bluuebunny/crossref_metadata_2025 huggingface dataset:
Myeloid leukaemic cells can lyse fibrin directly
Authors: Robbie Linda , Berry Susan , Moir Elaine , Booth Nuala A. , Culligan Dominic , Tighe Jane , Watson Henry , King Derek , Bennett Bruce
Published: July 2010

Purified preparations of circulating leukaemic blast cells from patients with acute myeloid (M1–7) or acute lymphoblastic leukaemia, and the myeloid or lymphoid cells from patients with chronic myeloid or lymphocytic forms of leukaemia, were incorporated into clots prepared from fibrinogen and plasminogen. Patterns of lysis were followed and measured by light transmission in a microtitre plate reader. Mature polymorphonuclear and mononuclear cell fractions from normal individuals were studied concurrently for comparison. Blast cells from the myeloid forms of acute leukaemia (M2–4) and ‘myeloid’ cell fractions from patients with chronic myeloid leukaemia were capable of lysing plasminogen‐containing clots; this activity was neutralized by addition of immunoglobulin against urokinase plasminogen activator (u‐PA), but not by anti‐tissue plasmogen activator (t‐PA). Mature polymorphonuclear and mononuclear cells from normal individuals lacked lytic activity, as did the leukaemic cells from patients with acute lymphoblastic or chronic lymphocytic leukaemia. Lysed blast cells showed the presence of free plasminogen activator on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) with overlay zymography, also neutralized by anti‐u‐PA, whereas normal polymorphonuclear and mononuclear cells did not. These observations suggest that mechanisms underlying some forms of severe bleeding in acute myeloid leukaemias have a critical fibrinolytic component generated by the blast cells themselves.
Novel type of BCR–ABL transcript in a chronic myelogenous leukaemia patient relapsed after bone marrow transplantation
Authors: Roman Jose , Parziale Adele , Gottardi Enrico , De Micheli Daniela , Cilloni Daniela , Tiribelli Mario , Gonzalez Maria Gracia , Rodriguez Maria del Carmen , Torres Antonio , Saglio Giuseppe
Published: July 2010

We identified a novel BCR–ABL transcript in a chronic myelogenous leukaemia (CML) patient who relapsed after bone marrow transplantation (BMT), containing a fusion between part of BCR exon 3, 44 nucleotides derived from ABL intron 1b and ABL exon 2. The breakpoints were located within BCR exon 3 on chromosome 22 and within the ABL intron 1b on chromosome 9, and the transcript derives from a splicing of ABL exon 2 to a putative splicing acceptor site 44 nucleotides downstream to the breakpoint on chromosome 9. The patient's clinical course strengthens the idea that short forms of BCR–ABL transcripts are associated with a more aggressive disease.
Impaired expression of the CD3‐zeta chain in peripheral blood T cells of patients with chronic myeloid leukaemia results in an increased susceptibility to apoptosis
Authors: Chen Xiao , Woiciechowsky Anja , Raffegerst Silke , Schendel Dolores , Kolb Hans‐Jochem , Roskrow Marie
Published: July 2010

In patients with myeloid malignancies, cell‐mediated immunity is often suppressed, being most profound in those with advanced disease. Such immune dysfunction, as demonstrated in many patients with chronic lymphocytic leukaemia (CLL) and myelodysplastic syndrome (MDS), may, at least in part, be due to altered expression of the CD3‐zeta chain, which is an important component of the T‐cell receptor (TCR). We speculated that impaired expression of the TCR‐zeta chain would be evident in peripheral blood T cells of patients with chronic myeloid leukaemia (CML) and that such an abnormality would result in an increased ex vivo susceptibility to apoptosis. In this study, we demonstrated that, compared with normal controls, zeta chain expression was significantly downregulated in all of the T‐cell subsets (P < 0·009) in more than 90% of CML patients. In addition, there was a significantly lower expression of the CD3‐epsilon chain (P < 0·001) in patients than in controls. In those patients with abnormal zeta chain expression, the proportion of lymphocytes with spontaneous DNA fragmentation, as determined by terminal deoxynucleotide transferase‐mediated dUTP‐biotin nick‐end labelling (TUNEL) assays, was also significantly higher (P < 0·002) than controls. From all of the patients tested, it was possible to upregulate partially zeta chain expression and hence to reduce the susceptibility to apoptosis by cross‐linking the T cells with interleukin (IL)‐2, interferon (IFN)‐alpha or immobilized CD3. In addition, such cross‐linked T cells showed a significantly higher capacity to proliferate than the native CML T cells.
Molecular remission without blood product support using all‐trans retinoic acid (ATRA) induction and combined arsenic trioxide/ATRA consolidation in a Jehovah's Witness with de novo acute promyelocytic leukaemia
Authors: Kennedy Glen A , Marlton Paula , Cobcroft Ralph , Gill Devinder
Published: July 2010

Arsenic trioxide has recently been used in the treatment of both relapsed and de novo acute promyelocytic leukaemia (APML). Molecular remissions have been attained using arsenic trioxide with minimal associated haematological toxicity, making protocols utilizing this drug an attractive option for Jehovah's Witnesses with APML. A 62‐year‐old female Jehovah's Witness with de novo APML was treated with all‐trans retinoic acid induction followed by combined arsenic trioxide/ATRA consolidation, and achieved molecular remission with minimal haematological toxicity and no blood product support.
Bone sialoprotein mRNA and protein expression in human multiple myeloma cell lines and patients
Authors: Bellahcène A. , Van Riet I. , De Greef C. , Antoine N. , Young M. F. , Van Camp B. , Castronovo V.
Published: July 2010

Bone sialoprotein (BSP) is a glycoprotein essentially found in mineralizing connective tissues. We have recently demonstrated that BSP is ectopically expressed by carcinomas that metastasize to bone with high frequency. Multiple myeloma (MM) is characterized by the localization of tumour plasma cells in the bone marrow. In this study, BSP expression was evaluated in human myeloma cell lines and in bone marrow aspirates and one ascites fluid from MM patients. BSP was detectable in conditioned media of MM cell lines. Using FACS analysis and in situ hybridization, we demonstrated that tumour cells from all MM patients and cell lines analysed express BSP at both the protein and the mRNA level.
Risk factors for development of a second lymphoid malignancy in patients with chronic lymphocytic leukaemia
Authors: Maddocks‐Christianson Kami , Slager Susan L. , Zent Clive S. , Reinalda Megan , Call Timothy G. , Habermann Thomas M. , Bowen Deborah A. , Hoyer James D. , Schwager Susan , Jelinek Diane F. , Kay Neil E. , Shanafelt Tait D.
Published: July 2010

Previous studies suggested that patients with chronic lymphocytic leukaemia (CLL) are at a three‐ to fivefold increased risk of developing a second lymphoproliferative disorder (LPD). This observational cohort study used the Mayo Clinic CLL Database to identify factors associated with developing a second LPD. A second LPD was identified in 26 (2·7%) of 962 CLL patients during a median follow‐up of 3·3 years. Diffuse large B‐cell lymphoma was the most common subtype of secondary LPD (12 of 26 cases). Patients previously treated for CLL had a trend toward higher prevalence of second LPD (4%) compared with previously untreated patients (2%; P = 0·053). More strikingly, patients treated with purine nucleoside analogues (PNA) had a significantly increased risk of subsequent second LPD (5·2%) compared with patients who had not received PNA (1·9%; P = 0·008). No statistically significant association was observed between risk of second LPD and other CLL characteristics (ZAP‐70, CD38, IgVH mutation status or cytogenetic abnormalities). In this series, prior treatments with PNA or anthracyclines were the only significant factors associated with risk of developing a second LPD in patients with CLL. Physicians should strictly adhere to established criteria to initiate treatment for CLL patients who are not participating in clinical trials.
Influence of human leucocyte antigen disparity and graft lymphocytes on allogeneic engraftment and survival after umbilical cord blood transplant in adults
Authors: Van Heeckeren Willem J. , Fanning Laura R. , Meyerson Howard J. , Fu Pingfu , Lazarus Hillard M. , Cooper Brenda W. , Tse William W. , Kindwall‐Keller Tamila L. , Jaroscak Jennifer , Finney Marcie R. , Fox Robert M. , Solchaga Luis , Forster Margaret , Creger Richard J. , Laughlin Mary J.
Published: July 2010

The dose of graft‐nucleated cells and CD34+ haematopoietic progenitor cells are predictors of allogeneic engraftment and survival in umbilical cord blood (UCB) recipients. In this single institution prospective phase II trial, flow cytometric analyses of CD34+ progenitor and lymphocyte populations in unmodified single unit human leucocyte antigen (HLA)‐disparate UCB grafts infused into 31 consecutive adults (median age 41 years, range 20–64) receiving myeloablative conditioning were compared with clinical outcomes. Median infused UCB graft‐nucleated cells and CD34+ dose was 2·2 × 107/kg and 1·2 × 105/kg respectively. Day to absolute neutrophil count ≥0·5 × 109/l with full donor chimerism averaged 27 d (range 12–41). Univariate analyses demonstrated that UCB graft‐infused cell doses of CD34+ (P = 0·015), CD3+ (P = 0·024) and CD34+HLADR+CD38+ progenitors (P = 0·043) correlated with neutrophil engraftment. This same analysis did not demonstrate a correlation between CD34+ (P = 0·11), CD3+ (P = 0·28) or CD34+HLADR+CD38+ (P = 0·108) cell dose and event‐free survival (EFS). High‐resolution matching for HLA‐class II (DRB1) resulted in improved EFS (P = 0·02) and decreased risk for acute graft‐versus‐host disease (GVHD) (P = 0·004). Early mortality (prior to post‐transplant day +28) occurred in three patients, while 26 patients achieved myeloid engraftment. These results suggest that UCB graft matching at DRB1 is an important risk factor for acute GVHD and survival, while higher UCB graft cell doses of CD34+, committed CD34+ progenitors and CD3+ T cells favourably influence UCB allogeneic engraftment.
Familial myelodysplasia and acute myeloid leukaemia – a review
Authors: Owen Carolyn , Barnett Michael , Fitzgibbon Jude
Published: July 2010

Familial occurrence of myelodysplasia (MDS) and/or acute myeloid leukaemia (AML) is rare but can provide a useful resource for the investigation of predisposing mutations in these myeloid malignancies. To date, examination of families with MDS/AML has lead to the detection of two culprit genes, RUNX1 and CEBPA. Germline mutations in RUNX1 result in familial platelet disorder with propensity to myeloid malignancy and inherited mutations of CEBPA predispose to AML. Unfortunately, the genetic cause remains obscure in most other reported pedigrees. Further insight into the molecular mechanisms of familial MDS/AML will require awareness by clinicians of new patients with relevant family histories.
The influence of prednisolone on the recirculation of peripheral blood lymphocytesin vivo
Authors: BLOEMENA E , WEINREICH S , SCHELLEKENS P T A
Published: July 2010

In humans and in laboratory animals, administration of a single dose of glucocorticosteroids induces a rapid and transient decrease in the numbers of peripheral blood lymphocytes through a redistribution of circulating lymphocytes. This lymphocytopenic effect is an important factor in the immunosuppressive capacity of corticosteroids. We have investigated whether the redistribution is due to an increased influx of circulating lymphocytes from the blood into the lymphoid organs or to a decreased efflux from the organs. Fluoresceinated lymphocytes were injected intravenously in unrestrained normal and prednisolone-treated rabbits and the distribution curves in the peripheral blood were analysed on the basis of a two-compartment model (known from pharmacokinetic studies). In this way. it is shown that the rapid fall in peripheral blood lymphocyte numbers induced by prednisolone is due to a decreased efflux of circulating lymphocytes from the lymphoid organs. In addition, it is demonstrated that prolonged infusion of prednisolone leads to an altered distribution pattern of the circulating lymphocytes among the organs.
Haemorrhage produces depressions in alloantigen-specific immune responses in the mouse through activation of suppressor T cells
Authors: CHANG Y H , JAGELS M , ABRAHAM E
Published: July 2010

The development of cell-mediated and humoral immune responses in BALB/c mice (H-2d) directed toward El4 cells (H-2b) was suppressed (cell-mediated cytotoxicity, 40–50% of control; antibody titres 127±17 versus 287±17 in controls) following haemorrhage of 30% total blood volume. This haemorrhage-induced depression of immune response can be transferred to normal recipients with T cells (3 × 107) from haemorrhaged syngeneic donors. Flow microfluorimetry (FMF) analysis showed no shift in CD8:CD4 ratios following haemorrhage. These results suggest that haemorrhage suppresses immune response through activation of suppressor T cells.
T activation marker evaluation in ARC patients treated with AZT. Comparison with CD4+ lymphocyte count in non-progressors and progressors towards AIDS
Authors: LEVACHER M , TALLET S , DAZZA M C , DOURNON E , ROUVEIX B , POCIDALO J J
Published: July 2010

Reductions in the percentage and absolute number of CD4+ lymphocytes, as well as abnormally high levels of activated peripheral T lymphocytes (CD3+ HLA-DR+ phenotype) and an increased proportion of CD8+ cells coexpressing the CD57 surface antigen (involved in natural killer activity) have been reported in HIV infection and associated with disease progression. We prospectively measured these subsets of lymphocytes in 34 patients with advanced AIDS-related complex (ARC) treated with azidothymidine (AZT). Peripheral blood lymphocyte phenotyping was performed before treatment, then at weeks 12 and 24. A striking fall in the proportion of activated T lymphocytes from baseline was observed (P < 0.001) at week 24. In contrast, the percentage of CD4+ cells showed a slight and transient rise at week 12 (P<0.05). No modification in levels of CD8+ or CD8+ CD57+ cells was detected during the study. Of the 34 patients, 11 developed AIDS, and 23 remained AIDS-free during 51 weeks of follow-up. Similar patterns of change in CD4+ and HLA-DR+ CD3+ lymphocytes were found in the AIDS progressors and nonprogressors. Likewise, HIV p24 antigenaemia showed parallel decreases in both groups of patients. Although changes in CD4+ cells, p24 antigenaemia and HLA-DR-reactive T lymphocytes were not predictive of clinical outcome, large differences existed between the two groups prior to the initiation of therapy. The short-term onset of AIDS was associated with lower CD4+ cell numbers, higher levels of serum p24 antigen and a greater proportion of activated T lymphocytes. Our results suggest that the possible interest of T lymphocyte activation markers, in conjunction with conventional phenotyping, should be investigated further.
Antibody binding and inhibition of pyruvate dehydrogenase (PDH) in sera from patients with primary biliary cirrhosis
Authors: SUNDIN U
Published: July 2010

Autoantibodies reactive against mitochondria which are present in sera from patients with primary biliary cirrhosis (PBC) have been shown to bind to the lipoamidc acctyltransferase moiety (E2) of the pyruvate dehydrogenase enzyme complex (PDH). This newly described antigen has been shown to be identical to M2, a 70-kD antigen of the inner mitochondrial membrane. Sera from 10 patients with PBC, 11 patients with chronic active hepatitis (CAH), 20 healthy controls and patients with thyroiditis and systemic lupus erythematosus (SLE) were tested by ELISA for the presence of antibodies (IgG and IgM classes and IgG subclasses) reactive against PDH. The effect of serum and separated IgG and IgM on the PDH enzyme activity was measured spectrophotometrically. Nine out of 10 PBC sera were positive by immunofluorescence for mitochondria (M2 pattern). These nine sera reacted strongly with both IgG and IgM to PDH in the ELISA and also inhibited the enzyme activity by 80% (s.d. 25%). This was significantly different compared with the controls (4 ± 6%: P<0.001). Enzyme inhibition was mainly caused by IgG. Of all control sera (from healthy and patient individuals) only one patient with CAH reacted significantly in the tests. This CAH patient had a high antibody titre against mitochondria as measured by immunofluorescence.
Anti-cardiolipin antibodies and HIV infection
Authors: MACLEAN C , FLEGG P J , KILPATRICK D C
Published: July 2010

Anti-cardiolipin antibodies of IgG class were found in 48% of intravenous drug users, 38% of homosexuals and 14% of heterosexuals (with no other risk factor) infected with HIV. Anti-cardiolipin antibodies were not increased in HIV-negative heterosexual partners of HIV-infected patients, but mildly elevated levels were detected in HIV-negative drug users, relative to healthy controls unselected for HIV status. Among HIV infected drug users, anti-cardiolipin antibodies were more common in men (P=0.01). Anti-cardiolipin antibodies were not associated with thrombocytopenia, Pneumocystis carinii pneumonia, disease progression or clinical stage. Anti-cardiolipin antibodies appear to be another non-specific marker of HIV infection which may be particularly common in male intravenous drug users infected with the virus.
Immune activation and T cell subset abnormalities in circulation of patients with recently diagnosed type I diabetes mellitus
Authors: AL-KASSAB A S , RAZIUDDIN S
Published: July 2010

The peripheral blood lymphocytes from patients with insulin-dependent diabetes mellitus (IDDM) and healthy controls were analysed for the HLA-DR+, interleukin-2 receptor-positive (IL-2R+) activating antigens, and for CD45R+ and CDw29+ subsets from the purified CD4+ and CD8+ T cell populations. Patients with IDDM had an increased percentage of HLA-DR+ and IL-2R+ cells in both CD4+ and CD8+ T cells. However, the percentage of CD4+CD45R+ suppressor/inducer T cells were decreased and CD4+CDw29+ helper/inducer T cells increased in all patients with IDDM, compared with healthy controls. Thus, IDDM patients exhibit a deficiency in the CD4+CD45R+ suppressor/inducer T cell subsets, which is probably related to the autoimmune phenomenon in this disease. In contrast, the percentage of CD8+CD45R+ and CD8+CDw29+ T cell subsets showed no major differences between patients with IDDM and controls. An alteration in the CD4+CD45R+ and CD4+CDw29+ T cell subsets appears to be a characteristic feature, and may relate to the impaired cell-mediated immunity in IDDM. These data provide new evidence for T cell dysregulation in IDDM.
Characterization of the binucleated giant cells generated in the autologous mixed leucocyte reaction from patients with rheumatoid arthritis
Authors: GOTO M , TSAI V , ZVAIFLER N J
Published: July 2010

Binucleated giant cells several limes larger than lymphocytes or monocytes were generated in an autologous mixed leucocyte reaction (AMLR) independent of DNA synthesis in patients with rheumatoid arthritis (RA). The AMLR giant cells with multiple cytoplasmic granules were nonspecific esterase-staining positive, phagocytic, non-adherent, HLA-DR+, CD11b+, CD14+, 4F2+, CDW29+, and anti-transferrin receptor positive, but negative for T, B or NK markers. RA patients aged less than 60 years form more giant cells: 12.6 ± 13.5% (n= 33) as compared with 0.4±1.5% in age- and sex-matched normals (n= 38) (P<0.001). More giant cells were seen over age 60 in both groups: RA 20.1 ± 15.5% (n=5) and healthy controls 3.0 ±3.2% (n= 8) (P<0.01). Neither disease activity nor treatment appear to influence the result in RA. The giant cells that are probably derived from monocytcs in AMLR may explain the formation of the giant cells in rheumatoid granulation tissues.
Detection of transforming growth factor-beta in rheumatoid arthritis synovial tissue: lack of effect on spontaneous cytokine production in joint cell cultures
Authors: BRENNAN F M , CHANTRY D , TURNER M , FOXWELL B , MAINI R , FELDMANN M
Published: July 2010

The presence of transforming growth factor-beta (TGF-β) in inflammatory joint disease was investigated. Synovial fluid from patients with rheumatoid arthritis (RA) and patients with other non-autoimmune inflammatory joint diseases contained high levels of both active and latent TGF-β. Levels of active TGF-β did not correlate with drug regimen in either patient group or with the recovery period in the individuals with non-RA joint disease. Freshly isolated synovial cells from individuals with RA were shown by Northern blotting to express the mRNA for TGF-β1 and to secrete latent TGF-β protein which could be neutralized by antibodies to TGF-β1 and TGF-β2. Lipopolysaccharide-stimulated peripheral blood mononuclear cells from normal donors produced interleukin-l (IL-1) and tumour necrosis factor-alpha (TNF-α) which was inhibited by pretreatment of these cells with recombinant TGF-β. Cytokine production was not inhibited if the addition of TGF-β was used after the inducing stimulus, suggesting that in activated cells cytokine production cannot be inhibited. This was confirmed by the observation that neither TGF-β1 or TGF-β2 inhibited spontaneous IL-1 or TNF-α production by rheumatoid synovial mononuclear cells in culture. These findings show that despite the presence of active TGF-β in RA synovial joints and the spontaneous production of latent (potentially active) TGF-β by RA cells in culture, additional TGF-β did not inhibit ongoing cytokine synthesis in vitro. This suggests that TGF-β may not inhibit cytokine production in the rheumatoid joint although it cannot be ruled out that in rivo TGF-β already has an immunosuppressive effect which cannot be further increased in vitro by exogenous protein.
A primed state exists in vivo following histological regression of amyloidosis
Authors: HAWKINS P N , PEPYS M B
Published: July 2010

Using a sensitive, quantitative, and non-invasive in vivo method, based on the specific binding of serum amyloid P component to amyloid fibrils, we have directly documented the spontaneous resolution of AA amyloid deposits in mice, and the prolonged existence thereafter of a primed state of enhanced susceptibility to further amyloid deposition. These results may have important implications for understanding and management of amyloidosis in humans.
Sequential measurement of the murine acute-phase protein serum amyloid P component (SAP) as an indicator of graft-versus-host disease following allogeneic bone marrow transplantation in mice
Authors: BAYSTON K F , HUBY R , COHEN J
Published: July 2010

Murine models of bone marrow transplantation (BMT) are used commonly for studies of the pathogenesis and treatment of graft-versus-host disease (GVHD). We report here that the sequential measurement of the mouse acute-phase protein SAP can be used to provide a sensitive, quantitative index of the severity of GVHD. Thirty mice underwent allogeneic, and a further 30 syngeneic BMT. GVHD w as assessed in vivo by clinical appearances and weight change, and post mortem by histology and calculation of splenic indices. Blood was obtained twice/week for SAP measurement and blood culture. In all mice an initial rise in SAP levels due to irradiation was followed by a return to baseline. Thereafter in syngeneic marrow recipients levels remained low. In contrast, after allogeneic BMT SAP levels rose progressively as mice developed GVHD, reaching a peak of 135 μg ml prior to death, from a nadir at day 20 of 15 μg ml. Mice with high splenic indices and histological evidence of severe GVHD had significantly higher SAP levels than mice with mild GVHD (P=0.002). Elevation in SAP levels occurred independently of bacteraemia. We conclude that in murine BMT sequential measurement of SAP provides an objective means of assessing GVHD in tiro.
Graft-versus-host reactions in the rat mimic toxin-induced autoimmunity
Authors: TOURNADE H , PELLETIER L , PASQUIER R , VIAL M C , MANDET C , DRUET P
Published: July 2010

Gold salts, d-penicillamine or mercurials induce autoimmunity in Brown Norway (BN) rats and provoke an immunosuppression in Lewis (LEW) rats. It has been suggested that immunologically mediated manifestations induced by drugs could result from graft-verves-host (GVH) like reactions. We show that BN spleen cells transferred into (LEW × BN)FI hybrids induce a chronic GVH reaction (GVHR). This reaction led to an autoimmune disease quite similar to that induced by drugs in BN rats. In both situations, a common part of the B cell repertoire is triggered. In contrast. LEW spleen cells transferred into (LEW × BN)FI hybrids provoke a lethal GVHR. This is to be compared with the CD8-mediated immunosuppression observed in LEW rats injected with HgCI2. These findings are in agreement with the prediction that immune dysregulation induced by drugs leads to GVH-like reactions either stimulatory or suppressive depending upon the strain tested.
Monocyte disorder causing cellular immunodeficiency: a family study
Authors: PRIETO J , SUBIRÁ M L , CASTILLA A , CIVEIRA M P , SERRANO M
Published: July 2010

We report a familial type of monocyte dysfunction not recognized previously. This disorder was observed in a young adult man with a long clinical history of recurrent, sell-limited episodes of cryptogenic fever accompanied by digestive and respiratory symptoms and repeated oral and skin infections. Lectin-induced lymphocyte transformation was reduced and skin tests revealed anergy to tuberculin and candidin. Monocytes from this patient exhibited markedly diminished expression of cytoskeletal vimentin intermediate filaments. HLA-DR antigens and immunological receptors for IgG Fc and C3b. These abnormal monocytes demonstrated impaired phagocytosis and reduced accessory cell function on PHA-mediated lymphoeyte activation. Release of soluble lymphocyte-activating factors by these cells was found to be defective. Lymphocytes from the patient responded appropriately to lectin in the presence of normal monocytes. Two family members of the proband presented similar monocyte defects although they only manifested minor clinical symptoms. This syndrome underlines the interest of testing monocyte markers and function in subjects with clinical manifestations of immunodeficiency.
Effect of human intestinal macrophages on immunoglobulin production by human intestinal mononuclear cells isolated from patients with inflammatory bowel disease
Authors: WU K C , MAHIDA Y R , PRIDDLE J D , JEWELL D P
Published: July 2010

The effect of macrophages on spontaneous immunoglobulin production by isolated human intestinal mononuclear cells (MNC) is unknown. Depietion of macrophages by adherence to fibronectin or by panning with macrophage-specific monoclonal antibody 3C10 lead to a significant reduction in IgA, IgG and lgM production by intestinal MNC from both normal (n= 10) and inflammatory bowel disease (IBD) (n= 13) mucosa. The reduction in immunoglobulin produced by macrophage-depleted intestinal MNC was greater in IBD patients than in normal controls. There was a significant correlation (r=0·816, P<0·001) between the percentage of macrophages depleted by panning with 3C10 and the reduction in IgG produced by macrophage-depleted intestinal MNC. Addition of either fibronectin-adherent cells or the supernatant from these macrophage-enriched cells enhanced immunoglobulin production in a dose-dependent fashion. A greater increase in IgG production by macrophage-depleted cells was seen when cultured with supernatant from inflamed IBD mucosal cells, compared with that from normal mucosal cells. The soluble factor(s) responsible in the supernatant was acid and heat susceptible but was not affected by freezing and thawing. Addition of recombinant human interleukin-1β or human interferon-gamma to cell cultures did not influence immunoglobulin production. Thus, human intestinal macrophages enhance spontaneous immunoglobulin production by isolated intestinal MNC by secreting soluble factor(s) which remain to be fully characterized.
Immunoregulatory CD4+ CD45R+ suppressor/inducer T lymphocyte subsetsand impaired cell-mediated immunity in patients with Down's syndrome
Authors: RAZIUDDIN S , ELAWAD M E
Published: July 2010

The monoclonal antibodies 2H4 and 4B4 allow CD4+ and CD8+ T lymphocytes to be subdividedinto CD45R+ and CDW29+ functional subpopulations. The CD4+ CD45R+ lymphocytes aredesignated as suppressor/inducer and CD4+ CDW29+ as helper/inducer subsets. Peripheral bloodlymphocytes from 19 patients with Down's syndrome and 19 age- and sex-matched normal controlswere analysed for the CD45R+ and CDW29+ subsets from the CD4+ and CD8+ T lymphocytes. Thepercentage of CD4+ CD45R+ cells (suppressor inducer) was markedly increased and of CD4+CDW29+ cells (helper/inducer) decreased in all patients with Down's syndrome. In contract, thepercentage of CD8+ CD45R+ and CD8+ CDW29+ subsets showed no major differences betweenpatients with Down's syndrome and normal controls. Moreover, an alteration in the CD4+ andCD45R+ and CD4+ CDW29+ T cell subsets was accompanied by a markedly reduced proliferativeresponse to phytohaemagglutinin and concanavalin A stimulation of the CD4+ T lymphocytes. Thus, a deficiency exists in patients with Down's syndrome in the CD4+ CDW29+ helper/inducerT cell subset which may contribute to their impaired cell-mediated immunity.
Specificity andin vitrotransfer of the immunosuppressive effectof detergent-disrupted influenza virus vaccine
Authors: SMITH T L , JENNINGS R
Published: July 2010

Primed murine splenocytes give an in vitro antibody response to influenza whole virus vaccine(WVV), as measured by enzyme immunoassay (EIA). When subunit vaccine (SV) of cither influenzaA or influenza B virus was added to in vitro splenocyte cultures stimulated with WVV, the EIAantibody response to homologous WVV was reduced. This reduction in antibody response wasobserved when SV was prepared using zwitterionic detergent (empigen BB). non-ionic detergent(triton-X-100) or cationic detergent cetyl-trimethyl ammonium bromide (CTAB); it was found to beeffected only by SV of strains of the same virus subtype—when SVs prepared from a helerotypic(H3N2) strain, an H1N1 strain and an influenza B strain were added to splenocyte cultures in thepresence of WVV. When splenocytes from immunologically naive mice, exposed in vitro to SV, weretransferred to secondary cultures of primed splenocytes, the antibody response to WVV in thesecondary cultures was also reduced. Mechanisms that may suppress the in vitro antibody responseare discussed.
T lymphocyte interferon-gamma production induced byPlasmodium falciparumantigen is high in recently infected non-immune and low in immune subjects
Authors: CHIZZOLINI C , GRAU† G E , GEINOZ A , SCHRIJVERS D
Published: July 2010

Interferon (IFN) alpha and gamma were measured by radio-immunoassays in supernatanls fromcultures of peripheral blood mononuclear cells (PBMC) or purified T cell subsets incubated witheither Plasmodium falciparum schizonl-enriched malaria antigen (mAg). uninfected red blood cells(RBC) or pokeweed mitogen (PWM). Cell donors were 24 clinically immune, healthy African adultnative residents of a P. falciparum-endemic region, Haut-Ogooué, Gabon, and seven non-immune. European temporary residents with a history of a single to a few malaria infections during theprevious 1 to 9 months. When PBMC were cultured in medium alone or with RBC antigen no or lowtitres of IFN-γ were detected. PBMC proliferation and IFN-γ production observed in the presence ofmAg were dose dependent and significantly correlated. When cultured with mAg. PBMC from non-immune Europeans produced significantly higher levels of IFN-γ than did PBMC from clinicallyimmune Africans. No such difference was found when PBMC were cultured with PWM. The mAg-induced IFN-γ production was due mainly to CD4+ T cells and was not enhanced by CDS+ T celldepletion. No IFN-α was detected in culture supernatants. Thus. P. falciparum antigens are able toinduce in vitro production of IFN-γ by CD4+ Tcells; however, in this sample, individuals consideredlo be clinically resistant to malaria were low producers of IFN-γ.
Stimulation of FACS-analysed CD4+ and CD8+ human tumour-infiltrating lymphocytes with ionomycin + phorbol-12, 13-dibutyrate does not overcome their proliferative deficit
Authors: STOECK M , MIESCHER S , QIAO L , CAPASSO P , BARRAS C , VON FLIEDNER V
Published: July 2010

Human tumour-infiltrating lymphocytes (TIL) were prepared by enzyme digestion from a series of different tumours and were purified on a fluorescence-activated cell sorter (FACS II) according to their CD4+ and CD8+ phenotype. CD4+ and CD8+ TIL were stimulated separately in a low density microculture system with phytohacmagglutinin (PHA) or with ionomycin plus phorbol-12, 13-dibutyrate (PDBu). The PHA-induced proliferation of TIL was highly decreased when compared with control peripheral blood lymphocytes. A decreased proliferation of TIL was also observed when cells were stimulated with ionomycin plus PDBu, a combination which is thought to circumvent early events associated with lymphocyte activation. Some TIL were also plated in limiting dilution where they showed decreased frequencies of proliferating T cell precursors. The data suggest that one component of the inhibition of TIL must be acting ‘ownstream’ of the early events of lymphocyte activation.
Reduction of graft-versus-host disease in neonatal Fl hybrid mice
Authors: DEGIORGI L , HABESHAW J A , POVEY S , MATOSSIAN-ROGERS A
Published: July 2010

Pre-immunization of BALB/c (H-2d) mothers with C57BL/10 (H-2b) or CBA/H (H-2k) spleen cellspartially protected the Fl hybrid offspring of (BALB/c × C57BL/10) or (BALB/c × CBA/H) matingsfrom graft-versis-host-disease (GVHD) induced by neonatal intraperitoneal inoculation with spleencells of the paternal strain. The effects achieved were manifest as a reduction in mortality. Experiments to establish whether the phenomenon was antibody mediated were performed bypassive pre-immunization of BALB/c mothers with alloantisera obtained from BALB/c previouslyimmunized with C57BL/10 spleen cells. Alloantisera produced an equivalent reduction in GVHDmortality. Some of the F1 mice that survived challenge with paternal strain spleen cells were proven tobe haemopoietic chimaeras using immunofluorescence with anti-MHC monoclonal antibodies andpolymorphism of the enzyme glucose-phosphale-isomerase present in the strains used. The possiblemechanisms of protection from GVHD in our mouse model are discussed.
The level of mannan-binding protein regulates the binding of complement-derived opsonins to mannan and zymosan at low serum concentrations
Authors: SUPER M , LEVINSKY R J , TURNER M W
Published: July 2010

When sera diluted to 5% in a buffer containing calcium and magnesium were incubated with mannancoated ELISA plates, C4 fragments, properdin and factor B were bound to the plates as well as the expected opsonic C3 fragments, C3b and C3bi. The calcium-dependent lectin mannan-binding protein, which is structurally similar to C1q, was also shown to bind in this assay and analysis of sera from 179 healthy blood donors revealed that the binding levels of all these proteins were highly significantly correlated. Results obtained with a previously described C3b opsonic assay using zymosan also correlated with the mannan-binding levels. When the sera were diluted to 5% in the presence of Mg-EGTA there was no detectable binding of complement proteins to the mannan surface, confirming that no alternative pathway activation occurred at this serum concentration. When sera were diluted to 5% in a buffer containing EDTA in order to study immunoglobulin binding in the absence of complement activation, the levels of bound IgG1, IgG2, IgG3, IgA and IgM antibodies were found to be completely unrelated to the C3bi binding levels previously observed. The results suggest that in this experimental system using low concentrations of serum, mannan-binding protein initiates an antibody-independent mechanism of cleavage of the classical pathway component C4, which subsequently regulates the degree of cleavage of C3 and recruitment of alternative pathway proteins.
Differences between C4A and C4B in the handling of immune complexes: the enhancement of CR1 binding is more important than the inhibition of immunoprecipitation
Authors: GATENBY P A , BARBOSA J E , LACHMANN P J
Published: July 2010

There are two isotypes of C4—C4A and C4B—, encoded within the major histocompatibility complex with quite different properties. In this study we have compared purified C4A and C4B with regard to their ability to prevent immune complex precipitation and to enhance the binding of both preformed and nascent immune complexes to the receptor CR1 on red cells. C4A was modestly more effective than C4B at inhibiting immunoprecipitation, particularly in antibody excess. In the CRI binding assay C4A was markedly more effective than C4B in enhancing binding to CRI. This difference was seen with both preformed and nascent immune complexes at equivalence and antibody excess. Thus the major differences between C4A and C4B in regard to immune complex handling is at the level of CRI binding. Given the strong association of C4A* QO alleles with immune complexmediated diseases like systemic lupus erythematosus, these findings have important pathogenetic implications.
Depressed degranulation response of synovial fluid polymorphonuclear leucocytes from patients with rheumatoid arthritis to IgG aggregates
Authors: DULARAY B , DIEPPE P A , ELSON C J
Published: July 2010

No difference was found between the degranulation responses to FMLP of synovial fluid (SF) polymorphonuclear leucocytes (PMNL), from patients with rheumatoid arthritis (RA), and either paired blood PMNL or blood PMNL from a healthy donor. In contrast, the response of SF PMNL to heat-aggregated IgG was often reduced compared with autologous blood PMNL. Similarly, SF from some (35%) RA patients stimulated degranulation of PMNL but the response of SF-derived PMNL to autologous stimulatory SF was reduced compared with the response of blood PMNL. The stimulatory activity of the SF was removed by sepharose-protein A. These results were taken to suggest that the activity is due to immunoglobulin aggregates and that SF PMNL (from some RA patients) are tachyphylactic to stimulation by immunoglobulin aggregates as measured by degranulation because they have been stimulated by immunoglobulin aggregates in vivo. In other studies the concentration of myeloperoxidase (MPO) was measured enzymically in RA SF and was found to be present in varying amounts. However, only a weak relationship was found between MPO levels and either PMNL numbers or levels of complement-bearing IgG aggregates in SF. It is considered that the relationship between MPO and immunoglobulin aggregates levels is obscured by the presence of a peroxidase inhibitor in the fluids and/or because only aggregates bound to tissue stimulate degranulation in vivo.
Adsorption of anti-dsDNA antibodies by immobilized polyanionic compounds
Authors: AOTSUKA S , FUNAHASHI T , TANI N , OKAWA-TAKATSUJI M , KINOSHITA M , YOKOHARI R
Published: July 2010

Antibodies to dsDNA are characteristically present in serum from patients with systemic lupus erythematosus (SLE). and have been shown to have the capacity to react with various molecules bearing repeating negative charges. After a number of polymeric or monomeric molecules with differently charged groups and hydrophobic molecules had been coupled covalently as ligands on cellulose gel, the adsorption capacities of the ligands for anti-dsDNA antibodies were evaluated. It was found that gels coupled with polyanionic dextran sulphate (DXS) and polyacrylic acid (PA) and monoanionic sulphanilic acid (SA) absorbed anti-dsDNA antibodies effectively. DXS gel also adsorbed antibodies to ssDNA and heparan sulphate, antigens with repeating negatively charged moieties, while no ligand was able to adsorb anti-nRNP antibodies. The finding that DXS gels adsorbed anti-dsDNA antibody in proportion to their charge density, and that the interaction between anti-dsDNA and DXS gel is broken readily by an increase in ionic strength, indicated that the binding is ionic in nature. Moreover, virtually all F (ab′)2 anti-dsDN A became adsorbed onto the DXS gels, suggesting that the binding occurred via specific antigen-binding sites on the antibody molecule. Binding of these polyanion-binding autoantibodies with anionic sites in the glomerular basement membrane may therefore cause the tissue damage observed in SLE.
Quantitative study ofLeishmania braziliensis braziliensisreactive T cells in peripheral blood and in the lesions of patients with American mucocutaneous leishmaniasis
Authors: CONCEIÇÃO-SILVA F , DÓREA R C C , PIRMEZ C , SCHUBACH A , COUTINHO S G
Published: July 2010

A limiting dilution analysis (LDA) was utilized to estimate the frequency of L. braziliensis braziliensis reactive T cells (Lbb-T cells) in peripheral blood and in the lesions of patients with mild localized cutaneous leishmaniasis (LCL) or with severe mucosal leishmaniasis (MCL). The frequencies of Lbb-T cells in peripheral blood varied from 1:107 300 to 1:3587 and were not significantly different in MCL and LCL patients. However, a significant difference was encountered (P <0.02) between the T cells frequencies in cutaneous (1:748 to 1:45) and mucosal lesions (1:152 to 1:13). A positive correlation was also observed between these frequencies and the magnitude of delayed-type hypersensitivity (DTH) (P < 0.01) and the presence of fibrinoid necrosis and granulomatous reaction in the site of the lesions (P<0.05). The lack of correlation between the severity of disease (MCL or LCL) and the frequency of Lbb-T cells in peripheral blood gave no indications towards understanding the physiopathology of severe or mild disease. However, the correlation between high T cell frequencies in the site of the lesions, the magnitude of DTH, the fibrinoid necrosis and the severity of the disease (MCL lesions) points to the possibility that the presence of a strong T cell dependent cellular immune response in the site of the lesions may have a deleterious effect. However, a local well modulated T cell immune response might provide healing of the lesions.
Therapy of a fatal murine cytomegalovirus infection with thymic humoral factor (THF-γ2) treated immune spleen cells
Authors: RAGER-ZISMAN B , ZUCKERMAN F , BENHARROCH D , PECHT M , BURSTEIN Y , TRAININ N
Published: July 2010

Infection of mice with murine cytomegalovirus (CMV) presents a model for the study of the role of the immune system in the pathogenesis of human CMV. We performed adoptive transfer experiments to evaluate the prospects for enhancing the anti-viral potential of murine CMV immune spleen cells by THF-γ2. Adult BALB/c mice resistant to murine CMV become highly susceptible following immunosuppression by cyclophosphamide. Recipient mice were injected with murine CMV and cyclophosphamide concomitantly, and 24 h later adoptive transfers of syngeneic immune spleen cells were performed. We showed that passive transfers of murine CMV immune spleen cells prevented the development of a fatal disease in 38% of the recipient mice. Daily injections of murine CMV immune donor mice with THF-γ2 enhanced considerably (93%) the therapeutic potential of virus-specific immune cells. These experiments provide direct evidence for the antiviral capacity of THF-γ2 through its immunomodulatory effect on immune T cells.
Flare-up reaction of streptococcal cell wall induced arthritis in Lewis and F344 rats: the role of T lymphocytes
Authors: VAN DEN BROEK M F , VAN BRUGGEN M C J , STIMPSON S A , SEVERIJNEN A J , VAN DE PUTTE L B A , VAN DEN BERG W B
Published: July 2010

One i.p. injection of a sterile suspension of streptococcal cell walls (SCW) induces chronic erosive polyarthritis in susceptible Lewis rats, but not in resistant F344 or nude Lewis rats. Because continuous exacerbations may be one possible mechanism underlying chronic disease, we studied the mechanism of these flare-up reactions in Lewis and F344 rats. Injection of SCW into the right knee joint of rats induced a transient monoarthritis in both strains. Reactivation of the subsided arthritis by i.v. administration of the same antigen could be evoked only in the Lewis rat. Even repeated i.v. challenges with SCW failed to induce a flare-up reaction in the F344 rat, while the Lewis rat went through an exacerbation after every challenge. Removal of T lymphocytes by monoclonal antibodies before induction of an exacerbation rendered Lewis rats refractory to flare-up reactions, thus indicating the T cell-dependence of this reaction. Furthermore, when cell walls from heterologous bacteria were tested for their capacity to induce exacerbations of SCW-induced monoarthritis and to induce proliferation of SCW-specific T lymphocytes in vitro, a strong correlation between both features was found, again pointing to a role for SCW-specific T cells in exacerbations. Together, these data support our hypothesis that chronic arthritis is the result from repeated reactivations of a waning arthritis which are dependent on antigen-specific T lymphocytes.
Activation process of macrophages afterin vitrotreatment of mouse lymphocytes with dodecylglycerol
Authors: HOMMA S , YAMAMOTO N
Published: July 2010

Alkylglycerols, inflammation products of cancerous membrane lipids, efficiently activate macrophages. A brief in vitro treatment (30 min) of peritoneal cells (mixture of non-adherent and adherent cells) with a small amount (50 ng/ml) of synthetic dodecylglycerol (DDG) resulted in greatly enhanced Fc-receptor-mediated ingestion activity of macrophages. However, treatment of adherent cells (macrophages) alone with DDG produced no significant enhancement of macrophage ingestion activity, implying that macrophage activation requires a contribution of non-adherent cells. DDG-treated non-adherent cells were found to generate a macrophage-activating signal factor. Studies with a serum free-0.1 % egg albumin-supplemented RPMI 1640 medium revealed that a serum factor is essential for macrophage activation process. Time course analysis of stepwise transfers of conditioned media of DDG-treated or untreated B cells and T cells revealed that DDG-treated B cells rapidly transmit a factor to untreated T cells which yield the ultimate macrophage-activating factor. This signal transmission among these cells for the macrophage activation process is too rapid to allow time for synthesis of inducible gene products. Thus, we hypothesized that a serum factor is modified by the pre-existing function of DDG-treated B cells and further modified by the pre-existing function of untreated T cells to yield macrophage-activating factor. This hypothesis was confirmed by the demonstration that DDG-treated splenic non-adherent cell ghosts modify a serum factor to yield macrophage-activating factor.