摘要:

Abstract:To investigate the role of cytosolic free calcium ([Ca2+]iin interferon‐γ (IFN‐γ) pre‐activation (priming) of human neutrophilic granulocytes (PMN) we used three different fluorescence methods, i.e. digital imaging of single, adherent, Fura‐2 loaded cells, flow cytometric measurements of single, non‐adherent, Fluo‐3 loaded cells, and spectrofluorometry of Indo‐1 loaded PMN in suspension. IFN‐γ increased the [Ca2+]ilevel in single, adherent PMN during the second phase of the fMLP response. The bacterial peptide fMLP (N‐formyl‐L‐methionyl‐L‐leucyl‐L‐phenylalanine) is a known stimulant of the calcium/inositol phosphate system. The [Ca2+]iincrease was abolished in Ca2+‐free test buffer. Furthermore, the baseline [Ca2+]ilevel was found to be slightly increased in IFN‐γ primed PMN as analysed with flow cytometry. On the other hand, these [Ca2+]iresponses were not detectable with the other methods used. We suggest that IFN‐γ increases the plasma membrane permeability for calcium in PMN, and substantiate this by demonstrating compliance with a capacitative model for intracellular calcium regulation. Mathematical modeling also suggested that IFN‐γ primed human PMN may sequester 13% more Ca2+than unprimed cells in fMLP‐insensitive intracellular stores. Thus, the Ca2+responses to IFN‐γ are modest and not easily detectable with some of the methods currently in use. They nevertheless explain why fMLP elicit

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01867.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:We evaluated the utility of an immunocytochemical technique employing the commercially available anti‐CD56 monoclonal antibody, NKH 1. The utility and sensitivity of this technique in the detection of invasive neuroblastoma (NB) cells in the bone marrow were compared with those of Wright‐Giemsa staining. The correlation coefficient for the percent NB cells detected using Wright‐Giemsa staining with the percent NKH 1 immunoreactive cells was 0.78. In the analysis of specificity, this monoclonal antibody showed slight cross‐reactivity with normal bone marrow cells, including macrophages, lymphocytes and osteoblasts. In the evaluation of the sensitivity of the NKH 1 immunocytochemical technique, SK‐N‐DZ and SK‐N‐SH NB cell lines were added to morphologically normal bone marrow mononuclear cells from patients without NB to the final NB cell line at concentrations of 2%, 1% and 0.1%. NB cells at the final concentration of 0.1 % could be detected by the immunocytochemical technique. We conclude that the NKH 1 immunocytochemical staining technique is useful in the detection of metastatic NB cells

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01868.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:The intermediate filament proteins which include vimentin, desmin, and the keratins are one of three major classes of cytoskeletal proteins in eukaryotic cells. In this study we found that most of the vimentin of undifferentiated HL60 and cells induced to differentiate either along the monocytoid pathway by 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) or along the granulocytic pathway by retinoic acid was soluble in a buffer containing 1% Triton X‐100/0.6 mol/l KCl in which the intermediate filament proteins usually are not soluble. HL60 vimentin separated on Polyacrylamide gel electrophoresis into two proteins of Mr 55000 and 54000 that we detected by immunoblotting. The Mr 55000 species was the major form in undifferentiated HL60 cells and cells induced by retinoic acid. The distribution of both forms of vimentin changed during induction of differentiation by TPA and after 24 h the Mr 54000 species was predominant. After an additional 24 h exposure to TPA the relative levels of the two forms of vimentin approached equivalence and a high level of vimentin degradation products was seen. These results suggest that TPA may increase vimentin degradation along a pathway that has a Mr 54000 intermediate. In addition, the high levels of soluble vimentin in HL60 cells suggests that these cells may be a good model for studying components involved in vimentin

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01869.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:B‐cell non‐Hodgkin's lymphomas (B‐NHL) and B‐cell areas of reactive lymphadenopathies were investigated immunohistochemically for expression of two distinct ICAM‐1 epitopes, Me14/D12 and P3‐58, and the LFA‐1 α and β chains. Partial or total loss of expression of one or both epitope(s) and/or chain(s) was evident in all B‐NHL in function of increasing Working Formulation (WF) malignancy grade, with most defects in the high‐grade tumors, namely the lowest detectability of the ICAM‐1 Me14/D12 and LFA‐1 α chain, the lowest co‐expression of ICAM‐1 epitopes and LFA‐1 chains, and the most frequent simultaneous loss. The ICAM‐1 and LFA‐1 profiles overlapped within the low‐ and intermediate‐grades, whereas striking differences between the high‐grade subtypes were detected. Specifically, Burkitt's and lymphoblastic tumors always lost both epitopes and both chains. Large cell, immunoblastic tumors occasionally did so, and also showed either uncoordinated expression or co‐expression of these constituents. It is suggested that expression defects of this type may help differentiate malignant from benign lymphoproliferations, and also be involved in the progression of B‐NHL, since most are observed in high‐grade tumors, whose ICAM‐1 and LFA‐1 profiles indicate that their subtypes are the expressio

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01870.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Autologous bone marrow transplantation (ABMT) is frequently used in the treatment of neoplastic diseases. It involves several manipulationsin vitrothat can damage the stem cells responsible for grafting. Long‐term bone marrow cultures (LTBMC) reproduce the bone marrow microenvironment and support haematopoiesisin vitrofor several weeks. This technique provides information on the extent of the injury of stem cells during manipulations of bone marrow cellsin vitro.We studied the effect of the usual process of bone marrow cellsin vitroin autologous transplantation: cell concentration and cell cryopreservation. 28 bone marrows from healthy donors and 45 from patients who had undergone ABMT were assayed in LTBMC. The cultures were initiated after manual or automatic buffy coat cell separation or after density gradient or automatic mononucleated cells suspension. Bone marrows were studied pre‐ and post‐cryopreservation. The results show that several manipulations can disturb normal cell behaviour in LTBMC. Following automatic mononucleated cell separation and after thawing, the development of adherent cell layer in LTBMC is anomalous. These manipulations led to a delay in covering 50% of the flask surface, absence of adypocytes in adherent cell layer and absence of haemopoietic precursors in the supernatant at the 4thweek of culture. The results suggest that LTBMC can be used for control of the manipulationsin vitrorelated with

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01871.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Platelet refractoriness arising from HLA immunization develops in 20–50% of multitransfused patients with hematologic malignancies. We retrospectively analyzed the efficiency of leukocyte‐depleted blood components in preventing refractoriness in 140 adult patients with acute myeloid leukemia (AML), treated according to a standardized cytostatic protocol. Sixty‐eight patients received leukocyte‐depleted (L‐D) platelet concentrates (PCs) and red cells (RBCs), with fewer than 106leukocytes per unit, and 72 patients received standard (STD) blood components. Two of 67 (3%) evaluable patients in the leukocyte‐depleted group and 14 of 68 (21%) in the standard group became refractory during the median follow‐up time of 229 days. Nine of 24 (37%) previously pregnant women in the STD group but none of 17 in the L‐D group became refractory. Twenty patients were shifted during later treatment from L‐D to STD PCs; none became refractory. Corrected platelet increments (CI) at 18 hours were higher after STD PCs (6.5) than L‐D PCs (5.2), but more PCs and RBCs were transfused per patient in the STD group. It is concluded that effective leukocyte depletion prevents platelet refractoriness in patients with AML, even in those with previous immunization, and reduces the consumption

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01872.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:A serum‐free culture method was used to study the growth of megakaryocytic progenitor cells (CFU‐Meg) from patients with elevated platelet counts. The culture technique was combined with immunocytochemistry (APAAP, CD61) for the identification of CFU‐Meg derived cells in cytopreparations of cells eluted from the culture dishes. Twenty‐six patients with primary thrombocythaemia (14 untreated patients, UPT, 12 treated patients, TPT), 14 patients with reactive thrombocytosis (RT) and 9 normal individuals were studied. Unstimulated growth of CD61‐positive cells was detected in 8/14 UPT, 8/12 TPT, 12/14 RT and 5/9 normal subjects (with mean CD61‐positive cell counts of 75, 579, 236 and 7 per cytopreparation respectively). Cultures supplemented with interleukin 3 contained CD61‐positive cells in 11/14 UPT, 7/12 TPT, 14/14 RT and 5/9 normal subjects (with mean CD61‐positive cell counts of 157, 589, 250 and 7 per cytopreparation respectively). Thus, this serum‐free culture technique combined with sensitive positive identification of CFU‐Meg derived cells failed to discriminate between PT and RT. These results cast doubt on the usefulness of serum‐free culture assays for the detection of unstimulated CFU‐Meg growth in the differential diagnosis of patients with e

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01873.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Intravenous immunoglobulin replacement therapy reduces the number of bacterial infections in B‐cell chronic lymphocytic leukaemia (B‐CLL) patients. However, due to the complexity of immunodeficiency in B‐CLL and the cost‐effectiveness of replacement therapy, it is important to identify patients who are likely to benefit from the treatment and to investigate which dose should be used. 15 patients with hypogammaglobulinaemia and a history of recurrent infections received a fixed dose of 10 grams of gammaglobulin intravenously every 3 weeks. Serum IgG levels were significantly higher after three doses (p = 0.0002), and stabilized just above lower reference value after 11 doses. The total number of infection‐related events during 168 months before therapy was compared to the total number of infection‐related events in 169 months during therapy. The number of antibiotic prescriptions was reduced from 78 to 54 (N.S.), the number of admissions to hospital due to infections was reduced from 16 to 5 (p = 0.047) and the number of febrile episodes was reduced from 63 to 31 (p = 0.004). We conclude that a fixed low dose of gammaglobulin intravenously can restore normal serum IgG levels in hypogammaglobulinaemic B‐CLL patients, and leads to a decreased number of febrile episodes and admissions to hospital d

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01874.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01875.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0902-4441    

DOI:10.1111/j.1600-0609.1994.tb01876.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY