bluuebunny/crossref_metadata_2025 huggingface dataset:
Antisense oligonucleotides complementary to immunoglobulin sequences of BCL‐2/immunoglobulin fusion transcript induce apoptosis of t(14;18) lymphoma cells
Authors: Smith Mitchell R. , Xie Tao , Zhou Zhao‐zong , Joshi Indira
Published: July 2010

Antisense oligodeoxyribonucleotides directed at the bcl‐2 translational start site downregulate bcl‐2 and inhibit growth of the t(14;18)‐positive lymphoma line WSU‐FSCCL. Non‐specific downregulation of bcl‐2 expression is expected to be toxic to normal cells as well. The t(14;18) translocation results in a fusion transcript containing the entire bcl‐2 coding sequence with a 3′ breakpoint fused to the immunoglobulin JH region and the cμ heavy chain. We postulated that these immunoglobulin sequences would be a specific target for downregulation of the fusion gene. Here, we have demonstrated that antisense oligodeoxyribonucleotides targeted to immunoglobulin cμ sequences downregulate bcl‐2 protein expression and induce apoptosis of WSU‐FSCCL cells. Inhibiting growth of malignant cells by targeting non‐oncogenic sequences other than breakpoints of fusion transcripts expands the potential for tumour‐specific genetic manipulation.
Hypofibronectinaemia and severity of sickle cell anaemia in Nigeria
Authors: Emeribe A. O. , Udoh A. E. , Etukudoh M. H. , Okany C. C. , Kehinde M. , Catty D. , Raykundalia C.
Published: July 2010

Plasma fibronectin levels in sickle cell anaemia and apparently healthy Nigerians were investigated to determine any correlation with disease severity. A cheaper in‐house plasma fibronectin assay was also developed that could be adapted for use in Africa and elsewhere. Plasma fibronectin assay was concurrently carried out using the newly developed inhibition enzyme‐linked immunosorbent assay (ELISA) and a commercial competitive binding ELISA. The in‐house assay compared favourably with that of the commercial kit. The mean plasma fibronectin levels in sickle cell anaemia subjects were significantly lower than that of control subjects (P < 0·001). Plasma fibronectin concentration could therefore be useful in assessing the severity of sickle cell anaemia.
Primary human keratinocytes as targets in predicting acute graft‐versus‐host disease following HLA‐identical bone marrow transplantation
Authors: van Dijk Astrid M. C. , Kessler Floortje L. , Verdonck Leo F. , Stadhouders‐Keet Simone A. E. , van Lier Rene A. W. , de Gast Gijsbert C. , Otten Henny G.
Published: July 2010

Graft‐versus‐host‐disease (GVHD) remains a major problem following allogeneic bone marrow transplantation (BMT) and manifests itself mainly by damage to epithelial cells of the skin, gut and bile ducts. Reliable tests to predict GVHD are lacking. We developed an assay in which donor T cells are stimulated by patient keratinocytes (KCs), compared that with stimulation by patient peripheral blood mononuclear cells (PBMCs) and studied the relationship to GVHD. In 27 patients undergoing HLA‐identical BMT for haematological malignancies, donor T‐cell reactivity was determined as the helper T‐lymphocyte precursor (HTLp) frequency against host PBMCs (25 patient‐donor pairs) and host KCs (20 patient‐donor pairs). KCs were obtained by shave biopsies and cultured with interferon (IFN)‐γ to induce HLA class II expression. In assays using patient KCs and donor T cells, anti‐CD28 antibody was added to compensate for the lack of co‐stimulatory molecules on KCs. Results were related to the occurrence of GVHD. As BMTs were performed with partially T cell‐depleted grafts, GVHD was limited to grade 0 (five patients), grade I (seven patients) and grade II (12 patients). No differences were found in donor T‐cell reactivity to patient PBMCs, as expressed as HTLp frequency in patients with or without GVHD. However, significant differences (P < 0·01) were found in donor T‐cell reactivity to patient KCs when comparing patients with and without GVHD. Donor HTLp frequencies against patient KCs give a better prediction of GVHD than those against patient haemopoietic cells following HLA‐identical BMT, which may indicate that at least some minor non‐HLA histocompatibility antigens present on KCs are different from those on haemopoietic cells.
Phenotypic and molecular diversity of haemoglobin H disease: a Greek experience
Authors: Kanavakis Emmanuel , Papassotiriou Ioannis , Karagiorga Markissia , Vrettou Christina , Metaxotou‐Mavrommati Anna , Stamoulakatou Alexandra , Kattamis Christos , Traeger‐Synodinos Joanne
Published: July 2010

Haemoglobin H (Hb H) disease is the severest form of α‐thalassaemia compatible with post‐natal life and occurs when α‐thalassaemia mutations interact to reduce α‐globin synthesis to levels approximately equivalent to the output of a single α‐globin gene. Hb H disease has variable clinical expression, mainly related to underlying genotypes. The spectrum of α‐thalassaemia determinants in Greece appears greater than in any other population studied and, in 75 Greek Hb H disease patients, we found 12 α‐thalassaemia mutations interacting to produce 15 Hb H disease genotypes. Evaluation of haematological, biochemical and clinical findings, and correlation with genotypes, defined genetic predictors of disease severity and factors involved in disease progression. In accordance with previous reports, patients with non‐deletion α‐thalassaemia mutations had more severe clinical expression. Additionally, we found that all patients with the most severe phenotypes had α‐thalassaemic globin variants. Phenotypic severity was not simply related to the degree of α‐globin deficiency: high Hb H levels were found to exacerbate anaemia by negatively influencing tissue oxygenation, and both Hb H and α‐thalassaemic haemoglobin variants appear to reduce red cell survival within the bone marrow and circulation. Together with the long‐term follow‐up in many patients, this report provides comprehensive information for management of Hb H disease and appropriate family counselling.
Engraftment of T‐cell‐depleted allogeneic haematopoietic stem cells using a reduced intensity conditioning regimen
Authors: Craddock C. , Bardy P. , Kreiter S. , Johnston R. , Apperley J. , Marks D. , Huber C. , Kolbe K. , Goulding R. , Lawler M. , Goldman J. , Hughes T. , Derigs G.
Published: July 2010

Graft‐versus‐host disease (GVHD) remains a significant complication in patients undergoing allogeneic stem cell transplantation (SCT) using a reduced intensity conditioning regimen. Although T‐cell depletion (TCD) reduces the risk of GVHD after a myeloablative conditioning regimen, it is associated with an increased risk of graft failure. We have therefore examined whether TCD compromises engraftment using a fludarabine‐based conditioning regimen. Fifteen patients have been transplanted using such a regimen of whom 13 underwent ex vivo TCD. All but one patient demonstrated durable engraftment and no patient receiving a TCD product developed severe GVHD. Thus, TCD may play a role in GvHD prophylaxis using such regimens.
Oncogene mutation and prognosis in the myelodysplastic syndromes
Authors: Padua R. A. , West R. R.
Published: July 2010

In a series of myelodysplastic syndrome patients, mutational status (particularly RAS mutation) was found to be prognostic for survival, independently of four previously reported scoring systems (Bournemouth, Sanz, Lille, International).
Age‐dependent variations in the expression of PLC isoforms upon mitogenic stimulation of peripheral blood T cells from healthy donors
Authors: Di Pietro Roberta , Miscia Sebastiano , Cataldi Amelia , Rana Rosalba
Published: July 2010

The activation of phosphoinositide‐specific phospholipase C (PLC) is one of the early responses to various growth factors and it is known that these functions are altered with ageing. In the present investigation, the expression and cellular distribution of PLC isoenzymes in immunocompetent T lymphocytes from a test group of healthy individuals over 65 years old and a comparison group of healthy donors below 30 years old were compared using Western blot and confocal microscopy. All lymphocyte samples responded biochemically to phytohaemagglutinin (PHA) stimulation, as shown by at least 10‐fold increases in the β3 isoform and nuclear translocation of the β1, β2, β3, β4, γ1 and γ2 isoforms; however, consistent differences in the expression of the β2 isoform in unstimulated T cells and of the β2 and γ2 isoforms in stimulated T cells suggest that altered functions of the PLC pathway may have an age‐dependent impact on signal transduction events.
Is there a graft‐versus‐leukaemia effect in the absence of graft‐versus‐host disease in patients undergoing bone marrow transplantation for acute leukaemia?
Authors: Ringdén O. , Labopin M. , Gorin N. C. , Schmitz N. , Schaefer U. W. , Prentice H. G. , Bergmann L. , Jouet J. P. , Mandelli F. , Blaise D. , Fouillard L. , . Frassoni F ,   for the Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation
Published: July 2010

During a 13‐year period, 5200 autografts, 1039 HLA‐identical sibling transplants without acute or chronic graft‐vs.‐host disease (GVHD) and 67 twins were reported to the European Group for Blood and Marrow Transplantation EBMT. Follow‐up time was a median of 32 months. Diagnoses were acute myeloid leukaemia (AML, 4521) and acute lymphoblastic leukaemia (ALL, 1785) in first complete remission. The probability of relapse at 5 years was 51 ± 1% in the autografts, 45 ± 8% in the twins and 34 ± 2% among the HLA‐identical siblings (auto vs. sibs,P < 0·0001). In multivariate analyses, the following factors were significantly associated with an increased risk of relapse: ALL vs. AML M3 [relapse rate (RR) 2·29,P < 0·0001], AML non‐M3 vs. AML M3 (RR 1·8,P < 0·0001), autograft vs. sibling transplant (RR 1·76,P < 0·0001), interval diagnosis to transplantation < 261 d (RR 1·45,P < 0·001) and other conditioning vs. total body irradiation (RR 1·16,P = 0·001). Transplant‐related mortality was the same in the three groups at approximately 10% at 2 years. Five‐year leukaemia‐free survival was 42 ± 1% in the autografts, 44 ± 8% in the twins and 58 ± 2% among the siblings (auto vs. sibs,P < 0·0001). The factors significant for relapse were also significant in multivariate analyses for leukaemia‐free survival. In addition, children had a significantly better leukaemia‐free survival than adults (RR 0·82,P < 0·0001). Recipients of bone marrow from HLA‐identical siblings without GVHD had a lower risk of relapse and a better leukaemia‐free survival than recipients of autografts. This may be as a result of a graft‐vs.‐leukaemia effect in the absence of GVHD.
DDAVP (desmopressin; 1‐deamino‐cys‐8‐d‐arginine‐vasopressin) treatment in children with haemophilia B
Authors: Ehl S. , Severin T. , Sutor A. H.
Published: July 2010

We tested the response to desmopressin (1‐deamino‐cys‐8‐d‐arginine‐vasopressin; DDAVP) in four patients with haemophilia B [factor IX (F IX) at diagnosis 1·4–5%]. The activated partial thromboplastin time (aPTT) was significantly shortened in all patients. Although there was an up to 1·4‐fold increase in F IX levels in three patients, maximal F IX activity remained below 10%. Much more prominent were the increases in F VIII (three‐ to fourfold), in von Willebrand factor antigen (VWF:Ag; 2·5‐fold) and particularly in VWF collagen‐binding activity (VWF:CBA; fivefold). These changes were reflected by the prophylactic efficacy of DDAVP for dental surgery. After pretesting, DDAVP could be a useful drug for reducing the need for plasma products for prevention of minor surgical bleeding in patients with mild to moderate haemophilia B.
RUNX1 gene mutation in primary myelodysplastic syndrome – the mutation can be detected early at diagnosis or acquired during disease progression and is associated with poor outcome
Authors: Chen Chien‐Yuan , Lin Liang‐In , Tang Jih‐Luh , Ko Bo‐Sheng , Tsay Woei , Chou Wen‐Chien , Yao Ming , Wu Shang‐Ju , Tseng Mei‐Hsuan , Tien Hwei‐Fang
Published: July 2010

Mutations of Runt‐related transcription factor 1 (RUNX1) have been detected in patients with myelodysplastic syndrome (MDS). However, the prognostic implication of RUNX1 mutations in primary MDS is limited. The stage of the disease at which the mutations are acquired and whether they persist during the disease course also remain unclear. We analysed mutations of RUNX1 exons 3–8 in 132 patients with primary MDS and correlated the results with clinical features. Serial studies were performed during the follow‐up period. Sixteen patients (12%) had RUNX1 mutations at the time of diagnosis. All RUNX1 mutations that were detected at diagnosis remained unchanged during the clinical course. Two other patients acquired RUNX1 mutations at leukaemic transformation 34 months and 35 months after the diagnosis of MDS. Patients with RUNX1 mutations at diagnosis had higher neutrophil counts and higher frequency of −7/7q deletion than those without. Furthermore, RUNX1 mutation was closely associated with a short overall survival (P = 0·039). This is the first report to demonstrate that RUNX1 mutation can not only be detected early at diagnosis but also acquired during disease progression and is associated with poor prognosis in patients with primary MDS. It may play a role in the development and progression of a subset of primary MDS.
Ciclosporin and plasma exchange in thrombotic thrombocytopenic purpura: long‐term follow‐up with serial analysis of ADAMTS13 activity
Authors: Cataland Spero R. , Jin Ming , Lin Shili , Kennedy Melanie S. , Kraut Eric H. , George James N. , Wu Haifeng M.
Published: July 2010

We hypothesized that ciclosporin (CSA) as adjunct to plasma exchange (PE) improves the efficacy of PE in idiopathic thrombotic thrombocytopenic purpura (TTP) via suppression of the antibody inhibitor of ADAMTS13. Our preliminary findings with CSA and PE as the upfront treatment of TTP suggested that the addition of CSA to PE significantly decreased the exacerbation (disease recurrence within 30 d of the last PE) rates compared to a cohort that received corticosteroids and PE as their upfront therapy of TTP. We present an updated analysis with long‐term follow‐up of 18 patients with idiopathic TTP treated with concurrent CSA and PE with analysis of serial measurements of ADAMTS13 activity, antigen and inhibitor concentration in the context of clinical outcome data. Overall, 16/18 (89%) patients achieved remission, similar to historical remission rates in idiopathic TTP with PE with only one patient suffering an exacerbation. Clinical responses correlated with improvements in ADAMTS13 activity and suppression of the antibody inhibitor of ADAMTS13. These data suggest that the efficacy of CSA is at least in part related to its suppression of the antibody inhibitor of ADAMTS13 and a subsequent improvement in ADAMTS13 activity and antigen.
Developments in the therapeutic use of erythropoiesis stimulating agents
Authors: Jelkmann Wolfgang
Published: July 2010

Recombinant human erythropoietin (rHuEPO) has become the standard therapy for treatment of the anaemia of chronic kidney disease (CKD) during the past two decades. In addition, rHuEPO can be indicated for the treatment of cancer patients on chemotherapy and surgical patients to avoid allogeneic red blood cell transfusion. This review first describes recent attempts in developing rHuEPO congeners (mutated and pegylated rHuEPOs) and mimetics with prolonged half‐lives and improved application requirements. Secondly, the pathophysiological background of the regulatory guideline, that blood haemoglobin levels in anaemic CKD or cancer patients on chemotherapy should not be raised above the target value of 120 g/l, is discussed. Finally, potential novel indications are considered for the use of rHuEPO and its analogues as pleiotropic cytoprotectant agents for cardio‐, nephro‐, hepato‐ and neuroprotection.
Current status of globin gene therapy for the treatment of β‐thalassaemia
Authors: Lisowski Leszek , Sadelain Michel
Published: July 2010

β‐Thalassaemia major is a congenital anaemia for which there is presently no curative therapy other than allogeneic haematopoietic stem cell transplantation. This therapeutic option, however, is not available to most subjects for whom there is no available human leucocyte antigen‐matched bone marrow donor. The transfer of a regulated globin gene in autologous haematopoietic stem cells is therefore a direly needed alternative treatment. This strategy, simple in principle, raises major challenges in terms of controlling transgene expression, which ideally should be erythroid‐specific, differentiation‐ and stage‐restricted, elevated, position independent, and sustained over time. Using lentiviral vectors, it has been demonstrated that an optimized combination of proximal and distal transcriptional control elements permits lineage‐specific and elevated β‐globin expression in vivo, resulting in the correction of anaemia and secondary organ damage in β‐thalassaemic mice. Several groups have extended these findings to various models of β‐thalassaemia and sickle cell disease. Different globin vectors, however, do not express β‐globin at the same level, and accordingly require the delivery of markedly different vector copy numbers to correct anaemia. Insulators are under investigation to assess whether they might enhance globin gene expression or vector safety. Altogether, recent advances in globin vector design bode well for upcoming clinical trials to assess the therapeutic value of globin gene transfer.
Serological and histological characterization of the new mutant strain of lpr mice, CBA/KUms-lprcg/lprcg
Authors: KIMURA M , MOHRI H , SHIMADA K , WAKABAYASHI T , KANAI Y , MATSUZAWA A
Published: July 2010

CBA/KIJms-lprcg/lprcg mice with a novel mutation producing systemic lymphoproliferation wereinvestigated for their serological and histological characteristics. The mutant mice showed elevatedlevels of serum immunoglobulin, C1q-binding immune complexes and antibodies to nuclear antigenssuch as dsDNA and ssDNA and poly(ADP-ribose). In contrast, histopathological lesions, e.g.glomerulonephritis, vasculitis or interstitial pneumonitis, were not revealed by histological andimmunofluorescent examinations, except for lymphocytic infiltration in various organs. These resultssuggest that this mutant mouse strain may provide a new animal model For autoimmunity. However, further investigations are required to clarify whether this strain is unique as compared with otherwell-known lupus-prone strains of mice with respect to serological and histological abnormalities andbecome to be a new model of systemic autoimmune disease.
Cytokine regulation of intercellular adhesion molecule-1 (ICAM-1) expression on human glioblastoma cells
Authors: KUPPNER M C , MEIR E VAN , HAMOU M F , TRIBOLET N DE
Published: July 2010

Intercellular adhesion molecule-l (ICAM-1) has recently been identified as one of the ligands for lymphocyte function-associated antigen-1 (LFA-1). Immunohistochemical staining of frozen tissue sections using the ICAM-1 antibody RR1/1 demonstrated significant levels of ICAM-1 expression on human glioblastoma cells and on intratumoural vascular endothelial cells. ICAM-1 was weakly expressed or absent from low grade gliomas and absent from normal and fetal brain. ICAM-1 expression was similar to that of MHC class II, HLA-DR antigens. Glioblastoma cell lines constitutively expressed ICAM-1 to a minimal or moderate extent. Surface antigen expression of ICAM-1 and ICAM-1-specific mRNA could be significantly increased by incubating glioblastoma cells with interleukin-1β (IL-lβ), tumour necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ). IL-2, IL-4, IL-6 and transforming growth factor β2 (TGF-β2) had no significant effect on surface antigen expression. Significant enhancement of ICAM-1 expression was obtained using TNF-α and IL-β at 1 -10 U/ml and at 500 U/ml of IFN-γ. Induction of ICAM-1 specific mRNA was observed 4 h after cytokine treatment and decreased by 24 h. Surface antigen expression of ICAM-1 increased for up to 48 h after treatment.
Kupffer cell numbers during human development
Authors: COPE E M W , DILLY S A
Published: July 2010

Immunohistological assessment of Kupffer cells was made using the antibody MAC387 and an antibody to lysozyme. Autopsy liver samples from 13 fetuses aged from 17 weeks gestation to term, and from 10 neonates and children aged 1 day to 18 months, were studied. For comparison, 10 normal adult autopsy liver specimens were included. The number of positively staining cells per unit area was counted for periportal sinusoids (zone 1) and centrilobular sinusoids (zone 3). No difference was found between zone 1 and zone 3 macrophage numbers with either antibody at any stage of development. Hepatic sinusoidal macrophage numbers were low during early gestation but increased during intra-uterine life to reach approximately normal adult values in the neonatal period. The numbers of cells staining with MAC387 or lysozyme were similar in each case except for hepatic sinusoidal macrophages in fetuses of less than 30 weeks gestation. Here anti-lysozyme stained significantly fewer cells, suggesting that lysozyme production may be low in immature fetuses. No difference was found between infants of similar maturity who had died immediately or had lived for more than 48 h and hence been exposed to gut antigens.
Presence of antibodies against endothelial cells in the sera of patients with episodic angioedema and hypereosinophilia
Authors: LASSALLE P , GOSSBT F , GRUART V , PRIN L , CAPRON M , LAGRUE G , KUSNIERZ J P , TONNEL A B , CAPRON A
Published: July 2010

We reported three additional cases of a newly described syndrome called episodic angioedema with hypereosinophilia. In order to investigate its pathophysiological mechanisms, four parameters were concurrently investigated, including blood eosinophil density, serum chemoattractant activity, serum major basic protein (MBP) levels and the presence of anti-endothelial cell antibodies. Distribution of eosinophils through a metrizamide density gradient showed a preferential sedimentation of blood eosinophils in intermediate layers, clearly different from the hypodense cells (low-density layers) identified in a group of seven patients with idiopathic hypereosinophilic syndrome (HES). In two of the three patients with cyclic angioedema. a chemotactic activity towards eosinophils was detected in the serum (30 ± 6 and 42 ± 12 eosinophils per high-power field; P < 0.05 compared with a control group). Serum MBP levels were at 1524, 619 and 1200 pg/ml. All three patients had circulating anti-endothelial cell antibodies, predominantly of the IgG isotype, in contrast to controls (P < 0.01) or to patients with HES (P < 0.01). Specificity of the antibody for endothelial cells was demonstrated in the three patients studied by the absence of binding to various blood cells, including monocytes, lymphocytes, eosinophils and platelets. In one case (patient 2), the levels of anti-endothelial cell antibodies, as well as the serum chemoattractant activity to eosinophils varied according to the successive acute phases of the disease. Although further investigations are needed to clarify the exact pathophysiology of this syndrome, and especially the possible participation of the anti-endothelial cell antibodies in the cutaneous lesions, these data suggest that angioedema observed in this syndrome could result from the combined effects of activated eosinophils and of immunologically induced endothelial lesions.
Activation of lymphocytes after platelet allotransfusion possessing only class I MHC product
Authors: PÓCSIK É , MIHALIK R , GYÓDI É , RÉTI M , PÁLÓCZI K , PETRÁNYI G G , BENCZÚR M
Published: July 2010

After platelet allotransfusion, we found a characteristic increase in the expression of interleukin-2 receptor, dipeptydilpeptidase IV (CD26), activation-inducer molecule (AIM, CD69) and transferrin receptors (CD71) on day 3 indicating that important functional molecules expressed on the activation of lymphocytes by allogeneic platelets. At the same time, no consistent increase of other activation molecules such as Ki-1 (CD30), intercellular adhesion molecule (ICAM-1, CD54) and Ki-24 (CDw70) antigen expression was detected, probably as a result of the selective activation of some lymphocyte subsets. In order to obtain further evidence for the in vivo activation triggered by allogeneic platelets, a subsequent step of T cell activation towards differentiation was investigated with monoclonal antibodies to leucocyte common antigens. A sharp expression of the UCHL1, coupled with a decrease of the CD45R molecule was detected on day 7 or 14, suggesting a T cell priming.
Flow cytometric and functional analysis of mononuclear cells infiltrating the liver in experimental autoimmune hepatitis
Authors: KOHDA H , SEKIYA C , KANAI M , YOSHIDA Y , UEDE T , KIKUCHI K , NAMIKI M
Published: July 2010

Experimental autoimmune hepatitis was produced by immunizing Wistar rats with syngeneic liver proteins. Mononuclear cells infiltrating the liver tissue were identified by immunohistochemical techniques using monoclonal antibodies specific for subpopulations of rat lymphocytes. The strong infiltration of CD8+ cytotoxic T lymphocytes (CTL) were found in the portal areas. Subpopulations of mononuclear cells infiltrating the liver, spleen cells and peripheral blood lymphocytes were identified by flow cytometry. Flow cytometric analysis revealed the presence of CD5- and CD8+ lymphocytes in the liver tissues. Mononuclear cells infiltrating the liver were isolated from Wistar rats having autoimmune hepatitis to determine whether those exhibit cytotoxicity against syngeneic hepatocytes; they exhibited cytotoxicity against isolated syngeneic hepatocytes, but failed to lyse K562 cells, syngeneic concanavalin A-activated splenocytes and allogeneic hepatocytes. Depietion of CD8+ T cells significantly reduced the cytotoxic ability of mononuclear cells infiltrating into the liver against syngeneic hepatocytes. These findings support the idea that liver cell injury in experimental autoimmune hepatitis may at least in part be mediated by CTL.
Increased serum neopterin in patients with HIV-1 infection is correlated with reducedin vitrointerleukin-2 production
Authors: FUCHS D , SHEARER G M , BOSWELL R N , CLERICI M , REIBNEGGER G , WERNER E R , ZAJAC R A , WACHTER H
Published: July 2010

Recently we have observed that the CD4+ T cell response of peripheral blood mononuclear cells (PBMC) to soluble antigens is the first to be lost in the course of HIV-1 infection followed by the loss of response to HLA alloantigens. In this study we compared serum neopterin concentrations of individuals with early stages of HIV-1 infection (stages WR1 and WR2, Walter Reed staging system) with in vitro interleukin-2 (IL-2) production of PBMC in response to stimulation with soluble antigens (influenza A virus and tetanus toxoid) and alloantigens. Neopterin concentrations were significantly higher in HIV-l-seropositive individuals who showed deficient IL-2 production in response to recall antigens only or to all of the stimuli tested in vitro, compared with HIV-l seropositive individuals who exhibited no CD4+ T cell defects. No difference in serum neopterin concentrations was observed between the group that was functionally deficient to soluble antigens only versus those who were unresponsive to both types of stimuli. It appears that the selective loss of the MHC self-restricted CD4+ T cell function is associated with an increase in serum neopterin levels. Neopterin concentrations are an estimate of the activation status of macrophages. We conclude that defective in vitro production of lymphokines by T lymphocytes is associated with activated macrophages in vivo.
Serum sickness and acute renal failure after streptokinase therapy for myocardial infarction
Authors: DAVIES K A , MATHIESON P , WINEARLS C G , REES A J , WALPORT M J
Published: July 2010

A patient developed serum sickness and acute renal failure following therapy with streptokinase for myocardial ischaemia. There was a previous history of a cellulitic infection of the leg. and antibodies to streptokinase were measurable in a serum sample taken from the patient before therapy. A cryoglobulin was detected at the lime of presentation with serum sickness. This contained polyclonal IgG (with anti-streptokinase activity), streptokinase, and C3. Circulating immune complexes were demonstrated by C1q-binding assay. Deposition of C3 was observed in skin and renal biopsies, and bound to erythrocytes. Renal histology, however, showed acute tubular necrosis, with no vasculitis or inflammatory cell infiltrate. This case provides an unusual example of the characterization of an immune complex comprising a specific antibody and an exogenous antigen, and has clinical implications for the use of streptokinase.
Suppression of HLA class II expression on thyrocytes by interferon-alpha 1
Authors: GUERIN V , TODD I , HAMMOND L J , BOTTAZZO G F
Published: July 2010

Inappropriate expression of HLA class II molecules by human thyroid epithelial cells (thyrocytes) is commonly associated with autoimmune thyroid disease. HLA class 11 expression can be modulated in thyrocytes in vitro by a variety of substances: in particular, it is readily induced by interferon-gamma (IFN-γ). Here we show that recombinant IFN-α1 (rIFN-α1) does not induce HLA class II expression by thyrocytes. but rather it suppresses the induction of such expression by rIFN-γ. Similar effects were observed with IFN-α derived from a lymphoblastoid cell line. The effect of rIFN-α1 on thyrocytes differs from its effect on human monocytes, reported by others, in which it was found to enhance the expression of HLA class II. Thus, rIFN-α1 appears to have a differential effect on HLA class II expression, depending on the cell type involved.
IgA Polyspecific autoantibodies in IgA nephropathy
Authors: MATSIOTA P , DOSQUET P , LOUZIR H , DRUET E , DRUET P , AVRAMEAS S
Published: July 2010

The specificity of circulating and kidney-bound IgA during IgA nephropathy is suit a matter of discussion. In the present study, high levels of IgA antibodies directed against a panel of self and non-out of the seven kidney biopsies studied. After immunoedsorption of pooled selected serum samples on TNP and getin-coated columns, polyspecific IgA antibodies were elated. This supports the hypothesis that IgA-bearing B cells clones most probably producing polyspecific antibodies are a major feature of human IgA nephropathy. These findings also suggest that it may be hazardous to draw conclusions from the findings of apparent monospecific IgA antibodies in this condition.
Functional characterization of SV40-transformed adherent synovial cells from rheumatoid arthritis
Authors: GOTO M , OKAMOTO M , SASANO M , NISHIZAWA K , AOTSUKA S , YAMAGUCHI N , OBINATA M , IKEDA K
Published: July 2010

A total of 14 transformed cell clones were obtained by micro-injecting origin-defective SV40 DNA into three types of cloned adherent synovial cells (ASC) (dendritic cells (DCs), macrophage-Iike cells (MCs), and fibroblast-like cells (FCs)) from two rheumatoid arthritis patients (five DC clones (SV40-DCs), five MC clones (SV40-MCs) and four FC clones (SV40-FCs)). All the transformed cell nuclei expressed SV40-specific T antigen. The cells which formed a colony had a few times shorter doubling time than the original cells. IL-1α, IL-lβ and prostaglandin E2 were detected in the culture supernatant from the unstimulated transformed cells like untransformed cells. The SV40-DCs showed the most potent accessory cell function in oxidative mitogenesis assay among the three types of SV40-ASCs. Granulocyte macrophage colony stimulatory factor (GM-CSF) was detected only in the culture supernatant from the SV40-MCs without stimulation. Extensive phenotypic analysis revealed relatively cell-specific markers. SV40-DCs were HLA-DP+ and glial fibrillary acidic protein positive. SV40-MCs stained positive for 5′-nucleotidase and nonspecific esterase. These transformed ASCs retained much of the original cellular physiology of rheumatoid arthritis (RA) ASCs and may be a useful tool for characterizing the role of ASCs in the pathogenesis of RA.
FcγRII restriction fragment length polymorphism (RFLP): analysis in systemic lupus erythematosus and scleroderma and evidence of an alpha gene duplication
Authors: JAZWINSKA E C , OLIVE C , HOGARTH P M , GATENBY P A , SERJEANTSON S W
Published: July 2010

The characteristic finding of high levels of circulating immune complexes in patients with the autoimmune connective tissue diseases systemic lupus erythemalosus (SLE) or scleroderma has raised the possibility that these patients may have a primary defect in immune complex clearance. The Fc receptor for IgG (FcγR) plays a central role in the phagocytosis of antibody complexes. We have analysed FcγR (type II) RFLPs identified in TaqI-and MspI-restrictcdgenomic DNAand found that their distribution in SLE and selerodenna did not differ significantly from controls. Hybridization with specific regions of the FcγRII cDNA clone indicate that part of the FcγRIIα locus is duplicated in some individuals. A further FcγRII gene has recently been identified (FcγRIIα). This gene shows > 95% homology with FcγRIIα and may thus be the candidate gene for the apparant α duplication seen in some individuals. It is possible that an individual may possess one. two, three or four TaqI FcγRIIα/α′alleles. correlating with incidence and numerical heterogeneity in FcγRIIα and α′. The physiological effects of this numerical heterogeneity remain to be investigated.
Anti-idiotypes against anti-neutrophil cytoplasmic antigen autoantibodies in normal human polyspecific IgG for therapeutic use and in the remission sera of patients with systemic vasculitis
Authors: ROSSI F , JAYNE D R W , LOCKWOOD C M , KAZATCHKINE M D
Published: July 2010

Anti-neutrophil cytoplasmic antigen (ANCA) activity was inhibited in 15 out of 21 sera from patients with acute systemic vasculitis following incubation with normal polyspecific IgG for therapeutic use (IVIg). ANCA antibodies reacted with IVIg through idiotypic-anti-idiotypic interactions, as shown in competitive binding assays using F(ab′)2 fragments from IVIg and affinity chromatography of ANCA IgG on Scpharose-bound F(ab′)2 fragments from IVIg. Co-incubation of sera from patients with acute systemic vasculitis with paired autologous remission stage sera also resulted in inhibition of ANCA activity in acute sera. Remission sera contain IgM and IgG capable of interacting with β and/or α idiotypes of ANCA IgG from acute sera. Anti-idiotypic IgM may account for the lack of expression of ANCA activity in whole serum from patients in remission from systemic vasculitis. which were found to contain high litres of ANCA IgG. These observations suggest that remission of systemic vasculitis is associated with the generation of anti-idiotypes against autoantibodies rather than the suppression of production of ANCA autoantibodies. IVIg may modulate the activity of systemic vasculitis in vivo.
Induction of suppressor cells byMycobacterium paratuberculosisantigen in inflammatory bowel disease
Authors: EBERT E C , BHATT B D , LIU S , DAS K M
Published: July 2010

We studied the M. paratuberculosis-induced proliferation and suppressor cell generation by peripheral blood lymphocytes from patients with inflammatory bowel disease. Peripheral blood lymphocytes were separated from 33 patients with Crohn's disease, 18 with ulcerative colitis, nine with other intestinal diseases, and five with autoimmune disorders. Proliferation of peripheral blood lymphocytes from normal individuals in response to 10 μg/ml of M. paratuberculosis antigen was reduced by depletion of CD4+ T cells. The ability of M. paratuberculosis antigen to suppress concanavalin A-induced proliferation (expressed as a percentage suppression) was reduced by depletion of CD8+ T cells. This suppression was the same whether peripheral blood lymphocytes were from normal individuals, patients with intestinal diseases other than inflammatory bowel disease, or patients with autoimmune disorders (47 ± 14%, 44 ± 24%, and 30 ± 26%, respectively). In contrast, the suppression induced by M. paratuberculosis for patients with Crohn's disease and ulcerative colitis (66 ± 22% and 67 ± 22%) was much greater than that for normal individuals (P < 0.001). In particular, lymphocytes from patients with active Crohn's disease demonstrated little proliferation in response to this antigen but marked suppressor activity (79 ± 13%). How the immunomodulatory effects of this antigen relate to the pathogenesis of the inflammatory bowel diseases remains to be determined.
Plasma membrane association of primary biliary cirrhosis mitochondrial marker antigen M2
Authors: SUNDIN U , SUNDQVIST K G
Published: July 2010

Antibody reactivity against the ‘mitochondrial M 2 antigen’ was determined in sera from 10 patients with primary biliary cirrhosis (PBC). using Western blotting after SDS-PAGE separation of rat liver mitochondria (RLM) and plasma membrane proteins. The molecular weights of the major M2 antigens in rat liver mitochondria were 67 and 50 kD Two of the 10 PBC patients did not react to any of these major antigens, eight reacted to the 67-kD and four of those also to the 50-kD antigen. The 67- and 50-kD antigens were present in both plasma membrane and RLM and had affinity to concanavalin A. Antibody reactivity against the 67-kD antigen could be detected in both IgG and IgA as well as in the IgM class. The reactive IgG subclasses to both types of antigen preparations were mainly of the GI and G3 isotypes. This reactivity was always stronger with antigens from the plasma membrane preparations. Sera from two patients with high antibody litres against mitochondria also reacted with IgG2 against the 50-kD antigen from plasma membrane, but not to the corresponding antigen in mitochondria. Reactivity of antibodies in PBC sera to the periphery of viable hepatocytes and radioactive surface labelling of the 50-kD component are both consistent with a plasma membrane localization of M2. Serum from healthy controls and several patients with different diseases did not contain antibodies reactive against any of the antigens described. We suggest that antigens, partly identical to the mitochondrial M2, are located in the plasma membrane compartment. The PBC pathogenetical consequences of these findings arc discussed.
Kupffer cell depletionin vivoresults in clearance of large-sized IgA aggregates in rats by liver endothelial cells
Authors: BOGERS W M J M , STAD R K , JANSSEN D J , PRINS F A , VAN ROOIJEN N , VAN ES L A , DAHA M R
Published: July 2010

We investigated the clearance kinetics and tissue distribution of different sized IgA in normal and macrophage-depleted rats Rats were injected iv with liposomes containing dichloromethylene diphosphonate (DMDP). DMDP treatment resulted in complete depletion of liver macrophages 24-48 h after administration. Normal and macrophage depleted rats were injected intravenously with monomeric, dimeric, polymeric or aggregated polymeric IgA (AIgA) and assessed for blood clearance and tissue distribution. In normal rats, clearance of IgA was size dependent, i.e. a faster clearance with increasing size. No differences in clearance kinetics were observed of the different sized IgA between normal and DMDP-treated rats. TCA non-precipitable radioactivity, a measure for degradation of IgA, was found in the circulation of normal and DMDP-treated rats after AIgA administration. The liver was the main organ responsible for the clearance of IgA in normal and DMDP-treated rats. Immunofluorescence studies on liver biopsies indicated that AIgA was associated with Kupffer cells in normal rats. Electronmicroscopical studies revealed that the AIgA was internalized and located in vesicles in Kupffer cells. In DMDP-treated rats the AIgA was associated with endothelial cells and electron microscopy studies showed that this AIgA was taken up by endothelial cells. These data show that rat liver endothelial cells are able to bind, internalize and degrade AIgA in situations where Kupffer cells are absent, and that these cells may play an important role in the handling of AIgA and IgA-immune complexes.
T and B cell human responses to European bat lyssavirus after post-exposure rabies vaccination
Authors: HERZOG M , FRITZELL C , LAFAGE M , MONTAÑO HIROSE J A , SCOTT-ALGARA D , LAFON M
Published: July 2010

T and B cell human responses lo European bat lyssavirus (EBL1) induced by post-exposure rabies vaccination (PM virus vaccine) were evaluated by measuring plasmatic titres of EBL1-specific neutralizing antibodies; specific EBL1-binding antibodies; and proliferation indices of peripheral blood lymphocytes stimulated in vitro with EBL1. These parameters for vaccination efficacy were compared with those obtained with vaccine-related viruses (CVS and ERA) and with a non-vaccine-related virus, Mokola virus, the last implicated in vaccination failures. Twenty-two patients exposed to rabies risk who received a reduced rabies post-exposure vaccination were involved in the study. On day 21, vaccine induced CVS-specific neutralizing antibodies in all patients; but EBL1-spccific neutralizing antibodies were induced in only 73% of patients. No vaccinee had Mokola-specific neutralizing antibodies. Patients having EBL1-specific neutralizing antibodies were usually those in whom vaccination induced high titres of CVS-specific neutralizing antibodies. On day 21, peripheral blood lymphocytes of 86% of patients could be restimulaled in vitro with vaccine, 43% with EBL1 and 45% with Mokola. Patients exhibiting a high vaccine-specific proliferation response more likely developed an EBL1-or a Mokola-specific proliferalive response. No correlation was found between T and B cell responses. Rabies vaccination induced neither T nor B cell EBL1-specific responses in 22% of patients.
Identification of cells responsible for the suppression of anti-beta-cell cytotoxicity in type I (insulin-dependent) diabetes mellitus
Authors: KRUG J , BIERWOLF B , LAMPETER E , VERLOHREN H-J , LOHMANN D
Published: July 2010

Peripheral blood mononuclear cells (PBMC) from newly diagnosed type I diabetic patients induced a significantly higher cytotoxic insulin leakage from isolated rat islets during a 20-h incubation, compared with PBMC from healthy controls (13.9±7.7 versus 2.6±3.2 ng insulin/islet/20 h). The addition of PBMC from healthy subjects in a ratio of 1:4 suppressed the cytotoxic effect (13.9±7.7 to 6.7±4.4 ng insulin/islet/20 h). Depletion of CD8+ T lymphocytes from the PBMC of healthy subjects abolished their supprcssive capacity, while depletion of CD4+ T lymphocytes and pre-incubation with CD3 monoclonal antibodies had no effect.
Induction of cutaneousgraft-versus-hostdisease by local injection of unprimed T cells
Authors: KAWAI K , MATSUMOTO Y , WATANABE H , ITO M , FUJIWARA M
Published: July 2010

The skin is a major target organ in human graft-versus-host disease (GVHD) after bone-marrow transplantation. GVHD can be induced in mice by i.v. injection of T cells into unirradiated semi-allogeneic or lethatly irradiated allogeneic recipients. However, in the murine systemic GVHD model, cutaneous lesions occur only in lethally irradiated recipients. Since lethal irradiation itself might induce the epidermal cell damage, several investigators have employed another murine model of cutaneous GVHD, in which cutaneous lesions were induced by intradermal injection of alloreactive T cell clones. Using this system, it has been reported that both MHC class I- and II-reactive T cell clones can induce cutaneous GVHD in non-irradiated or sublethally irradiated recipients. However, it has remained unknown whether or not freshly prepared T cells are able to induce cutaneous GVHD after local injection into non-irradiated recipients. We show that unprimed T cells can induce cutaneous GVHD after local injection into unirradiated MHC class II- or I + II-disparate recipients. In contrast to alloreactive T cell clones, unprimed T cells could elicit only mild cutaneous lesions in MHC class I-disparate recipients. Since sublethal irradiation of MHC class I-disparate recipients did not result in the manifestation of cutaneous lesions after injection of unprimed T cells, host anti-donor responses by radiosensitive cells could not be responsible for this phenomenon. This experimental system provides a useful model for analysis of the regulation mechanisms in the induction of GVHD by unprimed T cells.
Expression of adhesion molecules in human intestinal graft-versus- host disease
Authors: NORTON J , SLOANE J P , AL-SAFFAR N , HASKARD D O
Published: July 2010

The distribution of three cellular adhesion molecules, ICAM-1. ELAM-1 and VCAM-1, was studied in normal rectal mucosa and in graft-Mrms-host disease (GvHD) using immunohistological and morphometric techniques. In normal controls, ICAM-1 was demonstrable on endothelial cells and leucocytes within the lamina propria, ELAM-I on endothelial cells only and VCAM-1 on lamina propria leucocytes, many of which exhibited long dendritic processes surrounding the glands. In GvHD, the enterocytes became positive for ICAM-1 and this was often associated with the presence of intra-epithelial LFA-1* lymphocytes and macrophages, ihe latter containing debris of apoptotic cells. The staining was. however, restricted to the luminal membrane of the epithelial cells, raising doubts about the role of ICAM-1 as a ligand for LFA-1 on mucosal leucocytes in rectal GvHD. ELAM-1 expression was increased in GvHD both in terms of the length of positive endothelium and staining intensity. VCAM-1 was increased on endothelial cells but not leucocytes in the lamina propria in contrast to our previous findings in cutaneous GvHD where VCAM-+ dendritic cells were increased and endothelial cells remained negative. Normal patterns of adhesion molecule staining were seen in two biopsies exhibiting no morphological evidence of GvHD, from patients who had strong clinical evidence of the disease, indicating that immunostaining for these molecules is unlikely to be of help in improving the sensitivity of histological diagnosis. However, the possibility that adhesion molecule staining may be useful in improving diagnostic specificity by helping to distinguish GvHD from identical histological changes produced by irradiation and cytotoxic drugs is worthy of further investigation.
Antibodies against C1q in anti-glomerular basement membrane nephritis
Authors: COREMANS I E M , DAHA M R , VAN DER VOORT E A M , MUIZERT Y , HALMA C , BREEDVELD F C
Published: July 2010

The prevalence of antibodies against the collagen-like region of the subcomponent of the first component of complement, Clq, was investigated in 11 patients with anti-glomerular basement membrane (GBM) nephritis. Anti-C1q antibodies (anti-ClqAb) were detected in seven patients. IgG anti-ClqAb were found in four and IgA anti-ClqAb in five patients. During follow up of the patients a relationship was observed between the levels of IgG anti-ClqAb and the levels of anti-GBM antibodies (anti-GBMAb). Gelfiltration experiments indicated that both IgG anti-ClqAb as well as IgG anti-GBMAb were monomeric and that binding also occurred with the F(ab′)2 fragments of the antibodies. Although anti-ClqAb and anti-GBMAb are both directed against a collagen-like structure, it was demonstrated by means of inhibition experiments that anti-ClqAb and anti-GBMAb are directed against different antigenic sites. Comparison of patients with anti-GBM nephritis with and without anti-ClqAb revealed that there were no differences in disease activity or disease severity. Therefore, the results of this study suggest that anti-ClqAb do not play a direct pathogenetic role in anti-GBM nephritis.