bluuebunny/crossref_metadata_2025 huggingface dataset:
Impaired high‐shear‐stress‐induced platelet aggregation in patients with chronic renal failure undergoing haemodialysis
Authors: Yoshida Eri , Fujimura Yoshihiro , Ikeda Yasuo , Takeda Ichiroh , Yamamoto Yasuhiro , Nishikawa Kiyoshi , Miyataka Kazuhiko , Oonuki Masahiro , Kawasaki Tomihisa , Katayama Masahiko , Narita Nobuhiro
Published: July 2010

We investigated shear‐induced platelet aggregation (SIPA) in 30 patients with chronic renal failure (CRF) undergoing haemodialysis. 26 patients showed a significant decrease in SIPA at high shear stress but no change in SIPA at low shear stress. The former reaction reflects the interaction between plasma von Willebrand factor (vWF) and its platelet receptors, glycoprotein (GP) Ib‐IX and lIb/ IlIa complex, whereas the latter is assumed to involve the binding of plasma fibrinogen to GP IIb/IIIa complex. These SIPA profiles in CRF patients after haemodialysis showed almost no change compared to those before haemodialysis.

The ratio of ristocetin cofactor/vWF antigen in plasma was slightly lower in CRF patients than in controls (P<0.01). However, the level of GPIb antigen in the platelets of these patients was significantly reduced (42.1±20‐3% of normal platelets), with partial destruction of GPIb antigen. The number of vWF receptors on the GPIb molecule was quantitated using the GPIb‐binding protein alboaggregin‐B (AL‐B), purified from the snake venom of Trimeresurus albolabris. AL‐B bound to GPIb at a total of 48 760±9944 molecules per normal platelet and a Kd of 85.44±15.70 nM at saturation. In contrast, binding in CRF platelets was 22 980±6395 molecules per platelet and Kd was 50.08±13.83 nM. Taking these results together, we conclude that the impaired SIPA found in CRF patients is due to both abnormalities in plasma vWF and in its platelet GPIb receptor.
A possible anti‐emetic role for sodium valproate in cytotoxic chemotherapy
Authors: Aiken T. C. , Collin R. C.
Published: July 2010

We report two patients with acute leukaemia who received emetogenic cytotoxic drugs and were on therapeutic doses of sodium valproate for epilepsy. Neither patient reported significant nausea nor vomited at any time during the chemotherapy, at times requiring no anti‐emetic treatment whatsoever. We suggest that this absence of nausea was due to an anti‐emetic effect of sodium valproate.
Detection of NPM‐ALK DNA rearrangement in CD30 positive anaplastic large cell lymphoma
Authors: Waggott W. , Lo Y.‐M. D. , Bastard C. , Gatter K. C. , Leroux D. , Mason D. Y. , Boultwood J. , Wainscoat J. S.
Published: July 2010

CD30 positive anaplastic large cell lymphoma (ALCL) is a type of non‐Hodgkin's lymphoma associated with a specific chromosome translocation between chromosomes 2 and 5. Recent molecular characterization of the translocation breakpoint has identified a gene fusion between NPM (nucleophosmin) and ALK (anaplastic lymphoma kinase). Using a DNA hybridization technique, the NPM rearrangement was found among 5/5 ALCL samples. We have developed a PCT methodology which has enabled the detection of the NPM‐ALK rearrangements amongst seven t(2; 5)(p23; q35) ALCL cases based on a long‐range PCR of genomic DNA. The rapidity and robustness of this method may have diagnostic applications for ALCL.
Expression of interleukin (IL) 1 type I and type II receptors in megakaryocytic cells and enhancing effects of IL‐1β on megakaryocytopoiesis and NF‐E2 expression
Authors: Yang Mo , Li Karen , Chui Cecilia Mei Yan , Yuen Patrick Man Pan , Chan Po Kwok , Chuen Carmen Ka Yee , Li Chi Kong , Fok AndTai Fai
Published: July 2010

Megakaryocytopoiesis is regulated by thrombopoietin (TPO) and cytokines such as interleukin 3 (IL‐3), IL‐6 and IL‐11. This study investigated the in vitro effects of IL‐1β on megakaryocytopoiesis and the expression of IL‐1 type I and type II receptors (IL‐1 RI and RII) on megakaryocytic cell lines and primary cells. Our results demonstrated that IL‐1β alone or in combination with TPO induced megakaryocyte colony forming units (CFU‐MK) from murine and human haematopoietic cells. Using reverse‐transcription polymerase chain reaction (RT‐PCR) and Southern hybridization techniques, the mRNA of IL‐1β, IL‐1 RI, IL‐1 RII and the transcription factor NF‐E2 were detected in CD61+CD41+ cells cultured from cord blood and four megakaryocytic cell lines, Meg‐01, DAMI, CHRF‐288–11 and M‐07e. The expression of IL‐1 RI and RII proteins was confirmed by flow cytometry and immunofluorescence staining. In Meg‐01 cells, the expression of NF‐E2 was increased at both mRNA and protein levels after treatment with IL‐1β for 4 h. This study demonstrated for the first time the presence of IL‐1 receptors on megakaryocytic cells and the induction of NF‐E2 by IL‐1β. The mitogenic effect of IL‐1β on this lineage could be mediated through IL‐1 receptors and the activation of NF‐E2.
Contrasting in vitro effects for the combination of fludarabine, cytosine arabinoside (Ara‐C) and granulocyte colony‐stimulating factor (FLAG) compared with daunorubicin and Ara‐C in P‐glycoprotein‐positive and P‐glycoprotein‐negative acute myeloblastic leukaemia
Authors: Higashi Y. , Turzanski J. , Pallis M. , Russell N. H.
Published: July 2010

It has been suggested that the FLAG remission induction regimen comprising fludarabine (F‐ara), cytosine arabinoside (Ara‐C) and granulocyte colony‐stimulating factor (G‐CSF) may be capable of overcoming P‐glycoprotein (P‐gp)‐related multidrug resistance (MDR) in patients with acute myeloblastic leukaemia (AML). We have investigated the in vitro response of P‐gp‐positive and ‐negative AML clones to FLAG and compared this with their response to treatment with Ara‐C and daunorubicin (DNR). Twenty‐four cryopreserved samples from patients with AML were studied using a flow cytometric technique for the enumeration of viable (7‐amino actinomycin D negative) cells. Samples consisted of 12 P‐gp‐positive and 12 P‐gp‐negative cases, as measured by the MRK16 antibody. The results were analysed by calculating the comparative drug resistance (CDR), i.e. the percentage cell death caused by Ara‐C + DNR subtracted from the percentage cell death, caused by FLAG after 48 h incubation in suspension culture. P‐gp‐positive clones were shown to have a significantly higher CDR than P‐gp‐negative clones (P = 0·001). Furthermore, a significant positive correlation (r2 = 0·40, P < 0·01) was found between P‐gp protein expression and CDR. However, P‐gp function, measured using cyclosporin modulation of rhodamine 123 (R123) uptake, was not associated with the CDR, demonstrating that there are other properties of P‐gp, besides its role in drug efflux, that modulate the responsiveness of AML blasts to chemotherapy. These results are consistent with a potential benefit for FLAG in P‐gp‐positive AML, but not P‐gp‐negative AML, compared with standard anthracycline and Ara‐C therapy.
Elevated serum and bronchoalveolar lavage fluid levels of interleukin 8 and granulocyte colony‐stimulating factor associated with the acute chest syndrome in patients with sickle cell disease
Authors: Abboud Miguel R. , Taylor Ellen C. , Habib David , Dantzler‐Johnson Terrie , Jackson Sherron M. , Xu Fushen , Laver Joseph , Ballas Samir K.
Published: July 2010

The role of cytokines in the development of acute chest syndrome (ACS) in patients with sickle cell disease (SCD) was studied. Serum interleukin 8 (IL‐8) levels were elevated in 14 episodes and undetectable in six out of 20 episodes of ACS in 19 patients with SCD. In contrast, IL‐8 levels were undetectable in the sera of 29 control patients with SCD studied during routine clinic visits or hospitalization for vaso‐occlusive crises. The differences in mean IL‐8 levels and the proportion of patients with detectable levels between the two groups were highly significant (P < 0·0001 and 0·04 respectively). The mean IL‐8 level in bronchial fluid samples from children with ACS was also significantly higher than that in sickle cell patients undergoing elective surgery (5500 ± 1400 pg/ml vs. 1900 ± 470 pg/ml, P = 0·03). Granulocyte colony‐stimulating factor (G‐CSF) (2000 ± 1700 pg/ml) was present in five out of six samples of bronchial fluid, but not serum, from children with ACS. All but one of the patients with ACS studied were negative for the Duffy red cell antigen, which is a receptor that binds and inactivates IL‐8 and other chemokines. These findings suggest that IL‐8 and G‐CSF may play a role in the development of the ACS and the complications associated with it.
Single nucleotide polymorphisms of the factor IX gene for linkage analysis in the southern Chinese population
Authors: Chan Vivian , Yam Irene , Yip B. , Au P. , Shing M. K. , Li C. K. , Chan T. K.
Published: July 2010

Carrier detection and prenatal testing for haemophilia B in Oriental populations have been hampered by the lack of informative markers within the factor IX (FIX) gene. We detected a T/C nucleotide variation at nucleotide 32770 in the poly‐A region of the FIX gene in the mother of a haemophilia B child. Analysis of 139 unrelated alleles revealed a heterozygosity rate of 0·193, thus offering an additional marker for linkage analysis. Together with two other polymorphic sites (5′MseI and 3′HhaI) found in Chinese and Thai populations, these polymorphisms were useful in 66% of the families studied.
Bone mineral metabolism in adults with β‐thalassaemia major and intermedia
Authors: Pollak Rivka Dresner , Rachmilewitz Eliezer , Blumenfeld Anat , Idelson Maria , Goldfarb Ada W.
Published: July 2010

Bone disease is an important cause of morbidity in older patients with β‐thalassaemia major and intermedia. We studied 27 women and 23 men with β‐thalassaemia major (37) and intermedia (13) whose mean age was 32·3 ± 9·7 years. Bone mineral density (BMD) of the lumbar spine, femoral neck and distal radius was determined by dual‐energy X‐ray absorbiometry (DXA). The longitudinal change in BMD over a mean of 5·6 years was determined in 19 patients. Serum 25‐hydroxyvitamin D, insulin growth factor‐1 (IGF‐1), bone formation markers bone‐alkaline phosphatase, osteocalcin and the resorption marker urinary N‐telopeptide cross‐linked type 1 collagen (NTx) were determined. The BsmI vitamin D receptor (VDR) gene polymorphism was analysed. Reduced BMD (Z‐score < −2) was present in 89%, 62% and 73% of patients in the spine, hip and radius respectively. Vitamin D deficiency was found in 62%, decreased IGF‐1 in 72% and increased urinary NTx in 84% of patients. Serum IGF‐1 correlated with spine and hip BMD (r = 0·4, r = 0·39, P < 0·01 respectively), and NTx correlated with the hip BMD Z‐score (r = 0·35 P < 0·05). The mean annual percentage change in spine BMD was −1·36%. Patients with the VDR BB genotype had lower spine BMD than patients with the bb genotype. In conclusion, bone loss continues in adult thalassaemia patients and is associated with increased bone resorption and decreased IGF‐1. The BsmI VDR gene polymorphism is associated with osteopenia in thalassaemia.
Effect of the angiotensin‐converting enzyme gene deletion polymorphism on the risk of venous thromboembolism
Authors: Jackson Andrew , Brown Karen , Langdown Jonathan , Luddington Roger , Baglin Trevor
Published: July 2010

Allele frequencies for the insertion/deletion (I/D) polymorphism of the angiotensin‐converting enzyme (ACE) gene were determined in a large case–control study of 517 unselected patients with venous thromboembolism and 478 blood donors. The D allele frequency was 0·53 [95% confidence interval (CI) 0·50–0·56] in patients and 0·54 (95% CI 0·50–0·57) in controls, giving an odds ratio for the D allele of 0·97 (95% CI 0·81–1·16). In the same population, the odds ratio for the factor V Leiden mutation (F5G1691A) was 6·9 (95% CI 4·0–11·9). Therefore, the ACE I/D polymorphism is not a risk factor in a representative group of unselected patients with venous thromboembolism. The possibility that the I/D polymorphism is a risk factor for venous thromboembolism specifically after hip replacement cannot be excluded.
Is it necessary to administer anti‐D to prevent RhD immunization after the transfusion of RhD‐positive platelet concentrates?
Authors: Atoyebi W. , Mundy N. , Croxton T. , Littlewood T. J. , Murphy M. F.
Published: July 2010

Serology for the presence of anti‐D after RhD‐incompatible platelet transfusions was performed in 24 RhD‐negative patients with haematological disease and 59 RhD‐negative patients with non‐haematological disease. None of the patients were given prophylaxis with anti‐D to prevent RhD immunization. Eight out of 59 (13·5%) non‐haematology patients developed detectable anti‐D, whereas 0 out of 24 (0%) of the haematology patients formed anti‐D (P = 0·06). The risk of alloimmunization after RhD‐incompatible platelet transfusions using platelet concentrates prepared by modern technical methods appears to be small in patients with haematological disease, but is significant in non‐immunocompromised patients.
A 1063G→A mutation in exon 12 of glycoprotein (GP)IIb associated with a thrombasthenic phenotype: mutation analysis of [324E]GPIIb
Authors: Tao Jianming , Arias‐Salgado Elena G. , González‐Manchón Consuelo , Iruín Gemma , Butta Nora , Ayuso Matilde S. , Parrilla Roberto
Published: July 2010

We report the molecular, genetic and functional analysis of a case of thrombasthenic phenotype. The proband showed absence of platelet glycoprotein (GP)IIb and very low content of GPIIIa, and both his parents showed a marked reduction in the levels of platelet GPIIb‐IIIa. Single‐stranded conformational polymorphism‐polymerase chain reaction (SSCP‐PCR) analysis and direct sequencing of PCR‐amplified GPIIb exon‐12 revealed the presence of a G→A transition at position 1063 with the expected substitution of glutamate 324 with lysine (K). This mutation did not alter the level of GPIIb mRNA. Co‐expression of normal or mutant [324K] GPIIb with normal human GPIIIa in Chinese hamster ovary (CHO) cells failed to show surface exposure of [324K]GPIIb‐IIIa complexes. Pulse‐chase and immunoprecipitation analysis demonstrated that [324K]GPIIb cDNA was translated into proGPIIb, but neither mutant GPIIb heavy chain (GPIIbH) nor [324K]GPIIb‐GPIIIa complexes were detected, suggesting that this mutation is the underlying molecular basis for the thrombasthenic phenotype. Mutation analysis demonstrated that 324E of GPIIb could be replaced by other negatively charged or polar amino acids (AAs) without impairing the surface expression of GPIIb‐IIIa. However, substitution of 324E of GPIIb for a positively charged AA other than K prevented the expression of GPIIb‐IIIa complexes. These observations suggest that a domain encompassing 324E of GPIIb is essential for heterodimerization with GPIIIa and its substitution for a positively charged residue precludes normal subunit association.
The platelet thrombopoietin receptor number and function are markedly decreased in patients with essential thrombocythaemia
Authors: Li Junzhi , Xia Yuping , Kuter David J.
Published: July 2010

Essential thrombocythaemia (ET) is a relatively common myeloproliferative disorder characterized by an elevated platelet count. As thrombopoietin (TPO) and the TPO receptor (c‐mpl) regulate platelet production in normal physiology, their role in ET was investigated. A well‐characterized cohort of 23 ET patients was evaluated and followed for 3 years. The TPO levels in these ET patients (189 ± 131 pg/ml) were the same as in normal subjects (179 ± 112 pg/ml) and TPO was not produced by ET platelets. There were 5·6 ± 5·5 TPO binding sites/ET platelet vs. 56 ± 17 TPO binding sites/normal platelet and this was associated in ET patients with normal‐sized platelet c‐mpl protein and mRNA, but a 10‐fold reduction in platelet c‐mpl mRNA. The Kd for the TPO receptor on ET platelets was 66 ± 30 pmol/l vs. 163 ± 31 pmol/l on normal platelets, but the c‐mpl cDNA had a normal nucleic acid sequence. The decreased number of ET platelet TPO receptors resulted in a fourfold decrease in the platelet‐dependent TPO clearance (0·30 ± 0·14 ml/h/109 ET platelets vs. 1·24 ± 0·38 ml/h/109 normal platelets) at a time when the platelet count in ET patients was 2·7‐fold above normal. The fourfold decrease in the TPO clearance, elevated platelet mass and resulting normal total TPO clearance explain the normal TPO levels. These results also suggest that the thrombocytosis in ET may be attributed to an alteration of the normal feedback interaction between TPO and its receptor and not as a result of any defect in the structure of TPO or c‐mpl.
Clinical course and predictive factors for cyclosporin‐induced autologous graft‐versus‐host disease after autologous haematopoietic stem cell transplantation
Authors: Baron Frédéric , Gothot André , Salmon Jean‐Paul , Hermanne Jean‐Philippe , Pierard Gérald E. , Fillet Georges , Beguin Yves
Published: July 2010

The administration of cyclosporin A (CyA) after autologous haematopoietic stem cell transplantation (HSCT) induces a systemic autoimmune syndrome mimicking graft‐vs.‐host disease (GVHD). This syndrome, termed autologous GVHD has notable anti‐tumour activity in animal studies. We intended to induce autologous GVHD with CyA in patients undergoing an autologous HSCT. We prospectively studied 118 patients with miscellaneous malignancies undergoing an autologous HSCT with low‐dose CyA to characterize the clinical syndrome, its frequency and clinical course, and to determine the factors affecting its incidence. Patients received CyA from d −1 through to d 28, first starting at 2 mg/kg intravenously and then orally as soon as feasible. The dose was adjusted to achieve pre‐dose blood levels around 100 ng/ml. A skin biopsy was performed when a skin rash was observed. Thirty‐three percent of the patients developed clinical GVHD: clinical stage 1 in 21 patients, stage 2 in seven patients, and stage 3 in three patients. Although total body irradiation (TBI) or high‐dose cyclophosphamide were previously thought to be needed, autologous GVHD occurred in five out of 12 patients (42%) after a preparative regimen with high‐dose melphalan alone. Autologous GVHD was significantly more frequent in patients older than 33 years, in patients who had received high doses of granulocyte‐macrophage colony forming units (CFU‐GM) and in those with a diagnosis of myeloid malignancy, compared with those with lymphoid malignancies or solid tumours. A significant negative association was also found with HLA‐DR6. In lymphoma patients, GVHD occurred more frequently in advanced disease than in first or second complete remission (CR1–2) patients. All other factors studied were not predictive for GVHD. In conclusion, CyA‐induced GVHD is reproducibly and safely induced with doses of CyA adapted to achieve blood levels around 100 ng/ml. In retrospective analysis, there was no survival advantage for patients with GVHD. Phase III trials with this approach are needed to evaluate its anti‐tumoral effect.
Cytogenetic responses in high‐risk myelodysplastic syndrome following low‐dose treatment with the DNA methylation inhibitor 5‐aza‐2′‐deoxycytidine
Authors: Lübbert Michael , Wijermans Pierre , Kunzmann Regina , Verhoef Gregor , Bosly Andre , Ravoet Christophe , Andre Marc , Ferrant Augustin
Published: July 2010

Decitabine (5‐aza‐2′‐deoxycytidine) acts as a powerful demethylating agent in vitro. Clinically, low‐dose decitabine ameliorates cytopenias including induction of trilineage responses in ≈50% of patients with high‐risk myelodysplastic syndrome (MDS). We examined the incidence and kinetics of cytogenetic responses to decitabine in these patients. Of 115 successfully karyotyped patients, 61 (53%) had clonal chromosomal abnormalities prior to treatment. Major cytogenetic responses were observed in 19 patients (31% of those with abnormal cytogenetics, 17% of all patients by intention‐to‐treat) after a median of three courses (range, 2–6) until best cytogenetic response. Progressive decrease of the abnormal clone over time was also determined using fluorescence in situ hybridization (FISH) analysis in two patients. Median duration of cytogenetic responses was 7·5 months (range, 3–15). Analysis of response by the International Prognostic Scoring System (IPSS) cytogenetic risk groups revealed three out of five cytogenetic responses (60%) in the IPSS ‘low‐risk’ group, 6 out of 30 with ‘intermediate risk’ (20%) and 10 out of 26 in the ‘high‐risk’ group (38%). Median survival in these cytogenetic subgroups was 30, 8 and 13 months respectively. The relative risk of death in patients achieving a major cytogenetic response was 0·38 (95% confidence interval 0·17–0·88) compared with patients in whom the cytogenetically abnormal clone persisted (P = 0·0213). In conclusion, repeated courses of low‐dose decitabine induce cytogenetic remissions in a substantial number of elderly MDS patients with pre‐existing chromosomal abnormalites; these are associated with improved survival compared with patients in whom the cytogenetically abnormal clone persists. Patients with ‘high‐risk’ chromosomal abnormalities may particularly benefit from this treatment.
Fibrinogen γ′ chain carboxy terminal peptide selectively inhibits the intrinsic coagulation pathway
Authors: Lovely Rehana S. , Boshkov Lynn K. , Marzec Ulla M. , Hanson Stephen R. , Farrell David H.
Published: July 2010

The minor γA/γ′ isoform of fibrinogen contains a high affinity binding site for thrombin exosite II that is lacking in the major fibrinogen isoform, γA/γA fibrinogen. The biological consequences of γ′ chain binding to thrombin were therefore investigated. Coagulation assays, thrombin activity assays, and a primate thrombosis model were used to characterize the biological effects of the γ′ 410–427 peptide. The γ′ peptide had little effect on thrombin cleavage of the small peptidyl substrate tosyl‐glycyl‐prolyl‐arginine‐4‐nitranilide acetate. However, in vitro assays demonstrated that the γ′ peptide inhibited thrombin cleavage of larger proteinaceous substrates, including fibrinogen and factor VIII. The γ′ peptide inhibited the activated partial thromboplastin time in plasma and showed greater inhibition of activated partial thromboplastin time assays than prothrombin time assays, consistent with the inhibition of factor VIII cleavage. Studies in a baboon thrombosis model showed that the γ′ 410–427 peptide inhibited fibrin‐rich thrombus formation (typical of venous thrombi) and, to a lesser extent, platelet‐rich thrombus formation (typical of arterial thrombi). These results indicate that binding of thrombin exosite II by the γ′ peptide has selective effects on the intrinsic pathway.
Identification and functional characterization of a polymorphic oestrogen response element in the human coagulation factor IX gene promoter
Authors: Adams B. , Western A. K. , Winship P. R.
Published: July 2010

The liver expressed procoagulant factor IX (FIX) shows inter‐individual variation in levels, some of which is heritable. Raised levels of FIX are associated with a thrombotic tendency. This study demonstrated that, in females but not males, part of this variation in FIX levels is due to polymorphic genotype at a locus in the factor IX gene (F9) promoter 698 bp upstream of the major transcription initiation site (−698C/T). The −698C allele (associated with higher FIX level) shows closer homology to a canonical ORE sequence and a higher binding affinity for oestrogen receptor α than the −698T allele. Reporter gene vectors were constructed with elements spanning residues −738 to +50 of the F9 promoter corresponding to wild type −698C and −698T alleles. A related series of vectors comprising three copies of the F9 ORE driving expression of a minimal synthetic promoter were also created. Transfection into the liver‐derived HepG2 and erythroleukaemic K562 cell lines demonstrated increased levels of expression in the presence of oestrogenic factors when compared to those found in their absence; this stimulation was more pronounced in the non‐liver derived K562 cell line and from the reporter vectors containing promoter elements corresponding to the −698C allele.
IgG subclass levels and immune reconstitution after T cell-depleted allogeneic bone marrow transplantation
Authors: KELSEY S M , LOWDELL M W , NEWLAND A C
Published: July 2010

Defects of humoral immunity are well documented after bone marrow transplantation (BMT). Immunoglobulin recovery can be impaired and selective deficiencies of IgG subclasses have been reported. The nature of these deficiencies may reflect patterns of infection in the post-BMT period. We studied immunoglobulin and IgG subclass recovery in 20 long term (> 100 days) survivors of T depleted allogeneic BMT. Although there was no fall in mean levels of IgG, IgM or IgA for the patient group, 14 patients (70%) developed a deficiency of one or more immunoglobulin isotype at some stage post-BMT. Eight patients (40%) had deficiency of IgG, IgA and IgM and six had selective deficiencies. When IgG subclasses were measured it was seen that mean levels of IgG2 and IgG4 fell post-BMT with trough levels occurring at around 120 days post-transplant. Sixty per cent of patients developed IgG2 subclass deficiency and of these patients 78% had an associated IgG4 deficiency. Deficiencies of IgG1 and IgG3 were less common and less prolonged than those of lgG2 and lgG4; in addition, mean levels of IgG1 and IgG3 showed a rise early post-BMT. In conclusion, a majority of our patients developed immunoparesis following BMT, usually at around 120 days after transplantation. IgG2 subclass deficiency, often in association with IgG4 deficiency, is common and may occur despite normal total IgG levels. Deficiencies of immunoglobulin and IgG subclasses may persist for longer than 1 year post-BMT. Differing profiles of immunoglobulin and IgG subclass recovery may help dictate patterns of infection in long-term survivors of BMT.
Modulation of leucocyte locomotion by interleukin-1
Authors: ZIELINSKI C C , PESAU B , KALINOWSKI W , MÜLLER C
Published: July 2010

The influence of interleukin-1 (IL-1) upon leucocyte locomotion in vitro was studied, using either casein or zymosan-activated serum (ZAS) as chemotaxigens. A pre-incubation of polymorpho-nuclear cells (PMNC) with ultrapure (purity 99%) human IL-1 (1 U/ml) for 2 and 5 h followed by a washing step resulted in a significant decrease in leucocyte locomotion against casein (P < 0.0005 and P<0.01, respectively), but not against ZAS (P>0.1). Moreover, the direct addition of this IL-1 preparation to leucocyte locomotion assays without pre-incubation produced a similar and significant inhibition of leucocyte locomotion directed against casein (P < 005). This inhibitory effect could not be augmented further by higher concentrations of ultrapure IL-1. In order to exclude the effect of possibly contaminating cytokines in the ultrapure IL-1 preparation used, additional assays with recombinant human IL-1 (rIL-1) alpha and rIL-1 beta (1 U/ml) were performed to investigate their influence upon leucocyte locomotion. It was found that both rIL-1α and rIL-1β inhibited leucocyte locomotion directed against casein significantly (P < 0.005). Similar to the previous experiments, leucocyte locomotion could not be further inhibited by higher concentrations (10 U/ml) of rIL-1. Thus, both ultrapure and rIL-1 were found to have the ability to inhibit leucocyte locomotion in vitro.
Oestrogen is a potent disease accelerator in SLE-prone MRL lpr/lpr mice
Authors: CARLSTEN H , TARKOWSKI A , HOLMDAHL R , NILSSON L-Å
Published: July 2010

The influence of oestrogen on the lupus disease in MRL/1 mice has been investigated. Adult, castrated male and female MRL/1 mice were administered with s.c. injections of 3–2 μg of 17β-oestradiol twice a week. The results clearly demonstrate that a relatively small dose of oestrogen is a potent accelerator of the lupus disease in this mouse strain. Thus, administration of oestrogen accelerates glomerulonephritis, lymphoproliferation and mortality. Our results also indicate that oestrogen exerts a dual effect on the immune system of MRL/1 mice by depression of antigen-specific and mitogen-induced T cell responses as well as enhancement of polyclonal Bcell activation and autoantibody formation. In addition, even short-term administration of oestrogen in the preclinical phase of the disease resulted in long-lasting effects as evaluated by reduced longevity and aggravation of renal disease.
Depletion of CD8+ T cells suppresses growth of Trypanosoma brucei brucei and interferon-gamma) production in infected rats
Authors: BAKHIET M , OLSSON T , VAN DER MEIDE P , KRISTENSSON K
Published: July 2010

Sprague-Dawley rats infected with Trypanosoma brucei brucei showed a strong and rapid induction of splenocyte IFN-γ (within 12 h post-infection) as measured by a single cell assay for IFN-γ secretion. Depiction of CD8+ cells in infected rats abrogated the IFN-γ production, suppressed parasite growth and increased survival of the animals. Induction of MHC class I antigens in the paraventricular and supra-optic hypothalamic nuclei caused by the trypanosome infection was also inhibited by the CD8+ cell depletion. It is suggested that the CD8+ cells are involved directly or indirectly in growth regulation of the parasite and that IFN-γ induced by the parasite may be one of the factors that trigger MHC expression and immunosuppression.
CD4+ T cells play a major role for IgM and IgG anti-DNA production in mice infected withPlasmodium yoelii
Authors: FOSSATI L , MERINO J , IZUI S
Published: July 2010

The infection by a non-lethal strain of Plasmodium yoelii induces the formation of autoantibodies such as anti-DNA and anti-Sm antibodies in mice. The extent of the relative increase in serum levels of IgM and IgG anti-DNA and anti-Sm antibodies and their kinetics were found to be similar to those of anti-hapten antibodies and of total IgM and IgG levels. This strongly suggested that anti-DNA and anti-Sm autoantibody responses observed in malaria-infected mice are a result of polyclonal activation of B cells. The analysis of the IgG subclasses reacting with DNA antigen showed significant levels of the T cell-dependent isotypes, IgG1 and IgG2. The role of T cells in the activation of autoreactive B cells was confirmed by using athymic nude mice. Indeed, BALB/c-nu/nu and C57BL/6-nu/nu mice failed to produce IgG anti-DNA antibodies after infection with P. yoelii. Moreover, the reconstitution of BALB/c nude mice with lymph node cells from congenic euthymic BALB-Igb mice showed the activation of autoreactive B cells in nude mice by T cells from euthymic mice. Studies in mice depleted of CD4+ T cells strongly suggested that malaria-induced anti-DNA antibodies were almost entirely dependent on the presence of CD4+ T cells, as this depletion significantly decreased IgM anti-DNA antibodies and completely abolished the IgG anti-DNA production, including the IgG3 subclass in infected mice. In contrast, depletion of the CD8+ T cell subset had no effect on the production of autoantibody in malaria-infected mice. Our results indicate that CD4+ T cells play a major role for both IgM and IgG anti-DNA production during the course of malaria infection.
Mononuclear cells in glomeruli and cytokines in urine reflect the severity of experimental proliferative immune complex glomerulonephritis
Authors: NOBLE B , REN K , TAVERNE J , DIPIRRO J , VAN LIEW J , DUKSTRA C , JANOSSY G , POULTER L W
Published: July 2010

Immunohistochemical methods were used to investigate the role of macrophages in the progression of proliferative immune complex glomerulonephritis. The mononuclear cell component of glomerular inflammation was analysed in three different stages of chronic serum sickness, each of which was clearly distinguished by criteria of kidney function. Urinary excretion of the macrophage secretory products interleukin-1 and tumour necrosis factor was also evaluated in relation to the functional severity of kidney disease. T lymphocytes and macrophages began to accumulate in glomeruli at the onset of proteinuria, but not before. Urinary excretion of interleukin-1 also began with proteinuria. Proteinuria increased in direct correlation with increases in the number of glomerular macrophages. Development of the most severe stage of glomerulonephritis, characterized by cachexia, declining kidney function, and necrotizing glomerular pathology, was accompanied by the disappearance of T cells from glomeruli and the expression of highly abnormal phenotypes by most macrophages. In addition, there was a switch from urinary excretion of interleukin-1 to excretion of tumour necrosis factor. The progression of proliferative immune complex glomerulonephritis was associated with qualitative as well as quantitative changes in glomerular macrophage populations. Differentiation and/or activation of those glomerular macrophages may have resulted from local T cell-mediated immunoregulation. Measurements of urinary cytokine excretion provided a reliable means of monitoring disease progression. The local action of tumour necrosis factor probably contributed to declining kidney function in the most severe stage of disease.
Isolation of a Trypanosoma cruzi antigen by affinity chromatography with a monoclonal antibody. Preliminary evaluation of its possible applications in serological tests
Authors: CARBONETTO C H , MALCHIODI E L , CHIARAMONTE M , DE ISOLA E DURANTE , FOSSATI C A , MARGNI R A
Published: July 2010

By affinity chromatography with a monoclonal antibody (163B6), obtained in our laboratory, we have isolated a T. cruzi antigen which could be useful for differential diagnosis of Chagas' disease from leishmaniasis. This antigen, a 52-kD protein, reacted with all sera from Chagas1 disease patients tested but not with sera from patients with leishmania, in ELISA. The 52-kD antigen is widely distributed in the Trypanosoma genus since the 163B6 monoclonal antibody reacts with T. rangeli and 8 strains and a clone of T. cruzi epimastigotes.
The specificity of anti-neutrophil cytoplasm autoantibodies in systemic vasculitides
Authors: LAI K N , JAYNE D R W , BROWNLEE A , LOCKWOOD C M
Published: July 2010

Autoantibodies to neutrophil cytoplasmic antigens are found in patients with small or medium-size vessel systemic vasculitis such as Wegcner's granulomatosis and microscopic polyartcritis. We studied the specificity of anti-neutrophil cytoplasm autoanlibodies (ANCA) by examining the binding of sera from patients with different forms of systemic vasculitis, to neulrophil extract size-fractionated by gel filtration for high performance liquid chromatography. Sequential fractions were collected for solid-phase radioimmunoassay. Patients with Wegcner's granulomatosis showed two peaks of activity over fractions of 12 and 2 kD. Patients with microscopic polyarteritis had two different peaks of aclivity over fractions of 100 and 25 kD. Patients with Takayasu's arteritis and dermal lymphocytic vasculitis were studied as controls (all were ANCA negative by indirect immunofluorescence studies and solid-phase radioimmunoassay). However, the patient with lymphoeytic vasculilis showed a distinct peak of activity over 22 kD and patients with Takayasu's arteritis had a single peak of activity over 200 kD. These results suggest the binding pattern of sera to fractionated neutrophil extract could be useful in differentiating between different diseases within the spectrum of systemic vasculilis. Cryptic antigen recognition in certain vasculitides may only be revealed by this procedure. We conclude that the association of ANCA and systemic vasculitis may not be limited to diseases of small or medium-size vessels, but could also include large vessel disease such as Takayasu's arteritis.
CD3-T cell receptor modulation is selectively induced in CD8 but not CD4 lymphocytes cultured in agar
Authors: OUDRHIRI N , FARCET J P , GOURDIN M F , M'BEMBA E , GAULARD P H , KATZ A , DIVINE M , GALAZKA A , REYES F
Published: July 2010

The CD3-T cell receptor (TcR) complex is central to the immune response. Upon binding by specific ligands, internalized CD3-TcR molecules increase, and either T cell response or unresponsiveness may ensue depending on the triggering conditions. Using semi-solid agar culture, we have shown previously that quiescent CD4 but not CDS lymphocytes generate clonal colonies under phyto-haemagglutinin stimulation. Here we have demonstrated that the agar induces selective CD3-TcR modulation in the CD8 and not in the CD4 subset. CDS lymphocytes preactivated in liquid culture and recultured in agar with exogenous recombinant interleukin-2 generate colonies with a modulated CD3-TcR surface expression. The peptides composing the CD3-TcR complex are synthesized in CD8 colonies as well as in CD4; however, the CD3 gamma chain is phosphorylated at a higher level in CD8 colonies. A component of the agar polymer, absent in agarose, appears to be the ligand that induces differential CD3-TcR modulation in the CD8 subset. In contrast to agar culture, CD8 colonies can be derived from quiescent CD8 lymphocytes in agarose. These CD8 colonies express unmodulated CD-TcR. CD3-TcR modulation with anti-CD3 monoclonal antibody prior to culturing in agarose inhibits the colony formation. We conclude that given triggering conditions can result in both CD3-TcR modulation and inhibition of the proliferative response selectively in the CD8 lymphocyte subset and not in the CD4.
Immunochemical characterization of human antibodies to lymphobiastoid interferon
Authors: THURMOND L M , REESE M J
Published: July 2010

Antibodies produced in recurrent respiratory papillomatosis (RRP) patients treated with Iymphoblastoid interferon (1yIKN) may neutralize antiviral activity or may only bind to 1yIFN. These antibodies were characterized for immunoglobulin class. IgG subclass. and light chain type by an indirect immunoassay. Serum dilutions were incubated on 1yIFN-coated plates and the presence of antibody detected using peroxidase-conjugated goat antibodies to each human immunoglobulin class and light chain isotype, or using MoAbs to each human IgG subclass. Neutralizing activity was measured as the inhibition of 1yIFN antiviral activity for Vervet monkey cells challenged with Semliki Forest Virus. Among antibody-positive patients, 12% produced IgM coincident with IgG, and 25% produced IgA coincident with IgG. Thus, antibody responses in patients treated with 1yIFN are not exclusively of IgG class. The predominant 1yIFN-specific subclasses were IgG1 and IgG3, which occurred in 70% and 83% of patients, respectively. An IgG4 response was detected in two patients who also had antibody of other isotypes; no IgG2 antibody was detected in any patient. Antibodies were not IgG subclass-restricted, a trend which was more pronounced in patients having neutralizing antibody than non-neutralizing antibody. Light chain molecules of 1yIFN-specific antibody were of both k and λ isotypes. with k chains occurring most frequently. Among patients having non-neutralizing antibodies, monotypic light chains occurred in 65% of the patients, whereas no patient with neutralizing antibody had monotypic light chain antibody. Sera from 599 normal human volunteers were assayed for antibody, and seven were found to be immunoreactive to 1yIFN. Only-one serum of the seven was positive for neutralizing activity.
Cytotoxic CD4+ T cells specific forFrancisella tularensis
Authors: SURCEL H M , TAPANINAHO S , HERVA E
Published: July 2010

The specific cell-mediated cytotoxicity of tularaemia-immune human T lymphocytes were studied in vitro. Peripheral blood mononuclear cells (PBMC) of six tularaemia-vaccinated healthy subjects were stimulated with F. tularensis LVS whole cell antigen for 6 days and used as effector cells in a conventional 4-h 5lCr release cytotoxicity assay. The target cells were phagocyting autologous monocytes, which were pulsed with F. tularensis or PPD antigen. The specific lysis of the F. tularensis pulsed cells (42.6%± 11.7) was significantly higher (P < 0.05) than that of the PPD-pulsed ones (22.2%± 8.3) or unpulsed control cells (15.9%± 5.2). The cytotoxicity was associated with CD4+F. tularensis-specific T cell clones (TLC), which killed 36.3%± 12.3 of the F. tularensis-pulsed targets but only 6.9%± 6.5 of the unpulsed control targets. Their lysing was inhibited by monoclonal anti-HLA-DR and anti-HLA-DQ antibodies, but not by CD 15 (monocyte/macrophage) antibody. The functional role of CD4+ lymphocytes in tularaemia immunity is discussed.
Cellular origins of human polymeric and monomeric IgA: enumeration of single cells secreting polymeric IgA1 and IgA2 in peripheral blood, bone marrow, spleen, gingiva and synovial tissue
Authors: TARKOWSKI A , MOLDOVEANU Z , KOOPMAN W J , RADL J , HAAIJMAN J J , MESTECKY J
Published: July 2010

Using modified ELISA and spot-ELISA, which permit the parallel determination of heavy chain subclass and the presence of covalently linked J chain, we analysed IgA found in cell culture supernatants or secreted by individual cells from peripheral blood, spleen, bone marrow, gingiva and synovial tissue, with respect to its polymeric or monomeric IgA form (plgA, mlgA) and IgA I or IgA2 subclass. The ELISA for determination of J chain in tissue culture supcrnalants was specific and highly sensitive (detection limit in pg). The results demonstrated that IgA1-producing cells predominated in the tissues examined, and that J chain could be detected in association with the majority of IgA1 and lgA2 secreted by individual cells. With respect to the frequency of cells secreting polymeric, J chain-containing IgA, only 20-30% of cells from the bone marrow were engaged in the synthesis of pIgA. In other tissues the frequency of cells secreting pIgA1 and plgA2 was considerably higher. Peripheral blood mononuclear cells secreting pIgA2 were easily inducible during stimulation with T cell-dependent pokeweed mitogen, whereas Epstein-Barr virus-transformed cells secreted preferentially mIgA1. When the frequencies of plgA-, pIgA 1 - or pIgA 2-secret ing cells (determined by spot-ELISA technique) from different tissues were correlated with the proportion of pIgA EG mIgA (and IgA subclasses) secreted in tissue culture supernatants. data obtained suggest that many individual IgA-producing cells could be engaged in simultaneous secretion of mIgA and pIgA.
Immunoregulatory effects of Sizofiran (SPG) on lymphocytes and polymorphonuclear leukocytes
Authors: YONEDA K , UETA E , YAMAMOTO T , OSAKI T
Published: July 2010

The effect of SPG on leukocytes has been studied in 20 patients with oral carcinoma and the actions have been analysed in vitro. SPG 1 mg/kg was administered intramuscularly twice weekly. Peripheral venous blood was collected before, and 1 week and 2 weeks after the initiation of SPG treatment. Both CD16+ CD57– and CD16–CD57+ cell populations were significantly increased after treatment, but no T cell subset varied. While enhancement of lymphokine-activated killer activity could not be found, an increase in natural killer (NK) activity was observed in 15 of the subjects, and the mean NK level was significantly increased from an initial 34.7 ± 18.7% to 46.4 ± 16.5% after two weeks of injections. O2-production by polymorphonuclear leukocytes (PMNL) was stimulated 6 h after SPG injection. When PMNL were treated in vitro with SPG 32 μg/ml. enhanced 02-generation was induced and protein kinase C (PKC) activity in a membrane fraction increased. SPG did not directly affect non-specific PMNL killing of K562 cells or antibody-dependent cell mediated cytotoxicity against Raji cells, but non-specific PMNL killing was enhanced by culture-conditioned medium from peripheral blood mononuclear cells (PBMC) containing 10 μg/ml SPG. Interleukin-1β, -3, -4, -6, tumour necrosis factor-α, granulocyte-macrophage colony stimulating factor and IFN-γ levels in the conditioned medium were not increased compared with medium from PBMC not treated with SPG. No clear increase of these cytokines was found in serum from the SPG-treated patients. From the above results, enhancement of PMNL O2-generation by SPG seems to be a direct action of SPG, but the mechanism of elevation of the non-specific killing activity of PMNL and NK cells is not known. Perhaps other cytokines than those assayed have participated in increasing non-specific cytotoxicity.
Concomitant augmentation of CD4+CD29+ helper inducer and diminution of CD4+CD45RA+ suppressor inducer subset in patients infected with human T cell lymphotropic virus types I or II
Authors: LAL R B , RUDOLPH D L , SCHMID D S , LAIRMORE M D
Published: July 2010

To examine the immunomodulatory effects of HTLV infection, lymphocyte subset analysis was performed on patients infected with human T cell lymphotropic virus type-I (HTLV-I, n= 6) or -II (HTLV-II, n= 12) and on normal blood donors (n= 16). The percentages of total B lymphocytes (CD19), natural killer (NK) cells (CDI6), T lymphocytes and their subsets (CD2, CD3, CD4, CD5, CD7, CDS), and IL-2R (CD2S) were found to be within the range found in normal donors. However, the expression of CD8+ HLA-DR+ increased significantly in patients with HTLV-I or HTLV-II infection (14.1 ± 3.9% and 9.7 ± 2.4% respectively; P<0.01) when compared with controls (3.2 ± 1.1%). In addition, there was a significantly greater proportion of CD4+ CD29+ T lymphocytes (29.3 ± 6.1% and 31.1 ± 9.0%; P<0.05) with concomitant diminution of CD4+CD45RA+ T lymphocytes (8.3 ± 3.3% and 11.4 ± 1.5%; P<0.01) in patients infected with HTLV-I or HTLV-II respectively, when compared with controls. The increased percentage of CD4+CD29+ subpopulations showed a direct correlation (rs=0.86; P<0.001) with HTLV-specific antibody production. No difference in the CD8 population coexpressing CD29 and S6F1 (an epitope of LFA-1) were observed in the HTLV-infcctcd group when compared with normal donors and functional analysis exhibited minimal cytotoxicity against lectin labelled heterologous target cells. Thus, the shift in the suppressor/cytotoxic to helper/inducer‘memory’ CD4+ may be associated with immunoregulatory abnormalities often found in persons infected with HTLV-I or HTLV-II.
Effect of synthetic thymic humoral factor (THF-γ2) on T cell activities in immunodeficient ageing mice
Authors: GOSO C , FRASCA D , DORIA G
Published: July 2010

Immunodeficient ageing (C57BL/10 × DBA/2)F1 mice were treated by a single injection of synthetic thymic hormones and 4 days later their thymus and spleen cells were assayed in vitro for T cell activities. A few nanograms of THF–γ2 were found to raise the frequency of mitogen-responsive T cells in thymus and spleen cell populations as well as the frequency of cytokine-producing splenic T cells, up to the levels observed in young mice. Moreover, injection of THF-γ2 was found to restore T cell growth factor (TCGF) production by mitogen-stimulated spleen cells. Also, the helper activity of spleen cells was enhanced by this treatment and increased with increasing the THF-γ2 dose over a wide range. Similarly, the effects of thymopentin and thymosin–α1 on T helper cell activity increased with increasing the injected dose, but the efficiencies of THF-γ2 and thymopentin were, respectively. 400-fold and eight-fold greater than that of thymosin-α1.
Prolonged clinically asymptomatic evolution after HIV-1 infection is marked by the absence of complement C4 null alleles at the MHC
Authors: HENTGES F , HOFFMANN A , DE ARAUJO F OLIVEIRA , HEMMER R
Published: July 2010

The length of time after which persons infected with HIV-1 progress to AIDS is variable. Certain alleles at the MHC have been shown to influence negatively the clinical outcome of HIV-1-infected persons and to be associated with special clinical manifestations. We investigated the MHC class I, class II and class III antigens in 54 Caucasian HIV-1-infected persons. The MHC profile of individuals with a prolonged period before AIDS is marked by a lower frequency of C4 null alleles.