bluuebunny/crossref_metadata_2025 huggingface dataset:
Primary malignant ‘triton’ tumour of the lung
Authors: MORAN C.A. , SUSTER S. , KOSS M.N.
Published: November 2003

Two cases of malignant ‘triton’ tumour arising within lung parenchyma are described. The patients were a three‐year‐old child and a 53‐year‐old man. Both patients presented with shortness of breath and a large intrapulmonary mass on chest X‐ray. Neither patient had a history of von Recklinhausen's neurofibromatosis. The lesions were treated by pneumonectomy. Grossly, both tumours presented as large, soft and gelatinous intraparenchymatous masses measuring 130 mm and 80 mm, respectively. Histologically, they were characterized by an atypical spindle cell proliferation embedded in an abundant myxoid stroma. Focal areas of rhabdomyoblastic differentiation characterized by large cells with abundant eosinophilic cytoplasm and occasional cytoplasmic cross‐striations could be seen admixed with the atypical spindle cell elements. Immunohistochemical studies showed a focal positive reaction for S‐100 protein in the atypical spindle cells embedded within the myxoid stroma, and a strong positive reaction for desmin and myoglobin in the rhabdomyoblastic areas. The child died three months after diagnosis with extension of the tumour into the thoracic cavity. The second patient has been lost to follow‐up. Although rare, malignant ‘triton’ tumour should be considered in the differential diagnosis of primary spindle cell sarcomas of the lung.
Failure to detect colonic mucosal hyperproliferation in mutation positive members of a family with hereditary non‐polyposis colorectal cancer
Authors: JASS J.R. , AJIOKA Y. , RADOJKOVIC M. , ALLISON L.J. , LANE M.R.
Published: November 2003

Mucosal proliferation was studied in biopsies obtained from the ascending colon from four mutation positive members of a hereditary non‐polyposis colorectal cancer family (group I) and compared to subjects with some positive family history of colorectal cancer but lacking the clinical and pathological features of hereditary non‐polyposis colorectal cancer (group II). Labelling indices were derived through immunohistochemical staining of the cell cycle associated nuclear proteins proliferating cell nuclear antigen (PCNA) and Ki–67. Only perfectly longitudinally sectioned crypts were assessed and good intra‐ and interobserver reproducibility was demonstrated. The labelling indices and proliferative compartment locations were similar in both groups. The mean Ki‐67 derived labelling indices (group I 18.0% and group II 17.5%) were similar to values obtained for normal subjects in other studies. PCNA derived labelling indices (group I 58.5% and group II 57.0%) were high, but probably reflect optimization of staining through methacarn fixation. The negative findings do not fit with most published data but critical appraisal of the literature indicates that they are likely to be correct.
CD44H expression in reactive mesothelium, pleural mesothelioma and pulmonary adenocarcinoma
Authors: ATTANOOS R.L. , WEBB R. , GIBBS A.R.
Published: November 2003

Malignant mesotheliomas are known to produce hyaluronic acid, in contrast to most pulmonary adenocarcinomas which produce neutral mucin. CD44H is the major cell surface receptor for hyaluronic acid. The aim of this study was to investigate immunohistochemically the expression of this antigen in reactive mesothelium, pleural mesothelioma and pulmonary adenocarcinoma and to assess its diagnostic utility in distinguishing the two tumours. Diffuse and intense membranous CD44H immunoreactivity was seen in 15 of 20 (75%) mesotheliomas and in all 20 biopsies of reactive mesothelium. In contrast, focal (<10% tumour) expression of CD44H was seen in only three of 20 (15%) pulmonary adenocarcinomas. We advocate the use of CD44H as a positive mesothelial marker for incorporation alongside other established immunohistochemical markers used to distinguish mesothelioma from adenocarcinoma.
Erdheim–Chester disease with prominent pulmonary involvement associated with eosinophilic granuloma of mandibular bone
Authors: KAMBOUCHNER M. , COLBY T.V. , DOMENGE C. , BATTESTI J.P. , SOLER P. , TAZI A.
Published: November 2003

We report a patient with eosinophilic granuloma localized to the left mandible who was subsequently shown to have Erdheim–Chester disease involving the lower extremities, omentum and lung. The diagnosis of eosinophilic granuloma was based on the presence of typical CD1a+ Langerhans' cell granulomas in a biopsy of mandible. The diagnosis of Erdheim–Chester disease was established on the basis of the pattern of radioisotopic uptake by long bones, seen on a technetium bone scan, and the presence of characteristic histopathological features in biopsies of lung and peritoneum. The pathological findings in lung were compatible with the abnormalities observed by tomodensitometry, but strikingly different from those seen in Langerhans' cell granulomatosis. The differences in the histological features of pulmonary involvement seen in the two diseases, and the possible relationship between Langerhans' cell granulomatosis and Erdheim–Chester disease, are discussed.
Synovial sarcoma of the heart
Authors: Nicholson A.G. , Rigby M. , Lincoln C. , Meller S. , Fisher C.
Published: November 2003

Primary synovial sarcomas of the heart are exceptionally rare tumours, only five being previously documented. We report a sixth case and review published data. Ages ranged from 13–53 (mean 40.8) years. There were four men and two women who characteristically presented with syncope or dyspnoea. Most of the tumours arose in the right atrium where they formed pedunculated or polypoid masses. In the previously documented cases, all patients died within 9 months. However in our case, local excision was felt to have been complete and the patient remains disease free at 10 months.
Immunohistochemical detection of the Wilms’ tumour gene WT1 in desmoplastic small round cell tumour
Authors: CHARLES A.K. , MOORE I.E. , BERRY P.J.
Published: November 2003

The desmoplastic small round cell tumour (DSRCT) is a rare, highly malignant neoplasm usually presenting in the abdomen of adolescent males. A characteristic translocation between the Ewing’s sarcoma gene on chromosome 22 and the Wilms’ tumour gene WT1 on chromosome 11 has been described, producing a fusion gene with expression of the DNA binding area of WT1. Some Wilms’ tumour antibodies recognize epitopes of this part of the WT1 protein. All four cases of DSRCT examined showed strong staining of the tumours with an anti‐WT1 antibody, suggesting this may be useful in the diagnosis of these tumours.
Incidence of apoptosis in HIV‐myelopathy, myelodysplastic syndromes and non‐specific inflammatory lesions of the bone marrow
Authors: THIELE J. , ZIRBES T.K. , WIEMERS P. , LORENZEN J. , KVASNICKA H.M. , FISCHER R.
Published: November 2003

Myelodysplastic syndromes, AIDS‐related myelopathy and non‐specific inflammatory reactions (mostly rheumatoid myelitis) are characterized by normo‐ to hypercellular bone marrow, but frequently display cytopenias in the peripheral blood. In the current study we have addressed the question whether this situation reflects an increased programmed cell death in haemopoiesis. For this purpose, the in situ end‐labelling technique was applied to formalin‐fixed and paraffin‐embedded trephine biopsies derived from patients and a control group without any haematological disorder. Results were evaluated by morphometry. Significantly more apoptotic cell death was observed in the haemopoietic marrow of patients with either disease. Using double‐immunohistochemistry with the monoclonal antibody PG‐M1 (CD68), we were able to demonstrate that approximately one third of the apoptotic cells were ingested by macrophages. Our findings are in keeping with previously published data that postulated increased frequencies of macrophages in these disorders as well as raised serum levels of TNF‐α.
HLA‐DR4‐restricted T‐cell epitopes from the mycobacterial 60 000 MW heat shock protein (hsp 60) do not map to the sequence homology regions with the human hsp 60
Authors: MUSTAFA A. S. , LUNDIN K. E. A. , MELOEN R. H. , SHINNICK T. M. , COULSON A. F. W. , OFTUNG F.
Published: November 2003

The mycobacterial 60 000 MW heat shock protein (hsp 60) is a major antigen recognized by mycobacteria‐reactive human CD4+ T cells with lymphokine profiles and effector functions consistent with protective immunity. In addition, the presence of a large number of T‐cell epitopes presented by several HLA class II molecules makes this antigen relevant to subunit vaccine design. However, the results from animal models as well as human studies suggest that the mycobacterial hsp 60 may induce T‐cell‐mediated autoimmune conditions. In humans, the expression of HLA‐DR4 represents a risk factor for some autoimmune diseases. These observations suggest that the epitopes from the mycobacterial hsp 60 presented to T cells in the context of HLA‐DR4 could be relevant to autoimmunity. This is the first report on identification of HLA‐DR4‐restricted T‐cell epitopes from the mycobacterial antigen hsp 60. In total, five epitopes recognized in the context of HLA‐DR4 by the M. leprae hsp 60‐reactive CD4+ T‐cell clones from a subject immunized with M. leprae were defined by synthetic peptides. Two of the epitopes were M. leprae‐specific (aa 343–355, aa 522–534), whereas three epitopes were common to M. leprae and M. tuberculosis (aa 331–345, aa 441–455, aa 501–515). However, all of these epitopes belong to the regions that are highly divergent between the mycobacterial hsp 60 and the homologous human hsp 60 sequence, suggesting that the T cells recognizing the mycobacterial hsp 60 in the context of HLA‐DR4 may not necessarily induce autoreactivity.
T‐cell‐receptor dose and the time of treatment during murine retrovirus infection for maintenance of immune function
Authors: LIANG B. , ARDESTANI S. , MARCHALONIS J. J. , WATSON R. R.
Published: November 2003

C57BL/6 mice were injected with different doses of human T‐cell receptor (TCR) Vβ8.1 CDR1 peptide at different times after murine retrovirus (LP‐BM5) infection. Injection with TCR Vβ8.1 CDR1 peptide largely prevented the retrovirus‐induced reduction in B‐ and T‐cell proliferation, and T‐helper 1 (Th1) cytokines [interleukin‐2 (IL‐2) and interferon‐γ (IFN‐γ)] secretion. It also suppressed T‐helper 2 (Th2) cytokines (IL‐6 and IL‐10) production, which was stimulated by retrovirus infection. These effects were accomplished using at least 100 μg of peptide per mouse and the most effective dose of peptide had to be given within 4 weeks after retrovirus infection. Immunization with doses above 100 μg/mouse as long as 4 weeks postinfection maintained natural killer (NK) cell activity during retrovirus infection. Reducing the dose of peptide or delaying it until the disease progressed towards early murine acquired immune deficiency syndrome (AIDS) allowed development of immune dysfunction. These studies provide data suggesting that immune dysfunction, induced by murine retrovirus infection, was largely prevented by TCR Vβ CDR1 peptide injection.
Developmental heterogeneity of Vγ1.1 T cells in the mouse liver
Authors: KODAIRA Y. , YOKOMURO K. , TANAKA S. , MIYAZAKI J.‐I. , IKUTA K.
Published: November 2003

Modifications at V–(D)–J junctions increase the diversity of T‐cell receptors (TCR). It has been shown that the levels of N‐nucleotide insertion at the V–(D)–J junction in TCR transcripts are different between fetal and adult stages. To clarify developmental stages and pathways of γδ T cells in the liver, we analysed the nucleotide sequence of Vγ1.1–Jγ4 junctions of intra‐hepatic lymphocytes (IHL), spleen cells and developing thymocytes from normal and athymic nude mice. The level of N‐insertion increased in thymocytes during ontogeny. The percentage of Vγ1.1–Jγ4 transcripts with N‐insertion was 3% at day 16 of gestation, 42% at newborn, and 89% at 7 weeks. Transcripts from normal IHL showed intermediate levels of N‐insertion between those of newborn and adult thymocytes. In contrast the percentage of N‐insertion in nude IHL was 47%, and this value was comparable to that of newborn thymocytes. Among the transcripts of normal IHL, the sequences common with nude IHL showed a newborn level of N‐insertion (38%), and the remaining sequences showed an adult level (89%). These results suggested the possibility that Vγ1.1‐expressing T cells in IHL might be a heterogeneous population consisting of the cells developed extrathymically as well as the cells developed intrathymically. The Vγ1.1–Jγ4 junctions from spleen cells showed less variability than those from IHL and adult thymocytes. It suggested that γδ T cells bearing specific Vγ1.1 TCR develop and/or home in the spleen.
Role of intracellular calcium as a priming signal for the induction of nitric oxide synthesis in murine peritoneal macrophages
Authors: PARK Y. C. , JUN C. D. , KANG H. S. , KIM H. D. , KIM H. M. , CHUNG H. T.
Published: November 2003

Because the role of intracellular Ca2+ in the two‐signal process for the induction of nitric oxide (NO) synthesis is controversial, this study was undertaken to examine the role of Ca2+ in the transcriptional regulation of inducible NO synthase (iNOS) in murine peritoneal macrophages. Treatment of the cells with thapsigargin (TG) or 2,5‐di‐(t‐butyl)‐1,4‐benzodihydroquinone (tBuBHQ), which are the specific and potent Ca2+‐ATPase inhibitors of endoplasmic reticulum (ER), showed modest effects on tumoricidal function, whereas TG or tBuBHQ in combination with interferon‐γ (IFN‐γ) or lipopolysaccharide (LPS) showed marked effects on tumoricidal function of the cells. The tumoricidal effects of the activated macrophages were correlated with the amount of NO synthesis, and totally abrogated by the use of NOS inhibitor, N G‐monomethyl‐l‐arginine (NGMMA). The increases in NO synthesis was reflected as increased amounts of iNOS mRNA by Northern blotting. To confirm that iNOS induction was due to the changes in the intracellular Ca2+ level, the acetoxymethyl ester of 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid (BAPTA‐AM), an intracellular Ca2+ chelator, was used. Blocking the increase of cytosolic free Ca2+ significantly decreased the induction of NO synthesis. To demonstrate that intracellular Ca2+ acts as a ‘priming’ signal rather than a ‘triggering’ signal on the induction of NO synthesis by murine peritoneal macrophages, we designed several experiments. When the cells were treated with TG 6 hr after the treatment with IFN‐γ, there was no increase in NO synthesis. In addition, when the cells were treated with TG or LPS 6 hr after treatment with tBuBHQ, a synergistic increase on NO synthesis was shown only in the case of LPS. When phorbol 12‐myristate 13‐acetate (PMA), a protein kinase C (PKC) activator, was added to the cells 6 hr after the treatment with TG, there was a marked co‐operative induction of NO synthesis, even though PMA alone has no effect. Based on the results obtained in this study, we suggest that cytosolic Ca2+ might be enough for the expression of iNOS gene as a priming signal and PKC might be involved in the induction of NO synthesis as a triggering signal by post‐transcriptional modification of iNOS mRNA or iNOS itself in the activated murine peritoneal macrophages.
Regulation of T‐cell activation in the lung: alveolar macrophages induce reversible T‐cell anergy in vitro associated with inhibition of interleukin‐2 receptor signal transduction
Authors: STRICKLAND D. , KEES U. R. , HOLT P. G.
Published: November 2003

Alveolar macrophages (AM) are recognized as archetypal ‘activated’ macrophages with respect to their capacity to suppress T‐cell responses to antigen or mitogen, and this function has been ascribed an important role in the maintenance of local immunological homeostasis at the delicate blood : air interface. The present study demonstrates that this suppression involves a unique form of T‐cell anergy, in which ‘AM‐suppressed’ T cells proceed normally through virtually all phases of the activation sequence including Ca2+ flux, T‐cell receptor (TCR) modulation, cytokine [including interleukin‐2 (IL‐2)] secretion and IL‐2 receptor (IL‐2R) expression. However, the ‘suppressed’ T cells fail to up‐regulate CD2, and do not re‐express normal levels of TCR‐associated molecules after initial down‐modulation; moreover, they are unable to transduce IL‐2 signals leading to phosphorylation of IL‐2R‐associated proteins, and remained locked in G0/G1. The induction of this form of anergy is blocked by an NO‐synthase inhibitor, and is reversible upon removal of AM from the T cells, which then proliferate in the absence of further stimulation. We hypothesize that this mechanism provides the means to limit the magnitude of local immune responses in this fragile tissue microenvironment, while preserving the capacity for generation of immunological memory against locally encountered antigens via clonal expansion of activated T cells after their subsequent migration to regional lymphoid organs. In an accompanying paper, we demonstrate that a significant proportion of T cells freshly isolated from lung exhibit a comparable surface phenotype.
Induction of single and dual cytotoxic T‐lymphocyte responses to viral proteins in mice using recombinant hybrid Ty–virus‐like particles
Authors: LAYTON G. T. , HARRIS S. J. , MYHAN J. , WEST D. , GOTCH F. , HILL‐PERKINS M. , COLE J. S. , MEYERS N. , WOODROW S. , FRENCH T. J. , ADAMS S. E. , KINGSMAN A. J.
Published: November 2003

The induction of cytotoxic T‐lymphocyte (CTL) responses to viral proteins is thought to be an essential component of protective immunity against viral infections. Methods for generating such responses in a reproducible manner would be of great value in vaccine development. We demonstrate here that the recombinant antigen‐presentation system based on the yeast transposon (Ty) particle‐forming p1 protein is a potent means of inducing CTL responses to a variety of viral CTL epitopes, including influenza virus nucleoprotein (two epitopes), Sendai virus and vesicular stomatitis virus nucleoproteins, and the V3 loop of human immunodeficiency virus type‐1 (HIV‐1) gp120. CTL were primed by hybrid Ty–virus‐like particles (VLP) carrying the minimal epitope or as much as 19 000 MW of protein. Ty–VLP carrying two different epitopes (dual‐epitope Ty–VLP) were capable of priming CTL responses in two different strains of mice or against two epitopes in the same individual. Furthermore, co‐administration of a mixture of two different Ty–VLP carrying single epitopes could induce responses to both epitopes in the same individual. Ty–VLP appear to represent a reproducible and flexible system for inducing CTL responses in mice, and warrant further evaluation in primates.
Human γ/δ T‐cell response to Listeria monocytogenes protein components in vitro
Authors: MUNK M. E. , ELSER C. , KAUFMANN S. H. E.
Published: November 2003

Listeria
monocytogenes is a facultative intracellular pathogen that replicates inside mononuclear phagocytes and induces specific cellular immunity. Listeriosis encompasses many clinical syndromes and meningitis is the most frequent clinical manifestation. Human α/β and γ/δ T cells have been shown to respond to L.monocytogenes antigens and to play an important role in resistance against listerial infection. We investigated the nature of listerial ligands and the influence of the major virulence factor, listeriolysin (hly), on the stimulation of human γ/δ T cells from healthy individuals. We found that a listerial somatic protein ligand, which is sensitive to proteinase treatment, stimulated γ/δ T cells in vitro; the majority of Listeria‐responsive γ/δ T cells expressed Vγ9Vδ2 T‐cell receptor chains and human leucocyte antigen‐DR molecules; γ/δ T‐cell responses to hly+ and hly−Listeria strains were comparable; L. monocytogenes strains of different virulence stimulated γ/δ T cells equally. Thus, protein components of L. monocytogenes unrelated to virulence activate human γ/δ T cells in vitro
CD8‐deficient mice exhibit augmented mucosal immune responses and intact adjuvant effects to cholera toxin
Authors: HÖRNQUIST E. , GRDÌC D. , MAK T. , LYCKE N.
Published: November 2003

We used normal, CD4 and CD8 gene‐targeted mice to investigate the role of CD4+ and CD8+ T cells in the regulation of gut mucosal immune responses following oral immunizations with cholera toxin (CT) adjuvant. Phenotypic analysis of mucosa‐associated tissues revealed normal CD3+ T‐cell frequencies in CD4−/− and CD8−/− mice such that in CD4−/− mice the CD8+ and double‐negative (DN) T cells were increased. In CD8−/− mice the CD4+ T cells were increased, with the exception that in the intraepithelial compartment the CD3+ T cells were predominantly DN γδ T‐cell receptor (TCR)+ T cells. All mice, normal and deficient, failed to respond to oral immunization with the antigen, keyhole limpet haemocyanin (KLH), alone. In the presence of CT adjuvant, however, CD8−/− mice consistently exhibited three‐ to fivefold stronger gut mucosal responses compared to normal C57Bl/6 mice. By contrast, no difference was observed for systemic responses between CD8−/− and normal mice. Thus the up‐regulation selectively affected mucosal responses, suggesting that, contrary to the CD8−/− mouse gut, the normal gut mucosa may host CD8+ T cells that exert a local suppressive effect on T‐ and B‐cell responses. The magnitude of the enhancing effect of CT was comparable in CD8−/− and normal mice, clearly demonstrating that the adjuvant mechanism of CT does not require CD8+ T cells. On the other hand, the adjuvant effect of CT required CD4+ T cells, because no or poor anti‐KLH responses were observed in CD4−/− mice. In both normal and CD8−/− mice CT adjuvant promoted KLH‐specific CD4+ T‐cell priming without any selective effect on the differentiation towards a T‐helper type‐1 (Th1) or Th2 dominance. Furthermore, CT adjuvant increased the frequency of CD4+ T cells expressing a memory phenotype, i.e. CD44high, LECAM‐1low and CD45RBlow. We have shown, using gene‐targeted mice, that CD8+ T cells are not required for the adjuvant effect of CT, and that CD8+ T cells may exert local mucosal down‐regulation of intestinal immune responses.
Retrovirus‐elicited interleukin‐12 and tumour necrosis factor‐α as inducers of interferon‐γ‐mediated pathology in mouse AIDS
Authors: GIESE N. A. , GAZZINELLI R. T. , ACTOR J. K. , MORAWETZ R. A. , SARZOTTI M. , MORSE III H. C.
Published: November 2003

Spleen cells from mice resistant or sensitive to mouse acquired immune deficiency syndrome (MAIDS) were examined for cytokine mRNA. In MAIDS‐resistant BALB/c mice, cytokine transcripts peaked at 1 week after infection with Type 1 cytokines [interleukin‐2 (IL‐2), tumour necrosis factor‐α (TNF‐α), interferon‐γ (IFN‐γ), IL‐12], dominating over Type 2 cytokines (IL‐4, IL‐10). Expression of cytokines other than IL‐2 later declined to levels seen in uninfected mice. In MAIDS‐sensitive B6 mice, transcripts for all cytokines were increased at 1 week and, except for IL‐2, increased progressively. Spontaneous production of IFN‐γ protein was associated with enhanced mRNA expression at 1 week after infection of either strain, but none was detectable in association with even higher levels of transcripts at later times after infection of B6 mice. Spleen cells from longer‐term‐infected B6 mice, however, produced substantial amounts of IFN‐γ following treatment with lipopolysaccharide (LPS) or IL‐12. Inclusion of anti‐IL‐12 or anti‐TNF‐α antibodies blocked induction of IFN‐γ by LPS. Induction of IFN‐γ by IL‐12 was potentiated by TNF‐α following stimulation of intact spleen cells and purified CD4+ or CD8+ T cells, as well as negatively selected CD4−8− cells from infected B6 mice. Further studies showed that IFN‐γ knockout mice on a B6 background developed MAIDS with a prolonged time–course, whereas BALB/c knockout mice were unchanged in their resistance to MAIDS. These studies suggest that continuing low‐level expression of IFN‐γ, stimulated by IL‐12 and TNF‐α, contributes to the susceptibility of B6 mice to MAIDS but is not required for the resistance of BALB/c mice to disease.
Differential regulation of tumour necrosis factor‐α mRNA degradation in macrophages by interleukin‐4 and interferon‐γ
Authors: SUK K. , ERICKSON K. L.
Published: November 2003

Interferon‐γ (IFN‐γ) and interleukin‐4 (IL‐4) have been reported previously to mediate similar as well as antagonistic effects on murine macrophage functions. One effect common to both is the enhancement of tumour necrosis factor‐α (TNF‐α) secretion in macrophages. To assess further the effects of these two lymphokines on macrophage TNF‐α production, we investigated the role of these lymphokines in the induction and stability of TNF‐α messages along with interleukin‐1 (IL‐1) as a comparison. IFN‐γ and IL‐4 increased lipopolysaccharide (LPS)‐induced TNF‐α, IL‐1 steady‐state message levels. In contrast to IL‐1 messages, whose degradation was not significantly affected by either lymphokine, the stability of TNF‐α messages differed after IFN‐γ and IL‐4 treatment. Although IL‐4 treatment increased the TNF‐α transcription rate, an increase in the degradation rate of TNF‐α mRNA in the IL‐4‐treated cells resulted in a lower level of steady‐state mRNA than in the IFN‐γ‐treated cells. Additionally, a 18 000 MW cytoplasmic factor was found to have specific binding activity to the AU‐rich sequences of the TNF‐α message in peritoneal macrophages. Although the binding activity of this factor was not affected by either IFN‐γ or IL‐4, the binding of the factor to AU‐rich sequences appeared to be important in the rapid degradation of TNF‐α messages. Thus IFN‐γ and IL‐4 may differentially affect the post‐transcriptional control of TNF‐α gene expression. And this lymphokine‐mediated post‐transcriptional control of the TNF‐α gene does not appear to involve the alteration of binding activity of the 18 000 MW AU‐rich sequence binding factor.
In vivo treatment with anti‐interleukin‐13 antibodies significantly reduces the humoral immune response against an oral immunogen in mice
Authors: BOST K. L. , HOLTON R. H. , CAIN T. KINCY , CLEMENTS J. D.
Published: November 2003

Interleukin‐13 (IL‐13) is a cytokine which significantly enhances the proliferation and differentiation of B lymphocytes. We therefore evaluated its role in the formation of a humoral immune response in vivo. Upon oral immunization with the B subunit of Escherichia coli heat‐labile enterotoxin (LT‐B), rapid up‐regulation of IL‐13 mRNA expression in the mesenteric lymph nodes of LT‐B intubated mice occurred. This result suggested that IL‐13 might be involved in the formation of a mucosal antibody response against LT‐B if this cytokine was in fact secreted. To test this possibility, the coding region for murine IL‐13 was cloned into the pFLAG‐1 expression vector. Recombinant murine IL‐13 was purified from bacterial lysates and used as an immunogen to produce polyclonal anti‐IL‐13 antibodies. Groups of BALB/c mice treated in vivo with anti‐IL‐13 antibody 2 days before and on the day of oral immunization with LT‐B had significantly reduced intestinal IgA and serum IgG and IgA anti‐LT‐B antibody responses when compared to mice treated with control antibody. Furthermore, groups of mice primed with LT‐B and then treated with anti‐IL‐13 antibody prior to oral immunization with a second dose of LT‐B also had significantly reduced intestinal IgA and serum IgG and IgA anti‐LT‐B antibody titres compared to controls. In vitro LT‐B restimulation experiments using splenic mononuclear leucocytes isolated from LT‐B primed mice treated with anti‐IL‐13 antibody demonstrated decreased expression of IL‐4 and IL‐13 mRNA and decreased IL‐4 secretion when compared to controls. Together these results demonstrate an important role for IL‐13 in the formation of a humoral immune response at mucosal surfaces.
Acceleration of IgE responses by treatment with recombinant interleukin‐3 prior to infection with Trichinella spiralis in mice
Authors: KORENAGA M. , WATANABE N. , ABE T. , HASHIGUCHI Y.
Published: November 2003

Treatment of mice with recombinant interleukin‐3 (rIL‐3) accelerated an IL‐4‐dependent IgE production following infection with Trichinella spiralis. When mice were treated with a total of 1.5 × 104 units rIL‐3 for 5 days before infection with 400 muscle larvae, the serum IgE level increased prominently on day 5. Acceleration of IgE responses was dependent on the dose of rIL‐3 injected. Treatment of mice with a total of 103 units rIL‐3 could accelerate IgE responses. IgE responses were detected by a sandwich enzyme‐linked immunosorbent assay at least from day 3 in mice treated with rIL‐3. Acceleration of IgE responses was inhibited by anti‐IL‐4 monoclonal antibody. In contrast to this, IgG1 and IgG2a responses were not suppressed by the anti‐IL‐4 treatment. IL‐3 treatment could up‐regulate IgE and IgG1 responses but not the IgG2a response. IL‐3 treatment could also accelerate IgE responses in W/Wv mice infected with the parasites. These results suggest that IL‐3 is involved in regulation of IgE responses in mice and that mast cells do not play an essential role in acceleration of IgE responses induced by rIL‐3 treatment in this system.
HIV‐1 upregulates Fas ligand expression in CD4+ T cells in vitro and in vivo : association with Fas‐mediated apoptosis and modulation by aurintricarboxylic acid
Authors: MITRA D. , STEINER M. , LYNCH D. H. , STAIANO‐COICO L. , LAURENCE J.
Published: November 2003

CD4+ T‐lymphocyte apoptosis has been associated with human immunodeficiency virus (HIV)‐1 infection in vitro, paralleling the expression of Fas (APO‐1, CD95) on peripheral blood mononuclear cells from patients with HIV disease. However, the link between Fas induction, T‐cell activation, and cell death is unclear. We document, for the first time, marked upregulation of expression of mRNA for the ligand for Fas in peripheral blood mononuclear cells from HIV seropositive individuals, and demonstrate the ability of HIV infection to induce such expression in CD4+ T cells in vitro. We also define the relevance of this expression to HIV‐mediated CD4+ T cell death. Our ability to downregulate Fas ligand message and suppress HIV‐mediated apoptosis with aurintricarboxylic acid, a clinically used protease inhibitor with known activity against programmed cell death in other systems, may open up a new area of therapy for HIV infection.
Ligation of CD40 with soluble CD40 ligand reverses anti‐immunoglobulin‐mediated negative signalling in murine B lymphoma cell lines but not in immature B cells from neonatal mice
Authors: MARSHALL‐CLARKE S. , OWEN G. , TASKER L.
Published: November 2003

Ligation of surface immunoglobulin (sIg) on certain murine B‐lymphoma lines has been shown to initiate a programme leading to growth arrest and death of the cells by apoptosis. The cell lines WEHI 231 and CH33 which respond in this way to receptor cross‐linking have phenotypic characteristics resembling those of immature normal B cells, and their responses have been taken to model those responsible for clonal deletion or anergy. Cross‐linking of sIg on normal neonatal B cells has also been shown to inhibit their responsiveness to polyclonal activators. We have examined the ability of various co‐stimuli to modify the response of growth‐inhibitable B lymphoma lines to sIg cross‐linking. Our findings indicate that cell–cell contact between cells of the WEHI 231 or CH33 lines and activated T cells rescues these cells from growth arrest and apoptosis. Cell‐free supernatants from some T‐cell lines were also protective although recombinant IL‐4 had no effect. Analysis of the most effective signals and timing for inducing this protection suggested that it might, in part, be mediated by CD40 ligand (CD40L) expressed on or secreted by activated T cells. Using a soluble recombinant CD40L‐CD8 fusion protein we have now shown that co‐ligation of CD40 is sufficient to rescue WEHI 231 and CH33 cells from anti‐Ig‐induced apoptosis. In contrast, the inhibitory effect of anti‐Ig antibodies on the lipopolysaccharide (LPS)‐driven proliferation of neonatal B cells was not relieved by co‐ligation of CD40 with CD40L. These findings bring into question the usefulness of ‘immature’ B‐cell lines as models for tolerance induction.
A sigma ligand, SR 31747A, potently modulates Staphylococcal enterotoxin B‐induced cytokine production in mice
Authors: BOURRIÉ B. , BENOÎT J.‐M. , DEROCQ J.‐M. , ESCLANGON M. , THOMAS C. , LE FUR G. , CASELLAS P.
Published: November 2003

Sigma receptors originally described in distinct regions of the central nervous system are expressed on cells of the immune system. A sigma ligand, SR 31747A, was observed here to inhibit in vitro the Staphylococcal enterotoxin B (SEB)‐driven lymphocyte proliferation. In mice, the drug confers a potent protection against the lethality induced by SEB, stimulates the SEB‐induced serum release of interleukin (IL)‐10 and inhibits at the same time the systemic release of IL‐2, IL‐4, granulocyte–macrophage colony‐stimulating factor (GM‐CSF), IL‐6 and tumour necrosis factor‐α (TNF‐α). The enhancement of IL‐10 production by this compound is also effective in nude mice treated with SEB, indicating that IL‐10 of T‐cell origin is not involved in this process. The finding that a sigma ligand protects against the SEB‐induced toxicity provides insights into the clinical use of this family of compounds, particularly in food poisoning and septic shock where Staphylococcal enterotoxins are involved. The observation that this compound stimulates IL‐10 synthesis indicates that it could be a potent regulatory agent of chronic inflammatory diseases.
Reversal of mucosal tolerance by subcutaneous administration of interleukin‐12 at the site of attempted sensitization
Authors: CLAESSEN A. M. E. , VON BLOMBERG B. M. E. , DE GROOT J. , WOLVERS D. A. E. , KRAAL G. , SCHEPER R. J.
Published: November 2003

Oral feeding of proteins causes peripheral T‐cell tolerance, as revealed by reduced delayed‐type hypersensitivity (DTH) reactivity after immunization. This type of tolerance can be due both to passive T‐cell anergy and active immunosuppression. Using ovalbumin‐fed mice we studied whether putatively immunostimulatory cytokines could break this state of mucosal tolerance. Cytokines were administered locally at the site of attempted sensitization. It was found that neither interleukin‐2 (IL‐2), interferon‐γ (IFN‐γ) nor granulocyte–macrophage colony‐stimulating factor (GM‐CSF) could restore the response to immunization. In contrast, local administration of IL‐12 at the site of attempted immunization resulted in full recovery of DTH reactivity. The dichotomy between the two Th1 stimulatory cytokines IFN‐γ and IL‐12 was also reflected by different effects on ovalbumin‐specific antibody isotypes. Although both IFN‐γ and IL‐12 downregulated serum IgG1‐levels in tolerant mice, suggesting decreased ovalbumin‐specific Th2 function, only local administration of IL‐12 led to increased serum IgG2a levels. These results support the view that potentiation of Th1 effector function is critical for reversal of mucosal tolerance.
Signal transduction through μκ B‐cell receptors expressed on pre‐B cells is different from that through B‐cell receptors on mature B cells
Authors: NAKAMURA T. , KOYAMA M. , YONEYAMA A. , HIGASHIHARA M. , KAWAKAMI T. , YAMAMURA H. , SADA K. , OKUMURA K. , KUROKAWA K.
Published: November 2003

We introduced κ light chain genes into pre‐B cells to increase the surface μ HC expression, and established transfectants expressing mature type of B‐cell receptors (BCR) on pre‐B‐cell surfaces. Since the cytoplasmic conformations of the reconstituted BCR and intrinsic pre‐B‐cell receptor (pre‐BCR) are identical, they would be connected with the identical signal transduction pathways in pre‐B cells. By using the transfectants, we revealed that the reconstituted BCR on pre‐B cells was functionally equivalent to BCR on mature B cells in terms of the induction of intracellular Ca++ mobilization. However, we found that the signal‐transduction pathways through BCR on pre‐B cells were quantitatively different from those of BCR on mature B cells in two ways. First, cross‐linkage of the reconstituted BCR on pre‐B cells induced preferential tyrosine phosphorylation of p120 and p100, which was not observed when BCR on mature B cells was cross‐linked. Second, BCR in pre‐B cells was physically associated with a larger amount of phosphatidylinositol‐3 kinase (PI‐3K) than BCR in mature B cells in spite of the fact that both pre‐B and B cells expressed a similar amount of PI‐3K in cytoplasm. Signals through pre‐BCR and BCR are known to cause distinct biological effects in B‐cell development. The biochemical features in the downstream of reconstituted BCR on pre‐B cells, which we revealed in this study, will be of help in understanding the mechanism of functional differences between pre‐BCR and BCR.
Activation by mitogens and superantigens of axolotl lymphocytes: functional characterization and ontogenic study
Authors: SALVADORI F. , TOURNEFIER A.
Published: November 2003

Urodele amphibians have weak and slow immune responses compared to mammals and anuran amphibians. Using new culture conditions, we tested the ability of lymphocytes of a well‐studied salamander, the Mexican axolotl (Ambystoma mexicanum) to proliferate in vitro with diverse mitogenic agents. We demonstrated that the axolotl has a population of B lymphocytes that proliferate specifically and with a high stimulation index to the lipopolysaccharide (LPS) known as a B‐cell mitogen in mammals. This proliferative capacity is observed without significant changes throughout ontogenesis. In the presence of LPS, axolotl B lymphocytes are able to synthesize and secrete both isotypes of immunoglobulin described in this species, IgM and IgY. Moreover, a distinct lymphocyte subpopulation is able to proliferate significantly in response to the mitogens usually known as T‐cell specific in mammals, phytohaemagglutinin (PHA) and concanavalin A (Con A). The activated cells are T lymphocytes, as shown by depletion experiments performed in vitro with monoclonal antibodies, and in vivo by thymectomy. Splenic T lymphocytes of young axolotls (before 10 months) do not have this functional ability, which suggests maturation and/or migration phenomena during T‐cell ontogenesis in this species. Axolotl lymphocytes are able to proliferate in vitro with a significant stimulation index to staphylococcal enterotoxins A and B (SEA and SEB). These products act on mammalian lymphocytes as superantigens: in combination with products of the major histocompatibility complex (MHC), they bind T‐cell receptors with particular Vβ elements. The fact that these superantigens are able to activate lymphocytes of a primitive vertebrate suggests a striking conservation of molecular structures implied in superantigen presentation and recognition.
Differential effects of osteopontin on the cytotoxic activity of macrophages from young and old mice
Authors: ROLLO E. E. , DENHARDT D. T.
Published: November 2003

Osteopontin (OPN) is a secreted phosphoprotein found in body fluids (e.g. plasma, urine, milk) and in mineralized tissues. Its expression is increased in many transformed cells and in normal cells exposed to various cytokines. When stimulated with the inflammatory mediators lipopolysaccharide and interferon‐γ, mouse macrophages secrete nitric oxide (NO) as a cytotoxic agent effective against microbial invaders and tumour cells. This report documents (1) that thioglycollate‐elicited peritoneal macrophages, activated with the inflammatory mediators, produced less NO and exhibited reduced cytotoxicity towards target cells when they were obtained from old animals than when they were obtained from young animals; and (2) that OPN was able to inhibit both the induced NO synthesis and cytotoxicity, but more effectively in macrophages from the young animals than those from the old animals. This may be due to the observed higher level of OPN expression in macrophages from old animals.
Crystallization of a complex between the Fab fragment of a human immunoglobulin M (IgM) rheumatoid factor (RF–AN) and the Fc fragment of human IgG4
Authors: SOHI M. K. , CORPER A. L. , WAN T. , STEINITZ M. , JEFFERIS R. , BEALE D. , HE M. , FEINSTEIN A. , SUTTON B. J. , TAUSSIG M. J.
Published: November 2003

Rheumatoid factors (RF) are the characteristic autoantibodies found in patients with rheumatoid arthritis. They recognize epitopes in the Fc region of immunoglobulin G (IgG) and are often of the IgM isotype. In order to analyse the nature of RF–Fc interactions, we have crystallized a complex between the Fab fragment of a human monoclonal IgM rheumatoid factor (RF–AN) and the Fc fragment of human IgG4. The stoichiometry of the complex within the crystals was found to be 2 : 1 Fab : Fc. The crystals diffracted X‐rays to 0.3 nm resolution, and the space group was C2, with cell dimensions a = 16.03 nm, b = 8.19 nm, c = 6.42 nm, β = 98.3°. We have also determined the sequence of the variable region of the RF–AN light chain, not hitherto reported. This belongs to the VλIII‐a subgroup and is closely related to the germline gene Humlv318, from which it differs in three amino acid residues. This is the first reported crystallized complex between a human autoantibody and its autoantigen.
Analysis of peptide‐binding motifs for two disease associated HLA‐DR13 alleles using an M13 phage display library
Authors: DAVENPORT M. P. , QUINN C. L. , VALSASNINI P. , SINIGAGLIA F. , HILL A. V. S. , BELL J. I.
Published: November 2003

Major histocompatibility complex (MHC) molecules bind peptides bearing an appropriate ‘sequence motif’ for MHC binding. The use of phage display libraries exploits the ability of MHC class II molecules to exchange peptides in solution and thus select out peptide sequences with high‐affinity binding from a large array of random peptides. We have analysed the peptide binding motifs of HLA‐DRB1*1301 and *1302 using affinity purified HLA‐DR13 molecules to purify sequentially HLA‐DR13‐binding peptides from a large random library of M13 phage containing nonamer inserts in the pIII coat protein. These DR13 alleles differ only at position 86 of the HLA‐DRβ chain, where they contain valine and glycine residues respectively. These alleles were chosen because of their association with protection from severe malaria and chronic hepatitis B virus infection in West Africa. Analysis of the phage bound to these DR molecules suggests binding motifs. We compare the results derived from the use of the phage display library with results obtained from analysis of eluted peptides and peptide‐binding studies. This analysis shows that although there is a common theme to motifs derived using different methods, there are also subtle variations between them.
Development of CD8α/β+ TCRαβ intestinal intraepithelial lymphocytes in athymic nu/nu mice and participation in regional immune responses
Authors: EMOTO M. , EMOTO Y. , KAUFMANN S. H. E.
Published: November 2003

On the basis of the CD8 coreceptor expression, T‐cell receptor (TCR)αβ‐bearing intestinal intraepithelial lymphocytes (i‐IEL) segregate into two populations. The CD8αα+ TCRαβ i‐IEL develop thymus independently, whereas the CD8αβ+ TCRαβ i‐IEL are generally considered to be thymus dependent. Flow cytometry analysis revealed a distinct population of CD8αβ+ TCRαβ i‐IEL in individual athymic nu/nu mice. The i‐IEL encompassing CD8αβ+ TCRαβ cells expressed potent cytolytic and interferon‐γ‐producing activities. These findings demonstrate that CD8αβ+ TCRαβ i‐IEL can develop in nu/nu mice independently from a functional thymus and suggest that these cells, directly or indirectly, perform biological functions in the gut.
Granulocyte–macropha ge colony‐stimulating factor down‐regulates CD14 expression on monocytes
Authors: KRUGER M. , VAN DE WINKEL J. G. J. , DE WIT T. P. M. , COOREVITS L. , CEUPPENS J. L.
Published: November 2003

CD14 is a differentiation‐stage‐linked glycosyl‐phophatidyl‐inositol‐linked glycoprotein on human peripheral blood monocytes and tissue macrophages, which functions as a receptor for lipopolysaccharide. Here, the effects of granulocyte–macrophage colony‐stimulating factor (GM‐CSF), a cytokine with proliferation‐ and differentiation‐inducing properties on myeloid lineage cells, were studied on CD14 expression by peripheral blood cells. GM‐CSF down‐regulated the membrane expression of CD14 on monocytes, while it up‐regulated expression on neutrophils. GM‐CSF also decreased the spontaneous release of CD14 in monocyte culture supernatants. Down‐regulation of CD14 expression and release was accompanied by a decrease in the mRNA transcript for CD14, suggesting that it most likely reflects an effect on the transcriptional level. The functional significance of this phenomenon, and its potential relation to the terminal differentiation of monocytes, are discussed.
Mouse models of autoimmune disease suggest that self‐tolerance is maintained by unresponsive autoreactive T cells
Authors: TAGUCHI O. , TAKAHASHI T.
Published: November 2003

Multiple organ‐localized autoimmune diseases, such as thyroiditis and gastritis, spontaneously develop in BALB/c nu/nu (nude) mice receiving embryonic rat thymus grafts (TG) under their renal capsules (TG nude mice). When thyroid was grafted into the rat thymus of TG nude mice, development of autoimmune thyroiditis, but not other diseases, was completely prevented. However, when such mice received thyroid antigen plus complete Freund's adjuvant (CFA), severe autoimmune thyroiditis developed, suggesting that some thyroid‐specific autoreactive T cells migrate into the periphery, but remain unresponsive. Development of autoimmune diseases, including thyroiditis, in TG nude mice was prevented by a single intraperitoneal injection of splenic CD4+ cells from normal BALB/c mice and also from mice with intrathymic thyroid grafts, indicating that thyroid‐specific suppressor T cells are present in normal mice and that such T cells are neither deleted nor inactivated by the intrathymic thyroid grafts, in contrast to autoreactive T cells. Thus clonal deletion in the thymus, and clonal anergy and/or ignorance in the periphery, of autoreactive cells is important to maintain immune tolerance to organ‐specific antigen, but CD4 suppressor T cells may play a more important role in tolerance, and the failure of education of this population may cause autoimmune diseases in the TG nude mouse model.
Generation of MHC class I‐restricted cytotoxic T lymphocytes by expression of a viral protein in muscle cells: antigen presentation by non‐muscle cells
Authors: ULMER J. B. , DECK R. R. , DEWITT C. M. , DONNELLY J. J. , LIU M. A.
Published: November 2003

Expression of reporter genes in muscle cells has been achieved by intramuscular (i.m.) injection of plasmid DNA expression vectors. We previously demonstrated that this technique is an effective means of immunization to elicit both antibodies capable of conferring homologous protection and cell‐mediated immunity leading to cross‐strain protection against influenza virus challenge in mice. These results suggested that expression of viral proteins by muscle cells can result in the generation of cellular immune responses, including cytotoxic T lymphocytes (CTL). However, because DNA has the potential to be internalized and expressed by other cell types, we sought to determine whether or not induction of CTL required synthesis of antigen in non‐muscle cells and, if not, whether transfer of antigen to antigen‐presenting cells from muscle cells may be involved. In the present study, we demonstrate that transplantation of nucleoprotein (NP)‐transfected myoblasts into syngeneic mice led to the generation of NP‐specific antibodies and CTL, and cross‐strain protective immunity against a lethal challenge with influenza virus. Furthermore, transplantation of NP‐expressing myoblasts (H‐2k) intraperitoneally into F1 hybrid mice (H‐2d × H‐2k) elicited NP CTL restricted by the MHC haplotype of both parental strains. These results indicate that NP expression by muscle cells after transplantation was sufficient to generate protective cell‐mediated immunity, and that induction of the CTL response was mediated, at least in part, by transfer of antigen from the transplanted muscle cells to a host cell.