bluuebunny/crossref_metadata_2025 huggingface dataset:
Autoimmunity against blood cells in human immunodeficiency‐virus (HIV) infection
Authors: van der Lelie J. , Lange J. M. A. , Vos J. J. E. , van Dalen C. M. , Danner S. A. , von dem Borne A. E. G. Kr.
Published: July 2010

Summary. In persons with AIDS or at risk from AIDS, autoantibodies against platelets and granulocytes were frequently detected. Platelet‐bound immunoglobulins were demonstrated by immunofluorescence in all 16 patients with AIDS, in five out of seven patients with AIDS‐related complex/persistent generalized lymphadenopathy (ARC/PGL) and even in seven of 10 healthy sexually active homosexual men. Granulocyte‐bound immunoglobulins were found by immunofluorescence in 12 of the 16 AIDS patients, five of the seven patients with ARC/PGL and two of the 10 symptomless men. Red cell bound immunoglobulins were not detected. All patients with AIDS and ARC/PGL and three of the symptomless men were seropositive for human immunodeficiency virus (HIV).

The platelet‐ and granulocyte‐bound immunoglobulins could be eluted in 93% and 67% of the cases, respectively. This indicates that specific autoantibodies, rather than circulating immune complexes, which were frequently increased, accounted for the findings.

There was no relation between the serological findings and the platelet and granulocyte counts. We conclude that autoantibodies against platelets and granulocytes are common in patients with AIDS and those at risk.
Myelodysplastic syndromes: evolution of overt leukaemia by one or several steps of transformation
Authors: Dörmer P. , Hershko C. , Voss R. , Wilmanns W.
Published: July 2010

The evolution of leukaemia was studied prospectively in 29 patients with myelodysplastic syndrome (MDS) followed for 2–6 years by sequential blast counts, cell kinetics derived from quantitative14C‐autoradiography and karyotype analysis. Overt leukaemia developed in seven patients. Two distinct patterns of leukaemic evolution were identified. The first was characterized by a gradual increase in blast cell count and in the frequency of labelled blasts, and a corresponding reduction in myeloid maturation index indicating increased intracompartmental myeloblast divisions and premature myeloid cell death. A second pattern of leukaemic evolution was marked by a sudden rise in the blast cell population in a previously stable MDS. This rise was attributed both to an increased rate of blast proliferation, and the accumulation of non‐proliferating blasts. In an additional patient with smouldering ANLL and multiple karyotype abnormalities, transient clinical remission took place following prednisone and oxymetholone therapy, characterized by a sideroblastic morphology, normal karyotype, and persistence of a highly abnormal myeloid maturation index. The sudden emergence of overt leukaemia in previously stable MDS in some of our patients and the temporary reversal of overt leukaemia into sideroblastic anaemia in one case, lend support to the notion of leukaemic evolution by several steps of transformation. On the other hand, the gradual transition of MDS into overt leukaemia in other patients is compatible with a single step leukaemic transformation, although the possibility of clonal disease prior to the development of MDS cannot be excluded with certainty.
Acquired loss of red cell Kell antigens
Authors: Vengelen‐Tyler V. , Gonzalez B. , Garratty G. , Kruppe C. , Johnson C. L. , Mueller K. A. , Marsh W. L.
Published: July 2010

Summary. A 19‐year‐old patient with a long history of idiopathic thrombocytopenic purpura developed a potent antibody against a high‐incidence antigen in the Kell blood group system. The direct antiglobulin test on his red cells was negative. His cells exhibited profound depression of Kell blood group antigens, but antigens of other blood groups were normal. Transfusion of incompatible blood was well tolerated and differential agglutination tests, using selected Rh antisera, showed in vivo survival of the transfused red cells for more than 8 weeks. However, the transfused red cells also showed acquired loss of Kell antigens. Five months after the initial findings, Kell‐related antibody disappeared and Kell antigens reappeared on his red cells. The patient's serum stored from the initial investigation now reacted with his freshly collected red cells. These data suggest that an environmental agent in the patient's plasma was responsible for the temporary loss of Kell antigens from red cells in his circulation.
Interaction of urokinase with specific receptors abolishes the time of commitment to terminal differentiation of murine erythroleukaemia (Friend) cells
Authors: Rosso Mario Del , Pucci Marco , Fibbi Gabriella , Dini Germana
Published: July 2010

We have shown the presence of specific receptors for human urokinase on the surface of mouse erythroleukaemic cells. The receptor number increases when undifferentiated cells undergo hexamethylene‐bisacetamide induced differentiation, while affinity between receptor and ligand does not change. A monoclonal antibody against the 17 500 mol wt fragment of the non‐catalytic chain of urokinase (A chain) inhibits the specific binding, as we have previously shown in A431 cells. We have found that, upon the simultaneous addition of both urokinase or the A chain and hexamethylenebisacetamide, commitment is initiated immediately without the lag shown by control cultures of undiffer‐entiated cells treated with the low molecular weight inducer alone. A series of mechanisms possibly involved in the action of urokinase on mouse erythroleukaemic differentiation are also discussed.
Modulation of human marrow haematopoiesis by leucotrienes in vitro
Authors: Estrov Zeev , Halperin Daniel S. , Coceani Flavio , Freedman Melvin H.
Published: July 2010

The influence of products derived from the arachidonic acid 5‐lipoxygenase (5‐LO) pathway on normal human marrow haematopoiesis was studied using clonogenic assays. The leucotrienes (LT) B4 and C4 caused a reduction in granulocyte‐macrophage (CFU‐GM) as well as erythroid (CFU‐E and BFU‐E) colony numbers in a dose‐dependent manner. Conversely, nordihydroguaiaretic acid (NDGA), a LT synthesis inhibitor, promoted proliferation of these colonies. The inhibitory effect of LTC4 but not that of LTB4 was abolished by FPL‐55712, a LT antagonist; the enhanced colony growth induced by NDGA was reversed by both LTB4 and LTC4. LTD4 had no significant effect on colony growth despite its structural similarity to LTC4, which implies that the haematopoietic suppression by LTB4 and LTC4 is specific for these compounds. Depletion of marrow T‐cells or removal of adherent cells, or both, did not alter the response to LTB4 and LTC4, suggesting that LT action is exerted directly on progenitor cells and probably is not mediated by other cell populations. Our studies show that the 5‐LO pathway is functional in these culture systems and yields products with inhibitory properties. The 5‐LO pathway may have an important regulatory function in normal marrow haematopoiesis.
Presence of antibody to A‐ and B‐transferases in minor incompatible bone marrow transplants
Authors: Barbolla L. , Mojena M. , Boscá L.
Published: July 2010

The contribution of the bone marrow to plasma A‐ or B‐transferase activities has been studied in patients who underwent incompatible bone marrow transplantation (BMT). As deduced from major incompatibility (group O recipient/A donor), the contribution of the marrow to these plasma activities was c. 5–10% of the total activity. In cases of minor incompatible transplants (A recipient/O donor), normal plasma activity was present in two patients, while no activity was found in a further two in whom a potent anti‐transferase was detected. The antibody inhibited both A‐ and B‐transferase activities to a high titre. The patients in whom this antibody arose exhibited severe graft‐versus‐host disease.
Long‐term survival and late relapses in acute leukaemia in adults
Authors: Brincker H. , Christensen B. E.
Published: July 2010

34 out of 403 apparently unselected adult patients with acute leukaemia referred to a single department from 1970 through 1989 survived more than 3 years. The cumulative rate of relapse after 3 years was 39% in patients with acute myeloblastic leukaemia (AML) and 74% in patients with acute lymphoblastic leukaemia (ALL). The latest relapse was observed 75 months after diagnosis in AML and 98 months after diagnosis in ALL. 65% of the long‐term survivors were able to undertake normal physical activity, 26% had decreased activity, and 9% were unable to work.

5‐year survival for all patients, whether treated or not, during two successive decades was 16% versus 18% and 5% versus 6%, respectively, for ALL and AML. The departmental results were identical with population‐based national results. Only in patients 15–49 years of age with AML was there evidence that more intensive treatment had led to better survival.
Clinical and laboratory evaluation of National Health Service factor VIII concentrate (8Y) for the treatment of von Willebrand's disease
Authors: Cumming A. M. , Fildes S. , Cumming I. R. , Wensley R. T. , Redding O. M. , Burn A. M.
Published: July 2010

This study was carried out to assess the efficacy of NHS 8Y concentrate in the treatment of patients with von Willebrand's disease (vWD). Eight patients (two type I vWD, one type IIA vWD, two type IIB vWD, and three type III vWD) were treated on a total of 10 occasions with 8Y. Following each treatment episode there was a temporary correction of patients’bleeding time (BT) measurements. Other laboratory parameters—von Willebrand factor ristocetin cofactor activity (vWf:RiCo), vWf antigen (vWf:Ag) levels, and factor VIII coagulant activity (factor VIII:C)—were also corrected. Plasma vWf multimers temporarily reflected those present in the infused concentrate. An effective clinical response was observed in each case despite, as revealed by autoradiography and scanning densitometry of SDS‐agarose electrophoresis gels, a reduction in the concentration of the largest vWf multimers in 8Y compared with normal plasma. Overall, the clinical effectiveness of 8Y in vWD was comparable to that seen with cryoprecipitate. We conclude that NHS 8Y concentrate may be used as an alternative to cryoprecipitate for the treatment of vWD.
Cytokine gene expression in peripheral blood mononuclear cells during graft‐versus‐host disease after allogeneic bone marrow transplantation
Authors: Tanaka Junji , Imamura Masahiro , Kasai Masaharu , Masauzi Nobuo , Matsuura Atsushi , Ohizumi Hiroko , Morii Ken , Kiyama Yoshio , Naohara Tohru , Saitho Masao , Higa Toshio , Honke Koichi , Gasa Shinsei , Sakurada Keisuke , Miyazaki Tamotsu
Published: July 2010

Summary. Cytokine gene expression in peripheral blood mononuclear cells during the development of graft‐versus‐host disease (GVHD) in patients who underwent allogeneic bone marrow transplantation (allo BMT) was analysed using a semiquantitative reverse‐transcriptase polymerase chain reaction (RT‐PCR). The expression of interleukin (IL)‐lβ, IL‐6, and tumour necrosis factor (TNF)‐α mRNA was increased during the development of GVHD and the degree of this increment depended on the severity of the disease. IL‐2 expression was not detected at all and interferon‐γ expression was not much changed during GVHD. In patients with hepatic veno‐occlusive disease (VOD), another transplantation‐related complication, the expression of IL‐1β and TNF‐a mRNA was increased but IL‐6 mRNA expression showed little increase. These findings suggest that IL‐lβ, IL‐6 and TNF‐α produced by peripheral blood mononuclear cells play an important role in the development of GVHD. Furthermore, liver dysfunction due to GVHD or VOD may be distinguishable by this type of cytokine analysis. Analysis of cytokine mRNA expression in peripheral blood mononuclear cells after allogeneic bone marrow transplantation may provide important information concerning the immune response and the cytokine network system in marrow transplant patients.
Effect of different human immunodeficiency virus type‐1 (HIV‐1) isolates on long‐term bone marrow haemopoiesis
Authors: Cen Dazhi , Zauli Giorgio , Szarnicki Robert , Davis Brian R.
Published: July 2010

Summary. Haemopoietic cytopenias are a frequent occurrence in human immunodeficiency virus type‐1 (HIV‐1) induced disease. In order to examine the possible direct inhibition of marrow haemopoiesis by HIV‐1, we have investigated the effect of HIV‐1 infection on myelopoiesis in long‐term bone marrow cultures. In vitro exposure of normal marrow cultures to three different lymphocytotropic HIV‐1 isolates resulted in productive infection, as demonstrated by a progressive increase of gag p24 antigen. In these experiments, ICR‐3 isolate, but not LAV' or NL4–3 isolates, accelerated the loss of non‐adherent cells. A differential ability of these HIV‐1 isolates to suppress myelopoiesis was confirmed in long‐term cultures in which virus was added continuously. In these cultures, ICR‐3, and to a lesser extent also NL4–3, but not LAV', induced a progressive decrease in the number of total non‐adherent cells as well as non‐adherent colony forming units‐granulocyte/macrophage (CFU‐GM). Furthermore, exposure of normal purified CD34+ cells to ICR‐3 induced defects in their ability to form haemopoietic colonies: this inhibitory effect was significantly relieved by pretreatment of ICR‐3 with an anti‐gp 120 antibody. Similar exposure of CD34+ cells to LAV' and NL4–3 induced no such defects.

These data indicate that some HIV‐1 isolates can impair bone marrow haemopoiesis in a dose‐dependent fashion, acting, at least in part, at the level of haemopoietic stem/ progenitor cells.
Thalassaemic osteoarthropathy treated with radiotherapy
Authors: Davies A. N.
Published: July 2010

Summary. Patients with beta thalassaemia may develop a specific osteoarthropathy involving the feet. A number of different treatments for this condition have been tried, including rest, analgesia and hypertransfusion. We report a case of a patient with thalassaemic osteoarthropathy who responded to radiotherapy after failing conventional treatment.
B lymphocytes with latent EBV infection appearing in long‐term bone marrow cultures (HLTBMCs) from haematological patients induce lysis of stromal microenvironment
Authors: Pavlova Borislava G. , Muuhlberger Helmut H. , Strobl Herbert , Gill Renate , Haslberger Alexander , Varga Franz , Auer Herbert , Heinz Renate , Salamon Julius , Stacher Alois , Karlic Heidrun
Published: July 2010

Human long‐term bone marrow cultures (HLTBMCs) are a valuable in vitro model for studying the role of the haemopoietic microenvironment. Here we report the spontaneous appearance of EBV‐positive B cells in 6/40 HLTBMCs from patients with various haematological diseases after 3‐5 months of culture.

After subcultivation of these cells, a novel type of cell line could be characterized, which displayed surface markers and morphological features typical for EBV transformed B‐cell lines.

As the deproteinized and ultrafiltrated culture supernatants of these cell lines were found to contain an agent with stroma toxic properties, they were termed SSB lines (stroma‐toxic‐agent‐secreting B‐cell lines). This agent also exhibited a colony‐inhibitory activity on in vitro myelopoiesis and erythropoiesis. These properties are typical for the two polyamines spermine and spermidine which were detected at elevated levels in the culture supernatants of SSB lines.

The hypothesis that latent presence of EBV in bone marrow may induce an increased synthesis of spermine and spermidine, which are known to be associated with malignant haematological diseases and bone marrow aplasia, is discussed.
A specific allele of the histidine‐rich glycoprotein (HRG) locus is linked with elevated plasma levels of HRG in a Dutch family with thrombosis
Authors: Hennis B. C. , Van Boheemen P. A. , Koeleman B. P. C. , Boomsma D. I. , Engbsser L. , Van Wees A. G. M. , Novakova I. , Brommer E. J. P. , Klufft C.
Published: July 2010

Recent studies describe families with both elevated plasma HRG levels and thrombosis. In order to study the possibility that allelic variants of the HRG locus are associated with differences in HRG level, we studied linkage between HRG levels and a dinucleotide repeat polymorphism in a Dutch family which was selected on the presence of both thrombosis and elevated plasma HRG levels. No other known risk factors from thrombosis were found in this family. Linkage was calculated between the dinucleotide repeat and the HRG level considering the HRG level as a quantitative phenotype assuming a population prevalence of elevated HRG of 5%. Two classes of HRG levels were defined by a mean and a variance: one class with normal HRG levels and a second class with high HRG levels. Using a mean HRG level of 99% for individuals with a normal HRG level and 145% for individuals with high HRG, a maximum lod score of 4.17 (odds in favour of linkage of 22 000:1) was found at a recombination fraction of 0, indicating linkage. Considering the pedigree, an association was found between the presence of a specific allele (no. 6) of the dinucleotide repeat polymorphism and plasma HRG levels. Family members carrying allele 6 were found to have higher HRG plasma levels compared with family members lacking allele 6 (149% v 109% respectively). We conclude that in this family, linkage is found between the HRG locus and the HRG level, and that a HRG gene coupled to allele 6 of the dinucleotide polymorphism is associated with elevated plasma HRG levels. No evidence was found for a causal relationship between elevated plasma HRG levels and thrombosis in this family.
Spectrin Anastasia (αI/78): a new spectrin variant (α45 Arg Thr) with moderate elliptocytogenic potential
Authors: Perrotta Silverio , Iolascon Achille , Angelis Filomena De , Pagano Leonilde , Colonna Giovanni , Cutillo Stefano , Del Giudice Emanuele Miraglia
Published: July 2010

We describe a white Italian kindred in which hereditary elliptocytosis (HE) is associated with abnormal level of α1/78 peptide in spectrin digest. Clinical phenotype varied among the family members ranging from asymptomatic to mild haemolytic HE. The original mutation responsible is a G‐C substitution of the spectrin α‐gene: α45 Arg Thr (AGG ACG). The corresponding spectrin is designated spectrin Anastasia. Utilizing a secondary structure predictive method we suggest that this mutation has a poor capability to induce conformational changes of the tetramerization site and thus shows a moderate elliptocytogenic potential.
Large‐scale feasibility of gene transduction into human CD34+ cell‐derived dendritic cells by adenoviral/polycation complex
Authors: Di Nicola Massimo , Carlo‐Stella Carmelo , Milanesi Marco , Magni Michele , Longoni Paolo , Mortarini Roberta , Anichini Andrea , Tomanin Rosella , Scarpa Maurizio , Gianni A. Massimo
Published: July 2010

With a view to using multiple injections of anti‐cancer dendritic cell (DC)‐based vaccines, we evaluated the feasibility of the adenoviral transduction of large amounts of human CD34+ cell‐derived DCs, and analysed the persistence of the transgene expression and the integrity of DC functional activity after the transduction/cryopreservation procedures. Mature DCs generated from highly enriched human CD34+ cells were transduced by a recombinant adenovirus (rAd‐MFG) that carried a modified, membrane‐exposed, alkaline phosphatase (AP) sequence as the reporter gene. Cationic lipids such as LipofectAmine or poly‐l‐lysine were mixed with the viral particles before the transduction of the target cells. The highest transduction efficiency was obtained at a multiplicity of infection (MOI) rate of 500 (AP + DCs: 50 ± 2%, viability = 95%) under both small‐ and large‐scale conditions. The addition of poly‐l‐lysine or LipofectAmine increased the percentage of transduced cells at an MOI of 500 (CD1a+/AP+ cells = 85 ± 3% and 80 ± 2% respectively). Polycations made it possible to reduce the amounts of viral particles, with high efficiency of transduction being achieved at a MOI of 100 with 10 μg/ml poly‐l‐lysine (CD1a+/AP+: 68 ± 9%) or 30 μg/ml LipofectAmine (CD1a+/AP+: 60 ± 7%). Evaluation of the immunophenotype of the transduced DCs showed that the lack of a DC subpopulation was more susceptible to adenoviral transduction. Cryopreservation of transduced DCs did not modify the viability or percentage of AP+ cells that maintain antigen‐presenting cell (APC) functions. These findings indicate the efficacy of this method for the transduction of large amounts of CD34+ cell‐derived DCs using small quantities of adenoviral vector mixed with polycations. Cryopreservation of transduced DCs did not damage their viability or APC functions, thus making it possible to plan multiple injections of engineered DC‐based vaccines.
Feasibility of peripheral blood stem cell rescue as intensification in elderly patients with acute myelocytic leukaemia: a pilot study from the Gimema Group
Authors: Montillo Marco , Tedeschi Alessandra , Pagano Livio , Venditti Adriano , Ferrara Felicetto , Fabris Piero , Martino Bruno , Musso Maurizio , De Rosa Gennaro , Specchia Giorgina , Monaco Michele , Sparaventi Giovanni , Spadea Antonio , Palmas Angelo , Deplano Wilma , Manna Annunziata , Melillo Lorella , Miraglia Eustachio , Mirto Salvatore , Mandelli for the Gimema Group Franco
Published: July 2010

Elderly patients with untreated acute myeloid leukaemia (AML, n = 47) tested the feasibility of out‐patient consolidation therapy and post‐consolidation treatment (for patients aged < 71 years) with autologous peripheral blood stem cell transplantation (APBSCT). Overall, 13 patients out of 24 (51%) who achieved complete remission (CR) were eligible for further treatment after consolidation. Five patients were primed with granulocyte colony stimulating factor (G‐CSF); a suitable number of CD34+ cells were harvested in three patients and were actually autotransplanted. The toxicity of APBSCT was negligible. Psychosocial problems impaired treatment of some patients on an out‐patient basis. Resistant disease, toxicity and logistic problems reduced the number of patients to whom this procedure could actually be applied.
Autologous T lymphocytes recognize the tumour‐derived immunoglobulin VH‐CDR3 region in patients with B‐cell chronic lymphocytic leukaemia
Authors: Rezvany Mohammad Reza , Jeddi‐Tehrani Mahmood , Rabbani Hodjattallah , Rudén Ulla , Hammarström Lennart , Österborg Anders , Wigzell Hans , Mellstedt Håkan
Published: July 2010

We have previously shown that autologous T cells recognize leukaemic cells from patients with chronic lymphocytic leukaemia (B‐CLL) in an MHC class I‐ and/or II‐restricted manner. A candidate recognition structure might be the tumour cell‐derived Ig VH complementarity‐determining region (CDR)3. Three patients with B‐CLL were analysed for the presence of autologous T cells recognizing the tumour‐specific VH‐CDR3 region. The VH region was shown to be mutated in all three patients. In two patients, a VH‐CDR3‐specific T‐cell response was detected by proliferation assay, as well as by γ‐interferon (IFN) production. The responses could be inhibited by monoclonal antibodies against MHC class II, but not MHC class I. In the third patient, a VH‐CDR3 proliferative response was detected, which could be inhibited by an anti‐MHC class I monoclonal antibody, but not by anti‐MHC class II antibodies. No γ‐IFN response could be detected in this patient. In no patient was an interleukin (IL)‐4 response noted. Thus, in patients with B‐CLL, naturally occurring T cells recognizing the tumour‐unique VH‐CDR3 region are present.
High bax expression is a good prognostic indicator in acute myeloid leukaemia
Authors: Ong Y. L. , McMullin M. F. , Bailie K. E. M. , Lappin T. R. J. , Jones F. G. C. , Irvine A. E.
Published: July 2010

Most cytotoxic drugs kill cells by instigating the process of apoptosis and it has been suggested that apoptotic markers may provide an indication of tumour chemosensitivity. The aim of this study was to determine if such a relationship exists in acute myeloid leukaemia (AML). The levels of spontaneous apoptosis, bcl‐2 and bax were evaluated in 56 newly diagnosed AML patients to determine if they correlated with a response to cytotoxic therapy. Spontaneous apoptosis was lower, but bcl‐2, bax and the bcl‐2/bax ratio were higher in AML compared with normal individuals. AML patients with high bax expression at diagnosis had significantly better prognosis for disease‐free survival, event‐free survival and overall survival (P = 0·016). In the standard risk group, high bax expression was in keeping with significantly improved survival. Multivariate analysis revealed bax to be an independent predictor of survival. There was a significant reduction in bcl‐2 and bax expression when AML patients entered complete remission and also in relapsed AML patients who entered a second remission. This study suggests that bax is a useful prognostic indicator in AML and may assist with therapeutic decision‐making for patients in the standard risk category.
Irreversible loss of donor blood leucocyte activation may explain a paucity of transfusion‐associated graft‐versus‐host disease from stored blood
Authors: Chang Hong , Voralia Michael , Bali Meenakshi , Sher Graham D. , Branch Donald R.
Published: July 2010

Transfusion‐associated graft‐versus‐host disease (TA‐GVHD) is usually a fatal outcome of blood transfusion therapy, caused by viable leucocytes contained in the donor blood. Most cases of TA‐GVHD occur when less than 4‐d‐old blood is transfused. We therefore examined the molecular changes that occur during storage that may account for the paucity of TA‐GVHD following infusion of older blood. Leucocyte number and viability were essentially unchanged from freshly obtained blood, but the expression of cell‐surface lymphocyte activation antigens (CD3, CD4, CD28, CD2, CD45) decreased rapidly within the first 24 h and continued to fall to less than 20% of original levels by d 9 of storage at 4°C. The decrease in CD antigen expression directly correlated with a decreasing ability to induce activation of the T‐lymphocyte cellular signal transduction pathway. As a result, cells became less responsive in a mixed lymphocyte culture (MLC) by d 3, with abrogation of the MLC responsiveness by d 5. Donor leucocytes stored for 4 d or less at 4°C were able to partially re‐express CD antigens and reconstitute their signalling pathway when placed at 37°C, whereas those stored for more than 4 d were not. These irreversible changes result from a permanent downregulation of donor cell protein synthesis. These findings provide a mechanism to explain the paucity of TA‐GVHD following transfusion of blood that is more than 4 d‐old. Further study may show that aged blood provides additional assurances for the prevention of TA‐GVHD; however, use of aged blood should not replace current protocols using irradiation.
Factor XIII Val34Leu and the risk of venous thromboembolism in factor V Leiden carriers
Authors: Franco Rendrik F. , Middeldorp Saskia , Meinardi Johan R. , Van Pampus Elisabeth C. M. , Reitsma Pieter H.
Published: July 2010

A mutation in factor XIII (Val34Leu) was reported to protect against venous thromboembolism. We evaluated the effect of Val34Leu on thrombotic risk in 352 factor V Leiden carriers who were first‐degree relatives of 132 thrombotic propositi carrying factor V Leiden. The total observation period was 2594 years in 92 Val34Leu carriers and 7444 years in 260 non‐carriers. The annual incidence of a first episode of venous thromboembolism was 0·31% in Val34Leu carriers and 0·44% in non‐carriers [relative risk (RR) for venous thromboembolism: 0·7, 95% CI 0·3–1·5]. Age‐specific RR for venous thromboembolism were (for Val34Leu carriers and non‐carriers respectively): 1·0 (95% CI 0·3–3·2) in the age group of 15–30 years, 0·4 (95% CI 0·05–3·0) in the age group of 30–45 years, 0·6 (95% CI 0·1–2·9) in the group aged 45–60 years and 0·5 (95% CI 0·06–4·5) in relatives older than 60 years. In conclusion, the impact of FXIII Val34Leu on the venous thromboembolic risk is modest, suggesting that screening for this mutation in factor V Leiden carriers is not justified.
Prevalence of the factor VR506Q mutation in two Irish control populations: use of a novel nested polymerase chain reaction approach
Authors: Livingstone W. J. , Keenan C. , White B. , Mynett‐Johnson L. , Lawler M. , Mayne P. , Smith O. P.
Published: July 2010

The prevalence of factor V (FV) Leiden among normal populations has primarily been determined using blood donors. This control group is carefully selected and therefore may not accurately reflect the true prevalence within the population. We assessed the prevalence of FV Leiden within the Irish population using Guthrie card samples randomly selected from all newborns. We compared this result with the prevalence of FV Leiden within blood donors. A novel nested polymerase chain reaction (PCR) method for FV Leiden was developed for analysis of the Guthrie card samples. There was no significant difference between the allele frequency within the Guthrie card samples and blood donors (2·07% vs. 2·35%, P = 0.66)
In vitro quantification of anti‐red blood cell antibody production in idiopathic autoimmune haemolytic anaemia: effect of mitogen and cytokine stimulation
Authors: Barcellini Wilma , Clerici Giuliana , Montesano Rosanna , Taioli Emanuela , Morelati Fernanda , Rebulla Paolo , Zanella Alberto
Published: July 2010

The immunopathogenic mechanisms underlying idiopathic autoimmune haemolytic anaemia (AIHA) are still unknown, although regulatory cytokines are thought to play an important role. We investigated cytokine production by mitogen‐stimulated whole blood cultures from 21 patients with AIHA and from 22 age‐ and sex‐matched controls. In parallel experiments, we studied the effect of mitogen and cytokine stimulation on anti‐red blood cell (RBC) IgG antibody production, assessed as both binding on autologous RBCs and secretion in culture supernatants. To quantify anti‐RBC antibody, we set up a sensitive and quantitative solid phase competitive immunoassay. The results showed that in AIHA patients production of interleukin (IL)‐4, IL‐6 and IL‐13 was significantly increased, whereas that of interferon (IFN)‐γ was reduced. Multivariate analysis showed that IFN‐γ was the only independent factor significantly associated with the reduced T‐helper‐1‐like cytokine profile. Patients with active haemolysis showed further reduction of IFN‐γ and IL‐2 production and increased secretion of transforming growth factor (TGF)‐β. In AIHA patients, mitogen stimulation, as well as IL‐6, significantly increased autologous anti‐RBC‐binding relative to unstimulated cultures. Mitogen stimulation and addition of IL‐4, IL‐6, IL‐10, IL‐13 and TGF‐β significantly increased both autologous anti‐RBC binding and antibody secretion in AIHA patients compared with controls. The results suggest that a reduced T‐helper‐1‐ and a predominant T‐helper‐2‐like profile and elevated TGF‐β levels might play a role in the immunopathogenesis of AIHA. Furthermore, our competitive anti‐RBC antibody was able to detect anti‐RBC antibody production in some direct antiglobulin test (DAT)‐negative AIHA patients.
Stroma‐derived factor 1α induces a selective inhibition of human erythroid development via the functional upregulation of Fas/CD95 ligand
Authors: Gibellini Davide , Bassini Alessandra , Re Maria Carla , Ponti Cristina , Miscia Sebastiano , Gonelli Arianna , La Placa Michele , Zauli Giorgio
Published: July 2010

CXC chemokine receptor 4 (CXCR4), the high‐affinity receptor for stroma‐derived factor 1α (SDF‐1α), shows distinct patterns of expression in human CD34+ haematopoietic progenitor cells induced to differentiate in vitro along the granulocytic and erythroid lineages. In serum‐free liquid cultures supplemented with stem cell factor (SCF), interleukin 3 (IL‐3) and granulocyte colony‐stimulating factor, the expression of surface CXCR4 progressively increased in cells differentiating along the granulocytic lineage. The addition in culture of 200 ng/ml of SDF‐1α, a concentration which maximally activated intracellular Ca2+ flux, only modestly affected the expression levels of CD15 and CD11b granulocytic antigens, as well as the total number of viable cells. On the other hand, in liquid cultures supplemented with SCF, IL‐3 and erythropoietin, SDF‐1α induced the downregulation of glycophorin A erythroid antigen, accompanied by a progressive decline in the number of viable erythroblasts. Moreover, in semisolid assays, SDF‐1α significantly reduced the number of plurifocal erythroid colonies (erythroid blast‐forming units; BFU‐E), whereas it did not affect granulocyte–macrophage colony‐forming units (CFU‐GM). We also demonstrated that the inhibitory effect of SDF‐1α on glycophorin A+ erythroid cell development was mediated by the functional upregulation of CD95L in erythroid cultures. These data indicate that SDF‐1α plays a role as a negative regulator of erythropoiesis.
Oxygen tension influences the differentiation, maturation and apoptosis of human megakaryocytes
Authors: Mostafa Sigma S. , Miller William M. , Papoutsakis E. Terry
Published: July 2010

Megakaryocytes (Mks) mature adjacent to bone marrow (BM) sinus walls and subsequently release platelets within the sinusoidal space or in lung capillaries. As the sites for platelet release have higher levels of oxygen tension (pO2) than the core of the BM where stem and progenitor cells reside, we investigated whether pO2influences Mk maturation. Mks were generated from CD34+cells (from mobilized peripheral blood from cancer patients) under 5% and 20% O2. At day 15, CD41+Mk expansion in 20% and 5% O2cultures was 85‐fold and 31‐fold respectively. Twenty percent O2cultures also had higher levels of high ploidy (≥ 8N, eightfold higher) and proplatelet‐forming (fivefold higher) Mks. At day 21, 20% O2cultures had a fivefold higher number of apoptotic Mks. In contrast, 5% O2promoted Mk colony‐forming unit (CFU‐Mk) generation and maintenance. Similar results were observed in cultures initiated with CD41+Mks, indicating that pO2directly affects Mks. The change from 20% to 5% O2on day 5 and day 7 delayed both maturation and apoptosis, suggesting that these two processes are closely linked. These results were confirmed in CD34+cultures from normal BM samples. These data may provide insights intoin vivoMk maturation, such as an explanation for hypoxia‐induced thrombocytopenia in animals.
Novel association of haemophagocytic syndrome with Kaposi's sarcoma‐associated herpesvirus‐related primary effusion lymphoma
Authors: Pastore Raymond D. , Chadburn Amy , Kripas Christopher , Schattner Elaine J.
Published: July 2010

Haemophagocytic syndrome (HPS) is a fulminant, often fatal, systemic illness that occurs in association with infection and malignancy. We provide the first report of HPS that heralded a primary effusion lymphoma (PEL), a rare neoplasm linked to Kaposi's sarcoma‐associated herpesvirus. The patient was a 38‐year‐old man with acquired immunodeficiency syndrome who presented with fever, sweats, lymphadenopathy, splenomegaly and refractory anaemia. Examination of the spleen demonstrated haemophagocytosis; analysis of ascites revealed PEL. Treatment with chemotherapy and ganciclovir resulted in complete remission of both conditions. This case illustrates the diagnostic challenges posed by HPS and supports the trial of antiviral agents in combination with chemotherapy in patients with PEL.
Heparin‐induced thrombocytopenia and thrombosis syndrome: in vivo cross‐reactivity with danaparoid and successful treatment with r‐Hirudin
Authors: Keng Tee Beng , Chong Beng H.
Published: July 2010

Heparin‐induced thrombocytopenia and thrombosis syndrome (HITTS) is an immune‐mediated drug reaction that occurs 5–14 d after initiation of heparin therapy and is a potentially life‐threatening thrombotic complication. The antibody–heparin–PF4 complexes cause platelet activation and generation of platelet microparticles. The need for anticoagulant treatment in asymptomatic thrombocytopenia is uncertain. However, treatment is warranted in HITTS, as illustrated in the case reported here. Danaparoid, r‐Hirudin and argatroban are effective drugs. Danaparoid has a 10–50% in vitro cross‐reactivity rate with the HIT antibodies, but has been proven to be clinically efficacious even in these cases. Here, we report a case of in vivo cross‐reactivity with danaparoid, the patient showed an excellent recovery with r‐Hirudin.
Flexible and dynamic organization of bone marrow stromal compartment
Authors: Anjos‐Afonso Fernando , Bonnet Dominique
Published: July 2010

The bone marrow mesenchymal compartment contains putative stem/progenitors of skeletal tissue components such as bone, cartilage, haematopoiesis‐supporting stroma and adipocytes. Previously appreciated as vital to the support of haematopoiesis, these cells have also been recently recognized as having significant immunomodulatory properties with implications for allogeneic haematopoietic cell transplantation. Despite having been studied for more than three decades and currently being used in different clinical settings, their biology remains elusive. The aim of this review is to critically analyse the field of mesenchymal stem/progenitor cell biology, in respect of their relationship with other mesenchymal cell‐types. Several issues concerning lineage commitment and inter‐conversion potential between different mesenchymal cell‐types are reviewed.
Phase I/II clinical trial of sequential subcutaneous and intravenous delivery of dendritic cell vaccination for refractory multiple myeloma using patient‐specific tumour idiotype protein or idiotype (VDJ)‐derived class I‐restricted peptides
Authors: Curti Antonio , Tosi Patrizia , Comoli Patrizia , Terragna Carolina , Ferri Elisa , Cellini Claudia , Massaia Massimo , D’Addio Alessandra , Giudice Valeria , Di Bello Cristiana , Cavo Michele , Conte Roberto , Gugliotta Gabriele , Baccarani Michele , Lemoli Roberto M.
Published: July 2010

Fifteen multiple myeloma (MM) patients who had failed maintenance therapy after tandem autologous stem cell transplantation underwent anti‐idiotype (Id) vaccination with dendritic cells (DCs). CD14+‐derived DCs were loaded with the autologous Id as whole protein (=6) or Id‐derived class I‐restricted peptides (=9) and keyhole limpet hemocyanin (KLH). Vaccination consisted of three subcutaneous (sc) and two intravenous injections of increasing DC doses at 2 weeks interval. DC therapy was well tolerated. Most patients developed both humoral and T‐cell responses to KLH, suggesting immunocompetence. Eight of 15 patients developed an Id‐specific T‐cell proliferative response, 8/15 increased interferon‐γ‐secreting T cells and 4/15 showed an Id‐positive delayed‐type hypersensitivity test. Anti‐Id cytotoxic T‐lymphocyte precursors increased after DC vaccination in 2/2 evaluable patients. A more robust T‐cell response was observed after sc DC injections and increased Id‐specific T‐cell proliferation was found up to 1 year after vaccination. VDJ‐derived peptides were as effective as the whole protein in stimulating T‐cell responses. Clinically, 7/15 patients have stable disease after a median follow‐up of 26 months, one patient achieved durable partial remission after 40 months, and seven patients progressed. In conclusion, sc injections of cryopreserved Id‐pulsed DCs were safe and, in contrast with intravenous administrations, induced anti‐MM T‐cell responses.
Increased elastase secretion by peripheral blood monocytes in cystic fibrosis patients
Authors: JONES M M , SEILHEIMER D K , PIER G B , ROSSEN R D
Published: July 2010

Morbidity and mortality in cystic fibrosis (CF) is predominantly due to destruction of pulmonary tissue. The host immune response may, in part, play a pathogenic role in pulmonary destruction in these patients. To further understand host immune response in CF, we examined the state of activation of peripheral blood monocyles in CF. Baseline elastase activity was 2.2-fold greater in the CF monocytes than in controls. Pseudomonas aeruginosa mucoid exopolysaccharide (MEP) and high molecular weight polysaccharide (HMP) increased elastase activity in both control and CF monocytes, with a greater absolute increase in the CF monocytes. There was no difference in baseline or MEP-stimulated secretion of interleukin-1 (IL-1) or interleukin-6 (IL-6) between CF and control monocytes. Ibuprofen enhanced both MEP and HMP-stimulated elastase activity, whereas dexamethasone suppressed both baseline and stimulated elastase activity > 20% in both CF and control monocytes. These results suggest that circulating monocytes in CF are stimulated in vivo, resulting in a remarkably elevated elastase activity in vitro. Elevated elastase release by peripheral blood monocytes as they enter the lung in response to chemotactic stimuli may contribute to lung destruction in CF.
Antibody responses reveal differences in oral tolerance to wheatand maize grain protein fractions
Authors: JOHNSON R B , LABROOY J T , SKERRITT J H
Published: July 2010

The influence of diet on humoral immune responses to gluten- and maize-derived proteins wasexamined using ELISA and protein blotting techniques. Mice raised on the maize-based (gluten-free)diet responded well to parenteral immunization with each of six gluten-derived protein preparations(whole gliadin, two omega-gliadin fractions, wheat salt-soluble proteins, a peptic-tryptic digest and asubtilisin digest of gluten), as serum antibody levels increased at least 300-fold in each case. Incontrast, mice raised on the wheat-based diet responded poorly to immunization with either wholegliadin or omega-gliadin and were virtually non-responsive to enzymic digest of gluten. Diet had littleeffect on the magnitude of the antibody response to wheat salt-soluble proteins, with both groupsshowing a 300-fold increase in titre. Similarly, tolerance to alpha-zeins, the alcohol-soluble proteinsof maize, did not occur on either diet. However, some oral tolerance was observed to maize glutelin. The specificity of the various antibody responses was then analysed by immunoblotting. Followingimmunization with gluten proteins or digests, antibodies from the maize-fed mice bound more or lessequally to each of the main gliadin bands and to the glutenins while the mice on the wheat-based diethad antibody specific for omega-gliadin proteins. Serum antibodies from the maize-fed mice, immunized with cither alpha-zein or maize glutelin, showed even labelling of the major maizeendosperm proteins while antibodies from mice on the wheat diet showed strong labelling of the M,27000 and 58000 bands. These results show that diet influenced the specificity, as well as themagnitude of serum antibody responses to cereal proteins. In addition, oral tolerance appeared toaffect the humoral response to some cereal proteins more than others. Both of these findings haveimportant implications for our understanding of coeliac disease.
Autoreactive cytotoxicity in HIV-infected individuals
Authors: ISRAËL-BIET D , VENET A , BELDJORD K , ANDRIEU J M , EVEN P
Published: July 2010

A possible role for autoimmunity in the pathogenesis of HIV infection has been suggested, based upon the certain degree of homology shared by HIV gp41 and MHC class II molecules. A number of humoral markers of autoimmunity have since been found in seropositive subjects. We have evaluated the cellular autoreactive response in HI V-infected individuals. Our study demonstrates the existence of a cytolytic activity, present in seropositive but not in seronegative subjects. This activity is mediated by CD3 T cells, which only occasionally express the CD8 or the CD4 surface markers. Effector cells do not appear to exert their activity in a MHC-restricted fashion, since allogeneic target cells could also be killed, recovered from allogeneic seropositive as well as from seronegative subjects. Several types of target cells were lysed: T cell blasts and Epstein-Barr virus (EBV) transformed B cells, suggesting that the target antigen is common to at least these two cell types. The fact that cells from seronegative individuals were lysed argues against the recognition of an HIV-specific antigen. The nature of the target determinants and the identity of the effector cells are discussed.
Multiple infusions of human intravenous immunoglobulin in chimpanzees do not lead to immune elimination
Authors: LEIBL H , WOLF H M , EDER G , MANNHALTER J W , EIBL M M
Published: July 2010

Administration of human i.v. immunoglobulins was shown to lead to a permanent increase in IgGl and IgG2 levels in chimpanzees. Half-lives of human IgG1 and IgG2 in chimpanzees were comparable to those found in humans, and no signs of immune elimination were observed. Furthermore, long-term treatment of chimpanzees had no effect on the percentage of immunoregulatory T cells (CD2+, CD4+ and CD8+ T cells) as determined by FACS analysis. In addition, serum IgM levels in chimpanzees were found to be comparable to those in humans, whereas the chimpanzees' IgG levels are slightly elevated due to higher concentrations of IgG2 and, in particular, IgG4.
Effects of FK506 on rat thymus: time-course analysis by immunoperoxidase technique and flow cytofluorometry
Authors: TAKAI K , JOJIMA K , SAKATOKU J , FUKUMOTO T
Published: July 2010

The effect of administration of FK506 at 1 mg/kg body weight for 14 days on rat lymphoid tissues, especially the thymus, and recovery after discontinuation of treatment, were investigated by the immunoperoxidase technique and flow cytofluorometry using monoclonal antibodies OX6, OX7, OX8, OX18 and W3/25, reactive with rat lymphocytes. Marked reduction of the thymic medulla upon treatment was clearly demonstrated by staining with OX18 and OX6. Changes produced by FK506 were also observed in the cortical area of the thymus, and were especially marked in the subcapsular area and around blood vessels. Eventually, the thymic cortex appeared patchy, this change being maximal 14 days after the start of administration. Obvious restitution of the thymic medulla was evident about 14 days after withdrawal of FK506. Flow cylometric analysis of the thymus showed that the percentages of cells labelled positively with OX7, OX8 and W3/25 were increased with FK506 treatment, and recovered to the normal levels soon after withdrawal. Furthermore, the peak of fluorescence intensity of OX7+, OX8+ and W3/25+ cells showed a temporary shift to the right during FK506 treatment; however, the peak of fluorescence intensity of OX18+ cells showed a temporary shift to the left. Treatment with FK506 also produced a significant change in 3H-thymidine uptake by thymocyte. These results suggest that FK506 may inhibit the proliferation, maturation and differentiation of thymocytes. However, thymocytes prepared from FK506-treated rats and labelled with FITC behaved similarly to rat thymocytes in normal recipient rats. This suggests that during FK506 treatment thymocytes may retain their potential for peripheral mobilization.