bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid flow cytometric test for the diagnosis of membrane cytoskeleton‐associated haemolytic anaemia
Authors: King May‐Jean , Behrens Judith , Rogers Chris , Flynn Clare , Greenwood David , Chambers Keith
Published: July 2010

The flow cytometric test measures the fluorescence intensity of intact red cells labelled with the dye eosin‐5‐maleimide, which reacts covalently with Lys‐430 on the first extracellular loop of band 3 protein. In this study, red cells from patients with hereditary spherocytosis (HS), congenital dyserythropoietic anaemia type II, South‐east Asian ovalocytosis and cryohydrocytosis have produced a greater degree of reduction of mean channel fluorescence readings than those for other patient groups and normal controls. The predictive value of this test for membrane abnormality was compared with the results obtained from the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) method, which is currently the reference laboratory test for the identification of membrane protein deficiencies in hereditary spherocytosis and for the detection of spectrin variants in hereditary elliptocytosis. The dye method is a reliable, speedy diagnostic test (2 h from sample collection to result) for HS with a sensitivity of 92·7% and a specificity of 99·1%. Thus, it will serve well as a first‐line screening test for the diagnosis of hereditary spherocytosis in routine haematology.
Increased apoptosis in acquired sideroblastic anaemia
Authors: Matthes Thomas W. , Meyer Geneviève , Samii Kaveh , Beris Photis
Published: July 2010

Idiopathic acquired sideroblastic anaemias (IASAs) form a subgroup of the myelodysplastic syndromes and are characterized by mitochondrial iron accumulation, bone marrow erythroid hyperplasia and decreased peripheral red blood cell counts. Increased intramedullary apoptosis of erythroid precursors is presumed to constitute the pathophysiological mechanism explaining this ineffective erythropoiesis, but if and how mitochondrial dysfunction is implicated in this process is currently unknown. We therefore studied bone marrow precursor cells obtained from nine patients with IASA for (i) caspase 3 activity, (ii) numbers of Annexin V‐ and 7‐amino‐actinomycin‐positive cells, (iii) numbers of cells with diminished mitochondrial membrane potential, ΔΨm, and (iv) numbers of cells producing reactive oxygen species (ROS), and we compared the results with those of five normal bone marrow samples. Compared with controls, we found increased caspase 3 activity in all IASA samples, which correlated with increased numbers of Annexin‐V‐positive cells (r = 0·7). Analysis of different subpopulations showed increased apoptosis in erythroid populations compared with myeloid and/or lymphoid populations in five out of nine cases, and increased apoptosis in the last two populations in four out of nine cases. As evidence of mitochondrial dysfunction, ΔΨm was found to be diminished in the erythroid subpopulations of all cases of IASA (66·6 ± 17% vs. 34·6 ± 12% in normals). ΔΨm decrease was correlated to Annexin V positivity (r = 0·7). Astonishingly, no difference was found between IASA and normal bone marrows with regard to the number of ROS‐producing cells. In fact, both groups exhibited a similar low proportion of ROS production (10·3 ± 7% in normals vs. 6·8 ± 5% in IASA). Taken together, our results show that mitochondria are clearly implicated in the apoptotic process in IASA patients. Whether this is a result of an intramitochondrial defect (e.g. Fe accumulation, secondary to mitochondrial or nuclear DNA mutations) or is secondary to an extracellular stimulus [e.g. tumour necrosis factor (TNF), Fas ligand (FasL)] remains to be determined.
Defibrotide for the treatment of hepatic veno‐occlusive disease: results of the European compassionate‐use study
Authors: Chopra R. , Eaton J. D. , Grassi A. , Potter M. , Shaw B. , Salat C. , Neumeister P. , Finazzi G. , Iacobelli M. , Bowyer K. , Prentice H. G. , Barbui T.
Published: July 2010

Severe hepatic veno‐occlusive disease (VOD) is a recognized complication of autologous and allogeneic stem cell transplantation (SCT) that is often fatal. Defibrotide (DF) is a polydeoxyribonucleotide that has been found to have anti‐thrombotic, anti‐ischaemic and thrombolytic properties without causing significant anticoagulation. Preliminary studies have demonstrated activity for DF in the treatment of VOD, with minimal associated toxicity. In the present study, 40 patients who fulfilled established criteria for VOD were treated with DF on compassionate grounds in 19 European centres; 28 patients met risk criteria predicting progression of VOD and fatality or had evidence of multiorgan failure (MOF), and were defined as ‘poor‐risk’. DF was commenced intravenously at a median of 14 d (range, −2 d to 53 d) post SCT at doses ranging from 10 to 40 mg/kg. The median duration of therapy was 18 d (range, 2–71 d). Twenty‐two patients showed a complete response (CR) (bilirubin < 34·2 μmol/l and resolution of signs/symptoms of VOD and end‐organ dysfunction) [CR = 55%, confidence interval (CI) 40–70%] and 17 patients (43%) are alive beyond d +100. Ten poor‐risk patients showed a complete response (CR = 36%, CI 21–51%). These results demonstrate that DF is an active treatment for VOD following SCT and a randomized trial is now underway in order to further evaluate its role.
The risk of venous thromboembolism in family members with mutations in the genes of factor V or prothrombin or both
Authors: Martinelli Ida , Bucciarelli Paolo , Margaglione Maurizio , De Stefano Valerio , Castaman Giancarlo , Mannucci Pier Mannuccio
Published: July 2010

Factor V Leiden and the G20210A mutation in the prothrombin gene are the most frequent abnormalities associated with venous thromboembolism. It is unknown whether the risks due to the presence of either mutation are of the same magnitude. We compared the prevalence and incidence rate of venous thromboembolism in relatives with either mutation or both. The finding of different rates might influence the strategies for primary prevention of thrombosis in carriers of these mutations. The study population included 1076 relatives of probands with the prothrombin gene mutation, factor V Leiden or both who underwent screening for inherited thrombophilia and were found to be carriers of single mutations or double mutations or who were non‐carriers. The prevalence of venous thromboembolism was 5·7% in relatives with the prothrombin gene mutation, 7·8% in those with factor V Leiden, 17·1% in those with both mutations and 2·5% in non‐carriers. Annual incidences of thrombosis were 0·13% [95% confidence interval (CI) 0·06–0·24], 0·19% (0·13–0·25), 0·42% (0·15–0·83) and 0·066% (0·03–0·11), respectively, and the relative risk of thrombosis was two times higher in carriers of the prothrombin gene mutation, three times higher in those with factor V Leiden and six times higher in double carriers than in non‐carriers. The incidence of venous thromboembolism in carriers of the prothrombin gene mutation is slightly lower than that observed in carriers of factor V Leiden, whereas in carriers of both mutations it is two or three times higher. These findings suggest that lifelong primary anticoagulant prophylaxis of venous thromboembolism is not needed in asymptomatic carriers of single or double mutations. Anticoagulant prophylaxis seems to be indicated only when transient risk factors for thrombosis coexist with mutations.
Clinical manifestations and complications of childbirth and replacement therapy in 385 Iranian patients with type 3 von Willebrand disease
Authors: Lak M. , Peyvandi F. , Mannucci P. M.
Published: July 2010

Type 3 is the most severe form of von Willebrand disease (VWD) transmitted as an autosomal recessive trait. We collected data on the clinical manifestations of type 3 VWD by examining 385 patients from 300 Iranian kindreds, who were compared with 100 age‐matched patients with severe haemophilia A. Joint and muscle bleeding was less frequent than in haemophiliacs, perhaps because factor VIII levels were in general higher (median value 4% vs. 1% or less). Mucosal tract haemorrhages such as epistaxis and menorrhagia were the most prevalent symptoms in VWD. Post‐circumcision and oral cavity bleeding occurred frequently when prophylactic replacement therapy was not carried out or was inadequate. The course of pregnancy was usually uneventful, but increased bleeding occurred at parturition when affected women were treated with replacement therapy for less than 3–4 d. Ten of 385 (2·6%) of these multitransfused patients developed an alloantibody to VWF and 55% are chronically infected with the hepatitis C virus.
Cyclosporin A response and dependence in children with acquired aplastic anaemia: a multicentre retrospective study with long‐term observation follow‐up
Authors: Saracco Paola , Quarello Paola , Iori Anna Paola , Zecca Marco , Longoni Daniela , Svahn Johanna , Varotto Stefania , Del Vecchio Gian Carlo , Dufour Carlo , Ramenghi Ugo , Bacigalupo Andrea , Locasciulli Anna ,   on behalf of the Bone Marrow Failure Study Group of the AIEOP (Italian Association of Paediatric Haematology Oncology)
Published: July 2010

Immunosuppressive therapy (IST) with antithymocyte globulin and cyclosporin A (CyA) is the standard treatment for children with acquired aplastic anaemia (AAA) lacking a matched donor. Survival rates of more than 80% at 5 years are achieved, but the response is drug‐dependent in 15–25% of cases. This study, of 42 consecutive children with AAA treated with IST, assessed the incidence of CyA‐dependence, CyA and granulocyte colony‐stimulating factor (G‐CSF) tapering schedules and the impact of drug accumulation on progression to myelodysplasia/acute myeloid leukaemia (MDS/AML). Overall survival was 83% at 10 years. CyA‐dependence without a predictive marker was observed in 18% of responders. Probability of discontinuing CyA was 60·5% at 10 years; a slow CyA tapering schedule was performed in 84% of patients; the cumulative incidence of relapse was 16% at 10 years. Relapse risk was significantly associated with rapid CyA discontinuation: 60% compared to 7·6% in the slow tapering group (P = 0·001). Cumulative incidence of MDS/AML was 8% at 10 years, with a significant correlation with both G‐CSF cumulative dose and second IST. This long‐term follow‐up of children with AAA shows that IST with a slow CyA tapering course is an effective treatment with a low‐relapse rate in these cases.
Considerations in the approach to invasive fungal infection in patients with haematological malignancies
Authors: Bow Eric J.
Published: July 2010

Invasive infections because of opportunistic yeasts and moulds have contributed significantly to the morbidity and mortality associated with potentially curative treatment for haematological malignancies. Many risk factors have been identified that permit the clinician to predict the likelihood of these infections. The diagnostic process involves maintaining a high index of suspicion based upon an understanding of the clinical circumstances under which invasive fungal infections occur, of the spectrum of fungal syndromes, and of the advantages and limitations of diagnostic testing strategies now available. Treatment strategies may be categorized as prophylactic, pre‐emptive, empiric, or directed based upon the circumstances. The therapeutic options have increased in recent years but are not applicable to all clinical circumstances. These considerations are discussed.
Genetic variation and susceptibility to infection: the red cell and malaria
Authors: Weatherall D. J.
Published: July 2010

There is now convincing evidence that the extremely high frequencies of certain genetic disorders of the red cell involving haemoglobin, the red‐cell membrane, or its metabolic pathways reflect relative resistance to malaria over thousands of years. At least some progress has been made towards an understanding of the cellular mechanisms involved, although much remains to be learnt. As well as the extremely valuable information that this field is providing about how exposure to infection has moulded the current structure of the human genome, recent research in this field is starting to provide some valuable new approaches to the better control of parasitic and other infections that remain a major global health problem.
An introduction to induced pluripotent stem cells
Authors: Hanley Joanna , Rastegarlari Ghasem , Nathwani Amit C.
Published: July 2010

Recent landmark studies show that it is now possible to convert somatic cells, such as skin fibroblasts and B lymphocytes, into pluripotent stem cells that closely resemble embryonic stem cells. These induced pluripotent stem (iPS) cells can be generated without using human embryos or oocytes, thus bypassing some of the ethical issues that have limited the use of human embryonic stems (hES) cells. Additionally, they can be derived from the patient to be treated, thereby overcoming problems of immunological rejection associated with the use of allogeneic hES cell derived progenitors. Whilst these patient‐specific iPS cells have great clinical potential, their immediate utility is likely to be in drug screening and for understanding the disease process. This review discusses the promise of iPS cells as well as the challenges to their use in the clinic.
Suppression of experimental allergic encephalomyelitis by intraventricular administration of interferon-gamma in Lewis rats
Authors: VOORTHUIS J A C , UITDEHAAG B M J , DE GROOT C J A , GOEDE P H , MEIDE P H VAN DER , DIJKSTRA C D
Published: July 2010

Experimental allergic encephalomyelitis (EAE) is an autoimmune inflammatory disease of the central nervous system (CNS) which causes paralysis. Several studies have reported the involvement of Ia antigen-expressing cells in the pathogenesis of EAE. Interferon-gamma (IFN-γ) can induce Ia antigen expression on a wide range of cells. We examined the effect of IFN-γ on EAE in Lewis rats. Systemically administered IFN-γ did not change the disease course of EAE, whereas IFN-γ applied locally into the ventricular system of the CNS resulted in complete suppression of clinical signs. Furthermore, we found that systemic administration of anti-IFN-γ just prior to the onset of clinical symptoms resulted in a more severe disease course. We conclude that IFN-γ is capable of exerting a suppressive action in EAE, possibly through induction of Ia antigen expression or through the induction of suppressive mechanisms locally in the CNS.
Autoantibodies to mitochondrial and centromere antigens in primary biliary cirrhosis and systemic sclerosis
Authors: MCHUGH N J , JAMES I E , FAIRBURN K , MADDISON P J
Published: July 2010

High levels of IgG antibodies to the multi-enzyme complex bovine pyruvate dehydrogenase (PDHC) were detected by a sensitive ELISA in all patients with histologically proven primary biliary cirrhosis (PBC), in a minority of patients with systemic sclerosis without overt PBC or with chronic liver diseases of another cause and in no rheumatoid arthritis or normal controls. Most (15/21) but not all of these positive patients recognized a 70-kD (E2) component of PDHC on immunoblotting. Additional recognition of a centromere-associated 140-kD polypeptide was associated with the presence of scleroderma-like features (Raynaud's phenomenon and sclerodactyly) in PBC, further emphasizing the serological and clinical overlap between these two conditions.
Recognition of synthetic peptides of Sm-D autoantigen by lupus sera
Authors: BARAKAT S , BRIAND J-P , WEBER J-C , VAN REGENMORTEL M H V , MULLER S
Published: July 2010

The reactivity of autoantibodies present in the serum of patients with systemic lupus erythematosus (SLE) was investigated by ELISA using seven overlapping synthetic peptides representing the entire sequence of the polypeptide D component of ‘Sm antigen’. Of the 165 SLE sera tested, 59% were found to contain IgG antibodies able to bind to peptide 1–20, while 37% of the sera reacted with peptide 44–67. All sera reacting with peptide 44–67 also reacted with peptide 1–20. These two peptides were only seldom recognized by the sera of 187 patients with other rheumatic autoimmune diseases or by 53 sera of normal individuals. In a parallel study using sera that reacted with the D band in immunoblotting, most of the sera recognized peptides 44–67 (89%) and 1–20 (67%), while 33% of them reacted with peptide 97–119. The use of these synthetic peptides in ELISA may be of considerable help for detecting anti Sm autoantibodies.
Cross-reactive idiotypes in sera from patients with leprosy, lupus and Lyme disease and from healthy individuals
Authors: MACKWORTH-YOUNG C G
Published: July 2010

Monoclonal IgM autoantibodies have previously been generated from a patient with lepromatous leprosy. Polyclonal anti-idiotypes raised against two of these monoclonal antibodies (8E7 and TH9) were used in an immunoassay to detect the presence of idiotype in human serum. The anti-idiotypes recognize different but overlapping sets of idiotypic determinants, some of which are present on antibodies which bind to Mycobacterium leprae. Sera were tested from 16 individuals with leprosy. 45 with systemic lupus erythematosus. 20 with Lyme disease, and 80 healthy subjects. Positive sera were detected in all groups (seven, two, three, and four, respectively). In most cases the serum bound to both anti-idiotypes, the idiotype being present in the IgM and/or IgG fraction. Levels of the two idiotypes varied independently of total serum IgG concentration and, in serial serum samples from one patient, independently of each other. The results indicate that 8E7 and TH9 may be representative of serum antibodies which are commonly expressed in leprosy, but may also be expressed in other diseases and in health; and they suggest that such scrum antibodies are encoded by a widely shared set of variable region genes.
Stimulation of the respiratory burst and promotion of bacterial killing in human granulocytes by intravenous immunoglobulin preparations
Authors: MARÓDI L , KALMÁR A , KARMAZSIN L
Published: July 2010

We have examined the effect of two i.v. immunoglobulin preparations on the metabolic and functional activities of neutrophil granulocytes from the peripheral blood. Production of superoxide anion (O2) by granulocytes was measured through superoxide dismutase inhibitable reduction of ferricytochrome C after incubation of cells for various times together with immunoglobulin (concentration ranging from 0.25 to 5.0 mg/ml). The results showed dose-dependent response of O2 production independent of the incubation time. Granulocytes containing ingested Staphylococcus aureus released a significantly (P < 0.001) larger amount of O2 and killed a higher number (P < 0.001) of viable bacteria in the presence of 5 mg/ml immunoglobulin than did cells incubated in the absence of extracellular i.v. immunoglobulin. These data raise the possibility that immunoglobulin concentrates for i.v. use may enhance the anti-bacterial activities of phagocytic cells through direct stimulation of the respiratory burst. Inflammatory reactions observed during i.v. immunoglobulin infusion in hypo- or agammaglobulinaemic patients may also be related to phagocytic cell activation.
Synovial fluid antigen-presenting cells unmask peripheral blood T cell responses to bacterial antigens in inflammatory arthritis
Authors: LIFE P F , VINER N J , BACON P A , GASTON J S H
Published: July 2010

We and others have previously shown that synovial fluid (SF) mononuclear cells (MC) from patients with both reactive arthritis and other inflammatory arthritides proliferate in vitro in response to bacteria clinically associated with the triggering of reactive arthritis. In all cases, such SFMC responses are greater than the corresponding peripheral blood (PB) MC responses, often markedly so, and the mechanism for this is unclear. We have investigated this phenomenon by comparing the relative abilities of irradiated non-T cells derived from PB and SF to support autologous T cell responses to ReA-associated bacteria. Seven patients whose SFMC had been shown previously to respond to bacteria were studied. We demonstrate antigen-specific responses of PB T cells to bacteria in the presence of SF non-T cells which are in marked contrast to the minimal responses of either unfractionated PBMC or PB T cells reconstituted with PB non-T cells. We also show that PB, but not SF T cells respond strongly to autologous SF non-T cells in the absence of antigen or mitogen. These findings demonstrate that SF antigen-presenting cells (APC) are potent activators of PB T cells. We conclude that the contrasting responses of SFMC and PBMC to bacterial antigens may be accounted for at least in part by an enhanced ability of SF APC to support T cell proliferative responses.
Repeated exposure to bacterial lipopolysaccharide interferes with disposal of pathogenic immune complexes in mice
Authors: CAVALLO T , GRANHOLM N A
Published: July 2010

Patients with systemic lupus erythematosus (SLE) experience clinical flares in association with superimposed bacterial infection. To investigate whether heightened immune phenomena during the course of bacterial infections were related to abnormal disposal of immune complexes, we administered bacterial lipopolysaccharide (LPS) to C57BL/6 mice for 5 weeks. Control mice received vehicle only. We then challenged the mice with a subsaturating dose of radiolabelled immune complexes intravenously and determined the localization of immune complexes in liver, spleen and kidney. In comparison to control mice, mice exposed to LPS developed features of polyclonal B cell activation, autoimmune phenomena, delayed removal of immune complexes from the circulation, diminished liver uptake of immune complexes, and enhanced localization of immune complexes in the kidneys. The findings could not be attributed to biological processes dependent on complement concentration. Instead, interferences with Fc receptor function, or with endocytosis of immune complexes may represent likely possibilities. Thus, clinical flares in patients with SLE, in the presence of a superimposed infection, may result from enhanced localization of immune complexes in organs due to altered mechanisms of their disposal.
Experimental studies of immunologically mediated enteropathy. V. Destructive enteropathy during an acute graft-versus-host reaction in adult BDF1 mice
Authors: MOWAT A McI , FELSTEIN M V
Published: July 2010

As a means of investigating further the pathogenesis of intestinal immunopathology, we have attempted to produce a destructive enteropathy by inducing an acute graft-versus-host reaction (GVHR) in mature, immunocompetent mice. Adult (C57B1/10 × DBA/2)F1 (BDF1) mice given C57B1/10 (B10) spleen cells develop a severe GVHR which is associated with marked weight loss and high mortality. In the intestine an initial phase of enteropathy characterized by intense crypt hyperplasia is replaced by more severe intestinal damage which includes villus atrophy and loss of intra-epithelial lymphocytes. These pathological alterations are paralleled by the generation of anti-host cytotoxic T lymphocytes (CTL). marked immunosuppression and the loss of natural killer (NK) cells. In contrast to these findings, adult BDF1 mice given DBA/2 donor cells do not develop an acute systemic GVHR and have no CTL or intestinal pathology, despite prolonged splenomegaly and enhanced NK cell activity. Thus, destructive enteropathy can be induced during a GVHR in intact hosts and our results confirm that this enteropathy has a biphasic pattern, with villus atrophy representing the progression of initial crypt hyperplasia in severe forms of disease associated with weight loss and specific CTL.
Human monoclonal IgM with autoantibody activity against two gangliosides (GM1 and GDlb) in a patient with motor neuron syndrome
Authors: JAUBERTEAU M O , GUALDE N , PREUD'HOMME J L , RIGAUD M , GIL R , VALLAT J M , BAUMANN N
Published: July 2010

Small amounts of oligoclonal immunoglobulins were detected by Western blotting in the serum from a patient with motor neuron syndrome. The prominent one. a monoclonal IgMλ, reacted strongly with the gangliosides GM1 and GDlb and more weakly with asialo GM1, as shown by immunoenzymatic staining of thin-layer chromatograms of gangliosides, ELISA on purified glycolipid coats and immunoadsorption with purified GM1. Affinity-chromatography with purified GMI resulted in the purification of monoclonal IgMλ. This purified IgM and its Fab fragments showed the same pattern of reactivity with gangliosides as that observed with whole serum. Such monoclonal IgM could be responsible for motor neuron diseases in some patients with overt or barely detectable monoclonal gammopathies.
Aberrant IgG subclass distribution to measles in healthy seropositive individuals, in patients with SSPE and in immunoglobulin-deficient patients
Authors: MATHIESEN T , HAMMARSTRÖM L , FRIDELL E , LINDE A , WIRSEN G , SMITH C I E , NORRBY E , WAHREN B
Published: July 2010

Sera from healthy seropositive donors, patients with acute measles, subacute sclerosing panencephalitis, common variable immunodeficiency, and CH gene deletions were analysed for anti-measles IgG1–4. Compared with other anti-viral immune responses of IgGl and IgG3, an unusual predominance of specific IgGl prevailed; only four out of a total of 68 patients showed anti-measles IgG3. Of the 17 healthy, measles seropositive serum donors, all showed specific IgGl, none showed IgG3 and six had IgG4. Eight out of 10 patients with SSPE showed an anti-measles IgGl and IgG4 response while IgG3 was not seen. The IgGl and IgG4 subclass patterns had some exceptions. Antimeasles IgG3 was found in five out of five patients with deletion of the γ-1 encoding gene segments and in four out of 15 patients with recent measles antigen stimulation. The subclass pattern was suggested to reflect the immunological compromise associated with measles infections.
Echinococcus granulosus: the potential use of specific radiolabelled antibodies in diagnosis by immunoscintigraphy
Authors: ROGAN M T , MORRIS D L , PRITCHARD D I , PERKINS A C
Published: July 2010

Diagnosis of hydatid disease in man is frequently dependent on the imaging of cysts in situ by techniques such as ultrasonography and CAT scans. Such methods are useful but are not specific and can lead to errors in diagnosis. The present work reports preliminary experiments on the development of a specific imaging technique for hydatid cysts using radiolabelled antibodies. A purified preparation of antigen B of hydatid fluid was used to raise polyclonal antisera in rabbits and the resulting affinity-purified IgG labelled with 131I. Gerbils with an established Echinococcus granulosus infection were injected intraperitoneally with the labelled antibody and imaged 48 h later with a gamma camera. Hydatid cysts could be identified within the peritoneal cavity and post-mortem assessment of activity showed the cysts to contain approximately four times as much activity as the surrounding organs thereby indicating successful targeting of the antibody to the cysts.
A phenotypic study of B lymphocyte subpopulations in human bone marrow
Authors: CHAPPLE M R , MACLENNAN I C M , JOHNSON G D
Published: July 2010

The regulatory mechanisms that monitor the size of the peripheral B cell pool and determine cell death or survival are poorly understood. In rodents B lymphopoiesis is maintained at a high rate throughout adult life, and under resting conditions there is little recruitment into the long-lived peripheral pool; it therefore follows that most newly formed B lymphocytes have a very short life-span. The maturation stages of B lymphopoiesis in humans and in experimental mammals appear to be similar. We have determined the phenotype of sIgM- and sIgD-expressing cells from normal adult human bone-marrow and peripheral blood by dual immunofluorescence with an extensive panel of monoclonal antibodies representative of major B cell clusters, in order to identify antigenic differences that may play a regulatory role. Antibodies of the CD21, CD22 and CD9 clusters, the unclustered restricted B antibody 7-F-7 and anti-IgD were reactive with different proportions of sIgM+ cells in blood and bone marrow; 29·5% (range 5·60%) of sIgM+ cells in marrow were sIgD−and most of these cells were also CD21− and CD22−, thus defining a unique marrow population. However, newly formed and mature re-circulating cells comprising the sIgM+ sIgD+ population could not be distinguished by the panel of antibodies.
Lymphocyte activation markers in idiopathic myositis: changes with disease activity and differences among clinical and autoantibody subgroups
Authors: MILLER F W , LOVE L A , BARBIERI S A , BALOW J E , PLOTZ P H
Published: July 2010

We studied the immunologic correlates of disease activity and differences among subgroups of patients with idiopathic inflammatory myopathy by analysing phenotypic and activation marker expression on peripheral blood mononuclear cells (PBMC). Compared with controls, myositis patients with clinically active disease (n = 51) had significantly lower proportions of CD8- cells and higher proportions of PBMC that expressed DR. CD3- DR, CD14- DR, interlcukin-2 receptors. and the late T cell activation markers CD26 and TLiSA1. TLiSA1 expression, a marker for cytotoxic differentiation, correlated significantly with both clinical activity indices and serum levels of muscle-associated enzymes. In serial studies of seven patients, the proportion of PBMC expressing MHC class II antigen and late T cell activation markers decreased as myositis disease activity decreased, independent of type of therapy. Among the clinical subgroups, polymyositis (n= 21) and inclusion body myositis (n=21) were virtually indistinguishable; dcrmatomyositis patients (n= 19) showed decreased proportions of CD3+DR+ and TLiSA1+ cells, and increased proportions of CD20+ and CD20+ DR+ cells compared with the other two groups. Patients with autoantibodies to histidyl-tRNA synthctasc (Jo-1 antigen. n= 11) had significantly lower proportions of CD3+ andCD4+ cells, lower CD4/CD8 ratios, and higher proportions of DR+ cells expressing CD20, compared with patients without anti-Jo-1 antibodies. These findings support the concept that activated lymphocytes, especially cells undergoing anamnestic responses and cytotoxic differentiation, are important in the pathogenesis of idiopathic myositis. Moreover, taken together with other studies, these data suggest that groups of patients segregated by clinical or autoantibody status have different mechanisms of systemic immune activation and immunopathology.
Autoantibodies against neutrophil cytoplasm components in systemic lupus erythematosus and in hydralazine-induced lupus
Authors: NÄSSBERGER L , SJÖHOLM A G , JONSSON H , STURFELT G , ÅKESSON A
Published: July 2010

Anti-neutrophil cytoplasm antibody (ANCA) has been shown to be no marker of systemic lupus erythematosus (SLE) including lupus nephritis or of progressive systemic sclerosis (PSS). Antibodies against myeloperoxidase (anti-MPO) and elastase, two granulocyte lysosomal enzymes, were found in patients with SLE but not in those with PSS, except for one patient who had anti-MPO. Anti-MPO was present in 21% of patients with SLE, and at low concentrations in about 80% of these cases. Anti-elastase was found in four patients with SLE. In another group of six patients with a SLE-like syndrome induced by anti-hypertensive treatment with the anti-hypertensive hydralazine, anti-MPO antibodies occurred in all six, and anti-elastase antibodies in five. Monitored during a 2-year follow-up period, anti-MPO antibodies were found to persist, whereas anti-elastase antibodies were rapidly eliminated, after withdrawal of the drug.
Participation of eosinophils in the toxic oil syndrome
Authors: TEN R M , KEPHART G M , POSADA M , ABAITUA I , SOLDEVILLA L , KILBOURNE E M , DUNNETTE S L , GLEICH G J
Published: July 2010

The participation of eosinophils in the Spanish toxic oil syndrome (TOS) was investigated. Eosinophil infiltration and degranulation in tissues from 52 patients with the TOS were examined by immunofluorescence staining for the eosinophil granule major basic protein (MBP). Serum MBP levels were determined in sera from 323 patients. Eosinophil infiltration and degranulation were found in several tissues, especially during the acute phase of the TOS, and serum MBP was significantly elevated during all phases of the disease, suggesting that eosinophils play a role in the pathogenesis of the TOS.
Immunologic abnormality in NZB/W Fl mice. Thymus-independent expansion of B cells responding to interleukin-6
Authors: MIHARA M , FUKUI H , KOISHIHARA Y , SAITO M , OHSUGI Y
Published: July 2010

We have previously reported that B cell abnormality in NZB/W Fl mice developed independently of thymus. Here we examined further whether B cells from NZB/W F1 mice respond to interleukin-6 (IL-6), a factor for terminal differentiation of B cells. When freshly isolated splenic B cells were incubated for 5 days in the presence of human IL-6, an increased production of both IgM and IgG, including anti-DNA antibody, was evident in NZB/W Fl mice; there was no increase in BALB/c mice. A magnitude of augmentation in IgG but not IgM production by IL-6 became more apparent in older NZB/W F1 mice. The increased immunoglobulin production seen with IL-6 was neutralized by treatment with rabbit anti-recombinant human IL-6 antibody. As B cells from athymic NZB/W Fl nude mice also responded to IL-6, it was suggested that B cells in NZB/W Fl mice differentiated into the IL-6-responding state in a thymus-independent manner. This B cell abnormality may be associated with the pathogenesis of autoimmune disease in NZB/W Fl mice.
HL-60 cells induced to differentiate towards neutrophils subsequently die via apoptosis
Authors: MARTIN S J , BRADLEY J G , COTTER T G
Published: July 2010

The human promyelocytic HL-60 cell line can be induced to differentiate to neutrophil-like cells in response to a variety of chemical stimuli. We have found that retinoic acid-treatment of HL-60 cells over a period of 6–8 days resulted in a progressive increase in the proportion of cells with mature neutrophil morphologies and was closely followed by an increase in the proportion of cells exhibiting the morphological characteristics of apoptosis, the non-pathological mode of cell death. Using Percoll step-density gradients we have demonstrated a marked increase in the buoyant density of these cells and have used this density difference to obtain enriched fractions of cells for more detailed study. Degradation of the nuclear DNA of these cells into integer multiples of about 200 base pairs, indicative of endogenous endonuclease activation a major characteristic of programmed cell death, was also demonstrated. From these observations we conclude that the mode of cell death in cultures of terminally differentiated HL-60 cells is that of apoptosis. These results parallel those of a recent report which has shown apoptosis to be the mode of cell death of ageing peripheral blood neutrophils. Because of this, we believe that our observations further validate the use of the HL-60 cell line as a model system for the study of human granulopoiesis in vitro and further, that this model system may be useful for gaining insight into the underlying mechanisms involved in apoptosis.
Antibody-mediated demyelination in experimental allergic encephalomyelitis is independent of complement membrane attack complex formation
Authors: PIDDLESDEN S , LASSMANN H , LAFFAFIAN I , MORGAN B P , LININGTON C
Published: July 2010

The effects of decomplementation by cobra venom factor (CVF) on the pathogenesis of inflammation and demyelination in experimental allergic encephalomyelitis (EAE) and acute antibody-mediated demyelinating EAE (ADEAE) have been quantified histologically and immunocytochcmically. In rats immunized with 50 μg of myelin basic protein in Freund's complete adjuvant containing 100μg heat-killed Mycobacterium tuberculosis H37Ra, clinical signs of EAE were completely suppressed by two injections of CVF given 9 and 12 days post-immunization. Suppression of clinical disease was associated with a dramatic reduction in peri-vascular inflammation in the CNS. although immunohistochemical staining identified small numbers of infiltrating T cells and macrophages. In contrast, CVF treatment had no significant effect on the clinical severity of ADEAE and although C9 deposition within the CNS was virtually abolished, there was no statistically significant decrease in the extent of demyelination or inflammation. These observations indicate that in the absence of complement components C3 and C5 an antibody-dependent cell-mediated cytotoxic response plays an important role in the pathogenesis of antibody-mediated demyelination. The major role of the complement cascade in EAE appears to be the generation of pro-inflammatory factors that enhance the inflammatory response within the CNS in animals facing a mild encephalitogenic challenge.
Cytokine-independent progression of immunoglobulin productionin vitroby B lymphocytes from patients with systemic lupus erythematosus
Authors: PELTON B K , SPECKMAIER M , HYLTON W , FARRANT J , DENMAN A M
Published: July 2010

B lymphocytes from patients with systemic lupus erythematosus (SLE) secreted high levels of immunoglobulin spontaneously when cultured in vitro. Addition of the cytokines interleukin-2, interleukin-4 and interleukin-6 either alone or in combination failed to augment spontaneous immunoglobulin synthesis. Percoll-separated low-density SLE B lymphocytes matured into immunoglobulin-secreting cells also independent of exogenous interleukins. During maturation these cells became enlarged and less dense, and began to express CD23. This was in contrast to normal B cells, which did not secrete immunoglobulin spontaneously but synthesized IgM after interleukin stimulation. These results indicate that in vitro immunoglobulin synthesis by SLE B cells is already initiated in these cells and progresses independently of further stimulatory manoeuvres.
Cyclosporin A in the prevention and treatment of experimental autoimmune glomerulonephritis in the brown Norway rat
Authors: REYNOLDS J , CASHMAN S J , EVANS D J , PUSEY C D
Published: July 2010

Experimental autoimmune glomerulonephritis (EAG) was induced in brown Norway (BN) rats by a single i.m. injection of collagenase-solubilized homologous glomerular basement membrane (GBM) in Freund’scomplete adjuvant. This model of anti-GBM disease is characterized by the development. over several weeks, of circulating and deposited anti-GBM antibodies, accompanied by albuminuria. We examined the effects of treatment with oral cyclosporin A (CsA) at different doses, starting at the time of immunization and during the course of the disease. Prctreatmcnt with CsA 5 mg/kg daily produced a moderate reduction in circulating anti-GBM antibody levels, reduced deposition of antibody on the GBM and decreased albuminuria. Doses of 10 and 20 mg/kg CsA produced a marked reduction in circulating antibody, absence of delectable deposited antibody and virtual absence of albuminuria. Renal function remained normal in CsA-treated and control animals. When CsA treatment was introduced at 2 or 4 weeks after immunization, there were significant effects on the subsequent autoimmune response and albuminuria at 10 and 20 mg/kg daily. These studies demonstrate that CsA in conventional doses has a therapeutic effect in this model of anti-GBM disease, and suggest a role for T lymphocytes in the palhogcncsis of EAG.
Clinical efficacy of intravenous immunoglobulin in patients with severe inflammatory chest disease and IgG3 subclass deficiency
Authors: BERNATOWSKA-MATUSZKIEWICZ E , PAC M , SKOPCYNSKA H , PUM M , EIBL M M
Published: July 2010

To investigate the efficacy of i.v. IgG treatment in pediatric patients with inflammatory lung disease, a prospective, controlled clinical trial was carried out over a 2-year study period. Patients were enrolled on the basis of severe clinical symptomatology. After 1 year of conventional treatment, the patients received 400 mg/kg per month of an i.v. IgG product containing only trace amounts of IgG3 in addition to their regular treatment throughout the second year. Significant clinical improvement, as documented by duration of hospital stay (first year 27.8 days, second year 4.9 days), use of antibiotics (132.8 versus 30.9 days) and use of steroids (21.4 versus 0.7 days) could be observed. Data obtained on a subgroup of patients with lgG3 deficiency were analysed separately. These results indicate that patients with severe chest disease who have lgG3 deficiency will also benefit from i.v. IgG treatment. The mode of action cannot be attributed to replacement of the respective isotypes, but is probably due to the effect of i.v. IgG in preventing repeated viral infections.
Human double-negative (CD4-CD8-) T cells bearing αβ T cell receptor possess both helper and cytotoxic activities
Authors: MATSUMOTO M , YASUKAWA M , INATSUKI A , KOBAYASHI Y
Published: July 2010

Expression of CD4 or CD8 on the cell surface is an important guide for discriminating the immunological functions of T cells. However, a minor T cell subset, which lacks both CD4 and CD8 molecules but bears the usual form of T cell receptor (TCR) αβ (CD4-CD8-TCRαβ+ T cells), has recently been found not only in mice but also in humans, and its role in immune response is now of considerable interest. In order to clarify the characteristics of this newly defined T cell subpopulation, we established five IL-2-dependent CD4-CD8-TCRαβ+ T cell clones from the peripheral blood of a healthy individual, and examined their various biological functions. It was found that all clones not only helped B cells in immunoglobulin production, but also exerted major histocompatibility complex-unrestricted cytotoxicity. Although their CD3/TCR complexes were functionally competent, the cytotoxicity seemed to be mediated via unknown molecules other than the CD3/TCR complex, as evidenced by the failure of CD3 MoAb to inhibit the cytotoxic activity. Our present findings showed that CD4-CD8-TCRαβ+ T cells possess potential bifunction, i.e. helper and cytotoxic activities. Their roles in the pathogenesis of immunodeficiency are discussed.
Intravenous immunoglobulins (IVIg) in the treatment of autoimmune diseases
Authors: KAVERI S-V , DIETRICH G , HUREZ V , KAZATCHKINE M D
Published: July 2010

Intravenous immunoglobulin (IVIg) therapy is increasingly used in autoimmune diseases. Although its efficacy has only been established in a few specific antibody-mediated autoimmune conditions, accumulating evidence on the regulatory role of circulating immunoglobulins in the selection of peripheral B cell repertoires makes it an attractive potential therapeutic option to clinical immunologists. This overview briefly discusses the current use of IVIg in human autoimmune diseases with a particular emphasis on the possible mechanisms by which IVIg could suppress pathological autoimmune responses.
Clonal analysis of liver-infiltrating T cells in patients with LKM-1 antibody-positive autoimmune chronic active hepatitis
Authors: LÖHR H , MANNS M , KYRIATSOULIS A , LOHSE A W , TRAUTWEIN C , BÜSCHENFELDE K-H MEYER ZUM , FLEISCHER B
Published: July 2010

Autoantibodies against microsomal antigen of liver and kidney (LKM-1) are diagnostic markers for a subgroup of autoimmune chronic active hepatitis (AI-CAH). Cytochrome P4S0dbl, now classified as cytochrome P450 IID6, is the major antigen of LKM-1 antibodies. Immunohistological studies suggest that hepatic injury in AI-CAH is mediated by liver-infiltrating T cells. In the present study the specificity and function of liver-infiltrating T cells was analysed at the clonal level. Phenotypical characterization of 189 T cell clones isolated from four liver biopsies of LKM-1 antibody-positive patients showed an enrichment of CD4+CD8- T cells. Five CD4+CD8- T cell clones proliferated specifically in the presence of recombinant human LKM-1 antigen (rLKM). This reaction was restricted to autologous antigen-presenting cells and to HLA class II molecules. In order to see whether rLKM was also recognized by peripheral blood T lymphocytes (PBL) we tested the proliferative response of PBL from several individuals. PBL from three of the four patients with LKM-1 antibody-positive AI-CAH proliferated to rLKM, whereas no response was seen with PBL from patients with LKM-1 antibody-negative chronic liver diseases and from healthy blood donors. These data demonstrate that the LKM-1 antigen is recognized by liver-infiltrating T cells in LKM-1 antibody-positive AI-CAH. For further functional characterization, liver-derived T cell clones were tested for their cytotoxic activity. In the presence of phytohacmagglutinin 24 out of 26 CD4-CD8+ but also 20 out of 63 CD4+CD8- T cell clones lysed autologous as well as allogenic EBV-transformed B cell lines or K562 cells. Five CD4-CD8+ T cell clones lysed autologous but not allogenic B cell lines spontaneously in a HLA class I-restricted manner. Although the antigen specificity of these clones is still unknown the data show the presence of autoreactive T cells at the site of inflammation that could contribute in the pathogenesis of AI-CAH.
Antibodies to acetylcholine receptors in myasthenia gravis.In vitrosynthesis by peripheral blood lymphocytes before and after thymectomy
Authors: KUKS J B M , LIMBURG P C , OOSTERHUIS H J G H , THE T H
Published: July 2010

Pokeweed antiogen (PWM)-d riven in vitro synthesis of antibodies to the acetylcholine receptor (PSA) was stucture. in non-thymoma patients with myasthenia gravis. In a group of 46 patients, the occurrence: of PSA was related to the presence of the thymus or, in operated patients, the absence of a clinical effect of thymectomy. Sixteen patients were followed before and soon after thymectomy. PSA disappeared in all patients, at least temporarily, between 6 weeks and I year afterwards, independent of the clinical course and eventual clinical effect of the operation. A recurrence was found only in one of the five patients who derived no benefit from the operation. These findings support the hypothesis that the therapeutic effect of thymectomy can be explained by removal of a source of autoreactive lymphocytes. There was no correlation between the changes in serum levels of a-AChR and clinical improvement, suggesting a minor role of circulating peripheral blood lymphocytes (PBL) and the thymus in the total production of a-AChR.
Phagocytosis ofGiardia lambliatrophozoites by cytokine-activated macrophages
Authors: BELOSEVIC M , DANIELS C W
Published: July 2010

Phagocytosis of Giardia lamblia trophozoites by cytokine-activaled and non-activated bone marrow-derived macrophages was examined in vitro. Macrophages treated with recombinant interferon-gamma (IFN-γ) and bacterial lipopolysaccharide (LPS) ingested a significantly higher number of in vitro-grown trophozoites than untreated macrophages. Maximal uptake of parasites occurred after 4 h and 6 h of incubation where 81.4% and 79.1% of macrophages were positive for trophozoites. Other cytokines tested, IL-2, IL-3, IL-4, IL-5, GM-CSF, CSF-I and tumour necrosis factor-alpha (TNF-α) either alone or in combination with LPS, failed to activate macrophages to phagocytose G. lamblia. The induction of this activated macrophage anti-microbial function was achieved pharmacologically using phorbol myristate acetate (PMA) and ionophore A23187. The giardicidal activity of macrophages activated with IFN-γ and LPS or that induced by PMA and A23187 was inhibited by H-7, indicating the role for protein kinase C in the intracellular events following activation.