bluuebunny/crossref_metadata_2025 huggingface dataset:
Single agent thalidomide in patients with relapsed or refractory acute myeloid leukaemia
Authors: Thomas Deborah A. , Estey Elihu , Giles Francis J. , Faderl Stefan , Cortes Jorge , Keating Michael , O'Brien Susan , Albitar Maher , Kantarjian Hagop
Published: October 2003

Summary. Thalidomide is a putative anti‐angiogenesis agent that has significant anti‐tumour activity in haematological malignancies with increased bone marrow angiogenesis, including multiple myeloma (MM) and myelodysplastic syndromes (MDS). Increased levels of the mitogen for angiogenesis, vascular endothelial growth factor (VEGF), correlate with worse survival in acute myeloid leukaemia (AML). A phase II trial of thalidomide was conducted in patients with relapsed‐ or refractory‐AML previously treated with cytarabine‐containing regimens. A total of 16 patients with refractory‐ or relapsed‐AML were treated with thalidomide 200–800 mg orally daily (median dose 400 mg daily) for a median of 27 d (range, 3–94 d). Overall, one patient (6%) achieved complete remission (CR) lasting for 36 months, and two patients had a transient reduction in marrow blasts from 8% and 7% to less than 5% in both cases. There was no correlation between reduction in levels of angiogenesis markers and response. Toxicities related to thalidomide were significant, and precluded dose escalation beyond 400 mg orally daily in most patients. Although there appears to be some evidence of biological activity, single agent thalidomide is not an optimal choice of therapy for salvaging patients with relapsed‐ or refractory‐AML. Thalidomide analogues with more potent immunomodulatory activities and more favourable toxicity profiles may offer more promise as anti‐AML therapy.
Bi‐allelic silencing of the Fanconi anaemia gene FANCF in acute myeloid leukaemia
Authors: Tischkowitz Marc , Ameziane Najim , Waisfisz Quinten , De Winter Johan P. , Harris Richard , Taniguchi Toshiyasu , D'Andrea Alan , Hodgson Shirley V. , Mathew Christopher G. , Joenje Hans
Published: October 2003

Summary. Fanconi anaemia (FA) is a chromosomal instability disorder associated with a high risk of acute myeloid leukaemia (AML). Previous work has shown that the AML cell line CHRF‐288, derived from a sporadic AML‐M7 patient, does not express FANCF protein and exhibits a cellular FA phenotype. We show that this phenotype is corrected by a FANCF‐expressing plasmid and that the absence of FANCF protein is explained by hypermethylation of the promoter region of the FANCF gene. As FANCF is localized in a hot‐spot region for somatic hypermethylation (11p15), FANCF silencing might be an early step in sporadic carcinogenesis, including leukaemogenesis.
The polycythaemia rubra vera‐1 gene is constitutively expressed by bone marrow cells and does not discriminate polycythaemia vera from reactive and other chronic myeloproliferative disorders
Authors: Bock Oliver , Serinsöz Ebru , Neusch Michael , Schlué Jerome , Kreipe Hans
Published: October 2003

Summary. Little is known about the expression of the polycythaemia rubra vera‐1 (PRV‐1) gene in bone marrow cells. To determine the expression level of PRV‐1 in the bone marrow, we analysed PRV‐1 quantitatively in Polycythaemia vera, other chronic myeloproliferative disorders, and reactive states. We demonstrated that PRV‐1 was constitutively expressed in both myeloproliferative and reactive states. We concluded that, rather than an upregulation of the PRV‐1 gene in the clonal haematopoiesis of polycythaemia vera, a failure to downregulate PRV‐1 in granulocytes emigrating from the bone marrow might be responsible for the increase of transcripts in peripheral cells.
Differential apoptosis and Fas expression on GPI‐negative and GPI‐positive stem cells: a mechanism for the evolution of paroxysmal nocturnal haemoglobinuria*
Authors: Ismail Medhat M. , Tooze Jennifer A. , Flynn Julie A. , Gordon‐Smith Edward C. , Gibson Frances M. , Rutherford Tim R. , Elebute Modupe O.
Published: October 2003

Summary. Paroxysmal nocturnal haemoglobinuria (PNH) has a dual pathogenesis. PIG‐A mutations generate clones of haemopoietic stem cells (HSC) lacking glycosylphosphatidylinositol (GPI)‐anchored proteins and, secondly, these clones expand because of a selective advantage related to bone marrow failure. The first aspect has been elucidated in detail, but the mechanisms leading to clonal expansion are not well understood. We have previously shown that apoptosis and Fas expression in HSC play a role in bone marrow failure during aplastic anaemia. We have now investigated apoptosis in PNH. Ten patients were studied. Apoptosis, measured by flow cytometry, was significantly higher among CD34+ cells from patients compared with healthy controls. Fas expression was also increased. Cells that were stained for CD34, CD59 and apoptosis showed a significantly lower apoptosis in CD34+/CD59− compared with CD34+/CD59+ cells from the same patient. In three patients, staining for CD34, CD59 and Fas revealed lower Fas expression on CD34+/CD59− cells. Differential apoptosis of CD34+/CD59− HSC may be sufficient in itself to explain the expansion of PNH clones in the context of aplastic anaemia. In addition to demonstrating a basic mechanism underlying PNH clonal expansion, these results suggest further hypotheses for the evolution of PNH, based on the direct or indirect resistance of GPI‐negative HSC to pro‐inflammatory cytokines.
Frequent aberrant promoter hypermethylation of O6‐methylguanine‐DNA methyltransferase and death‐associated protein kinase genes in immunodeficiency‐related lymphomas
Authors: Rossi Davide , Gaidano Gianluca , Gloghini Annunziata , Deambrogi Clara , Franceschetti Silvia , Berra Eva , Cerri Michaela , Vendramin Chiara , Conconi Annarita , Viglio Alessandra , Muti Giuliana , Oreste Pierluigi , Morra Enrica , Paulli Marco , Capello Daniela , Carbone Antonino
Published: October 2003

Summary.  Aberrant promoter hypermethylation is a mechanism of tumour suppressor gene inactivation. We explored aberrant promoter hypermethylation of multiple genes in 88 human immunodeficiency virus (HIV)‐non Hodgkin lymphomas (NHL), 25 post‐transplant lymphoproliferative disorders (PTLD) and five common variable immunodeficiency (CVI)‐related NHL. Twenty‐six of 79 (32·9%) HIV‐NHL, eight of 14 (57·1%) PTLD and two of five (40·0%) CVI–NHL showed aberrant hypermethylation of O6‐methylguanine‐DNA methyltransferase (MGMT). Aberrant hypermethylation of death‐associated protein‐kinase (DAP‐K) occurred in 70 of 84 (83·3%) HIV–NHL, 19 of 25 (72·0%) PTLD and three of five (60·0%) CVI–NHL. These data implicate MGMT and DAP‐K hypermethylation in lymphomagenesis of immunodeficient hosts. In particular, promoter hypermethylation of DAP‐K represents the most frequent molecular alteration yet identified in immunodeficiency‐related lymphomas.
Endothelial cell activation in thrombotic thrombocytopenic purpura (TTP): a prospective analysis
Authors: Romani de Wit Thalia , Fijnheer Rob , Brinkman Herm‐Jan , Kersting Sabina , Hene Ronald J. , Van Mourik Jan A.
Published: October 2003

Summary. It is generally assumed that endothelial cell injury is the primary event in the pathogenesis of thrombotic thrombocytopenic purpura (TTP). In this study, we have determined the extent of vascular perturbation during acute episodes of the disease. We performed a prospective, serial study of nine patients with relapsing TTP during hospitalization and treatment, and assessed the degree of endothelial cell involvement at admission, exacerbation and remission by measurement of von Willebrand factor (VWF) and VWF‐propeptide levels. Measurement of both VWF and its propeptide enabled discrimination between acute and chronic perturbation of the endothelium. Elevated levels of both VWF and propeptide were found at admission. These levels decreased immediately upon plasma exchange therapy. However, plasma VWF and propeptide concentrations did not change, even at the time of acute exacerbation. These observations suggest that endothelial cell activation is not the primary event leading to TTP. Vascular perturbation seems to be a consequence, rather than a cause, of the disease.
Survival after relapse following tandem autotransplants in multiple myeloma patients: the University of Arkansas total therapy I experience
Authors: Fassas Athanasios B.‐T. , Barlogie Bart , Ward Susan , Jagannath Sundar , Vesole David , Mattox Sandy , Siegel David , Muwalla Firas , Zangari Maurizio , Anaissie Elias , Rhee Frits Van , Thertulien Raymond , Lee Choon‐Kee , Desikan Raman , Arzumanian Varant , Mccoy Jason , Tricot Guido
Published: October 2003

Summary. Despite the superiority of high‐dose (compared with standard) treatment in multiple myeloma, relapses still occur. We evaluated relapse patterns, salvage treatments employed and outcome in patients given tandem transplants on our total therapy I protocol. We focused on 146 patients (of 231 enrolled) who received tandem autotransplants ≤12 months apart and survived ≥2 months after the second transplant. With a median follow‐up of 9 years after enrollment, 31 (21%) patients remain in complete or stable partial remission. Ninety‐five (65%) patients received therapy for relapsing myeloma. The median time from the first transplant to relapse was 2·9 years. The median overall survival from relapse was 2·4 years. In one‐quarter (23/95) of cases, the postrelapse interval exceeded the interval from the first transplant to relapse. On multivariate analysis, the presence of any cytogenetic abnormalities [P < 0·001, Hazard Ratio (HR): 3·84] and β‐2 microglobulin levels  > 4 mg/l at relapse (P < 0·001, HR: 2·87) were significant for poor survival after relapse. The median survival after relapse was 5·1, 1·3 and 0·7 years in patients with none (44%), one (46%) and two (10%) poor‐risk factors, respectively. In conclusion, a sizeable fraction of myeloma patients relapsing after tandem autotransplants without poor‐risk features enjoyed meaningful survival prolongation when appropriately treated.
Intracellular signal transduction of interferon on the suppression of haematopoietic progenitor cell growth
Authors: Kato Kouji , Kamezaki Kenjirou , Shimoda Kazuya , Numata Akihiko , Haro Takashi , Aoki Kenichi , Ishikawa Fumihiko , Takase Ken , Ariyama Hiroshi , Matsuda Tadashi , Miyamoto Toshihiro , Nagafuji Koji , Gondo Hisashi , Nakayama Kei‐Ichi , Harada Mine
Published: October 2003

Summary. Interferon (IFN)‐α and IFN‐γ suppress the growth of haematopoietic progenitor cells. IFN‐α activates Janus kinase‐1 (Jak1) and Tyrosine kinase‐2 (Tyk2), followed by the phosphorylation of the signal transducers and activators of transcription, Stat1 and Stat2. IFN‐γ activates Jak1 and Jak2, followed by the activation of Stat1. Activated Stats bind the promoter regions of IFN‐inducible genes. We evaluated the role of Tyk2 and Stat1 in the IFN‐mediated inhibition of haematopoietic progenitor cell growth. While IFN‐α (1000 U/ml) suppressed the number of granulocyte‐macrophage colony‐forming units (CFU‐GM) or erythroid burst‐forming units (BFU‐E) from wild‐type mouse bone marrow cells, this suppression was partially inhibited by a deficiency in Tyk2 and completely inhibited by a deficiency in Stat1. High levels of IFN‐α (10 000 U/ml) suppressed the CFU‐GM or BFU‐E obtained from Stat1‐deficient mice, but did not suppress this growth in cells from Tyk2‐deficient mice. Stat1 was phosphorylated by IFN‐α in Tyk2‐deficient cells, although the level of phosphorylation was weaker than that observed in wild type mice. Thus, the inhibitory signal on haematopoietic progenitor cells mediated by IFN‐α may be transduced by two signalling pathways, one regulated by Tyk2 and the other dependent on Stat1. IFN‐γ also suppressed the number of CFU‐GM or BFU‐E, and this pathway was mediated by IFN‐γ in a Stat1‐dependent manner, independently of Tyk2.
Lymphoproliferative disease in antibody deficiency: a multi-centre study
Authors:   ASSOCIATED STUDY GROUP, GOMPELS M M , HODGES E , LOCK R J , ANGUS B , WHITE H , LARKIN A , CHAPEL H M , SPICKETT G P , MISBAH S A , SMITH J L
Published: October 2003

We have undertaken a retrospective study of antibody deficient patients, with and without lymphoma, and assessed the ability of specific polymerase chain reaction (PCR) primers to determine if the detection of clonal lymphocyte populations correlates with clinical and immunohistochemical diagnosis of lymphoma. We identified 158 cases with antibody deficiency presenting during the past 20 years. Paraffin-embedded biopsy specimens or slides were available for analysis in a cohort of 34 patients. Of these patients, 29 had common variable immunodeficiency, one X-linked agammaglobulinaemia, one X-linked immunoglobulin deficiency of uncertain cause and three isolated IgG subclass deficiency. We have confirmed that lymphoma in antibody deficiency is predominantly B cell in origin. Clonal lymphocyte populations were demonstrated in biopsies irrespective of histology (16/19 with lymphoma and 11/15 without). Isolated evidence of clonality in biopsy material is therefore an insufficient diagnostic criterion to determine malignancy. Furthermore, our data suggest that clonal expansions are rarely the result of Epstein–Barr virus-driven disease.
Evidence for increased expression of regulatory cytokine receptors interleukin-12R and interleukin-18R in common variable immunodeficiency
Authors: McQUAID A , TORMEY V J , TRAFFORD B , WEBSTER A D , BOFILL M
Published: October 2003

We investigated the expression of T helper (Th)1/Th2 regulatory cytokine receptors on lymphocytes from patients with common variable immunodeficiency (CVID), a disorder associated with raised Th1 cytokine production, comparing the results with those from healthy individuals and atopic asthmatics, the latter generally considered to have a Th2-driven disease. We proposed that alterations in some of the relevant receptors might be related to the observed imbalances in Th1/Th2 cytokines. Cells from CVID patients showed an increase in the percentages of CD212 [interleukin (IL)-12Rβ1] cells within the CD4+ CD45RA+ and CD8+ CD45RA+ subsets (24% and 41%, respectively), as compared to CD4+ CD45RA+ and CD8+ CD45RA+ in healthy subjects (6% and 23%, respectivey). Approximately 21% of the CD4+ CD45RA+ naïve cells expressed IL-18Rα, compared with 11% in healthy subjects. In contrast, the cytokine-receptor expression in asthmatics was similar to that of controls. In spite of the above differences, after 72 h of stimulation with anti-CD3 and anti-CD28, cytokine receptor up-regulation was similar in all three groups, with up to 80% of both CD45RA+ and CD45RO+ lymphocytes expressing CD212 (IL-12Rβ1) and IL-18Rα. Approximately 50% of the ‘naïve’, and 25% of the ‘memory’ subpopulations up-regulated IL-12Rβ2. These findings provide further evidence of a polarization towards a Th1 immune response in CVID, the mechanism possibly involving up-regulation of IL-12-mediated pathways.
Interferon-γ and skin test responses of schoolchildren in southeast England to purified protein derivatives fromMycobacterium tuberculosisand other species of mycobacteria
Authors: WEIR R E , FINE P E M , NAZARETH B , FLOYD S , BLACK G F , KING E , STANLEY C , BLISS L , BRANSON K , DOCKRELL H M
Published: October 2003

The immune responses of schoolchildren in southeast England to Mycobacterium tuberculosis and other species of mycobacteria were studied prior to vaccination with bacille Calmette-Guérin (BCG). Data are presented for tuberculin (Heaf) skin test and interferon-γ (IFN-γ) responses to M. tuberculosis purified protein derivative (PPD), and IFN-γ responses to PPDs from eight other environmental mycobacteria, measured in 424 schoolchildren (13–15 years of age). Responses to M. tuberculosis PPD were detected in 27% of schoolchildren by in vitro IFN-γ response and in 20% by the Heaf test. IFN-γ responses were more prevalent to PPDs from species of mycobacteria other than M. tuberculosis, predominantly those of the MAIS complex and M. marinum (45–60% responders). Heaf test and IFN-γ responses were associated (P < 0·001) for M. tuberculosis, MAIS and M. marinum. These findings have implications for appropriate implementation of vaccination against tuberculosis.
Significant elevation of serum levels of eotaxin-3/CCL26, but not of eotaxin-2/CCL24, in patients with atopic dermatitis: serum eotaxin-3/CCL26 levels reflect the disease activity of atopic dermatitis
Authors: KAGAMI S , KAKINUMA T , SAEKI H , TSUNEMI Y , FUJITA H , NAKAMURA K , TAKEKOSHI T , KISHIMOTO M , MITSUI H , TORII H , KOMINE M , ASAHINA A , TAMAKI K
Published: October 2003

Atopic dermatitis (AD) is a chronic and relapsing inflammatory skin disease characterized by the predominant infiltration of T cells, eosinophils and macrophages in lesional skin. Recently, eotaxin-2/CCL24 and eotaxin-3/CCL26 were identified as CC chemokines that signal exclusively via the CCR3 receptor and have eosinophil-selective chemoattractant activity, as does eotaxin/CCL11. We previously reported that serum levels of thymus and activation-regulated chemokine (TARC)/CCL17 and macrophage-derived chemokine (MDC)/CCL22 were correlated with the severity of AD. In this report, we investigated the participation of eotaxin-2/CCL24 and eotaxin-3/CCL26 in AD, first measuring the serum levels of eotaxin-2/CCL24 and eotaxin-3/CCL26 in 30 patients with AD, 20 patients with psoriasis vulgaris and 20 healthy controls. The serum levels of eotaxin-3/CCL26 (but not eotaxin-2/CCL24) were significantly higher in patients with AD than in either healthy controls or patients with psoriasis vulgaris; furthermore, the eotaxin-3/CCL26 levels in patients with moderate and severe AD were significantly higher than eotaxin-3/CCL26 levels in patients with mild AD. The serum eotaxin-3/CCL26 levels tended to decrease after treatment, but there was no significant difference between groups. Moreover, the serum eotaxin-3/CCL26 levels were significantly correlated with the serum TARC/CCL17 and MDC/CCL22 levels, eosinophil numbers in peripheral blood and the scoring AD (SCORAD) index. Our study strongly suggests that serum levels of eotaxin-3/CCL26, but not of eotaxin-2/CCL24, have a notable correlation with disease activity of AD and that eotaxin-3/CCL26, as well as TARC/CCL17 and MDC/CCL22, may be involved in the pathogenesis of AD.
Autoantibodies against mannose-binding lectin in systemic lupus erythematosus
Authors: SEELEN M A , TROUW L A , VAN DER HOORN J W A , FALLAUX-VAN DEN HOUTEN F C , HUIZINGA T W J , DAHA M R , ROOS A
Published: October 2003

In systemic lupus erythematosus (SLE), autoantibodies directed against complement components of the classical pathway, especially against C1q, are associated with severe disease and are of prognostic value for flares of lupus nephritis. Mannose-binding lectin (MBL), the recognition unit of the MBL pathway of complement activation, has structural similarities to C1q. Deficiencies of MBL have been shown to predispose to the development of SLE and to influence the course of the disease. We hypothesized that the presence of autoantibodies to MBL, analogous to autoantibodies to C1q in patients with SLE, may contribute to disease development. The occurrence of anti-MBL autoantibodies was assessed by enzyme-linked immunosorbent assay (ELISA) of 68 serum samples from 20 patients with SLE and in serum from 70 healthy controls. Levels of antibodies directed against MBL were significantly higher in patients with SLE compared to healthy subjects. No significant difference was found between patients with active disease compared to those with inactive disease. While the occurrence of anti-C1q autoantibodies was associated with renal involvement, no such relationship was found for anti-MBL autoantibodies. A significant correlation was found between anti-MBL and anti-C1q antibody levels. The level of anti-MBL antibodies was negatively correlated with MBL–complex activity of circulating MBL. Anti-MBL autoantibodies were of the immunoglobulin G (IgG) isotype and the binding site of IgG anti-MBL was located in the F(ab′)2 portion. We conclude that anti-MBL are present in sera from SLE patients and influence the functional activity of MBL.
Langerhans’ cells in the murine oral mucosa in the inductive phase of delayed type hypersensitivity with 1-chloro-2, 4-dinitrobenzene
Authors: OKAMURA T , MORIMOTO M , YAMANE G , TAKAHASHI S
Published: October 2003

We created a murine model of delayed-type hypersensitivity (DTH) to 1-chloro-2, 4-dinitrobenzene (DNCB). Using this murine model, we compared oral mucosal sensitization and skin sensitization for the difference in reaction during the elicitation phase. Evaluation of sensitizability, using the mouse ear swelling test (MEST) after oral mucosal or skin sensitization, showed that the ear swelling response peaked 24 h after challenge. The optimal induction concentration was 1·0% in both oral mucosal and skin sensitization, resulting in a positive reaction rate of 100%. However, the ear swelling response 24 h after challenge with the optimal concentration of DNCB (1·0%) was significantly lower in oral mucosal than in skin sensitization. We compared the oral mucosal and skin sensitization sites for the number of Langerhans’ cells (LC) and the antigen-presenting capability in the induction phase. The numbers of F4/80+ major histocompatibility complex (MHC) class II+ LC before induction did not differ significantly between the oral mucosa and the skin. After induction, F4/80+ MHC class II+ LC increased in number, but the increase was significantly smaller in the oral mucosa than in the skin. MEST on anti-CD86 antibody-administered mice showed that ear swelling was similarly suppressed after oral mucosal or skin sensitization. In murine models of DTH after oral mucosal sensitization, the number of F4/80+CD86+ LC increased after induction, but the increase was significantly smaller than that in murine models of DTH after skin sensitization. This study showed that, in murine models of DTH, oral mucosal sensitization elicited a weaker reaction than skin sensitization. This was presumably because oral mucosal sensitization induced fewer LC, resulting in lower antigen-presenting capability.
Clinical course and immune response of a renal cell carcinoma patient to adoptive transfer of autologous cytotoxic T lymphocytes
Authors: KAWAI K , SAIJO K , OIKAWA T , MORISHITA Y , NOGUCHI M , OHNO T , AKAZA H
Published: October 2003

The cytotoxic T lymphocyte (CTL) is a promising candidate for an effector cell in adoptive immunotherapy for renal cell carcinoma (RCC). Here we report the clinical course and in vivo immune responses of a RCC patient with bulky retroperitoneal lymph node (RPLN) metastases who received adoptive autologous CTL therapy. A 56-year-old woman diagnosed with RCC with multiple RPLN metastases underwent unilateral nephrectomy. Autologous RCC cells were primary-cultured from surgical specimens. Before addition of peripheral blood mononuclear cells (PBMC) for CTL induction, subconfluent RCC cells were irradiated with 50 Gy. The PBMCs were then cultured on RCC cells in the induction medium supplemented with four kinds of interleukins. The induced CTLs showed the potent killing activity against autologous RCC cells in a typical MHC-class I-restricted manner. The patient received three courses of CTL therapy with a total of 10·2 × 109 cells, and the RPLN mass decreased markedly in size after the second course. Eosinophilia and enhanced CTL inducibility from peripheral blood were observed after CTL administrations. The patient was progression free without further treatment; however, she developed rapidly progressive glomerulonephritis more than 1 year after the last treatment. The patient died of newly developed metastases 27 months after the start of CTL therapy. At autopsy, viable RCC cells were found in multiple metastatic sites. However, only diffuse fibrous tissue was observed in the responding RPLN mass. Apparent histological divergence was observed between primary and metastatic sites.
Involvement of Fcγ receptors andβ2 integrins in neutrophil activation by anti-proteinase-3 or anti-myeloperoxidase antibodies
Authors: Reumaux D , Kuijpers T W , Hordijk P L , Duthilleul P , Roos D
Published: October 2003

We previously described the requirement of tumour necrosis factor-alpha (TNF-α) and the role of β2 integrins in the Fc-gamma receptor IIa (FcγRIIa)-mediated mechanism of neutrophil activation by antiproteinase-3 (anti-PR3) or anti-myeloperoxidase (anti-MPO) antibodies. In the present study, we assessed the involvement of FcγRIIIb by studying the respiratory burst activation of completely FcγRIIIb-deficient neutrophils primed by TNF-αand exposed to anti-PR3 or anti-MPO. Activation of the NADPH oxidase occurred normally in these neutrophils, which indicates that engagement of FcγRIIIb is not essential in our model. Experiments performed with neutrophils from severe leucocyte adhesion deficiency (LAD) patients confirmed that β2 integrins play a pivotal role in this activation. We next studied whether adhesion per se, β2-integrin-mediated adhesion, or β2-integrin ligation without adhesion is necessary or sufficient for this activation. Anti-PR3 or anti-MPO induced an FcγRIIa-dependent burst in TNF-primed neutrophils incubated in wells coated with poly-l-lysine, known to induce β2-integrin-independent adhesion, but this reaction was still inhibited by blocking CD18 antibodies. In a system with granulocyte-macrophage colony-stimulating factor (GM-CSF)-primed neutrophils, which did not enhance adhesion, we measured a similar activation by anti-PR3 or anti-MPO and inhibition by CD18. We also noticed that treatment with the β2-integrin-activating CD18 MoAb KIM185 per se is insufficient for neutrophil activation by anti-PR3 or anti-MPO. We therefore conclude that ligation of β2 integrins rather than adherence per se is essential for this activation, and that TNF-α or GM-CSF is needed for priming but not for adherence.
A recombinant fragment of human SP-D reduces allergic responses in mice sensitized to house dust mite allergens
Authors: STRONG P , TOWNSEND P , MACKAY R , REID K B M , CLARK H W
Published: October 2003

C57Bl6 mice sensitized to Dermatophagoides pteronyssinus and challenged with D. pteronyssinus allergen extract given intranasally followed by treatment with intranasal applications of a 60-kDa truncated, trimeric recombinant form of human SP-D (rfhSP-D) showed a significant reduction in serum IgE, IgG1, peripheral blood eosinophilia and airway hyperresponsiveness compared to saline or bovine serum albumin-treated controls. Intracellular cytokine staining of lung and spleen homogenates showed increases in interleukin (IL)-12 production in lung tissue and normalization of IL-12 and interferon (IFN)-γ in spleen tissue. In previous studies we demonstrated the effectiveness of native SP-D and rfhSP-D in down-regulating allergic responses to allergens of Aspergillus fumigatus. The results reported here indicate that rfhSP-D can suppress the development of allergic symptoms in sensitized mice challenged with allergens of the common house dust mite.
Anti-annexin V antibodies in Takayasu's arteritis: prevalence and relationship with disease activity
Authors: TRIPATHY N K , SINHA N , NITYANAND S
Published: October 2003

Annexin V has an important role in the regulation of apoptosis and antibodies directed against it have been shown to lead to apoptosis of vascular endothelial cells. To evaluate the role of anti-annexin V antibodies (AA5A) in Takayasu's arteritis (TA), we investigated these antibodies in the sera of 66 TA patients, 50 healthy controls and in the follow-up sera of 12 active TA patients by enzyme-linked immunosorbent assay. The AA5A-positive patients were analysed further for the presence of anti-endothelial cell antibodies (AECA) and anticardiolipin antibodies (ACLA) to determine the relationship of AA5A with these autoantibodies. AA5A were observed in 36% (24/66) of the patients versus 6% (3/50) of the controls (P < 0·001) and in 53% (19/36) of patients with active TA versus 17% (5/30) of those with inactive disease (P < 0·01). Levels of AA5A were also observed to be significantly higher in patients with TA compared to controls (0·557 ± 0·362 versus 0·259 ± 0·069; P < 0·0001) and in patients with active disease compared to those with inactive disease (0·700 ± 0·403 versus 0·385 ± 0·205; P < 0·0001). In the follow-up study, 6/12 patients who became inactive during follow-up also showed normalization of AA5A levels. AECA and ACLA were detected in 54% (13/24) and 12% (3/24) of the AA5A-positive patients, respectively. Our results show that a significant proportion of TA patients have AA5A, which exhibit an association with AECA and because they have a correlation with disease activity thus appear to be involved in the disease pathogenesis.
Modulation of the expression of M150 on macrophages by Th1/Th2 cytokines and co-stimulatory molecules CD40, B7-1, B7-2 and ICAM-1
Authors: SUVAS S , VOHRA H , AGREWALA J N
Published: October 2003

M150 is an 150-kDa protein associated with the surface of macrophages and is responsible chiefly for the activation of Th1 cells. It is a unique subset of the lysosome-associated membrane protein-1 glycoprotein and its co-stimulatory activity depends on its post-translational modification, which has a distinct glycosylation pattern restricted to macrophages. In the present study, we have observed that M150 is expressed constitutively on peritoneal but not splenic macrophages isolated from mice of different genetic backgrounds: Balb/c, C57BL/6 and C3He. However, M150 was expressed not only on peritoneal but also on splenic macrophages of non-obese diabetic (NOD) mice. Expression on splenic macrophages was induced by culture with lipopolysaccharide (LPS). Expression could also be significantly up-regulated by interferon (IFN)-γ and granulocyte-macrophage colony stimulating factor (GM-CSF) but was inhibited by interleukin (IL)-10; IL-4 exhibited no effect. Further, cross-linking of B7-2, CD40, ICAM-1 but not B7-1 enhanced the level of M150 significantly. IFN-γ and GM-CSF acted synergistically with CD40. The significance of these findings is that cytokines IFN-γ, GM-CSF and IL-10 and the co-stimulatory molecules B7-2, CD40 and ICAM-1 can regulate the expression of M150 on macrophages.
No strong relationship between mannan binding lectin or plasma ficolins and chemotherapy-related infections
Authors: KILPATRICK D C , MCLINTOCK L A , ALLAN E K , COPLAND M , FUJITA T , JORDANIDES N E , KOCH C , MATSUSHITA M , SHIRAKI H , STEWART K , TSUJIMURA M , TURNER M L , FRANKLIN I M , HOLYOAKE T L
Published: October 2003

Chemotherapy causes neutropenia and an increased susceptibility to infection. Recent reports indicate that mannan-binding lectin (MBL) insufficiency is associated with an increased duration of febrile neutropenia and incidence of serious infections following chemotherapy for haematological malignancies. We aimed to confirm or refute this finding and to extend the investigation to the plasma ficolins, P35 (L-ficolin) and the Hakata antigen (H-ficolin). MBL, L-ficolin and H-ficolin were measured in 128 patients with haematological malignancies treated by chemotherapy alone or combined with bone marrow transplantation. Protein concentrations were related to clinical data retrieved from medical records. MBL concentrations were elevated compared with healthy controls in patients who received chemotherapy, while L-ficolin concentrations were decreased and H-ficolin levels were unchanged. There was no correlation between MBL, L-ficolin or H-ficolin concentration and febrile neutropenia expressed as the proportion of neutropenic periods in which patients experienced fever, and there was no relation between abnormally low (deficiency) levels of MBL, L-ficolin or H-ficolin and febrile neutropenia so expressed. Patients with MBL ≤ 0·1 µg/ml had significantly more major infections than no infections within the follow-up period (P < 0·05), but overall most patients had signs or symptoms of minor infections irrespective of MBL concentration. Neither L-ficolin nor H-ficolin deficiencies were associated with infections individually, in combination or in combination with MBL deficiency. MBL, L-ficolin and H-ficolin, independently or in combination, did not have a major influence on susceptibility to infection in these patients rendered neutropenic by chemotherapy. These results cast doubt on the potential value of MBL replacement therapy in this clinical context.
Enhancement of Th2 pathways and direct activation of B cells by the gingipains ofPorphyromonas gingivalis
Authors: YUN L W P , DECARLO A A , COLLYER C , HUNTER N
Published: October 2003

Porphyromonas gingivalis cysteine proteinases (gingipains) have been associated with virulence in destructive periodontitis, a disease process that has been linked with Th2 pathways. Critical in maintaining Th2 activity is the response of B lymphocytes to environmental interleukin (IL)-4, a cytokine that also counteracts Th1-cell differentiation. Here we demonstrate that while the gingipains effectively degrade interleukin (IL)-4 under serum-free conditions, limited hydrolysis was observed in the presence of serum even after prolonged incubation. Gingipains up-regulated CD69 expression directly in purified peripheral blood B cell preparations. Further, the induction of IL-4 receptor expression on B cells by gingipains correlates with B cell activation, which is also manifested by a mitogenic response. These results suggest that the gingipains of P. gingivalis act during the early stage of B-cell growth as a competence signal, whereby sensitized B cells might become more responsive to further challenge in the disease-susceptible individual.
Contrasting activity of cytosin–guanosin dinucleotide oligonucleotides in mice with experimental colitis
Authors: OBERMEIER F , DUNGER N , STRAUCH U G , GRUNWALD N , HERFARTH H , SCHÖLMERICH J , FALK W
Published: October 2003

Intestinal inflammation in inflammatory bowel disease (IBD) and experimental models of colitis is characterized by a dysregulated intestinal immune response with elevated levels of Th1 cytokines. The luminal flora has been implicated as a major factor contributing to the initiation and perpetuation of inflammation in experimental colitis by mechanisms not known. Bacterial DNA contains unmethylated cytosin–guanosin dinucleotides (CpG) which strongly activate Th1-mediated immune responses. To test whether these CpG-motifs modulate intestinal inflammation we treated mice with dextran sulphate sodium (DSS)-induced colitis with CpG-containing oligodeoxynucleotides (CpG-ODN). CpG-ODN given after the onset of DSS colitis aggravated the disease, as indicated by a significantly increased loss of body weight and a 30% increase of the histological score. Further, we found a severe increase of proinflammatory cytokines (interleukin (IL)-6: 40-fold; interferon (IFN)-γ : 11-fold). In a pretreatment setting CpG-ODN reduced weight loss significantly and reduced intestinal inflammation by 45%. Colonic IFN-γ and IL-6 mRNA levels were reduced by 75%, and IL-10 was elevated by 400% compared to controls. The prophylactic CpG-effect was not imitated by IL-12 because IL-12 pretreatment was not protective. In time-course experiments, CpG-ODN pretreatment over 5 days resulted in a tolerance effect concerning its IFN-γ-inducing quality, and during the following days of colitis induction IL-10 secretion from mesenterial lymph node cells was elevated compared to controls. Therefore, the prophylactic effect of CpG-ODN might be explained by its tolerizing effect and/or the increased ability for IL-10 production during the consecutive intestinal inflammation.
HLA-B27-restricted T cells from patients with ankylosing spondylitis recognize peptides from B*2705 that are similar to bacteria-derived peptides
Authors: Frauendorf E , Von Goessel H , May E , Märker-Hermann E
Published: October 2003

Ankylosing spondylitis (AS) is an inflammatory systemic disease affecting the spine, sacroiliacal and peripheral joints. Although the aetiology of AS remains unknown, the strong association with the HLA-B27 allele might reflect directly a detrimental effect of the HLA-B27 molecule itself, resulting from its potential capability to present ‘arthritogenic’ peptides to CD8+ T cells. Because some forms of SpA are triggered by enterobacterial infection, such arthritogenic peptides might originate from autologous and/or bacterial proteins triggering cross-reactive CD8+ T cell clones. Intriguingly, two peptides from the second extracellular domain of HLA-B*2705 share sequence homologies with several enterobacterial antigens, exhibit the HLA-B27-binding-motif, and are presented by HLA-B*2705 itself. The objective of this study was to examine the clonal T cell reactivity against these peptides in patients with AS. To this end, we screened peripheral blood lymphocytes (PBL) of 26 patients with AS and 24 healthy donors for TNF-α-producing cells using ELISPOT assays. PBL and synovial fluid-derived lymphocytes (SFL) of peptide-responsive patients were then stimulated and cultured with the relevant peptide and control peptides in vitro. Antigen-specific T cell lines (TCL) were identified by standard chromium release assays. Clonal analysis was performed subsequently applying TCRB-CDR3 spectratyping. Among eight peptides tested, only the HLA-B27 168–176 peptide LRRYLENGK was recognized by PBL from B27+ AS patients but not from B27+ healthy controls (P = 0·001). LRRYLENGK-specific T cell clones used preferentially the TCRBV5S1 and the BV14 segment. These results suggest that an HLA-B27-derived peptide with homology to bacterial peptides may play a role in AS.
The immunosuppressive drug mycophenolate mofetil impairs the adhesion capacity of gastrointestinal tumour cells
Authors: LECKEL K , BEECKEN W-D , JONAS D , OPPERMANN E , COMAN M C , BECK K-F , CINATL J , HAILER N P , AUTH M K H , BECHSTEIN W O , SHIPKOVA M , BLAHETA R A
Published: October 2003

Immunosuppression correlates with the development and recurrence of cancer. Mycophenolate mofetil (MMF) has been shown to reduce adhesion molecule expression and leucocyte recruitment into the donor organ. We have hypothesized that MMF might also prevent receptor-dependent tumour dissemination. Therefore, we have investigated the effects of MMF on tumour cell adhesion to human umbilical vein endothelial cells (HUVEC) and compared them with the effects on T cell–endothelial cell interactions. Influence of MMF on cellular adhesion to HUVEC was analysed using isolated CD4+ and CD8+ T cells, or WiDr colon adenocarcinoma cells as the model tumour. HUVEC receptors ICAM-1, VCAM-1, E-selectin and P-selectin were detected by flow cytometry, Western blot or Northern blot analysis. Binding activity of T cells or WiDr cells in the presence of MMF were measured using immobilized receptor globulin chimeras. MMF potently blocked both T cell and WiDr cell binding to endothelium by 80%. Surface expression of the endothelial cell receptors was reduced by MMF in a dose-dependent manner. E-selectin mRNA was concurrently reduced with a maximum effect at 1 µm. Interestingly, MMF acted differently on T cells and WiDr cells. Maximum efficacy of MMF was reached at 10 and 1 µm, respectively. Furthermore, MMF specifically suppressed T cell attachment to ICAM-1, VCAM-1 and P-selectin. In contrast, MMF prevented WiDr cell attachment to E-selectin. In conclusion, our data reveal distinct effects of MMF on both T cell adhesion and tumour cell adhesion to endothelial cells. This suggests that MMF not only interferes with the invasion of alloactivated T cells, but might also be of value in managing post-transplantation malignancy.
Circulating lymphocyte subsets linked to intracellular cytokine profiles in normal humans
Authors: MATSUI M , ARAYA S , WANG H-Y , ONAI N , MATSUSHIMA K , SAIDA T
Published: October 2003

To determine whether there is an association between intracellular cytokine profiles and the expression of surface antigens, we performed a simultaneous flow cytometric analysis of these laboratory parameters in 11 healthy volunteers. Peripheral blood lymphocytes were double-stained for CD4 or CD8, as well as CD11a, CD25, CD26, CD29 and CD45RA or the chemokine receptors CCR3, CCR4, CCR5 or CXCR3. Portions of the cell samples were cultured for 4 h in the presence of 1 µm monensin and 20 µg/ml brefeldin A with or without stimulation by phorbol myristate acetate plus ionomycin for the detection of intracellular interferon-γ (IFN-γ), interleukin-2 (IL-2), tumour necrosis factor (TNF)-α, and IL-4. As a result, CD4+CD29high helper inducer T cells were closely associated with IFN-γ and TNF-α producing CD4+ cells, while CD4+CXCR3+ cells showed a negative correlation with IL-4-producing cells, suggesting that both of these CD4+ subsets consist mainly of Th1 cells. In contrast, CD4+CD45RA+ cells were correlated inversely with IFN-γ and TNF-α-producing cells, and CD8+CD11ahigh killer effector and total CCR5+ cells showed an inverse correlation with IL-2 producing cells, suggesting an immunoregulatory role for these three subsets in non-pathological conditions. Therefore, monitoring of lymphocyte subsets that express functional surface antigens could provide additional information concerning immune deviation, as assessed by the production of Th1/Th2 type cytokines. Further, this type of combined study may provide clues for the pathogenesis of immune-mediated disorders.
Effects of streptozotocin on autoimmune diabetes in NOD mice
Authors: KOULMANDA M , QIPO A , AUCHINCLOSS H , SMITH R N
Published: October 2003

Non-obese diabetic (NOD) mice develop autoimmunity that destroys their native beta cells causing diabetes. Their autoimmunity will also destroy syngeneic transplanted islets and transfer both autoimmunity and diabetes via spleen cells to non-diabetic mice. In this report, we studied the effects of streptozotocin (STZ) on the autoimmune diabetes in NOD mice. We transplanted NOD.SCID islets into three groups of NOD mice: (1) spontaneously diabetic NOD mice (NOD-sp.); (2) prediabetic NOD mice made diabetic by streptozotocin (NOD-stz); and (3) diabetic NOD mice also treated with streptozotocin (NOD-sp./stz). In the first group, the transplants were rejected within 3 weeks. In the second and third groups, the transplants survived indefinitely. Alloxan, a drug similar to streptozotocin, did not have the same effect as streptozotocin. The ability of streptozotocin to prevent diabetes in young NOD mice was reversed by anti-CD8 antibody treatment but not by anti-CD4 treatment. Streptozotocin also made spleen cells from diabetic NOD mice less effective transferring diabetes. These results indicate that streptozotocin treatment both prevents and reverses the islet destructive autoimmunity in NOD mice. We postulate that the effects of streptozotocin treatment may be mediated in part by regulatory T cells.
Role of IFN-α/βand IL-12 in the activation of natural killer cells and interferon-γproduction during experimental infection withTrypanosoma cruzi
Authors: UNE C , ANDERSSON J , ÖRN A
Published: October 2003

Control of Trypanosoma cruzi infection depends largely upon the production of interferon (IFN)-γ. During experimental infection this cytokine is produced early, mainly by natural killer (NK) cells and later by T cells. As NK cells have been reported to participate in defence against T. cruzi, it is of importance to study the regulation of NK cell functions during infection with the parasite. Several innate cytokines regulate NK cell activity, among them being interferon (IFN)-α and IFN-β (type 1 IFNs) and interleukin (IL)-12, which have all been reported to be involved in protection against T. cruzi. The role of these cytokines in regulation of NK cell functions and disease outcome were studied by infection of mutant mice lacking the IFN-α/β receptor (IFNα/βR–/–) or IL-12 (IL-12–/–) with T. cruzi. IFNα/βR–/– mice were unable to activate the cytotoxic response but produced IFN-γ, and were not more susceptible than controls. IL-12–/– mice were extremely susceptible and failed to produce T cell-derived IFN-γ and nitric oxide (NO), although NK cytotoxicity was induced. The results indicate that IL-12 protects against T. cruzi by initiating T cell-mediated production of IFN-γ, but that endogenous IFN-α/β and NK cell cytotoxicity are not of major importance in defence.
Increased serum transforming growth factor-β1 in human colorectal cancer correlates with reduced circulating dendritic cells and increased colonic Langerhans cell infiltration
Authors: HUANG A , GILMOUR J W , IMAMI N , AMJADI P , HENDERSON D C , ALLEN-MERSH T G
Published: October 2003

Cancer-related cytokines may interfere with the differentiation and migration of dendritic cells (DCs) and with the associated up-regulation of co-stimulatory molecules in vitro. We determined whether cytokines affected the distribution and activation of DCs in patients with colorectal cancer by measuring the levels of serum cytokines [transforming growth factor (TGF)-β1 and vascular endothelial growth factor (VEGF)], DC numbers and phenotype from peripheral blood and mesenteric lymph nodes draining the cancer, and the infiltration of DCs into colorectal cancer. A significant increase in the serum level of TGF-β1 correlated with a significant reduction in the level of circulating DCs in cancer patients that was associated with an increased infiltration of Langerhans cells into colorectal mucosa. The prevalence but not intensity of co-stimulatory molecule expression in circulating and mesenteric lymph node DCs was reduced in patients with colorectal cancer compared to patients with inflammatory bowel conditions. There was no correlation between co-stimulatory molecule expression and serum TGF-β1. Thus the circulating DC depletion in colorectal cancer could be explained by a TGF-β1-related DC redistribution from the circulation into the colorectal cancer and adjacent mucosa where DC levels were increased. There was an impairment of DC activation within colorectal cancer that was not related to serum level of cytokines.
Avidity progression of dietary antibodies in healthy and coeliac children
Authors: SAALMAN R , DAHLGREN U I , FÄLLSTRÖM S P , HANSON L Å , AHLSTEDT S , WOLD A E
Published: October 2003

In most individuals minute amounts of food proteins pass undegraded across the intestinal mucosa and trigger antibody formation. Children with coeliac disease have enhanced antibody production against gliadin as well as other dietary antigens, e.g. β-lactoglobulin, in cow's milk. Antibody avidity, i.e. the binding strength between antibody and antigen, often increases during antibody responses and may be related to the biological effectiveness of antibodies. The aim of the present study was to determine the avidity of serum IgG antibodies against β-lactoglobulin and gliadin in healthy children during early childhood and compare these avidities to those found in children with coeliac disease. The average antibody avidity was analysed using a thiocyanate elution assay, whereas the antibody activity of the corresponding sera was assayed by ELISA. The avidity of serum IgG antibodies against β-lactoglobulin as well as gliadin increased with age in healthy children, even in the face of falling antibody titres to the same antigens. Children with untreated coeliac disease had IgG anti-β-lactoglobulin antibodies of significantly higher avidity than healthy children of the same age, and the same trend was observed for IgG antigliadin antibodies. The present data suggest that the avidities of antibodies against dietary antigens increase progressively during early childhood, and that this process seems to be accelerated during active coeliac disease.