bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow cytometry analysis of dual red blood cell populations after bone marrow transplantation
Authors: Blanchard D. , Bruneau V. , Germond‐Arnoult F. , Bernard D. , Gourbil A. , David B. , Muller J. Y. 
Published: July 2010

Flow cytometry represents an alternative method to agglutination assays for the accurate quantification of mixed field populations of erythrocytes observed after bone marrow transplantation. Murine monoclonal antibodies directed against the blood group ABH antigens were selected and processed in order to prepare ready‐to‐use fluorescent reagents. Anti‐A (NaM87‐1F6; IgG3), anti‐B (NaM9‐2E11; IgG3) and anti‐H (NaM19‐7E11; IgM) were purified, labelled with fluorescein isothiocyanate, and used in a direct flow cytometry assay. Anti‐A1 (NaM1‐1C9; IgG3) was no longer active after FITC‐labelling and then was used in an indirect assay. The agglutination was prevented by formaldehyde pretreatment of erythrocytes.Using artificially‐made double populations of erythrocytes, measured values with mixtures of 1‐100% of cells were very closely related to expected values, showing both the sensitivity and the accuracy of the method. From careful investigation of a series of bone‐marrow transplanted patients, we conclude that engraftments could be demonstrated earlier by flow cytometry than by agglutination, because minor populations (1‐10%) of cells could be determined accurately only with labelled reagents. In addition, the disappearance of the donor cells on a long‐term follow‐up of patients enabled an earlier detection of graft failure in one case. The proposed method provides appreciable help to follow engraftment in patients and may have more general applications for the study of other haemopoietic chimaeras.
Bone marrow pathology in relapsing polychondritis: high frequency of myelodysplastic syndromes
Authors: Diebold J. , Rauh G. , Jäger K. , Löhrs U. 
Published: July 2010

Haemopathologic changes were studied in 19 patients (13 male, six female, age 33‐85 years, mean 56 years) with relapsing polychondritis (RP). Anaemia was found in eight, thrombocytopenia in two and splenomegaly in three patients. A total of 17 bone marrow biopsies were obtained from seven individuals. Bone marrow evaluation revealed myelodysplastic syndromes (MDS) with marked trilineage hyperplasia and dysplasia in three cases. Since an excess of myeloblasts or an increase of CD34 positive progenitor cells was not seen, the disorders were designated as‘refractory anaemia’or with regard to the dysplastic megakaryopoiesis‘MDS, unclassifiable'. Two of the three patients died after 10 and 55 months of follow‐up due to infectious complications. In a further patient, bone marrow analysis repeatedly showed an unexplained granulopoietic hyperplasia, which, however, was not dysplastic enough to allow a diagnosis of MDS. The remaining patients had clearly reactive changes.Our findings support the notion that RP is a heterogenous disorder and suggest that RP may at times represent a paraneoplastic phenomenon of an underlying MDS. Since HLA typing revealed a significantly increased frequency of the antigen DR4 (10/17 patients positive = 59%), we hypothesize that immunological imbalances due to the MDS in conjunction with a specific immunogenetic background may play key roles in the pathogenesis of RP in these patients.
Assessment of intestinal blood‐flux by laser Doppler fluxmetry in mice with altered intestinal iron absorption
Authors: Raja K. B. , Simpson R. J. , Peters T. J. 
Published: July 2010

Laser Doppler fluxmetry (LDF) has been used to assess mucosal blood‐flux in the intestine of normal CD1 mice and groups of animals with altered iron metabolism (i.e. iron‐deficient, hypoxic and hypotransferrinaemic [hpx/hpx]). All experimental animals showed a 2‐3‐fold increase in duodenal iron absorption, mainly due to changes in the ‘mucosal transfer’phase. However, only the hypoxic mice exhibited any increases in duodenal blood‐flux. In the hpx/hpx group, blood flux was maintained at control levels despite the anaemia and without any significant alterations in red blood cell velocity. Moreover, these two groups demonstrated a further capacity to increase blood‐flux above basal levels during onset of acute hypoxia (inhalation of 10% 02). Changes in duodenal blood‐flux were apparent on both the mucosal and serosal surfaces, suggesting that the LDF signal reflects flux throughout the thickness of the mouse intestine.Iron absorption is likely to be more related to plasma flux changes than to blood‐flux changes. Deduction of plasma flux changes from blood‐flux changes is complicated by vasodilation effects on capillary haematocrits. It is clear from the data, however, that plasma flux changes do not parallel changes in iron absorption in the experimental models.The lack of correlation between the duodenal blood‐flux and iron absorption values in the experimental models argues against the possible involvement of blood‐flux in the control of duodenal iron absorption.
CHO expression of a novel human recombinant IgG1 anti‐RhD antibody isolated by phage display
Authors: Miescher Sylvia , Zahn‐Zabal Monique , De Jesus Maria , Moudry Radmila , Fisch Igor , Vogel Monique , Kobr Michel , Imboden Martin A. , Kragten Eddy , Bichler Johann , Mermod Nicolas , Stadler Beda M. , Amstutz Hanspeter , Wurm Florian 
Published: July 2010

Replacement of the hyperimmune anti‐Rhesus (Rh) D immunoglobulin, currently used to prevent haemolytic disease of the newborn, by fully recombinant human anti‐RhD antibodies would solve the current logistic problems associated with supply and demand. The combination of phage display repertoire cloning with precise selection procedures enables isolation of specific genes that can then be inserted into mammalian expression systems allowing production of large quantities of recombinant human proteins. With the aim of selecting high‐affinity anti‐RhD antibodies, two human Fab libraries were constructed from a hyperimmune donor. Use of a new phage panning procedure involving bromelin‐treated red blood cells enabled the isolation of two high‐affinity Fab‐expressing phage clones, LD‐6‐3 and LD‐6‐33, specific for RhD. These showed a novel reaction pattern by recognizing the D variants DIII, DIVa, DIVb, DVa, DVI types I and II, DVII, Rh33 and DFR. Full‐length immunoglobulin molecules were constructed by cloning the variable regions into expression vectors containing genomic DNA encoding the immunoglobulin constant regions. We describe the first, stable, suspension growth‐adapted Chinese hamster ovary (CHO) cell line producing a high affinity recombinant human IgG1 anti‐RhD antibody adapted to pilot‐scale production. Evaluation of the Fc region of this recombinant antibody by either chemiluminescence or antibody‐dependent cell cytotoxicity (ADCC) assays demonstrated macrophage activation and lysis of red blood cells by human lymphocytes. A consistent source of recombinant human anti‐RhD immunoglobulin produced by CHO cells is expected to meet the stringent safety and regulatory requirements for prophylactic application.
Abdominal venous thrombosis in neonates and infants: role of prothrombotic risk factors – a multicentre case–control study
Authors: Heller Christine , Schobess Rosemarie , Kurnik Karin , Junker Ralf , Günther Gudrun , Kreuz Wolfhart , Nowak‐Göttl Ulrike ,   for the Childhood Thrombophilia Study Group
Published: July 2010

The factor V (FV) G1691A mutation, the prothrombin (PT) G20210A variant, the methylenetetrahydrofolate reductase (MTHFR) T677T genotype, together with fasting homocysteine (HCY) concentration, lipoprotein (Lp)(a), anti‐thrombin (AT), protein C (PC), protein S (PS) and anti‐cardiolipin antibodies were investigated in 65 consecutively recruited infants (neonate to < 12 months) with renal venous thrombosis (RVT; n = 31), portal vein thrombosis (PVT; n = 24) or hepatic vein thrombosis (HVT n = 10), and 100 age‐ and sex‐matched healthy controls. FV G1691A was found in 14 babies (heterozygous: RVT n = 9, PVT n = 4; homozygous HVT n = 1) and five controls, the MTHFR TT677 genotype together with increased HCY in four infants with thrombosis (RVT n = 2; PVT n = 1; HVT n = 1) compared with one control, and the PT G20210A variant was present in one control only. PC type I deficiency was diagnosed in three patients (RVT n = 2; PVT n = 1) and AT deficiency in two patients (RVT n = 1; PVT n = 1). Three neonates with spontaneous thrombosis showed FV G1691A combined with Lp(a) and the FV G1691A was combined with the PT G20210A genotype in two infants. Additional triggering factors were reported in 27 patients (41·5%). The overall odds ratios (ORs) and 95% confidence intervals (CIs) with respect to the different thrombosis locations were: RVT (OR/CI: 10·9/3·85–31·1; P < 0·0001), PVT (5·47/1·7–17·6; P < 0·0007) and HVT (3·3/0·58–18·7; P = 0·18). The data presented here suggest that genetic prothrombotic risk factors also play an important role in abdominal venous thrombosis during infancy.
The reconstitution of CD45RBhiCD4+ naive T cells is inversely correlated with donor age in murine allogeneic haematopoietic stem cell transplantation
Authors: Hirayama Masahiro , Azuma Eiichi , Jiang Qi , Kobayashi Michihiro , Iwamoto Shotaro , Kumamoto Tadashi , Kisenge Rodrick , Yamamoto Hatsumi , Komada Yoshihiro 
Published: July 2010

A high incidence of opportunistic infections after unrelated bone marrow transplantation has been reported. Delayed lymphocyte recovery may be associated with opportunistic infections. Immune reconstitution is influenced by recipient age and graft‐vs‐host disease (GVHD). In fact, children develop GVHD less frequently than adults. However, the role of donor age is largely unknown. We examined the effect of donor age on lymphocyte reconstitution after transplant. Three‐month‐old BALB/c recipient mice were lethally irradiated and transplanted with allogeneic haematopoietic stem cells from A/J donor mice of different ages, ranging from 0 d to 12 months. The recovery of absolute lymphocyte counts and those of CD3+ T cells, CD4+ T cells and CD45RBhi CD4+ naive T cells in the early post‐transplant period correlated inversely with donor age. Recipient mice transplanted with haematopoietic stem cells from younger donors showed significantly higher survival rates and mitogenic responses than adult donors. As T cells, especially CD4+ naive T cells, play an important role in host defence, faster recovery of CD4+ naive T cells in younger donors may contribute to reduced mortality in the early post‐transplant period. The results suggest that it could be better to choose a younger donor if sufficient cell dose is available.
Expression and production of interleukin 10 in human myeloma cell lines
Authors: Otsuki Takemi , Yamada Osamu , Yata Kenichiro , Sakaguchi Haruko , Kurebayashi Junichi , Yawata Yoshihito , Ueki Ayako 
Published: July 2010

Recent investigations of the cytokine network surrounding myeloma cells have disclosed the importance of gp130‐related cytokines including interleukin (IL)‐6 for myeloma cell survival and proliferation, identification of IL‐10 as a growth factor for myeloma cells, the close relationship between IL‐10 and the receptors for gp130‐related cytokines, and the growth enhancement effect of IL‐11 and IL‐7 on myeloma cells. In this study, IL‐10 production was observed in three out of seven human myeloma cell lines examined and five (including three producing lines) out of 10 lines exhibited mRNA expression of IL‐10. The IL‐10 mRNA expression was also enhanced in approximately one third of primary specimens, whereas the IL‐10 receptor (R) expression was not changed compared with that of normal component marrow controls. However, reverse transcription‐polymerase chain reaction (RT‐PCR) assay of various cytokines and their receptors showed no particular association with IL‐10‐producing myeloma lines compared with non‐producing lines. Supplementing exogenous IL‐10 or neutralization of the IL‐10 signal by anti‐IL‐10 monoclonal antibody (mAb) in a culture conditions did not significantly affect myeloma cell growth regardless of expression of IL‐10 or its receptor (IL‐10R). However, supplement of anti‐IL‐10 mAb caused upregulation of certain genes such as IL‐11, leukaemia inhibitory factor receptor (LIFR) and syndecan‐1 in IL‐10R‐expressing cell lines. These findings indicate that the cytokine network surrounding myeloma cells is complicated and variable. In addition, IL‐10 may modify this network and the cellular biological properties of myeloma cells rather than cell proliferation.
Ribosomal dysfunction and inherited marrow failure
Authors: Ganapathi Karthik A. , Shimamura Akiko 
Published: July 2010

SummaryImpairment of ribosome biogenesis or function characterizes several of the inherited bone marrow failure syndromes: Diamond‐Blackfan anaemia, dyskeratosis congenita (DC), Shwachman‐Diamond syndrome and cartilage‐hair hypoplasia. These syndromes exhibit overlapping but distinct clinical phenotypes and each disorder involves different aspects of ribosomal biogenesis. The clinical characteristics of each syndrome are briefly reviewed. Molecular studies of ribosome biogenesis and function in each of these syndromes are discussed. Models of how impairment of ribosomal pathways might affect haematopoiesis and tumorigenesis are explored.
The role of macrophage activation and ofBcg-encoded macrophage function(s) in the control ofMycobacterium aviuminfection in mice
Authors: APPELBERG R , SARMENTO A M 
Published: July 2010

SUMMARYFollowing the intraperitoneal inoculation of 2.5 × 108 colony-forming units of Mycobacterium avium strain ATCC 25291, there was bacillary growth in the liver, spleen and peritoneal cavity of C57BL/6, C57BL/10, DBA/1 and BALB/c mice whereas DBA/2, C3H/He, CBA/Ca and CD-1 mice controlled the infection showing constant or slightly decreasing numbers of viable bacteria in the liver and spleen and effective clearance of the bacilli from the peritoneal cavities. The acquisition of non-specific resistance (NSR) to Listeria monocytogenes during the infection by M. avium was high in C57BL/6, BALB/c and C3H/He mice and negligible in DBA/2 and CD-1 mice. The magnitude of the acquisition of NSR was reduced in T cell-deficient mice and was directly proportional to the dose of the inoculum of M. avium. The production of hydrogen peroxide by phorbol myristate acetate-stimulated peritoneal macrophages of M. avium-infected mice was higher in C57BL/6 and BALB/c mice than in CD-1, DBA/2 and C3H/He animals. BALB/c. Begr (C.D2) mice, unlike their congenic strain BALB/c, restricted bacterial growth following the intravenous inoculation of 2.5 × 108 CFU of M. avium as efficiently as DBA/2 mice. C.D2 and BALB/c peritoneal macrophages from infected mice produced similar amounts of H2O2 but BALB/c mice developed higher levels of NSR to listeria than C.D2 mice. The production of nitrite by peritoneal macrophages from infected mice was found to be enhanced in DBA/2 and C3H/He but not in BALB/c, C57BL/6, CD-1 and C.D2 mice. Resident peritoneal macrophages from C.D2 mice were more bacteriostatic in vitro for M. avium than macrophages from BALB/c mice. The same relative differences between the two macrophage populations were observed when the cells were activated with lymphokines. The results show that the resistance to M. avium infection in mice is under the control of the Bcg gene and that susceptibility may be due to some defect in macrophage antibacterial function not completely overcome by the activation of this phagocyte in the susceptible strains of mice.
Human IgG anti-F(ab′)2 antibodies possess rheumatoid factor activity
Authors: GERARDO S HUNT , PERSSELIN J E , STEVENS R H 
Published: July 2010

SUMMARYIndividual preparations of affinity purified anti-F(ab′)2 antibodies and anti-Fc antibodies isolated from the sera of patients with rheumatoid arthritis (RA), were examined for reactivity with the Fab and Fc fragments of human IgG. Western blot assays demonstrated specific interaction of affinitypurified anti-Fab antibodies with both Fab and Fc molecules. Approximately one-half of the anti-Fab antibody preparations studied contained IgG antibodies reactive with Fab and Fc fragments in ELISA, suggesting the existence of naturally occurring epibody-like autoantibodies in these patients. Thirteen of 14 affinity-purified anti-Fc antibody preparations contained IgG cross-reactive with Fab molecules in ELISA. Double-adsorption assays on affinity columns demonstrated that a minimum of 14%, and possibly as much as 50%, of the IgG anti-Fab antibodies reacted with the Fc of IgG. Conversely, a minimum of 12%, and possibly as much as 70%, of the IgG anti-Fc antibodies reacted with IgG Fab molecules. Anti-Fab antibodies isolated from non-RA individuals also exhibited anti-Fc reactivity in ELISA. demonstrating the presence of these dual-reactive antibodies in other autoimmune and normal individuals. These studies establish the presence of naturally occurring IgG autoantibodies reactive with both the Fab and Fc fragments of human IgG. Their existence emphasizes the potential of anti-immunoglobulin antibodies to recognize a multiplicity of antigens, possibly including other members of the immunoglobulin supergene family.
Immunosuppressive activity of FK-506 in rats: flow cytometric analysis of lymphocyte populations in blood, spleen and thymus during treatment and following drug withdrawal
Authors: WOO J , ROSS C S K , MILTON J I , THOMSON A W 
Published: July 2010

SUMMARYRats were immunized systemically with sheep red blood cells (SRBC) and given either FK-506 (I mg/ kg) or drug vehicle by i.m. injection for 7 days. In animals receiving FK-506, there was suppression (87%) of the splenic plaque-forming cell response on day 4 and marked reductions in the serum antibody litre throughout the 3-week period following immunization. Sequential flow cytometric analyses of blood lymphocytes revealed statistically significant attenuation, by FK-506. of the increase in relative numbers of OX-12+ (B) cells between days 4 and 7. Following drug withdrawal, OX-12 values remained elevated, whereas in control animals a decline was observed. These changes were reflected in concomitant increases in the relative numbers of OX-19+ (CD3+), W3/25+ (CD4+) and OX-8+ (CD8+) T cells; however, due to an overall reduction in lymphocytes by day 7, absolute values were not significantly affected compared with controls. The pattern of changes in OX-6+ (MHC class II+) cells in blood was similar to that observed for B cells. FK-506 also suppressed increases in the small proportion of blood-borne OX-40+ (activated CD4+) cells and OX-39+ (interleukin-2 receptor+) cells in the 7 day period following immunization; thereafter values for activation marker expression between treatment and control groups were similar. In the spleen, there were fewer significant differences between FK-506 and control groups in the incidences of cells expressing the above markers. OX-8+ cells, however, were significantly higher in drug-treated animals on day 7. and there were also reductions in the small proportions of OX-39+ and OX-40+ cells when compared with controls. In the thymus, reversible medullary atrophy induced by FK-506 was accompanied on day 7 by increases in the incidence of CD4+ and CD8+ cells and by a concomitant reduction in OX-44+ mature, medullary thymocytes. Two weeks after drug withdrawal, the phenotypic marker expression profile had been restored to normal in blood, spleen and thymus. These data provide new information on the apparent capacity of FK-506 to interfere with T cell maturation and its influence on lymphocyte activation in vivo.
Autoimmune syndrome after induction of neonatal tolerance to alloantigens: analysis of the role of donor T cells in the induction of autoimmunity
Authors: MERINO J , SCHURMANS S , WEN L , BRIGHOUSE G , LUZUY S , LAMBERT P-H 
Published: July 2010

SUMMARYThe injection of (C57BL/6 × BALB/c) F1 spleen cells into BALB/c newborn mice leads to activation of persisting F1 donor B cells and development of a lupus-like syndrome in tolerized BALB/c mice. This syndrome is characterized by hypergammaglobulinaemia, high levels of anti-DNA and anti-Sm antibodies, circulating immune complexes and deposits of immunoglobulin in renal glomeruli. The role of donor T cells in this model was investigated by injecting the newborn mice with F1 cells depleted in different T cell subsets by using specific monoclonal antibodies (MoAbs). Tolerance, as shown by an absence of H-2b-specific CTL alloreactivity and persistence of immunoglobulin bearing the donor allotype were observed in mice injected with F1 cells previously depleted in the CD4+ and/ or CD8+ T cell subsets as well as in those which received Thy-1+-depleted F1 spleen cells. In these mice, a typical autoimmune syndrome was found, including splenomegaly and lymphadenopathy, anti-ssDNA and anti-aortic myosin IgG antibodies and renal deposition of immunoglobulin. However, some quantitative changes were seen: the levels of anti-aortic myosin antibodies were lower in mice tolerized with CD4+-depleted F1 cells than in those receiving untreated F1 cells. Conversely, higher levels of these autoantibodies were observed in mice tolerized with CD8+-depleted F1 cells. These results suggest that mature donor T cells are not necessary neither for the establishment of neonatal tolerance to alloantigens nor for the activation of F1 donor B cells in the production of the autoimmune syndrome in tolerant mice, but they may contribute in the regulation of the expression of autoreactive B cell clones.
Increased production of the third component of complement (C3) by monocytes from patients with systemic lupus erythematosus
Authors: TSUKAMOTO H , UEDA A , NAGASAWA K , TADA Y , NIHO Y 
Published: July 2010

SUMMARYWe measured in vitro C3 production by peripheral blood monocytes from patients with systemic lupus erythematosus (SLE), and found it to be significantly greater than that from normal controls. We also found that monocytes from SLE patients with active disease produced a markedly larger amount of C3 than those from SLE patients with inactive disease. Production of C3 by monocytes correlated with serum levels of anti-dsDNA antibodies and inversely correlated with serum C3 levels in SLE patients. Serial measurement of C3 in the culture supernatant from each SLE patient showed that C3 production by monocytes fell in parallel with a decrease of disease activity. The effect of corticosteroids was ruled out as there was no relation between the level of C3 production by monocytes and the dose of prednisolone. This seems to be the first study in which the C3 production was assayed at a cellular level in SLE patients, and this study suggests that the local C3 production is increased in SLE patients.
A human monoclonal IgM with autoantibody activities against heparan sulphate and the mitotic spindle
Authors: ROUSSEL B , ARVIEUX J , JACOB M C , LORIMIER P , CAVIGIOLI E , MICOUIN C 
Published: July 2010

SUMMARYA monoclonal IgM κ from a patient with Waldenström's macroglobulinaemia (IgM-Rod) was found to react at temperatures below 28°C with all tissue basement membranes and the cell coat of non-haematopoietic cells. IgM-Rod antibody was directed against heparan sulphate side chains of heparan sulphate proteoglycans as shown by binding in a solid-phase ELISA to heparan sulphate glycosaminoglycans but not to other purified subcomponents of the extracellular matrix; and by specific inhibition of the observed reactivity by heparitinase treatment. IgM-Rod showed crossreactivity by indirect immunofluorescencc with an as yet unidentified structure expressed in the nucleus during cell division and becoming associated with the mitotic spindle apparatus. The coexistence of both binding activities for heparan sulphate and nuclei determinants in the same IgM molecule was deduced from adsorption-elution experiments and from the inhibitory effect of a mouse monoclonal anti-idiotypic antibody directed against the paratope of IgM-Rod.