bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of shear stress and a stable prostaglandin I2 analogue on adhesive interactions of colon cancer cells and endothelial cells
Authors: Yoshida N , Yoshikawa T , Nakagawa S , Sakamoto K , Nakamura Y , Naito Y , Kondo M 
Published: March 2003

SUMMARY
In the process of cancer metastasis, adhesion between cancer cells and endothelial cells is an important early step. In the present study, the effects of shear stress and the adhesion molecules responsible for cancer cell interactions with endothelial cells were investigated in a system similar to in vivo microcirculation. The effect of prostaglandin I2 (PGI2) also was determined. Human colon cancer cell line Colo 201 and human umbilical vein endothelial cells (HUVEC) were used. After HUVEC on a glass slide were incubated with IL-1β for 4 h, cancer cells in suspension were perfused on HUVEC at wall shear stresses of 5–40 μN/cm2. Experiments were videotaped, and the number of adherent cells were counted. Additionally, the effects of anti-sialyl Lewis a (SLea) MoAb, anti-E-selectin MoAb, and a PGI2 analogue were investigated. Expression of adhesion molecules on cancer cells and HUVEC was assessed using flow cytometry and enzyme immunoassay, respectively. Few cancer cells adhered to HUVEC without IL-1β; however, many cancer cells adhered to IL-1β-stimulated HUVEC at low shear stress (5–20 μN/cm2). Cancer cells did not migrate beneath HUVEC. The increased adhesion was inhibited by anti-E-selectin MoAb, anti-SLea MoAb, and a PGI2 analogue. In addition, the PGI2 analogue decreased the surface expression of SLea on Colo 201 cells. These results suggest that Colo 201 cells adhere to IL-1β-stimulated endothelial cells via SLea and E-selectin under low flow conditions; PGI2 analogues may protect against metastasis by inhibiting cancer cell–endothelial cell interactions.
Tumour necrosis factor (TNF) production by T cell receptor-primed T lymphocytes is a target for low dose methotrexate in rheumatoid arthritis
Authors: Hildner K , Finotto S , Becker C , Schlaak J , Schirmacher P , Galle P R , Märker-Hermann E , Neurath M F 
Published: March 2003

SUMMARYMethotrexate (MTX) is an effective immunosuppressive agent in various chronic inflammatory diseases such as rheumatoid arthritis (RA). However, its mechanisms of action are only partially understood. In this study, we assessed the effects of MTX on the differentiation of peripheral blood (PB) CD4+CD45RA ‘naive’ and CD4+CD45RO ‘memory’ T cells from healthy controls and patients with RA. Accordingly, purified T cells were primed and restimulated in vitro via the T cell receptor (TCR) in the presence of IL-2 to generate effector T cells secreting large amounts of Th1 and Th2 cytokines. We observed that low doses of MTX strongly suppress TNF and to a lesser extent interferon-gamma (IFN-γ) production by T cells from both healthy donors and RA patients when present during T cell priming via the TCR. Similar data were obtained for TCR-primed synovial fluid mononuclear cells in RA. In contrast, production of IL-4 by TCR-primed CD45RA T cells was significantly increased upon MTX treatment. Interestingly, MTX did not enhance IL-4 production when present during restimulation of effector CD45RO T cells, although it still suppressed TNF production. The results indicate that MTX effects depend on the stage of T cell activation and identify TNF production by TCR-primed T lymphocytes as a target for low-dose MTX treatment in RA. These findings could explain the delayed clinical effects of MTX and may contribute to its potent anti-inflammatory and immunoregulatory properties.
Turkeys are protected from infection withChlamydia psittaciby plasmid DNA vaccination against the major outer membrane protein
Authors: Vanrompay D , Cox E , Volckaert G , Goddeeris B 
Published: March 2003

SUMMARYPlasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydia psittaci serovar A strain has been tested for its ability to raise an immune response and induce protection against challenge with the same serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared with gene gun-based immunization. The gene gun delivery of pcDNA1/MOMP as well as the intramuscular–intranasal DNA delivery primed both T-helper and B cell memory, although rMOMP-expressing cells did not induce high antibody responses. Evidence for the priming of the memory was provided by the fact that the pcDNA1/MOMP inoculations raised antibodies belonging to the IgG and not IgM isotype. However, in response to challenge only five out of 15 vaccinated turkeys showed four-fold increases in serum IgG after challenge. By contrast, evidence for the priming of T cell memory in response to challenge was found in all vaccinated turkeys, as shown by the significantly heightened proliferative responses of peripheral blood lymphocytes following vaccination. Both immunization methods produced similar serological and lymphocyte proliferative responses. Notwithstanding the immunization method, a significant level of protection was observed in all pcDNA1/MOMP-immunized turkeys. The efficacy of MOMP-based DNA vaccination as a means of preventing severe clinical signs, lesions and chlamydia excretion in a turkey model of C. psittaci infection was demonstrated.
The Fas and Fas ligand pathways in liver allograft tolerance
Authors: Pan T L , Goto S , Lin Y C , Lord R , Chiang K C , Lai C Y , Chen Y S , Eng H L , Cheng Y F , Tatsuma T , Kitano S , Lin C L , Chen C L 
Published: March 2003

SUMMARY
The Fas and Fas ligand (Fas/FasL) pathways may play a central role in cytotoxicity or immunoregulation in liver transplantation. Here, in an attempt to examine the role of Fas/FasL on drug-free tolerance, we measured mRNA levels of Fas/FasL in livers by reverse transcriptase-polymerase chain reaction (RT-PCR), and also protein levels of Fas/FasL in livers by immunohistochemistry and in serum by dot blot assay. PVG recipients bearing DA livers showed serious rejection between post-operative (POD) days 7 and 14 , but this rejection was naturally overcome without any immunosuppression. Fas gene and protein products were expressed on almost every cell in livers taken from naive rats, and at any time point in both syngeneic and allogeneic orthotopic liver transplantation (OLT) rats. In contrast, FasL mRNA in DA livers was detectable at POD 2, peaked at POD 14, and declined at POD 63 in allogeneic OLT (DA-PVG). Although the FasL gene was detectable in isografts at POD 14, its expression was much lower than in allografts. The time course and localization of FasL expression indicated that the expression of FasL gradually switched from infiltrating cells to hepatocytes when the rejection was naturally overcome and tolerance was induced in this OLT model. Soluble Fas could constitutively be detected at any time point in the serum of the tolerogenic OLT (DA-PVG) rats and was not diminished during the rejection phase. Soluble FasL peaked at POD 14 in allogeneic OLT, while sFasL was significantly lower in the serum of normal and syngeneic OLT rats. These findings suggest that the Fas and FasL pathways, including soluble forms, may contribute to the control of the immune response in this drug-free tolerance OLT model.
Induced nitric oxide (NO) synthesis in heterologous nephrotoxic nephritis; effects of selective inhibition in neutrophil-dependent glomerulonephritis
Authors: Waddington S N , Mosley K , Cattell V 
Published: March 2003

SUMMARYIncreased NO synthesis, due to inducible NO synthase (iNOS) activity, is found in macrophage-associated glomerulonephritis. Little is known about NO in neutrophil-dependent immune complex inflammation, and its role remains controversial. We therefore studied early phase heterologous nephrotoxic nephritis (HNTN) induced in rats by nephrotoxic globulin and the effects of selective iNOS inhibition of this model. At 2 h of the model iNOS mRNA was induced and nitrite (NO−2) was generated in glomeruli incubated ex vivo (5.2 ± 1.0 nmol/2000 glomeruli per 24 h). There were 14.7 ± 2.2 polymorphonuclear neutrophils (PMN)/glomerulus (normal controls 0.1 ± 0.1). At 8 h urinary protein was 69 ± 15.3 (normal controls 0.6 ± 0.2 mg/24 h). Peritoneal PMN expressed iNOS and produced significant NO−2 (basal 11.2 ± 0.3 nmol/106 cells per 24 h). Selective iNOS inhibition with L-N6-(1-iminoethyl)-lysine (L-NIL) in vitro inhibited nephritic glomerular and PMN NO−2 synthesis. In HNTN L-NIL in vivo significantly suppressed elevated plasma NO−2/NO−3 levels (representative experiment: 17 ± 2 μm, untreated 40 ± 4 μm,P = &lt; 0.01, normal control 18 ± 2 μm). This inhibition did not affect leucocyte infiltration into glomeruli or induce thrombosis. There was no consistent effect on proteinuria. This is the first demonstration of glomerular iNOS induction and high output NO production in the acute phase of PMN-dependent acute immune complex glomerulonephritis. Selective iNOS inhibition does not affect the primary mechanism of injury (leucocyte infiltration) in this model.
Differences in cytokine production by peripheral blood mononuclear cells (PBMC) between patients with atopic dermatitis and bronchial asthma
Authors: Kimura M , Tsuruta S , Yoshida T 
Published: March 2003

SUMMARYIt is widely accepted that type 2 helper T (Th2) lymphocytes play a crucial role in the pathogenesis of atopic dermatitis (AD) as well as bronchial asthma (BA). We measured the amounts of IL-5 and interferon-gamma (IFN-γ) produced by PBMC upon stimulation with house dust mite (HDM) or Candida albicans (CA) in 17 children (3–15 years) with AD, and compared these values with those of 16 children with BA. Although IL-5 production by PBMC upon stimulation with HDM in patients with AD was significantly higher than that in 13 non-atopic controls (geometric mean = 23.4 pg/ml versus 5.9 pg/ml, P &lt; 0.05), it was significantly lower than that in patients with BA (177.8 pg/ml, P &lt; 0.001). The amount of IL-5 produced by PBMC upon stimulation with CA was also significantly lower in patients with AD than in those with BA (7.2 pg/ml versus 100.0 pg/ml, P &lt; 0.001). The production of IFN-γ by PBMC stimulated with HDM or CA was also significantly lower in patients with AD than in those with BA (HDM 4.3 pg/ml versus 12.6 pg/ml, P &lt; 0.05; CA 6.5 pg/ml versus 60.3 pg/ml, P &lt; 0.001). Consequently the ratio of IL-5 to IFN-γ production was high not only in patients with BA but also in those with AD. These findings suggest that there are some differences in the regulation of in vivo cytokine production between patients with AD and those with BA, although a Th2-dominant profile is common to both.
Expression of the BLM gene in human haematopoietic cells
Authors: Kaneko H , Matsui E , Fukao T , Kasahara K , Morimoto W , Kondo N 
Published: March 2003

SUMMARYBloom’s syndrome (BS) is a rare autosomal recessive disorder characterized by stunted growth, sun-sensitive erythema and immunodeficiency. Chromosomal abnormalities are often observed. Patients with BS are highly predisposed to cancers. The causative gene for BS has been identified as BLM. The former encodes a protein, which is a homologue of the RecQ DNA helicase family, a family which includes helicases such as Esherichia coli RecQ, yeast Sgs1, and human WRN. WRN is encoded by the gene that when mutated causes Werner’s syndrome. The function of BLM in DNA replication and repair has not yet been determined, however. To understand the function of BLM in haematopoietic cells and the cause of immunodeficiency in BS, expression of the BLM gene in various human tissues and haematopoietic cell lines was analysed and the involvement of BLM in immunoglobulin rearrangement examined. In contrast to WRN, BLM was expressed strongly in the testis and thymus. B, T, myelomonocytic and megakaryocytic cell lines also expressed BLM. All of the examined sequences at the junction of the variable (V), diversity (D) and joining (J) regions of the immunoglobulin heavy-chain genes were in-frame, and N-region insertions were also present. The frequency of abnormal rearrangements of the T cell receptor was slightly elevated in the peripheral T cells of patients with BS compared with healthy individuals, whereas a higher frequency of abnormal rearrangements was observed in the cells of patients with ataxia-telangiectasia (A-T). In DND39 cell lines, the induction of sterile transcription, which is required for class switching of immunoglobulin heavy-chain constant genes, was correlated with the induction of the BLM gene. Taking into consideration all these results, BLM may not be directly involved in VDJ recombination, but is apparently involved in the maintenance of the stability of DNA.
Soluble platelet selectin (sP-selectin) and soluble vascular cell adhesion molecule-1 (sVCAM-1) decrease during therapy with benznidazole in children with indeterminate form of Chagas’ disease
Authors: Laucella S A , Segura E L , Riarte E , Sosa Estani S 
Published: March 2003

SUMMARYThe immune response against Trypanosoma cruzi infection has been associated with both protection and pathogenesis. Central events in host defence system- and immune-mediated damage are tightly regulated by cell adhesion molecules (CAM). Levels of sP-selectin and sVCAM-1 were measured in sera from 41 children with the indeterminate phase of Chagas’ disease. Simultaneously, levels of soluble adhesion molecule were also quantified in Chagas’ disease children undergoing specific chemotherapy with benznidazole. Levels of sP-selectin and sVCAM-1 were found to be elevated in children with indeterminate Chagas’ disease before aetiologic therapy was started. However, a small group of patients showed sP-selectin and sVCAM-1 levels comparable to those of non-infected children. A positive correlation between levels of sVCAM-1 and sP-selectin in sera from Chagas’ disease patients was found. There was a significantly greater decrease in the titres of sP-selectin and sVCAM-1 in those children receiving benznidazole therapy compared with those children receiving placebo. Measurement of soluble adhesion molecules revealed differences in the activation of the immune system in children with the indeterminate form of Chagas’ disease. The early decrease of sP-selectin and sVCAM-1 levels after anti-parasitic treatment suggests that these molecules might be valuable indicators of effective parasitologic clearance.
T helper frequencies in peripheral blood reflect donor-directed reactivity in the graft after clinical heart transplantation
Authors: Vaessen L M B , Daane C R , Maat A P W M , Balk A H M M , Claas F H J , Weimar W 
Published: March 2003

SUMMARYWe describe the usefulness of a fast (48-h) limiting dilution assay (LDA) for the enumeration of human alloreactive helper T lymphocytes (HTL) in the peripheral blood, in relation to histologically defined rejection grades after heart transplantation. HTL frequencies (HTLf) in pretransplant samples varied from patient to patient, ranging from 106 to 625 HTL/106 peripheral blood mononuclear cells (PBMC). In the first week after heart transplantation (HTx), when immunosuppression was instituted, HTLf were significant lower (range 30–190 HTL/106). The level of HTL in the first week after HTx when rejection grade was 0 or 1A (ISHLT score) was considered to be the baseline frequency. This frequency did not correlate with the number of subsequent rejection episodes. During rejection (grade 3), donor-specific HTLf were increased above their baseline frequencies (P = 0.01). Expressed as percentage of baseline frequencies, HTLf increased significantly during acute rejection (AR) compared with 1–2 weeks before rejection (P = 0.003). The increase was specific, since viral infections did not result in a rise of donor-specific HTL, while also HTLf specific for third party HLA antigens were not elevated during rejection. Monitoring HTLf in peripheral blood with a shortened (48-h) assay may serve as a non-invasive method for detecting intragraft immunological reactivity. Demonstrating absence of donor-specific reactivity may limit the number of invasive endomyocardial biopsy (EMB) procedures and allow tapering of immunosuppressive treatment.
Melanoma antigen recognition by tumour-infiltrating T lymphocytes (TIL): effect of differential expression of Melan-A/MART-1
Authors: Ramirez-Montagut T , Andrews D M , Ihara A , Pervaiz S , Pandolfi F , Van Den Elsen P J , Waitkus R , Boyle L A , Hishii M , Kurnick J T 
Published: March 2003

SUMMARYWe have isolated, from an individual patient with metastatic melanoma, a series of eight TIL clones capable of lysing autologous melanoma cell targets. Six of the eight clones expressed TCRAV2S1 and lysed targets expressing HLA-A2 and the Melan-A/MART-1 peptide: AAGIGILTV. Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) analysis showed that the Melan-A/MART-1-specific clones were predominant in the bulk culture prior to cloning. However, the tumour progressed in vivo even in the presence of these tumour cell-lytic clones. Using the anti-Melan-A/MART-1 MoAb (A-103), we noted that Melan-A/MART-1 expression on three melanoma cell lines varied considerably during in vitro culture, in the absence of T cell immunoselection, relative to cell density. Tumour cells which spontaneously decreased Melan-A/MART-1 expression were less susceptible to specific TIL lysis. Melan-A/MART-1 expression and susceptibility to lysis increased in cells cultured at lower density. These data suggest that modulation of tumour antigen may account for tumour progression in the presence of tumour cell-lytic T lymphocytes. The observations suggest a possible explanation for the common finding of Melan-A/MART-1-specific lytic TIL in clinically progressing melanomas, as well as a possible pathway for therapeutic intervention.
Experimental lupus nephritis in severe combined immunodeficient (SCID) mice: remodelling of the glomerular lesions by bystander IgM antibodies
Authors: Ito M R , Terasaki S , Kondo E , Shiwaku H , Fukuoka Y , Nose M 
Published: March 2003

SUMMARYMRL/Mp-lpr/lpr (MRL/lpr) mice develop glomerular lesions with regular variations in their histopathological manifestations, similar to those in lupus nephritis. These lesions are mainly either cell-proliferative or wire loop-like and are associated with glomerular deposits of immunoglobulins, most frequently IgG and IgM. We previously established a nephritogenic IgG3-producing hybridoma clone, B1, from an MRL/lpr mouse, which induces only a ‘wire loop-like’ type of glomerular lesion when injected into SCID mice. Injection of SCID mice with an anti-trinitrophenyl IgM antibody-producing hybridoma clone, Sp6, following injection of the B1 clone, however, resulted in the development of a ‘cell-proliferative’ type of glomerular lesion, associated with an accumulation of both antibodies in glomeruli. This accumulation occurred even though Sp6 IgM antibodies did not react with B1 IgG3 antibodies and vice versa. A mutant clone of Sp6, T/13μE/3.1, which produces antibodies deficient in C1q binding, produced a similar effect as that of the Sp6 clone, i.e. ‘cell-proliferative’ lesions. Again the B1 antibodies did not react with T/13μE/3.1-IgM antibodies and vice versa. We therefore conclude that bystander IgM antibodies contribute to the remodelling of glomerular lesions in situ, following glomerular injury by the nephritogenic antibodies.
Autoimmunity to glutamic acid decarboxylase in patients with autoimmune polyendocrinopathy–candidiasis–ectodermal dystrophy (APECED)
Authors: Klemetti P , Björses P , Tuomi T , Perheentupa J , Partanen J , Rautonen N , Hinkkanen A , Ilonen J , Vaarala O 
Published: March 2003

SUMMARY
Antibodies to glutamic acid decarboxylase (GAD) occur frequently in patients with APECED, although clinical insulin-dependent diabetes mellitus (IDDM) is seen only in a subgroup of the patients. We studied the cellular immunity to GAD, antibodies to GAD and their association with the HLA DQB1 risk alleles for IDDM in patients with APECED. Proliferation responses to GAD were enhanced in the patients with APECED when compared with the control subjects (P = 0·004), but autoimmunity to GAD was not associated with IDDM in APECED. The levels of interferon-gamma (IFN-γ) secreted by GAD-stimulated T cells were higher in the patients than in control subjects (P = 0·001). A negative correlation (r = − 0·436, P = 0·03) existed between the antibody levels and the stimulation indices (SIs) to GAD. In 14 non-diabetic patients no difference in insulin secretion was observed in intravenous glucose tolerance test (IVGTT) between the patients with and without T cell reactivity to GAD. We conclude that cellular immunity to GAD detected as T cell proliferation response to GAD or IFN-γ secretion by GAD-stimulated T cells was frequent in patients with APECED (69%) and was not restricted to the patients with clinically detectable β-cell damage.
Local challenge of oral mucosa with gliadin in patients with coeliac disease
Authors: Lähteenoja H , Mäki M , Viander M , Toivanen A , Syrjänen S 
Published: March 2003

SUMMARY
In coeliac disease, gluten-containing diet challenges over many years are sometimes required for diagnosis, especially if the initial diagnosis was equivocal. The rectal gluten challenge has been proposed to simplify coeliac disease diagnosis. We were interested in studying whether the oral mucosa could be used for local challenge with gliadin as an aid in finalizing the diagnosis of coeliac disease. The study groups consisted of 37 treated coeliac disease patients and 10 controls. The challenges on the oral mucosa were performed either supramucosally with gliadin powder (coeliac disease patients) or by submucosal injection of dissolved gliadin (10 μg/ml) (coeliac disease patients and controls). A control challenge with submucosal gliadin solvent was made in the coeliac disease patients. B and T cells, mast cells and T cell subsets were counted and HLA-DR expression was determined. Biopsies were taken from each provoked area 24 h post-challenge. A significant increase in the number of CD4+ lymphocytes in the lamina propria (observed in 27/37 patients), but a decrease in the number of mast cells was observed in treated coeliac disease patients after submucosal challenge with gliadin. Following supramucosal challenge with gliadin the counts of intraepithelial CD4+ (in 25/37 patients) and CD8+ T cells (in 27/37 patients) increased significantly and the number of CD4+ T cells in the lamina propria was also significantly increased. Control subjects were tested by submucosal gliadin challenge and no significant changes in the number of cells were observed. HLA-DR expression did not show increased positivity in coeliac disease patients on submucosal challenge. For the first time the oral mucosa has been used for immunological testing and shown to react to gliadin challenge in coeliac disease patients. Recruitment of T cells upon submucosal gliadin challenge occurred towards the lamina propria, whereas it occurred towards the epithelium in supramucosal gliadin challenge. The numbers of T cells increased in the lamina propria after submucosal challenge. The results suggest that local oral challenge with gliadin may be used as a diagnostic method in coeliac disease; however, further studies in untreated coeliac disease patients are needed to evaluate the usefulness of this method.