressed in malignant mesothelioma. Since eNOS did not associate with VEGF, FLK1 or FLT1, its synthesis seems not to be regulated through VEGF in malignant mesothelioma as has been shown in non‐neoplastic endothelial cells.
Diverse differentiation in malignant peripheral nerve sheath tumours associated with neurofibromatosis‐1: an immunohistochemical and ultrastructural study
Authors: Takeuchi A , Ushigome S
Published: March 2003

Diverse differentiation in malignant peripheral nerve sheath tumours associated with neurofibromatosis‐1: an immunohistochemical and ultrastructural study

Aims: The diverse histological features in malignant peripheral nerve sheath tumours (MPNSTs) associated with NF‐1 were investigated by immunohistochemical and electron microscopic analysis. Our study is focused on the differentiation of the tumour cells in the heterogeneous components.

Methods and results: Twenty‐three cases were classified as conventional type, epithelioid type, anaplastic type, and heterogeneous type, and divided into three groups by the presence of S100 protein (S100)‐positive cells in each tumour; Group A was defined as having >50% S100+ cells, Group B as having <50%, and Group C as cases with no positive cells. To investigate the differentiation of the tumour cells, the morphology and immunoreactivity for neural or mesenchymal markers among the three groups were compared. For the identification of Schwannian, perineurial, and endoneurial differentiation, markers for S100, EMA and CD34 were used, respectively. In three tumours of the Group A type, there were no cases showing differentiation towards perineurial or endoneurial cells, or formation of heterogeneous components. In nine tumours of the Group B type, one tumour expressed EMA and CD34, suggesting probable perineurial and/or endoneurial differentiation. One tumour showed rhabdomyoblastic differentiation. Three tumours showed cartilaginous or osteogenic differentiation, and one of the three also showed a focal vascular differentiation. The surrounding areas of the heterogeneous components were composed of mixed S100+ cells and S100– cells. S100– cells in the areas were positive for CD34 in one case. In 11 tumours of Group C type, one tumour expressed EMA and CD34 suggesting perineurial and/or endoneurial cell differentiation. Three tumours showed rhabdomyoblastic differentiation. The tumour cells around the heterogeneous components in the three cases were negative for EMA and CD34.

Conclusion: Our results suggest that tumour cells differentiating to Schwann cells are not the only component of MPNSTs. Furthermore, tumour cells other than Schwann cells are largely related to the formation of the heterogeneous components in MPNSTs associated with NF‐1.
Scoring nuclear pleomorphism in breast cancer
Authors: Dunne B , Going J J
Published: March 2003

Scoring nuclear pleomorphism in breast cancer

Aims: Nuclear grade has equal weight with mitotic index and acinus formation in grading breast cancer, but criteria for its assessment are less well defined. This study examines consistency of nuclear grading in breast cancer and whether improved nuclear grading criteria are required.

Methods and results: Photographic prints of haematoxylin–eosin sections of 100 unselected symptomatic breast cancers were circulated to histopathologists who assigned each carcinoma a nuclear pleomorphism score on a linear analogue scale 0–100 (0–33 equating to nuclear pleomorphism grade 1, 34–66 to grade 2, and 67–100 to grade 3). Seventeen histopathologists completed the exercise, including 11 breast specialists. While κ scores for the implied nuclear grades indicated ‘moderate’ or ‘good’ agreement between individuals and the group as a whole, seven pathologists allocated analogue scores significantly lower than the median score allocated by the group to each case, while five allocated significantly higher scores. The range was from analogue scores 11.3 units lower on average than the median (assigning 27% of carcinomas nuclear grade 1, 60% grade 2, and 13% grade 3) to scores 7.5 units higher on average than the median (assigning only 2% carcinomas nuclear grade 1, 46% grade 2, and 52% grade 3). Five of six non‐specialists allocated scores significantly lower than the group medians but only two of 11 specialists did so (P=0.018).

Conclusions: Systematic differences between pathologists in scoring nuclear pleomorphism in breast cancer potentially contribute to differences in allocating overall grade and confirm the need for improved nuclear grading criteria. Specialists tend to allocate higher pleomorphism scores than non‐specialists.
Psammomatous malignant melanoma arising in an intradermal naevus
Authors: Monteagudo C , Ferrández A , González‐Devesa M , Llombart‐Bosch A
Published: March 2003

Psammomatous malignant melanoma arising in an intradermal naevus

Aims: A wide variety of differentiation patterns may be found in malignant melanoma. Schwannian features are unusual, and mostly present in the desmoplastic variant. We report the first description of psammoma bodies in malignant melanoma.

Methods and results: A malignant melanoma arose in an intradermal naevus of the scalp in a 51‐year‐old woman, displaying focal neural‐like features in the form of rosette‐like pseudo‐meissnerian alveolar nests, as well as numerous psammoma bodies grouped in a few areas. Tumour cell immunostaining for S100, HMB45, NKI‐C3, and vimentin was detected. In addition, both malignant and benign melanocytic cells showed widespread MART‐1 immunoreactivity. Differential diagnosis with psammomatous melanotic schwannoma, a feature of Carney’s complex, is particularly emphasized, since dermal variants of this nerve sheath neoplasm have been described. In addition, its potential relationship with cutaneous malignant melanotic neurocristic tumour is discussed.

Conclusions: This is, to our knowledge, the first reported case of cutaneous malignant melanoma with psammoma bodies.
Immunohistochemical markers in the differentiation of thymic and pulmonary neoplasms
Authors: Pomplun S , Wotherspoon A C , Shah G , Goldstraw P , Ladas G , Nicholson A G
Published: March 2003

Immunohistochemical markers in the differentiation of thymic and pulmonary neoplasms

Aims: The histopathological features of some thymic neoplasms overlap with those of pulmonary squamous and large‐cell undifferentiated carcinomas, and identification of the primary site may be difficult on routine staining. We have assessed a panel of antibodies that may help to distinguish between neoplasms from these two sites.

Methods and results: Antibodies identifying cytokeratin 7 (CK7), CD5, CD10, CD1a and thyroid transcription factor‐1 (TTF‐1) were applied to a series of 20 thymic neoplasms (thymic carcinomas, atypical thymomas and thymomas), 10 primary squamous cell carcinomas of the lung and 10 large‐cell undifferentiated carcinomas of the lung. Staining for TTF‐1 was positive in 3/10 large‐cell undifferentiated carcinomas, but negative in all other tumours. CD5 showed strong membranous staining in 3/6 thymic carcinomas and 1/14 thymomas, but only focal staining in 1/20 pulmonary carcinomas. CD1a was consistently positive in thymic lymphocytes in both typical and atypical thymomas, but only focally in 1/6 thymic carcinomas. CD1a stained dendritic cells in 7/20 pulmonary carcinomas, but did not stain lymphocytes. Staining for CK7 and CD10 did not aid in differentiating between a pulmonary or thymic origin of the tumour.

Conclusion: Staining for TTF‐1, CD5 and CD1a have potential use in distinguishing between pulmonary and thymic neoplasms.
Expression of respiratory mucins in fatal status asthmaticus and mild asthma
Authors: Groneberg D A , Eynott P R , Lim S , Oates T , Wu R , Carlstedt I , Roberts P , McCann B , Nicholson A G , Harrison B D , Chung K F
Published: March 2003

Expression of respiratory mucins in fatal status asthmaticus and mild asthma

Aims: The airways of patients with asthma are characterized by chronic inflammatory changes comprising mainly T‐cells and eosinophils, and airway remodelling with goblet cell metaplasia and submucosal gland hyperplasia. Mucus hypersecretion is often a marked feature, particularly in status asthmaticus. The matrix of airway sputum consists of high molecular glycoproteins and mucins. In this study, the expression and distribution of the major gel‐forming mucins MUC5AC and MUC5B were studied in fatal status asthmaticus tissues and bronchial biopsies of mild asthmatic patients. The effect of inhaled corticosteroids on the expression of these mucins was also investigated.

Methods and results:  Polyclonal antibodies specific for MUC5AC and MUC5B, and a monoclonal antibody for MUC5B were used to stain lung tissues and airway mucosal biopsies obtained from patients who died of status asthmaticus (n=5) and from mild asthmatics (n=4), respectively. Immunohistochemistry for MUC5AC revealed abundant staining of goblet cells situated in the epithelial surface lining and glandular ducts of tissues from patients with fatal asthma. MUC5B immunoreactivity was restricted to mucous cells of submucosal glands and to epithelial cells. In mild asthmatics, large amounts of MUC5B, but not MUC5AC, positive extracellular mucus was found in the airway lumen as plugs, adjacent to the epithelial lining and in the necks of glandular secretory ducts of mild asthmatics. The distribution of MUC5AC and MUC5B in bronchial biopsies of mild asthmatics was similar before and after inhaled steroid treatment.

Conclusions: The expression of MUC5AC and MUC5B shares a similar distribution to normal airways in different states of asthma. The distribution is not affected by topical corticosteroid therapy.
Keratin expression in human tissues and neoplasms
Authors: Chu P G , Weiss L M
Published: March 2003

Keratin expression in human tissues and neoplasms

Keratin filaments constitute type I and type II intermediate filaments (IFs), with at least 20 subtypes named keratin 1–20. Since certain keratin subtypes are only expressed in some normal human tissues but not others, and vice versa, various tissues have been subclassified according to the pattern of keratin staining. Simple epithelia generally express the simple epithelial keratins 7, 18, 19, and 20, while complex epithelia express complex epithelial keratins 5/6, 10, 14, and 15. When an epithelium undergoes malignant transformation, its keratin profile usually remains constant. The constitution and expression patterns of keratin filaments in human epithelial neoplasms are complex and often distinctive. In this article, we first briefly review the molecular and cell biology of keratin filaments. We then focus on the expression patterns of keratin filaments in various human neoplasms.
Mixed ductal–pancreatic polypeptide‐cell carcinoma of the pancreas
Authors: Chatelain D , Parc Y , Christin‐Maitre S , Parc R , Flejou J‐F
Published: March 2003

Mixed ductal–pancreatic polypeptide‐cell carcinoma of the pancreas

Aims:  Mixed ductal‐endocrine carcinomas of the pancreas are rare tumours with 10 cases reported in the English literature. We report the first case with a polypeptide‐cell component.

Methods and results:  The tumour was fortuitouslydiscovered in a 72‐year‐old woman during the exploration of an endometrial adenocarcinoma. It measured 100 mm and was located in the tail of the pancreas. On microscopic examination two intermingled endocrine and exocrine components were present. The endocrine component consisted of trabeculae and solid nests composed of cells immunoreactive for chromogranin A, synaptophysin and pancreatic polypeptide, but negative for p53 and Bcl‐2 proteins. The exocrine component was composed of tubules lined by atypical cylindrical cells immunoreactive for CK19, CEA, p53 and Bcl‐2. The stroma of the endocrine component contained amyloid deposits.

Conclusion:  Mixed ductal–endocrine carcinomas of the pancreas are often described in middle‐aged patients. The tumours are usually large and located in the head of the pancreas. An endocrine syndrome is rare and the prognosis is often unfavourable. We report the first case of mixed endocrine–exocrine carcinoma of the pancreas with a pancreatic polypeptide‐cell component. The histogenesis of mixed carcinoma of the pancreas is still uncertain but the over‐expression of p53 and Bcl‐2 could play a major role in the neoplastic progression of the ductal component.
Expression of cytokeratins 7 and 20 in carcinomas of the gastrointestinal tract
Authors: Kende A I , Carr N J , Sobin L H
Published: March 2003

Aims:  The differential expression of cytokeratin (CK) 7 and 20 by carcinomas may help in determining the primary site of a metastatic tumour. The aim of this study was to extend the published data on CK7 and CK20 expression in epithelial neoplasms of the gastrointestinal tract by considering the degree of differentiation and including some unusual neoplasms.

Methods and results:  Cases referred to the Armed Forces Institute of Pathology were studied prospectively for immunohistochemical expression of CK7 and CK20. Lesions from 105 patients were analysed. Adenocarcinomas of the upper gastrointestinal tract were positive for both CK7 and CK20 in 78% of cases; only poorly differentiated lesions were CK7–. Well‐differentiated and moderately differentiated adenocarcinomas of the large intestine, including appendix, were CK7–/CK20+ in the great majority of cases, as were goblet cell carcinoids, but half of the poorly differentiated adenocarcinomas exhibited aberrant expression, as did most of the mixed goblet cell carcinoid/adenocarcinomas. All five high‐grade neuroendocrine carcinomas were negative for both CK7 and CK20.

Conclusions:  Not only the site but also the grade and histological type of a gastrointestinal carcinoma should be considered when assessing cytokeratin phenotype.
Alteration of E‐cadherin‐mediated adhesion protein is common, but microsatellite instability is uncommon in young age gastric cancers
Authors: Lim S , Lee H S , Kim H S , Kim Y I , Kim W H
Published: March 2003

Aims:  Gastric adenocarcinoma in young patients has been considered to differ in many ways from gastric carcinoma in older patients. This study was designed to determine the clinicopathological features and molecular mechanisms.

Methods and results:  Based on 4123 patients of gastric cancer in Seoul National University Hospital, 135 patients (3.3%) were chosen by the age of 30 years or younger. Expression of E‐cadherin, β‐catenin, p53 and Epstein–Barr virus (EBV) was analysed using the tissue array method in formalin‐fixed paraffin‐embedded specimens and microsatellite instability (MSI) was determined. As a control, 320 cases of older patients were compared. Gastric adenocarcinoma of young patients revealed significant female predominance, type IV gross type, proximal location, diffuse type and frequent lymph node metastasis. In‐situ hybridization for EBV showed higher positivity in young patients (9/78, 11.5%) than in older ones, but not statistically significant. In EBV+ cases, p53 over‐expression was significantly higher in young patients than older patients (P < 0.05). Alteration of E‐cadherin or β‐catenin was significantly higher in younger patients than in older patients (P < 0.05). Overall survival was significantly poorer in younger patients than older ones. The frequency of MSI was rare (1.3%, P < 0.05) in young patients compared with older patients (9.3%).

Conclusions:  These data indicate that gastric adenocarcinoma of young patients has a poor prognosis, possesses aggressive histopathological features, exhibits reduced expression of E‐cadherin and β‐catenin, and demonstrates lower MSI than tumours in older patients.
The effects of monocytes on the transendothelial migration of T lymphocytes
Authors: Lidington E. A. , Mccormack A. M. , Yacoub M. H. , Rose M. L.
Published: March 2003

In vivo cell‐mediated immune reactions are characterized by mixtures of monocytes and T cells. The purpose of this study was to investigate the role of monocytes on T‐cell migration and induction of endothelial adhesion molecules. The in vitro model consisted of adding peripheral blood mononuclear cells (PBMC), T cells or mixtures of monocytes and T cells, to endothelial cells on a porous membrane and using flow cytometry to distinguish between the monocyte and lymphocyte components. PBMC and PBMC supernatants were highly potent at upregulating intercellular adhesion molecule‐1 (ICAM‐1) and inducing expression of vascular cell adhesion molecule‐1 (VCAM‐1) and E‐selectin. Induction by supernatants was inhibited by antibodies to tumour necrosis factor‐α (TNF‐α) and interleukin‐1 (IL)‐1β. Using monocyte‐enriched populations, as few as one monocyte to 100 endothelial cells was sufficient to upregulate adhesion molecules. Fixed monocytes also induced adhesion molecules and expressed surface‐bound cytokines. In contrast, highly purified unstimulated T cells were not found to induce adhesion molecules at 4, 6, 24 or 48 hr of coculture. Purified T cells showed low‐level migration through resting (VCAM‐1 negative) endothelium, which was approximately doubled by addition of small numbers of monocytes or TNF‐α. In conclusion, monocytes, via cell surface or released cytokines play an essential role in allowing large‐scale recruitment of T cells to inflammatory sites in vivo.
A 24 000 MW Trypanosoma cruzi antigen is a B‐cell activator
Authors: Cordeiro Da Silva A. , Espinoza A. G. , Taibi¶ A. , Ouaissi A. , Minoprio P.
Published: March 2003

Trypanosoma cruzi, the causative agent of Chagas’ disease, is a protozoan parasite that infects humans and other mammals in Central and Latin America. Several alterations of the immune response after infection have been described, such as severe immunosuppression of both cellular and humoral responses and massive polyclonal B‐ and T‐cell activation, including the expansion of self‐reactive clones. We have investigated the effects of the intraperitoneal injection of a recombinant 24 000 MW T. cruzi‐specific antigen (rTc24) on the immune response of normal and deficient strains of mice. We analysed the in vivo and ex vivo levels of lymphocyte activation and the proliferative responses to rTc24 by determining the expression of CD69 activation marker and the levels of thymidine incorporation by spleen cells. The numbers of antibody‐producing cells were determined by ELISPOT and the levels of immunoglobulin in the sera by isotype‐specific enzyme‐linked immunosorbent assay. We observed an increased [3H]thymidine ([3H]TdR) incorporation by spleen cells after rTc24 stimulation in vivo and in vitro. This proliferative activity induced by rTc24 was independent of the mouse strain used in the experiments (including C3H/HeJ mice) and ruled out the possibility that rTc24 preparations were contaminated by lipopolysaccharide. The injection of rTc24 protein induced preferentially the activation of B cells, as determined by the increased expression of CD69 molecules on IgM+ spleen cells. Considerable increases of IgM‐secreting B cells were determined in both athymic and euthymic BALB/c mice. Mice that are deficient in B cells (BALB.Xid) responded to rTc24 but to a lesser extent. These increases in IgM B‐cell numbers were accompanied by elevated levels of IgM immunoglobulins in the sera of injected animals. Our results suggest a role for rTc24 in B‐cell activation.
Prostaglandin and fatty acid modulation of Escherichia coli O157 phagocytosis by human monocytic cells
Authors: Davidson J. , Kerr A. , Guy K. , Rotondo D.
Published: March 2003

Phagocytosis by human monocytes is an important primary survival mechanism particularly during bacterial infection. However, the processes that control the events and mediators involved in the activation of monocytes and their impact on the phagocytosis of bacteria are poorly understood. The effect of bacterial endotoxin, interleukin‐1β (IL‐1β), fatty acids and prostaglandin E2 (PGE2) on the phagocytosis of fluoroscein isothiocyanate (FITC)‐labelled Escherichia coli (O157) by human blood monocytes and U937 cells was studied by flow cytometry. Endotoxin increased the phagocytosis of labelled bacteria by both monocytes and U937 cells. IL‐1β and the polyunsaturated fatty acids; dihomo‐γ‐linolenic and arachidonic acids also increased the phagocytic activity of both monocytes and U937 cells. In contrast, PGE2 suppressed phagocytosis in a concentration‐dependent manner. The cyclo‐oxygenase inhibitor, ketoprofen, further enhanced the increased phagocytic activity in the presence of endotoxin and interleukin‐1 (IL‐1) indicating suppression by endogenous prostaglandins. This was confirmed by the data which showed that lipopolysaccharide (LPS) and IL‐1 increased PGE2 release and ketoprofen inhibited release. Endotoxin and fatty acids increased IL‐1β release also, whereas PGE2 inhibited release. The data suggest that phagocytic activity may be linked to changes in IL‐1 levels. The data presented in this study also suggest that monocyte phagocytosis in the course of bacterial infection would be altered during pathophysiological events which result in elevation of extracellular fatty acids.
Granulocyte–macrophage colony‐stimulating factor induces the differentiation of murine erythroleukaemia cells into dendritic cells
Authors: CAO  , ZHAO  , YU  , WANG  , ZHANG  , ZHANG  , WANG
Published: March 2003

Dendritic cells (DC) are professional antigen‐presenting cells (APC) within the immune system and antigen‐pulsed DC can be used as an effective vaccine for active immunotherapy of cancer. Granulocyte–macrophage colony‐stimulating factor (GM‐CSF) plays an important role in the generation of DC. We previously showed that GM‐CSF can induce murine erythroleukaemia cells (FBL‐3) to differentiate into monocyte‐like cells. To develop a new vaccinating method to stimulate the host immune response to leukaemia, we further investigate whether FBL‐3 cells induced by GM‐CSF can differentiate into DC in the present study. After being treated with GM‐CSF, FBL‐3 cells expressed high levels of 33D1 and NLDC‐145, which are the specific markers of DC. The expression of MHC‐II, B7‐1, B7‐2 and vascular cell adhesion molecule‐1 (VCAM‐1) was up‐regulated markedly; the typical morphology of DC were also observed by electron microscopy. Functionally, the GM‐CSF‐induced FBL‐3 cells could apparently stimulate the proliferation of naive allogeneic and autologous T lymphocytes and induce the generation of specific CTL more efficiently than the wild‐type FBL‐3 cells. Mice immunized with GM‐CSF‐induced FBL‐3 cells could resist the subsequent challenge with the wild‐type FBL‐3 cells. Collectively, these data indicate that GM‐CSF differentiates murine erythroleukaemia cells into DC phenotypically, morphologically and functionally. FBL‐3‐derived DC can be used as a new type of vaccine. Our results may have important implications for the immunotherapy of leukaemia.
Lymphotactin: a key regulator of lymphocyte trafficking during acute graft rejection
Authors: WANG  , NONOMURA  , TAKAHARA  , LI  , AZUMA  , ICHIMARU  , KOKADO  , MATSUMIYA  , MIKI  , SUZUKI  , OKUYAMA
Published: March 2003

The attraction of leucocytes to allografts is essential for rejection. The prcocess is controlled by chemokines. In order to clarify the role of lymphotactin (a cytokine that represents a novel branch of the chemokine superfamily) in regulating leucocyte trafficking during graft rejection, we used rat renal transplantation models to examine its gene expression and the distribution of lymphotactin‐expressing cells in renal grafts. Lymphotactin mRNA was upregulated strongly in acutely rejecting renal allografts. The mRNA was undetectable in isografts, chronically rejecting renal allografts or normal kidney. Once lymphotactin was expressed, large numbers of infiltrating lymphocytes were seen. Moreover extended studies demonstrated that in cultured rat spleen cells the expression of lymphotactin mRNA was markedly induced by phytohaemagglutinin (PHA) or phorbol myristate acetate (PMA), and such induction was inhibited by the immunosuppressive drugs FK506 and cyclosporin. Collectively, these observations provide new evidence demonstrating that lymphotactin is a key regulator of lymphocyte motility and adhesiveness during acute allograft rejection. FK506 and cyclosporin inhibition of lymphotactin expression is likely to represent an important molecular mechanism of the action of the drugs.
Amidolytic and peptidolytic activities of immunoglobulin G present in sera from patients with rheumatoid arthritis, Sjogren’s syndrome and systemic lupus erythematosus
Authors: MATSUURA  , IKOMA  , SUGIYAMA  , FUNAUCHI  , SINOHARA
Published: March 2003

Polyclonal immunoglobulin G (IgG) from healthy subjects was found to be capable of hydrolyzing carbobenzoxy–Val–Gly–Arg–p‐nitroanilide (a synthetic chromogenic substrate for trypsin) and d–Pro–Phe–Arg–p‐nitroanilide (a substrate for plasma kallikrein). Statistically significant elevation of activity against the former substrate was found in patients with rheumatoid arthritis (RA), but not in patients with Sjogren’s syndrome (SjS) or systemic lupus erythematosus (SLE). On the other hand, IgG samples from the patients with these three autoimmune diseases showed reduced activity against d–Pro–Phe–Arg methylcoumarinamide, although the differences were not statistically significant. Preliminary studies have shown that two out of three IgG samples from RA patients exhibited the activity of cleaving a pentapeptide, Gln–Arg–Arg–Ala–Ala, whereas virtually no cleavage of the same peptide was observed with IgG from healthy controls or from patients with SjS or SLE.
Determination of cytokine co‐expression in individual splenic CD4+ and CD8+ T cells from influenza virus‐immune mice
Authors: FALCHETTI  , DI FRANCESCO  , LANZILLI  , GAZIANO  , CASALINUOVO  , PALAMARA  , RAVAGNAN  , GARACI
Published: March 2003

We have studied the patterns of interleukin‐2 (IL‐2), IL‐4 and interferon‐γ (IFN‐γ) co‐expression displayed by individual splenic CD4+ and CD8+ T cells in response to influenza virus immunization. Unseparated spleen cells obtained from mice intraperitoneally (i.p.) injected with A/PR8 (H1N1) influenza virus (PR8) were cultured for 24 hr in the presence of ultraviolet‐inactivated PR8. As controls, cultures of both naive spleen cells stimulated with PR8 or of immune cells lacking the inactivated virus were used. The frequencies of CD4+ and CD8+ T cells expressing IL‐2, IL‐4 and IFN‐γ were determined by three‐colour flow cytometric analysis of fixed and saponin‐permeabilized cells fluorescent‐stained for either CD4 or CD8 surface molecules and for one of the following combinations of two intracellular cytokines: IL‐2/IL‐4, IL‐2/IFN‐γ and IL‐4/IFN‐γ. The results showed that immunization with influenza virus induces in both CD4+ and CD8+ T cells a heterogeneity of cytokine response patterns that do not follow the type 1/type 2 polarized response model, but with substantial differences between the two populations. In fact, the analysis of the phenotypes of virus‐immune CD8+ T cells revealed similar significant proportions of cells either expressing any one of the three cytokines or co‐expressing combinations of them (i.e. IL‐4/IL‐2, IL‐4/IFN‐γ and IL‐2/IFN‐γ), whereas immune CD4+ T cells were seen to express almost exclusively a single cytokine per cell. The observed patterns of cytokine production suggest that influenza virus immunization induces the expression of a type 0 cytokine pattern at both population and single cell levels in CD8+ T cells and exclusively at the population level in CD4+ T cells.
Elevated major histocompatibility complex class I expression protects T cells from antibody‐ and macrophage‐mediated deletion
Authors: ORLIKOWSKY  , WANG  , DUDHANE  , MITTLER  , TREJO  , DANNECKER  , NIETHAMMER  , HOFFMANN
Published: March 2003

Macrophages are capable of destroying T cells with which they form cellular conjugates. The deletion can be prevented by the simultaneous transmission of costimulatory signals. We show here that T cells with elevated major histocompatibility complex (MHC) class I expression are resistant against macrophage‐mediated cytotoxicity. T cells that express the CD45RO isotype, considered memory T cells, exhibit MHC class I antigen at higher density than naive CD45RA T cells and upregulate MHC class I expression promptly when they form cellular conjugates with macrophages. We confirm previous observations that CD45RA T cells are more susceptible to antibody‐ and macrophage‐mediated deletion than memory CD45RO T cells. When MHC class I molecules are masked by specific monoclonal antibody or antibody Fab fragments, CD45RA T cells and CD45RO T cells exhibit equal susceptibility to macrophage cytotoxicity, demonstrating that the difference between CD45RA and CD45RO T cells in their sensitivity to macrophage cytotoxicity is determined by their MHC I expression. Separation of CD4 T cells from CD8 T cells deprives memory CD4 T cells of their resistance against macrophage cytotoxicity, suggesting that memory T cells’ resistance against destruction by macrophages is controlled by regulatory T cells.
The biological effects induced in mice by p36, a proteinaceous factor of virulence produced by African swine fever virus, are mediated by interleukin‐4 and also to a lesser extent by interleukin‐10
Authors: VILANOVA  , FERREIRA  , RIBEIRO  , ARALA‐CHAVES
Published: March 2003

We have previously presented indirect evidence that both specific immunosuppression and lymphocyte mitogenicity induced in mice by p36, a proteinaceous factor of virulence produced by porcine monocytes infected by African swine fever virus, were consistent with a Th2‐driven response. Here we show: (1) Interleukin‐4 (IL‐4) and interleukin‐10 (IL‐10) mRNA expression in the spleen and thymus of C57BL/6 mice were displayed early after p36 inoculation. The expression of thymic IL‐10 mRNA occurred, however, later than that of IL‐4 mRNA. (2) Increased serum levels of these two cytokines were also soon detected after the protein inoculation. (3) Both immunosuppressive and mitogenic effects of p36 were absent in IL‐4 gene‐targeted mice and partially abrogated in mice depleted of IL‐4 by neutralizing monoclonal antibodies. (4) IL‐10 depletion abrogated the immunosuppressive but not the p36 lymphocyte mitogenic biological effects. (5) The increase in the serum concentrations of both IL‐4 and IL‐10 were lower in thymectomized than in non‐thymectomized mice. (6) The expression of interferon‐γ (IFN‐γ) mRNA was weakly or not at all induced in p36‐treated mice. Taken together, these results are in agreement with the promotion of a Th2 immune response induced by p36.
Distribution of lymphocytes and adhesion molecules in human cervix and vagina
Authors: JOHANSSON  , RUDIN  , WASSÉN  , HOLMGREN
Published: March 2003

Knowledge of the histological distribution of leucocytes and adhesion molecules in the human genital tract is scarce although local immunity in this region is important. Using immunohistochemical methods, we here describe the organization of CD3+, CD8+ and CD4+ T cells, CD19+ B cells, CD38+ plasma cells, major histocompatibility complex (MHC) class II+ antigen‐presenting cells and CD14+ monocytes, as well as the expression of endothelial addressins in normal human ecto‐cervical and vaginal mucosa. T cells were clustered in a distinct band beneath the epithelium and were also dispersed in the epithelium and the lamina propria, whereas CD38+ plasma cells were present only in the lamina propria. MHC class II+ cells were numerous in the lamina propria and in the epithelium, where they morphologically resembled dendritic cells. Lymphoid aggregates containing CD19+ and CD20+ B cells as well as CD3+, CD4+ and CD8+ cells were also found in the cervix. The mucosal addressin cell adhesion molecule‐1 (MAdCAM‐1) was not expressed on the vascular endothelium in the cervical or vaginal mucosa. In contrast, intercellular adhesion molecule‐1 (ICAM‐1), vascular adhesion protein‐1 (VAP‐1) and P‐selectin were expressed in all tissue samples, and vascular cell adhesion molecule‐1 (VCAM‐1) and E‐selectin were found in four of seven samples. We conclude that the distribution of leucocytes and adhesion molecules is very similar in the ecto‐cervical and the vaginal mucosa and that the regulation of lymphocyte homing to the genital tract is different from that seen in the intestine. Our results also clearly suggest that the leucocytes are not randomly scattered in the tissue but organized in a distinct pattern.
Co‐immunization with DNA vaccines expressing granulocyte–macrophage colony‐stimulating factor and mycobacterial secreted proteins enhances T‐cell immunity, but not protective efficacy against Mycobacterium tuberculosis
Authors: KAMATH  , HANKE  , BRISCOE  , BRITTON
Published: March 2003

The development of more effective antituberculosis vaccines would assist in the control of the global problem of infection with Mycobacterium tuberculosis. One recent vaccination strategy is immunization with DNA plasmids encoding individual microbial genes. Using the genes for the M. tuberculosis‐secreted proteins, MPT64 (23 000 MW) and Ag85B (30 000 MW) as candidate antigens, we previously prepared DNA vaccines and demonstrated their ability to stimulate T‐cell responses and confer protection in a mouse model of aerosol tuberculosis (TB). The protective efficacy of the DNA vaccines was less than that promoted by the current vaccine Mycobacterium bovis bacille Calmette–Guèrin (BCG). To improve the immunogenicity and protective efficacy of these mycobacterial vectors, co‐immunization of a plasmid expressing granulocyte–macrophage colony‐stimulating factor (GM‐CSF) was investigated. Intramuscular immunization with DNA expressing MPT64 or Ag85B and GM‐CSF enhanced the antigen‐specific cellular immune response, with increased proliferative response and production of interferon‐γ (IFN‐γ). The titre of antimycobacterial protein immunoglobulin G (IgG) antibodies was unchanged. Mice immunized with DNA vaccines showed reduced pulmonary bacterial load following an aerosol challenge of M. tuberculosis, but codelivery of the plasmid expressing GM‐CSF did not increase the protective effect. Therefore, despite modifying the cellular immune response to DNA vaccines, GM‐CSF does not improve their protective efficacy at the peak of infection after an aerosol challenge with 100 c.f.u. of M. tuberculosis.
Integrin α1β1 (VLA‐1) mediates adhesion of activated intraepithelial lymphocytes to collagen
Authors: Roberts  , Brolin  , Ebert
Published: March 2003

Intraepithelial lymphocytes (IELs) from human intestinal epithelium are memory CD8+ T cells that bind to epithelial cells through human mycosal lymphocyte (HML)‐1 and to mesenchymal cells through very late activation antigen‐4 (VLA‐4). Their binding of extracellular matrix proteins and the mechanism involved were tested. Activated 51Cr‐labelled lymphocytes were incubated in protein‐coated microwells with various additives. After washing, the adherent cells were detected by radioactivity. The percentages of activated IELs that bound to collagen types I and IV were 20 and 31%, respectively; fewer bound to fibronectin or laminin. Compared to interleukin‐2‐activated peripheral blood CD8+ T lymphocytes, more IELs bound collagen IV and fewer bound fibronectin. IEL adhesion to collagen (but not fibronectin or laminin) was up‐regulated by antibody ligation of CD2 or by protein kinase C stimulation by phorbol ester; staurosporine reduced binding, while herbimycin, phytohaemagglutinin and CD3 ligation had no effect. Antibody‐blocking of integrin VLA‐1 subunits α1 (CD49a) and β1 (CD18) inhibited adhesion to collagen type I by 82±6% and to type IV by 94±1% (P<0·001), implicating VLA‐1 as the main collagen receptor for IELs. Cell adhesion was dependent on extracellular divalent cations, a characteristic event of VLA‐1 never before shown for IELs: manganese and magnesium ions supported binding in a dose‐dependent manner; calcium ions inhibited their effectiveness. Therefore, IELs bind collagen through integrin α1β1 after protein kinase C activation. Adhesion is modulated by divalent cations.
Role of interleukin‐4 in down‐regulation of contact sensitivity by γδ T cells from tolerized T‐cell receptor α−/− mice
Authors: Szczepanik  , Ptak  , Askenase
Published: March 2003

Contact sensitivity (CS) is a classical example of an in vivo T‐cell‐mediated immune response that is under regulation. Such down‐regulation can be mediated by αβ T cells in mice that are tolerized by prior exposure to high doses of antigen. In contrast, we demonstrated previously that such high‐dose antigen tolerance in T‐cell receptor (TCR) α−/− H‐2d mice induced antigen‐specific, apparently major histocompatibility complex‐unrestricted, CD4− CD8−γδ T cells, that also could down‐regulate CS responses antigen‐specifically in vivo, and also inhibited in vitro production of IFN‐γ. In the present experiments we employed H‐2b‐deficient TCRα−/− and TCRβ−/− mice, owing to different molecular constructs than were used previously, and confirmed that tolerized γδ T cells in these different H‐2bαβ TCR−/− mice down‐regulated CS. Thus, γδ T‐cell suppressor function was not limited to mice bearing a special transgenic TCRα−/− DNA construct. Furthermore, employing monoclonal antibody and complement depletion in vitro and adoptive transfer in vivo, characterized the phenotype of these γδ down‐regulatory T cells as: CD3+, CD28+, CD40‐ligand+, Fas+, FcγR+ and NK1.1−. Also, in vitro antigen desensitization of these trinitrophenyl (TNP)‐specific TCRγδ+ down‐regulatory cells was achieved with soluble TNP‐bovine serum albumin (BSA), but not with oxazolone‐BSA, showing that these suppressive γδ T cells have antigen‐specific receptors. Moreover, employing monoclonal antibody blocking of γδ suppressors in vitro, and of recipients in vivo, we showed that interleukin‐4 (IL‐4) was involved in this down‐regulation of CS by γδ T cells, while IL‐10 and transforming growth factor‐β2 were not. In summary, generation of antigen‐specific, double‐negative, γδ suppressor cells, by tolerance of high antigen doses in TCRα−/− mice, appears to be a general phenomenon, and IL‐4 production is involved in their down‐regulation of the T helper type 1 cells that mediate CS.
Ability of T cells from patients with rheumatoid arthritis to respond to immunoglobulin G
Authors: Lang  , Macht  , Kirwan  , Wraith  , Elson
Published: March 2003

The ability of T cells from rheumatoid factor (RF)‐positive patients with rheumatoid arthritis (RA) to respond to immunoglobulin G (IgG) was assessed. Peripheral blood mononuclear cells (PBMC) from RA patients and normal individuals were cultured with and without human IgG or Mycobacterium tuberculosis‐purified protein derivative (PPD) for 7 days and their proliferative response measured at intervals by their ability to take up tritiated thymidine. PBMC from 14/26 RA patients proliferated in response to IgG (taking a stimulation index of 3 or above as positive). The peak response varied between individuals but usually occurred on day 5, the same day, or 1 day later than the peak response to PPD. By contrast, PBMC from a significantly lower proportion (1/9) of normal individuals and patients with other arthritides (0/6) responded to IgG, although all responded to PPD. PBMC from 9/14 RA patients responded to Fab fragments of IgG but only 3/9 to the Fc fragment. Higher proliferative responses from RA PBMC were elicited by IgG aggregates than the original IgG preparation, but PMBC from 5/5 normal individuals and 5/6 patients with other arthritides failed to respond to the aggregates. The response to IgG was human leucocyte antigen (HLA)‐DR restricted and mediated by CD4+ T cells. It is considered that these results advance the hypothesis that IgG‐reactive T cells contribute to the initiation or perpetuation of RA.
Bacterial lipopolysaccharide acts as an adjuvant to induce autoimmune arthritisin mice
Authors: Yoshino S. , Sasatomi E. , Ohsawa M.
Published: March 2003

We investigated the ability of lipopolysaccharide (LPS) as an adjuvant to induce autoimmune arthritis. LPS from Escherichia coli was intraperitoneally injected into DBA/1J mice together with the joint cartilage component type II collagen (CII) on day 0. Thereafter, the injection of CII and LPS was continued every 2 weeks up to day 56. The results showed that mice injected with CII plus LPS had signs of arthritis on day 55 and the joint inflammation reached a peak on day 75. Injection of CII or LPS alone induced no arthritis. Histologically, marked oedema of synovium and intense infiltration of inflammatory cells, including neutrophils, were observed 3 days after the onset of joint inflammation. Twenty‐one days later, there were marked proliferation of synovial tissues with many mononulear cells and destruction of cartilage. Anti‐CII immunoglobulin G (IgG) and IgG2a antibodies were markedly produced in mice injected with CII plus LPS. Pronounced secretion of cytokines, including interleukins‐12 and ‐1β, interferon‐γ and tumour necrosis factor‐α, was also observed in these animals. Arthritis was passively transferred into naive syngeneic mice with sera but not with lymphoid cells from mice given CII with LPS. Other types of LPS from Salmonella enteritidis, Salmonella typhimurium and Klebsiella pneumoniae as well as lipid A from E. coli, induced inflammation in joints when administered with CII. Polymixin B sulphate mixed with LPS or lipid A blocked the induction of joint inflammation. These results indicate that LPS appears to play an important role as an adjuvant in the induction of arthritis in which autoimmunity to CII is involved.
Decreased expression of FcγRIII (CD16) by γδ T cells in patients withrheumatoid arthritis
Authors: Bodman‐smith M. D. , Anand A. , Durand V. , Youinou P. Y. , Lydyard P. M.
Published: March 2003

Some γδ T cells express a receptor for the Fc portion of immunoglobulin G (FcγRIII – CD16). The relevance of this Fc receptor to γδ T‐cell function is at present unclear. Our previous studies have shown that γδ T cells express activation markers in patients with rheumatoid arthritis (RA). In this study we have examined the relative proportions of CD16+γδ T cells in the blood and synovial fluid of these patients compared with control blood. CD16+γδ T cells from RA patients were significantly reduced in synovial fluid compared with the circulation. That this was due to blocking of antibody binding to CD16 was unlikely as treatment of blood γδ T cells with RA synovial fluid (known to contain immune complexes) failed to alter expression of CD16. Treatment of blood γδ T cells with phytohaemagglutinin in vitro, resulted in a time‐dependent decrease in expression of CD16, with a concomitant increase in expression of human leucocyte antigen‐DR, at the single cell level. We conclude that expression of CD16 by γδ T cells is lost in the synovial compartment as the result of activation.