bluuebunny/crossref_metadata_2025 huggingface dataset:
Metalloproteinase‐9 is increased after toluene diisocyanate exposure in the induced sputum from patients with toluene diisocyanate‐induced asthma
Authors: Park H.‐S. , Kim H.‐A. , Jung J.‐W. , Kim Y.‐K. , Lee S.‐K. , Kim S.‐S. , Nahm D.‐H.
Published: March 2003

Background and objective Persistent asthma symptoms are associated with airway inflammation and remodeling, which may be mediated through metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP). The aim of this study was to evaluate MMPs and TIMP involvement in toluene diisocyanate (TDI)‐induced asthma.

Materials and method Induced sputum was collected in eight newly diagnosed TDI‐induced asthma subjects (group I) before and 7 h after the TDI and placebo challenges and in 12 subjects with TDI‐induced occupational asthma diagnosed 5 years previously with persistent asthma symptoms (group II). Sera was collected in group I at diagnosis, and in group II, they were collected at the time of the study. 12 nonasthmatic healthy subjects were enrolled as controls. MMP‐9, MMP‐2 and TIMP‐1 levels in both sputum and serum were measured by enzyme‐linked immunosorbent assay (ELISA). Gelatinase activity in the sputum was confirmed by zymographic analysis.

Results The serum TIMP‐1 level was significantly higher in asthma patients than in healthy controls (P = 0.01), while MMP‐9 level was significantly lower in asthmatic patients (P = 0.03). There was no significant difference in MMP‐2 level (P = 0.27). MMP‐9 level in the sputum was significantly increased after the TDI challenges (P = 0.01). TIMP‐1 level in sputum tended to increase after TDI challenges, but no statistical significance was noted (P = 0.09). MMP‐9 and MMP‐9/TIMP‐1 levels in the sputum were significantly higher in group II than in group I (P = 0.04, P = 0.02) with no significant difference in TIMP‐1 level. Minimal amount of MMP‐2 was found in sputum. Zymography demonstrated that MMP‐9 level increased and active form of MMP‐9 was generated after the TDI bronchoprovocation test.

Conclusion TDI exposure leads to overproduction of MMP‐9, which may induce airway inflammation and remodeling, and then contribute to persistent asthmatic symptoms in TDI‐induced asthma.
Immunoglobulin E‐binding and skin test reactivity to hydrophobin HCh‐1 from Cladosporium herbarum, the first allergenic cell wall component of fungi
Authors: Weichel M. , Schmid‐Grendelmeier P. , Rhyner C. , Achatz G. , Blaser K. , Crameri R.
Published: March 2003

Background For many years, fungal spores have been recognized as potential causes of respiratory allergies. All fungal allergens cloned so far represent either secreted or cytoplasmatic proteins, but nothing is known about the involvement of fungal surface proteins in allergic diseases.

Methods A phage surface displayed cDNA‐library from the mould Cladosporium herbarum was constructed and phage displaying IgE‐binding proteins were selectively enriched with immobilized serum IgE from C. herbarum‐sensitized individuals. Inserts encoding putative allergens were sequenced, subcloned and used to produce recombinant proteins. Allergenicity of the proteins was evaluated by IgE binding in Western blots, enzyme‐linked immunosorbent assay (ELISA) and skin prick test in a total of 84 patients sensitized to either C. herbarum or Aspergillus fumigatus and three healthy controls.

Results After four rounds of affinity selection, the cDNA‐library was enriched for clones displaying IgE‐binding molecules. Sequencing of inserts showed that one clone contained an open reading frame predicting a protein of 105 amino acids and a calculated molecular weight of 10.5 kDa showing the classical signature of members of the hydrophobin family. The recombinant protein, termed HCh‐1, was able to bind IgE from six patients sensitized to fungi in vitro. Two of those patients were also included in a skin prick test survey and showed strong type I skin reactions to HCh‐1, demonstrating the allergenic nature of C. herbarum hydrophobin and indicating a prevalence of sensitization in the range of 8–9%. In contrast, the hydrophobin HYP1 from Aspergillus fumigatus was not recognized by the sera of the same patients and controls investigated with HCh‐1.

Conclusion C. herbarum hydrophobin represents the first component of the cell wall of fungi demonstrated to act as a rare but clinically relevant allergen in vitro and in vivo.
Suppression of allergic reaction by λ‐carrageenan: Toll‐like receptor 4/MyD88‐dependent and ‐independent modulation of immunity
Authors: Tsuji R. F. , Hoshino K. , Noro Y. , Tsuji N. M. , Kurokawa T. , Masuda T. , Akira S. , Nowak B.
Published: March 2003

Background Recognition of foreign substances by innate immunity through pattern recognition receptors (PRRs) regulates acquired immunity such as allergic reaction. Because PRRs recognize heterogeneous ligands, daily food intake can potentially regulate immune allergic reaction.

Objective Elucidation of the effect of λ‐carrageenan on allergic reactions was aimed.

Method IFN‐γ and IL‐4 was measured in in vitro T cell‐stimulated culture. Cytokine production from macrophages in response to λ‐carrageenan was measured as indicator for innate immunity activation. Mice were immunized with OVA in alum to induce specific IgE, and then histamine release was induced by systemic injection of OVA.

Results Activation of innate immunity by λ‐carrageenan is dependent on Toll‐like receptor‐4 (TLR4) and MyD88, in which induction of pro‐inflammatory cytokines such as TNF‐α and IL‐6 was largely impaired in macrophages from TLR4‐ and MyD88‐deficient mice. Footpad oedema, a model for in vivo inflammatory reactions, was significantly reduced in these mice. Similar to recent evidence showing a preference for the stimulation of Th1 via TLR/MyD88 signalling, λ‐carrageenan showed enhanced IFN‐γ and decreased IL‐4 in stimulated T cell cultures. Interestingly, increased IFN‐γ production was still seen in TLR4‐ and MyD88‐deficient splenocytes. Oral administration of λ‐carrageenan to immunized mice successfully decreased OVA‐specific IgE, and λ‐carrageenan was also effective in previously immunized mice. Further, serum histamine release upon systemic challenge of OVA was significantly inhibited. Neither OVA‐specific IgG1/IgG2a nor cytokine secretion from in vitro cultures were altered, suggesting the involvement of multiple PRRs as demonstrated by TLR4/MyD88‐independent IFN‐γ up‐regulation. The simultaneous feeding of OVA with lipopolysaccharide abrogated oral tolerance, but λ‐carrageenan was not only devoid of such an effect but was also found to promote oral tolerance in the absence of TLR4.

Conclusion λ‐Carrageenan was suggested to be a useful dietary supplement to ameliorate allergic reactions while maintaining oral tolerance‐dependent intestinal homeostasis.
Interleukin‐10 gene promoter region polymorphism is associated with eosinophil count and circulating Immunoglobulin E in adult asthma
Authors: Karjalainen J. , Hulkkonen J. , Nieminen M. M. , Huhtala H. , Aromaa A. , Klaukka T. , Hurme M.
Published: March 2003

Background IL‐10 has several functional effects relevant to asthma. It can modulate IgE production and induce apoptosis in eosinophils. Polymorphisms of IL‐10 gene have been shown to affect IL‐10 production.

Objective To establish whether IL‐10 polymorphisms are associated with asthma and phenotype‐related characteristics.

Methods The frequency of three single base exchange polymorphisms (at positions − 1082, − 819 and − 592) and corresponding haplotypes of the IL‐10 gene were analysed in 245 adult asthmatic subjects and 405 controls using PCR and restriction fragment length polymorphism (RFLP). The data were assessed for correlations with the eosinophil count, serum IgE and lung function.

Results The IL‐10 haplotype frequencies were similar in asthmatics and controls. Eosinophil count median was 2.0‐ to 3.2‐fold higher among asthmatics with rare ATA/ATA genotype than in asthmatics with other genotypes. No such difference was seen in the control group. When analysed by IL‐10 haplotype carrier state and gender, male asthmatics with ATA haplotype had 2.8‐fold higher serum IgE than those without ATA. A converse association was found in male controls with ATA haplotype, who had 1.9‐fold lower serum IgE than their ATA‐negative counterparts. The high IL‐10‐producing GCC haplotype was associated with impaired lung function in smoking male controls while in asthmatics no clear effect on lung function was found with any of the haplotypes studied.

Conclusion These results suggest that the eosinophil counts and serum IgE are differently regulated by IL‐10 genotype in asthmatic and in normal subjects. However, IL‐10 polymorphism is not related to susceptibility in asthma.
Familial and environmental determinants for wheezing and asthma in a case‐control study of school children in Palestine
Authors: El‐Sharif N. , Abdeen Z. , Barghuthy F. , Nemery B.
Published: March 2003

Background Our prevalence study on Palestinian school children aged 6–12 years showed lower rates for asthma and asthma symptoms than economically developed and industrialized countries. Reasons for such differences are largely unknown, and could possibly be related to different environmental and lifestyle factors.

Objective To investigate familial, early life exposures and indoor environmental determinants for asthma in children in Palestine.

Methods From the population of our previous study, a group of 273 children with wheeze in the past 12 months (of whom 99 children had physician‐diagnosed asthma) were matched with an equal number of non‐wheezing controls. This case‐control study involved a parental questionnaire; skin prick testing (SPT) with mixed house dust mites, cat and dog dander, mixed grass, mixed trees pollen, Alternaria tenuis, olive tree and cockroach extracts; and serum for total and specific IgE for the same eight allergens.

Results Paternal asthma and maternal hayfever significantly tripled the risk for their children to have wheezing. Previous diagnoses of bronchial allergy, bronchitis, pneumonia, or whooping cough, and positive SPT for house dust mites and cockroaches were significantly more likely among wheezing and asthmatic children than controls. Specific IgE levels for house dust mites and cat allergens showed significantly higher risk for reported wheezing. After adjustment for several environmental and sociodemographic factors using multivariate logistic regression analysis, paternal asthma, maternal hayfever, damp houses, cat and cockroach SPT positivity proved to be strong predictors for wheezing symptoms.

Conclusion Our study confirmed that familial ‘atopic’ diseases are significant predictors of childhood asthma in Palestinian children. Moreover, indoor environment such as presence of cats and domestic moulds also appear to play a role. Our findings are consistent with studies in Canada, New Zealand, Estonia and Sweden, and show promise to explore further gene–environment interaction in the genesis of asthma.
Pathogenesis of autoimmunity in αβ T cell-deficient lupus-prone mice
Authors: Peng S L , Cappadona J , Mcniff J M , Madaio M P , Owen M J , Hayday A C , Craft J
Published: March 2003

Murine lupus in MRL mice has been strongly attributed to αβ T cell-dependent mechanisms. Non-αβ T cell-dependent mechanisms, such as γδ T cells, have been shown to drive antibody and autoantibody production, but they have not been considered capable of inducing end-organ disease. Here, we have expanded upon the findings of such previous work by examining the mechanism and extent of end-organ disease attainable via γδ T cells and/or non-αβ T cell-dependent mechanisms, assessing two prototypical lupus lesions, renal and skin disease, in TCR α−/− MRL mice that possessed either functional or defective Fas antigen (Fas + or lpr). Observed to 1 year of age, TCR α−/− MRL mice developed disease characterized by increased mortality, overt renal disease and skin lesions. While delayed in onset and/or reduced in severity compared with TCR α+/+ MRL/lpr animals, renal and skin lesions in αβ T cell-deficient animals were clearly increased in severity compared with age-matched control non-autoimmune mice. In contrast to TCR α+/+ MRL mice, whose disease reflected pan-isotype immune complex deposition with significant complement fixation, renal disease in TCR α−/− MRL animals reflected predominantly IgG1 immune complex deposition, with poor complement fixation. Thus, this study demonstrates conclusively that non-αβ T cell-dependent mechanisms can induce renal and skin injury in murine lupus, but at least in the kidney, only via humoral autoimmunity of a relatively non-pathological isotype which results in the delayed onset of end-organ damage.
IL-15 mRNA expression is up-regulated in blood and cerebrospinal fluid mononuclear cells in multiple sclerosis (MS)
Authors: Kivisäkk P , Matusevicius D , He B , Söderström M , Fredrikson S , Link H
Published: March 2003

IL-15, produced by monocytes and epithelial cells, is a novel cytokine with actions similar to IL-2. IL-15 induces T cell proliferation, B cell maturation and natural killer (NK) cell cytotoxicity, and is a chemoattractant for T cells. We investigated the expression of IL-15 mRNA in blood and cerebrospinal fluid (CSF) mononuclear cells (MNC) in MS, an inflammatory disease of the central nervous system where cytokines are involved. MS patients had higher numbers of IL-15 mRNA-expressing blood MNC than patients with aseptic meningo-encephalitis (AM) and healthy controls. In CSF, MS patients had even higher numbers of IL-15 mRNA-expressing cells than in blood. This discrepancy between IL-15 mRNA expression between blood and CSF MNC was not seen in AM patients. Patients examined during the secondary chronic-progressive phase of MS had higher numbers of IL-15 mRNA-expressing blood MNC compared with patients examined during the relapsing-remitting phase. Levels of IL-15 mRNA-positive blood MNC were similar in patients with AM, myasthenia gravis, non-inflammatory neurological diseases and healthy controls. Taken together these data indicate that IL-15 mRNA expression is up-regulated in MS, further suggesting a role for proinflammatory cytokines in the pathogenesis of MS.
Th1/Th2 cytokine responses following HIV-1 immunization in seronegative volunteers
Authors:   THE AIDS VACCINE EVALUATION GROUP, Evans T G , Fitzgerald T , Gibbons D C , Keefer M C , Soucier H
Published: March 2003

The Th1/Th2 profile that follows human vaccination may profoundly influence the subsequent course of disease after infection. However, the ability to detect IL-4 has been limited outside trials of live vaccination. By using methods in which memory effector cells are allowed to antigenically expand by short term culture, followed by low-dose mitogenic stimulation, we have been able to follow the Th1/Th2 profile in HIV-1−volunteers enrolled in two phase I studies of HIV immunogens (a recombinant gp120 and a multivalent, octomeric V3 loop peptide). Antigen-specific interferon-gamma (IFN-γ) could be detected in primary stimulation, but IL-4 was observed only after antigenic expansion and restimulation. In both of these studies the responses after initial immunizations were dominated by IFN-γ, with IL-4 appearing only after multiple rounds of immunization, and IL-4 was temporally related to antibody production. Concomitant with the IL-4 production, the amount of supernatant IFN-γ declined. Antigen-specific IL-10 was not detected in either study. Such techniques, which have been shown to correlate with outcomes in immunotherapy, may prove useful as future surrogates of human vaccine response.
Prevalence and characterization of perinuclear anti-neutrophil cytoplasmic antibodies (P-ANCA) directed against HMG1 and HMG2 in ulcerative colitis (UC)
Authors: Sobajima J , Ozaki S , Uesugi H , Osakada F , Shirakawa H , Yoshida M , Nakao K
Published: March 2003

In a previous study, we reported that the high mobility group (HMG) non-histone chromosomal proteins HMG1 and HMG2 were novel target antigens of P-ANCA. In this study, we determined the immunodiagnostic value of anti-HMG1/HMG2 antibodies in patients with UC. Sixty sera from patients with UC were tested for reactivity with HMG1 and HMG2 by means of ELISA. Anti-HMG1 antibody was detected in 32% of patients (40% of P-ANCA+ patients). Anti-HMG2 antibody was detected in 33% (40% of P-ANCA+ patients). Thirty-five percent of sera were positive for antibody to either HMG1 or HMG2 (43% of P-ANCA+ patients). P-ANCA+ patients expressed anti-HMG1/HMG2 antibodies with significantly greater frequency compared with P-ANCA− patients. Furthermore, the anti-HMG1/HMG2 antibodies were significantly related to disease activity in UC. Sixteen of the 18 UC patients, who had high titres of anti-HMG1 or -HMG2 antibody during the active phase, showed lower titres in the inactive phase. Anti-HMG1/HMG2 antibodies appear to be useful as a marker for disease activity in UC.
Relationship between autoantibody epitopic recognition and immunoglobulin gene usage
Authors: Guo J , Mcintosh R S , Czarnocka B , Weetman A P , Rapoport B , Mclachlan S M
Published: March 2003

An immunodominant region recognized by serum autoantibodies has been defined on the autoantigen thyroid peroxidase (TPO) using recombinant human TPO-specific Fab or a panel of mouse MoAbs. We have now analysed the epitopic relationships between the four recombinant Fab that identify the A and B domains of the TPO immunodominant region and (i) the mouse TPO MoAb as well as (ii) nine new TPO-specific Fab isolated independently. Competition between mouse MoAbs and recombinant Fab for binding to 125I-TPO revealed three patterns. First, for MoAbs 15, 59, 64 and 18, TPO binding was virtually abolished (≈ 90%) by Fab which define the A domain of TPO, with less inhibition by B domain Fab. Second, for MoAbs 2, 9 and 47, the Fab competed much less for TPO binding, and, when detectable, inhibition was predominantly with B domain Fab (65–20%). Third, for MoAbs 53, 30, 1, 24 and 40, none of the Fab competed effectively for 125I-TPO binding. Thus, the epitopes for MoAbs 18, 59, 64 and 15 correspond to those of the A domain defined by the human Fab, and the epitopes for MoAbs 2, 9 and 47 correspond to those of the B domain. In the second part of the study, competition studies demonstrated that the epitopes of nine new Fab corresponded to those of the four Fab that define the immunodominant region. For four new Fab, TPO binding was inhibited to a greater extent by B- than by A- domain Fab (65–95% versus < 50%). In contrast, for five new Fab the A-domain Fab were more effective inhibitors (≈ 90%) than the B-domain Fab. In addition, consistent with previous observations, all five new Fab with 02/012 κ L chains, but none of the new Fab with non-O2/O12 l chains, interacted with A-domain epitopes. In conclusion, we have established the epitopic relationships between recombinant human Fab and mouse MoAbs that define the TPO immunodominant region on TPO. Further, analysis of recombinant TPO Fab isolated from patients on three continents strengthens the paradigm of a relationship between autoantibody epitopic recognition and immunoglobulin gene usage.
Regulation of secretion of IL-4 and IgG1 isotype by melatonin-stimulated ovalbumin-specific T cells
Authors: Shaji A V , Kulkarni S K , Agrewala J N
Published: March 2003

In the present study, we describe the potential role of melatonin, a pineal hormone, in regulating the activation of the antigen-specific T cell response. Melatonin encouraged the proliferation of Th cells and improved their ability to secrete IL-4, but down-regulated the levels of IL-2 and interferon-gamma (IFN-γ). Melatonin, however, could not exert any influence on the T cells of unprimed mice. On studying the regulation of subclass of IgG isotype, melatonin specifically enhanced the secretion of antigen-specific IgG1 antibodies and decreased the yield of IgG2a isotype. The results suggest that melatonin possibly acts by selectively activating a Th2-like immune response.
Capacity of murine T cells to retain long-term responsiveness to mycobacterial antigens is controlled by theH-2complex
Authors: Pichugin A V , Khaidukov S V , Moroz A M , Apt A S
Published: March 2003

It is firmly established that the allelic composition of the H-2 complex has a prominent impact on the course of tuberculosis (TB) infection in mice, including granuloma formation, mycobacterial spread in the lungs, and the dynamics of mortality. Although intuitively obvious, the role of long-term specific T cell responses in the expression of corresponding phenotypes is poorly understood. In this study we have compared polyclonal lymph node cell response (cell yield, proliferation, surface markers, IL-4/interferon-gamma (IFN-γ) production) to Mycobacterium tuberculosis H37Rv sonicate in repeated 10-day cycles of stimulation/rest between H-2 congenic IE-negative mouse strains, categorized on the basis of mortality following lethal challenge as TB-susceptible (C57Bl/6), TB-resistant (4R) and BCG non-protected (B10.M). The capacity to retain specific responsiveness to repeated stimulation by mycobacterial antigens depended upon both the H-2 haplotype of the host and the immunizing dose of the antigen. 4R lymph node cells following either 50 μg/mouse or 100 μg/mouse immunization constantly responded to sonicate, increased in numbers, and after the third stimulation/rest cycle developed into a stable CD3+ CD4+ cell line. B6 cells following either 50 μg/mouse or 100 μg/mouse immunization, and B10.M cells following 100 μg/mouse (but not 50 μg/mouse) immunization, lost the capacity to incorporate methyl-3H-thymidine during the second cycle, and died. Analogous results were obtained in the in vivo experiments, when the dynamics of the response over 12 weeks following a single immunization with the antigen was studied. In response to the antigen, cells from all three mouse strains produced significant amounts of IL-2 and IFN-γ, but not IL-4, indicating that they belong predominantly to the Th1-like subset. Among noteworthy differences between the mouse strains was a clear deficiency of CD8+ T cells in B6 cultures, and an unusually high proportion of CD3+ CD4− CD8− (double-negative) T cells in B10.M cultures following a high-dose immunization.
Regulation of the expression of aminopeptidase A, aminopeptidase N/CD13 and dipeptidylpeptidase IV/CD26 in renal carcinoma cells and renal tubular epithelial cells by cytokines and cAMP-increasing mediators
Authors: Kehlen A , Göhring B , Langner J , Riemann D
Published: March 2003

Aminopeptidase (AP) A is a transmembrane type II molecule widely distributed in mammalian tissues. Since APA expression may be absent in renal cell carcinoma (RCC), it is possible that there is an altered regulation or other defect of APA upon malignant transformation of proximal tubular cells. However, investigations into the regulation of APA on tumour cells are rare. We report, for the first time, that both transforming growth factor-beta 1 (TGF-β1) and tumour necrosis factor-alpha (TNF-α) down-regulate APA mRNA as well as protein expression in renal tubular epithelial cells and RCC cells in culture. In addition to this, both cytokines decrease dipeptidylpeptidase (DP) IV/CD26 mRNA, but not APN/CD13 mRNA expression. Otherwise, IL-4 and IL-13 increase CD13 as well as CD26 expression, but do not alter APA expression. Interferon-alpha (IFN-α), IFN-β and IFN-γ increase mRNA expression of all the three membrane ectopeptidases, whereas IL-1, IL-6, IL-7, IL-12 and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been found to be without any significant effect. Treatment of cultured cells with cAMP-increasing agents, such as 8-bromo-cAMP or A23187, results in an increase in APA and DPIV/CD26, but no change in APN/CD13 mRNA expression or even a decrease in it. Furthermore, AP inhibitors can influence APA mRNA expression, since bestatin causes an increase in APA expression in a time- and dose-dependent manner, whereas bestatin does not change CD13 or CD26 expression. No difference could be found with respect to the modulation by different mediators between RCC cells and renal epithelial cells, though permanent tumour cell lines such as Caki-1 and Caki-2 may have lost some of the normally expressed peptidases.
Differential inhibitory effects of indomethacin, dexamethasone, and interferon-gamma (IFN-γ) on IL-11 production by rheumatoid synovial cells
Authors: Taki H , Sugiyama E , Mino T , Kuroda A , Kobayashi M
Published: March 2003

IL-11, a member of the IL-6 type cytokines, has some biological activity related to the joint destruction in rheumatoid arthritis (RA), such as induction of osteoclast differentiation. However, its expression and regulation in rheumatoid inflamed joints has not been clarified. In the present study we examined the capacity of fresh rheumatoid synovial cells (fresh RSC) to produce IL-11, and the effect of indomethacin, dexamethasone and IFN-γ on IL-11 production. Fresh RSC obtained from eight patients with RA produced large amounts of IL-11, measured by ELISA, and showed strong expression of IL-11 mRNA, determined by Northern blotting. Indomethacin inhibited the production of IL-11 by about 55%. Prostaglandin E2 (PGE2) completely prevented the inhibition, suggesting that IL-11 production by fresh RSC was in part mediated by PGE2. Dexamethasone inhibited the production of IL-11 by more than 80%. Interestingly, the inhibition was not abolished by PGE2. IFN-γ inhibited the production of IL-11 from IL-1α-stimulated cultured rheumatoid synovial fibroblasts, although IFN-γ did not inhibit the production of IL-11 by fresh RSC. These results suggest that the production of IL-11 by rheumatoid synovia was differentially regulated by PGE2 and IFN-γ, and that treatment with indomethacin or dexamethasone decreased the level of IL-11 at inflammatory joints in patients with RA.
Transforming growth factor-beta 1 (TGF-β1) expression in ovine lentivirus-induced lymphoid interstitial pneumonia
Authors: Moreno B , Woodall C J , Watt N J , Harkiss G D
Published: March 2003

The aim of this study was to detect the localization of TGF-β1 protein expression in normal sheep lungs and lungs with interstitial pneumonia associated with infection with maedi-visna virus (MVV). Immunohistochemical localization of TGF-β1 was determined in 24 lungs of adult sheep naturally infected with MVV and six control lungs of seronegative sheep. The lungs of infected animals showed different lesional degrees: grade 0, no lesions; grade I, mild; grade II, moderate; grade III, severe. In normal lungs, TGF-β1 was primarily expressed in airway epithelium, bronchial cartilage and glands, endothelial cells and smooth muscle of blood vessels, alveolar macrophages and type II pneumocytes. No staining was observed in alveolar interstitium. In MVV-infected sheep an increased number of positive alveolar and interstitial macrophages and staining of alveolar interstitium was observed in grade I, grade II and some grade III lesions. In grade III lesions an inverse relationship was found between TGF-β1 staining and smooth muscle hyperplasia. Small lymphoid aggregates, in general, showed strong reactivity, whereas larger ones showed weak reactivity, mainly associated with follicular areas. No significant differences in the staining intensity of airways and blood vessels were observed between control and MVV lungs. The increased expression of TGF-β1 in early maedi lesions and its down-regulation in more advanced disease suggest the operation of a temporal regulatory mechanism whereby early expression may lead to the smooth muscle hyperplasia which develops during the disease. The striking inverse relationship between TGF-β1 expression and follicle organization is intriguing and warrants further investigation.
HIV-1 does not alterin vitroandin vivoIL-10 production by human monocytes and macrophages
Authors: Bergamini A , Bolacchi F , Faggioli E , Placido R , Vendetti S , Cappannoli L , Ventura L , Cerasari G , Uccella I , Andreoni M , Rocchi G
Published: March 2003

The present study analyses the ability of HIV-1 to modulate IL-10 production in cells of monocyte-macrophage lineage cultured in the presence of macrophage colony-stimulating factor (M-CSF). Both monocytes and macrophages spontaneously produced low amount of IL-10. Lipopolysaccharide (LPS) induced a strong IL-10 response in fresh monocytes and in M-CSF-treated macrophages. In contrast, macrophages cultured in the absence of M-CSF exhibited a marked decrease in their susceptibility to LPS stimulation. M-CSF increased the IL-10 response of macrophages to LPS by enhancing both the expression of membrane-bound CD14, the protein that serves as LPS receptor, and the sensibility of CD14-expressing cells to LPS stimulation. Neither spontaneous nor LPS-induced expression of IL-10 was modulated in monocytes and macrophages by infection with eight monocytotropic strains, as demonstrated by ELISA and cytofluorimetric analysis. In contrast, all the HIV-1 strains primed macrophages for an increased IL-6 response to LPS stimulation. To determine whether IL-10 production was associated with in vivo infection, monocytes from AIDS individuals were analysed for IL-10 production. We found that neither spontaneous nor LPS-induced IL-10 production were different between healthy controls and HIV-infected patients. Taken together, these data strongly suggest that HIV-1 infection of monocytes-macrophages does not play a significant role in the regulation of IL-10 in infected patients. This study also emphasizes the role of M-CSF activation in the regulation of the cytokine response in macrophages.
Elevated serum levels of soluble Fas/APO-1 (CD95) in patients with hepatocellular carcinoma
Authors: Jodo S , Kobayashi S , Nakajima Y , Matsunaga T , Nakayama N , Ogura N , Kayagaki N , Okumura K , Koike T
Published: March 2003

Fas (APO-1/CD95)-mediated apoptosis plays an important role in liver cell destruction in viral hepatitis. Using sandwich ELISA, we measured serum levels of soluble Fas (sFas) in patients with hepatocellular carcinoma (HCC) who were positive for hepatitis B surface antigen (HBsAg) or anti-hepatitis C virus (HCV) antibody. sFas levels were significantly higher in HCC patients (median 4·07 ng/ml; range 0·14–29·18 ng/ml) than levels in age-matched healthy donors (0·29 ng/ml; 0–4·90 ng/ml) (P < 0·0001) and HBsAg or anti-HCV antibody-positive patients with liver cirrhosis (LC) (2·16 ng/ml; 0·24–8·39 ng/ml) (P = 0·0015). An arbitrary cut-off level of 3·03 ng/ml (mean + 3 s.d. of controls) revealed the positive frequency of sFas in each group: 1·7% in healthy subjects, 25·9% in LC, and 59·0% in HCC (sensitivity 59·0% and specificity 74·1%). All HCC sera tested contained transmembrane-deleted sFas and some contained another sFas lacking the Fas C-terminal. The positive frequency of either sFas (59·0%) or α-fetoprotein (AFP) (57·4%) in HCC patients reached 77·0%. HCC patients with multiple tumour foci (7·53 ng/ml; 1·40–29·18 ng/ml) had significantly higher sFas levels than did patients with a solitary tumour (2·70 ng/ml; 0·14–19·0 ng/ml) (P = 0·003). In all of the sFas-positive patients with a solitary tumour, surgical removal of the tumour reduced sFas levels to the negative in the first post-op week. These findings suggest that sFas may be closely linked with HCC and may be a candidate for a clinical parameter for HCC.
Decay-accelerating factor (DAF) in stool specimens as a marker of disease activity in patients with ulcerative colitis (UC)
Authors: Inaba T , Mizuno M , Ohya S , Kawada M , Uesu T , Nasu J , Takeuchi K , Nakagawa M , Okada H , Fujita T , Tsuji T
Published: March 2003

Colonic epithelial cells of patients with UC express DAF in relation to the severity of mucosal inflammation. The aim of this study was to determine whether this factor in stool could be used as a marker of disease activity in UC patients. Stool DAF was measured by use of an immunoassay in 181 stool specimens obtained from 55 patients with UC of various levels of disease activity. Stool DAF concentrations in patients whose UC was active (0·0–785·6 ng/g stool; median 47·1 ng/g; n = 115) were significantly higher than concentrations in patients whose disease was inactive (0·0–48·6 ng/g; median 0·0 ng/g; n = 66) (P < 0·0001). Values in active UC patients also were higher than those in control patients with diarrhoea (0·0–30·0 ng/g; median 0·0 ng/g; n = 26) (P < 0·0001) and in control subjects without apparent colorectal disease (0–20·4 ng/g; median 0·0 ng/g; n = 44) (P < 0·0001). The elevated levels of stool DAF obtained from UC patients in relapse declined markedly in specimens collected after the disease went into remission following medical therapy. Stool DAF levels correlated with the severity of endoscopic and histological findings and the degree of DAF expression on the colonic epithelia. Our results suggest that the measurement of stool DAF is useful as a non-invasive means of monitoring intestinal disease activity in patients with UC.
Analysis of graft-versus-host disease (GVHD) and graft rejection using MHC class I-deficient mice
Authors: Shenoy S , Desch K , Duffy B , Thorson P , Mohanakumar T
Published: March 2003

GVHD is a major complication in allogeneic bone marrow transplantation (BMT). MHC class I mismatching increases GVHD, but in MHC-matched BMT minor histocompatibility antigens (mH) presented by MHC class I result in significant GVHD. To examine the modification of GVHD in the absence of cell surface MHC class I molecules, β2-microglobulin-deficient mice (β2m-/-) were used as allogeneic BMT recipients in MHC- and mH-mismatched transplants. β2m-/- mice accepted MHC class I-expressing BM grafts and developed significant GVHD. MHC (H-2)-mismatched recipients developed acute lethal GVHD. In contrast, animals transplanted across mH barriers developed indolent chronic disease that was eventually fatal. Engrafted splenic T cells in all β2m-/- recipients were predominantly CD3+ αβ TCR+ CD4+ cells (15–20% of all splenocytes). In contrast, CD8+ cells engrafted in very small numbers (1–5%) irrespective of the degree of MHC mismatching. T cells proliferated against recipient strain antigens and recognized recipient strain targets in cytolytic assays. Cytolysis was blocked by anti-MHC class II but not anti-CD8 or anti-MHC class I monoclonal antibodies (MoAbs). Cytolytic CD4+ T cells induced and maintained GVHD in mH-mismatched β2m-/- mice, supporting endogenous mH presentation solely by MHC class II. Conversely, haematopoietic β2m-/- cells were unable to engraft in normal MHC-matched recipients, presumably due to natural killer (NK)-mediated rejection of class I-negative cells. Donor-derived lymphokine-activated killer cells (LAK) were unable to overcome graft rejection (GR) and support engraftment.
Deoxyspergualin preferentially inhibits the growth and maturation of anti-CD40-activated surface IgD+ B lymphocytes
Authors: Morikawa K , Nemoto K , Miyawaki T , Morikawa S
Published: March 2003

Deoxyspergualin (DSG), an analogue of spermidin, is a potent immunosuppressive drug with an action quite distinct from that of cyclosporin, rapamycin, or FK506. In this study we investigated the effect of DSG and methyldeoxyspergualin (MeDSG) on the proliferation and differentiation of human B cells stimulated with anti-CD40 MoAb. Highly purified B cells obtained from tonsillar samples were used as target cells. Both agents inhibited the proliferative response of anti-CD40-stimulated B cells in the absence and presence of IL-4, IL-2 or IL-10 in a dose-dependent manner. This inhibitory effect differed markedly among cell populations based on surface IgD expression: strong inhibition of sIgD+ B cells but little inhibition of sIgD− B cells. The drugs also suppressed the production of IgG, IgM and IgA by unfractionated B cells, which suggests that DSG acts against post-switch (sIgD−) B cells. Although the drugs suppressed immunoglobulin synthesis by both sIgD+ and sIgD− B cells, the effect was more marked in the sIgD+ B cells. Analysis of the subclass of IgG secreted by slgD+ B cells revealed a decline in IgG1 and IgG3 in the presence of DSG. These results suggest that DSG preferentially inhibits the growth and maturation of sIgD+ naive B cells.
Neoplastic thymic epithelial cells of human thymoma support T cell development from CD4−CD8− cells to CD4+CD8+ cellsin vitro
Authors: Inoue M , Fujii Y , Okumura M , Takeuchi Y , Shiono H , Miyoshi S , Matsuda H , Shirakura R
Published: March 2003

Human thymoma is a thymic epithelial cell tumour which often contains a large number of immature T cells and is frequently associated with autoimmune diseases. Since thymic epithelial cells play key roles in the development and selection of T cells in the normal thymus, we hypothesized that the neoplastic thymic epithelial cells of thymoma may support T cell differentiation in the tumour. We characterized CD4−CD8− cells in thymoma and applied an in vitro reconstitution culture system using the CD4−CD8− cells and the neoplastic epithelial cells isolated from thymoma. CD34, a stem cell marker, was expressed on 29.9 ± 12.2% of CD4−CD8− cells in thymoma. TCRγδ was expressed on 27.4 ± 15.1% of CD4−CD8− cells and CD19, a B cell marker, was expressed on 14.1 ± 23.1% of CD4−CD8− cells. CD4−CD8− cells expressed both IL-7R α-chain and common γ-chain. Purified CD4−CD8− cells from thymomas were cultured with the neoplastic epithelial cells, and their differentiation into CD4+CD8+ cells via CD4 single-positive intermediates was observed within 9 days' co-culture in the presence of recombinant IL-7. Furthermore, we examined the reconstitution culture using CD34+CD4−CD8− cells purified from normal infant thymus. The CD34+CD4−CD8− cells in normal thymus also differentiated to CD4+CD8+ cells in the allogeneic co-culture with the neoplastic epithelial cells of thymoma. These results indicate that the tumour cells of thymoma retain the function of thymic epithelial cells and can induce differentiation of T cells in thymoma.
Analysis of orbital T cells in thyroid-associated ophthalmopathy
Authors: Förster G , Otto E , Hansen C , Ochs K , Kahaly G
Published: March 2003

Thyroid-associated ophthalmopathy (TAO) has a major effect on the two compartments of the retro-orbital (RO) space, leading to enlargement of the extraocular muscles and other RO tissues. T lymphocyte infiltration of RO tissue is a characteristic feature of TAO and there is current interest in whether these T cells are specifically and selectively reactive to RO tissue itself. We recently established 18 T cell lines (TCL) from RO adipose/connective tissue of six patients with severe TAO by using IL-2, anti-CD3 antibodies and irradiated autologous peripheral blood mononuclear cells (PBMC) to maintain the growth of T cells reactive to autologous RO tissue protein fractions. Here we report on the phenotype characteristics and cytokine gene expression profiles of these orbital TCL and on their immunoreactivity to the organ-specific thyroid antigens thyrotropin receptor (TSH-R), thyroidal peroxidase (TPO) and thyroglobulin (TG). Flow cytometry revealed that 10 TCL were predominantly of CD4+ phenotype, three being mostly CD8+ and five neither CD4+ nor CD8+. Analysis with reverse transcriptase-polymerase chain reaction (RT-PCR) of cytokine gene expression revealed both Th1- and Th2-like products in all TCL: IL-2 product (in 17 TCL), interferon-gamma (IFN-γ) (n = 10), tumour necrosis factor-beta (TNF-β) (n = 15), IL-4 (n = 12), IL-5 (n = 17), IL-6 (n = 13), TNF-α (n = 12) and IL-10 (n = 4). Reactivity to thyroid antigens was observed only in two TCL, the other 16 being uniformly unreactive. Although 10 out of 18 RO tissue-reactive TCL were predominantly CD4+ there were no significant relationships between TCL phenotype, cytokine gene profile, magnitude of reactivity to RO tissue protein or the (rare) occurrence of thyroid reactivity. The findings of both Th1- and Th2-like cytokine gene expression in all RO tissue-reactive TCL support the concept that TAO is a tissue-specific autoimmune disease, distinct immunologically from the thyroid, and involving both T cell and B cell autoimmune mechanisms in disease pathogenesis.
No evidence for abnormal immune activation in peripheral blood T cells in patients with hepatitis C virus (HCV) infection with or without cryoglobulinaemia
Authors:   MULTIVIRC GROUP, Cacoub P , Musset L , Hausfater P , Ghillani P , Fabiani F L , Charlotte F , Angevin E , Opolon P , Poynard T , Piette J-C , Autran B
Published: March 2003

The aim of this study was to investigate the peripheral blood lymphocyte (PBL) phenotypes and T cell repertoire in patients with HCV infection, with or without mixed cryoglobulinaemia (MC). The patients were: Group 1, 23 patients with HCV infection and MC; Group 2, 14 patients with HCV infection but without MC; Group 3, 10 patients with symptomatic essential MC. Twenty healthy blood donors were used as controls. Blood lymphocyte counts were determined, and flow cytometry was used to measure proportions of B cells (CD19+), natural killer (NK) cells (CD16+ CD56+), T cells (CD3+), CD4+ T cell subsets (memory CD4+ CD45RO+; naive CD4+ CD45RO−; Th0/Th2 CD4+ CD7−; activated CD4+ CD25+), and CD8+ T cell subsets (immunoregulatory CD8+ CD57+; cytotoxic CD8+ S6F1+, activated CD8+ CD25+). Bias in the usage of T cell receptor (TCR) Vβ chains was studied in a subgroup of 10 representative patients of Group 1 using a polymerase chain reaction (PCR) analysis of the Vβ segments with a series of 20 oligonucleotides specific for the Vβ families. The three groups were comparable for blood lymphocyte counts, and we observed no abnormal repartition of the following PBL subsets: T cells (CD3+), CD4+ and CD8+ subpopulations, B cells (CD19+), and the NK cells (CD16+ 56+). In none of the groups could we observe lymphocyte ex vivo activation as assessed by the normal expression of the activation cell markers: CD25 on CD4+ or CD8+ T cells, or CD5 on B cells. The repartition of naive and memory (CD45RO−/RO+) CD4+ T cells was normal and we did not observe any amplification of the CD4+ CD7− T cell subset differentiated in vivo in Th0/Th2 cells. There was no significant amplification of cytotoxic (SF6+) and immunoregulatory (CD57+) CD8+ T cells in HCV patients with or without MC. Finally, the usage of Vβ families in the TCR repertoire was normal in the patients tested. In patients with chronic HCV infection, with or without MC, we did not find any significant expansion or abnormal activation of T, B and NK cell subsets, dysbalance of the naive/memory subsets, or expansion of the Th0/Th2 subpopulation. These findings differ from the profound immune alterations that are observed in other chronic infections such as HIV or Epstein–Barr virus. Although this study was restricted to the peripheral blood, it suggests that in chronic HCV infection, MC is not the consequence of a chronic activation or dysregulation of the peripheral blood immune cells.
Expansion of CD56+ NK T and γδ T cells from cord blood of human neonates
Authors: Musha N , Yoshida Y , Sugahara S , Yamagiwa S , Koya T , Watanabe H , Hatakeyama K , Abo T
Published: March 2003

A particular T cell population expressing NK cell markers, CD56 and CD57, exists in humans. Many CD56+ T and CD57+ T cells (i.e. NK T cells) exist in the liver and increase in number in the blood with ageing. They may be a human counterpart of extrathymic T cells, similar to NK1.1+ CD3int cells seen in mice. We investigate here the existence of such NK T cells in human cord blood and the in vitro expansion of these cells by the stimulation of human recombinant IL-2 (rIL-2). There were very small populations (< 1.0%) of CD56+ T cells, CD57+ T cells, and γδ T cells in cord blood. However, all of these populations increased in number after birth and with ageing. When lymphocytes in cord blood were cultured with rIL-2 (100 U/ml) for 14 days, CD56+ T cells expanded up to 25% of T cells. CD57+ T cells were never expanded by these in vitro cultures. The expansion of γδ T cells (mainly Vγ9− non-adult type) also occurred in the in vitro culture. A considerable proportion of CD56+ T cells was found to use Vα24 (i.e. equivalent to invariant Vα14 chain used by murine NK T cells) for TCR αβ. These results suggest that neonatal blood contains only a few NK T cells but CD56+ NK T cells and γδ T cells are able to expand in vitro.
Effects of monoclonal antibodies to adhesion molecules on eosinophilic myocarditis inToxocara canis-infected CBA/J mice
Authors: Hokibara S , Takamoto M , Isobe M , Sugane K
Published: March 2003

Eosinophilic myocarditis followed by fibrosis of the cardiac muscle was observed in addition to peripheral blood eosinophilia in CBA/J mice infected with Toxocara canis. The infected mice were used as an experimental model of eosinophilic endomyocarditis associated with hypereosinophilic syndrome. Effects of in vivo treatment with MoAbs to adhesion molecules on eosinophilic myocarditis were examined using this experimental model. Expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells of capillaries in myocardium were increased 1 and 2 weeks after infection. Infiltration of very late antigen (VLA)-4+ and/or CD11a+ cells into the cardiac muscles was also observed 1 and 2 weeks after infection. Infiltration of eosinophils into the heart was significantly suppressed by anti-CD18 MoAb and anti-VLA-4 MoAb, and focal fibrosis of the cardiac muscle was also significantly suppressed by combined administration of anti-CD18 and anti-ICAM-1 MoAbs. These results indicate that adhesion molecules may play important roles in eosinophilic myocarditis, and that blockade of interaction between adhesion molecules and their ligands may help to control it.
FcγRIa–γ-chain complexes trigger antibody-dependent cell-mediated cytotoxicity (ADCC) in CD5+ B cell/macrophage IIA1.6 cells
Authors: Van Vugt M J , Van Den Herik-Oudijk I E , Van De Winkel J G J
Published: March 2003

Most receptors for immunoglobulins exist as multi-subunit complexes, with unique ligand binding α-chains, combined with accessory signalling (γ-, β-, or ζ-) chains. The myeloid class I receptor for IgG (FcγRIa) has been shown to be dependent on the FcR γ-chain for surface expression in vivo. In this study we assess the capacity of FcγRIa–γ-chain complexes expressed in IIA1.6 cells to trigger phagocytosis and ADCC. An intact immunoreceptor tyrosine-based activation motif (ITAM) signalling motif proved essential for triggering of biological function via the FcγRIa receptor complex. Both the FcR γ-chain and the FcγRIIa–ITAM proved active in directing phagocytosis of Staphylococcus aureus and ADCC of erythrocytes, triggered by the FcγRIa complex. The capacity of FcγRIa to trigger phagocytic and cytolytic activity by IIA1.6 cells, both considered ‘professional phagocyte’ functions, motivated us to re-evaluate the cell lineage and developmental stage of IIA1.6 cells. Although originally described as mouse B lymphocytes, the IIA1.6 cells proved positive for non-specific esterase activity and expressed the CD5 antigen. These combined characteristics place the IIA1.6 cells within a unique CD5+ B cell/macrophage lineage, optimally suited for cell biological analyses of phagocyte receptors.