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Biomedical subjects

M Arock

Publications and source records attributed to M Arock.

At least 73 records · Page 4Linked to original sources
PubMed1994

Haematological effects of postoperative autotransfusion in spinal surgery.

A prospective, randomized, controlled study was performed to determine the haematological and biochemical changes and clinical safety of postoperative autotransfusion (Solcotrans Orthopedic Plus system) in patients undergoing spinal surgery. Fifty patients were studied and were randomly allocated to Control (n = 25) and Solcotrans (n = 25) groups. Both groups had their postoperatively drained blood collected into the Solcotrans reservoir but only the Solcotrans group had this salvaged blood considered for reinfusion. After a 5-h postoperative collection period, analysis of the shed blood showed a haematocrit of 0.26 +/- 0.11, few platelets (80 +/- 63 10g.l-1), a fibrinogen level of less than 0.1 g.l-1 and a high level of D-dimers. The salvaged blood did not clot and aerobic and anaerobic culture produced no growth. The volume of blood collected was greater than 200 ml in 21 patients in the Solcotrans group who were autotransfused (384 +/- 101 ml, range 200-600 ml), and in 16 patients in the Control group. Within 15 min following completion of reinfusion of the salvaged blood there was a significant, but moderate decrease in platelet count (181 +/- 74 vs 223 +/- 90 10g.l-1, P < 0.001) and fibrinogen concentrations (2.1 +/- 0.8 vs 2.3 +/- 0.9 g.l-1, P < 0.02), and an increase in circulating D-dimers (P < 0.001) and plasma free haemoglobin concentrations (236 +/- 155 vs 82 +/- 79 mg.l-1, P < 0.001). Prothrombin time (PT) and activated partial thromboplastin time (APTT) did not increase, and potassium concentrations were not significantly affected.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood↗
PubMed1994

IgE-dependent activation of Fc epsilon RII/CD23+ normal human keratinocytes: the role of cAMP and nitric oxide.

Epidermal keratinocytes (EK) are exposed to multiple inflammatory stimuli and paracrine factors secreted by various dermal cells (lymphocytes, mast-cells, macrophages, fibroblasts) during wounding, cutaneous allergy and infections. We have previously demonstrated that following stimulation with interleukin-4 (IL-4) or interferon-gamma, human EK express the low affinity receptor for IgE (Fc epsilon RII/CD23) on their surface. In the present study, we showed that the ligation of CD23 by IgE/anti-IgE immune complexes or specific monoclonal antibody, induces a dose-dependent release of interleukin-6 and tumor necrosis factor-alpha from EK. CD23-ligation activates the nitric oxide-dependent pathway, as demonstrated by the high levels of nitrites released in cell supernatants, and the accumulation of intracellular cyclic nucleotides in EK. These second messengers are required for IgE-dependent stimulation of cytokine production by these cells, as this is completely abolished by cAMP or NO synthase antagonists. Human epithelial keratinocytes may thus participate in IgE-mediated immune responses, through their ability to express functional CD23 antigen.

Amino Acid Oxidoreductases↗
PubMed1994

Specific ligation of the HIV-1 viral envelope protein gp120 on human CD34+ bone marrow-derived progenitors.

The precise mechanisms of hematologic abnormalities observed during HIV infection remain unknown. In vitro experiments performed by various authors concerning the HIV toxicity on bone marrow-derived precursors did not allow them to determine whether this toxicity could be mediated through direct or non-direct effects, since it is today unclear if gp120 possesses a direct hematotoxic effect on human bone marrow progenies. The aim of our study was to determine whether labelled gp120 could specifically bind to the membrane of purified human normal CD34+ cells and to investigate the in vitro effect of the gp120 on their growth. To answer these questions, human CD34+ cells were purified from normal bone marrow samples, then labelled with monoclonal antibodies directed either against CD4 antigen or CD34 antigen and/or with FITC labelled gp120 and analyzed by FACS. Our results demonstrate the presence of about 5% of CD4+CD34+ cells and of nearly 12% of CD34+gp120+ precursors. Together with our results concerning the in vitro inhibitory effect of gp120 on the growth of the same purified CD34+ precursors, our data demonstrated the direct hematotoxic activity of HIV-derived gp120 and the possible HIV infection of hematopoietic progenitors through the interaction of gp 120 with CD34+ cell surface.

Antibodies, Monoclonal↗
PubMed1994

[Phenotypic and functional characterization of human prothymocytes].

We report herein the phenotypic and functional analysis of human bone marrow and thymus derived early T cells. Commitment to T cell lineage is acquired during CD7 antigen expression by CD34+ precursors in human bone marrow and before thymus colonization. Early thymocytes show similar phenotypic characteristics as bone marrow T cells. They rapidly acquire CD4 before the dual expression of CD4 and CD8. Their expansion and differentiation is regulated by two major factors: thymic stroma and cytokines produced by these stroma cells or by thymocytes themselves. Among cytokines, IL1 and sCD23 produced by thymic epithelial cells support in vitro early T cell development.

Adult↗
PubMed1993

IL-4 release by human leukemic and activated normal basophils.

Recently, authors have addressed the ability of human basophils to produce IL-4. We report here the detection of significant serum IL-4 levels in a case of acute transformation of chronic myelogenous leukemia with a predominant basophilic cell population. Leukemic basophils were isolated from patients' PBMC and assayed for their IL-4-mRNA expression and their ability to secrete this cytokine in vitro. Leukemic basophilic cells (> 90% toluidine blue positive) but not other PBMC expressed IL-4-mRNA, contained IL-4 protein, and secreted this cytokine. These cells had a spontaneous IL-4 secretion ability, without a need for an exogenous activator. Meanwhile, IL-4 release was significantly increased following leukemic cell activation through Fc epsilon RI-ligation or by Ca2+ ionophore. IL-4 and its mRNA were also detected in leukemic basophils from three other chronic myelogenous leukemia patients with moderate basophilia (13, 14, and 23% basophils in PBMC). To confirm these data in normal human cells, we have developed a method to obtain large numbers of purified basophils from human bone marrow cell cultures. In contrast to leukemic basophils, normal cells required in vitro activation through Fc epsilon RI ligation or by Ca2+ ionophore to express and secrete IL-4. Leukemic and normal basophils secreted histamine following in vitro activation, but were negative for tryptase. These data thus demonstrate the in vivo and in vitro ability of human basophils to produce IL-4.

Basophils↗
PubMed1993

[Effects of postoperative autotransfusion in prosthetic surgery of the hip with the ConstaVac device].

A new device for postoperative autotransfusion (ConstaVac, Stryker) was assessed after total hip replacement in 43 patients, mean age 63 +/- 13 years. Intraoperative blood was administered to 27 patients (63%), autologous blood only in 19, homologous blood only in 6 and both autologous and homologous blood in 2. The blood shed during an average 5-hour postoperative period had an haematocrit of 23 +/- 7% and included only few platelets (72 +/- 83 G.l-1). It did not clot as it contained there was less than 0.1 g.l-1 of fibrinogen and a high concentration of D-dimers. In 30 patients (70%), the amount of blood thus collected exceeded 200 ml. An average of 327 +/- 131 ml were subsequently retransfused to these patients. Postoperative autotransfusion induced a moderate but significant decrease in platelet counts (205 +/- 66 vs. 224 +/- 67 G.l-1, p < 0.02) and fibrinogen concentrations (2.3 +/- 0.7 vs. 2.4 +/- 0.6 g.l-1, p < 0.03), and an increase in circulating D-dimers (p < 0.001). Coagulation tests, free plasma haemoglobin and potassium concentrations were not significantly altered. Since the haematocrit of the blood lost was lower than that of the patients', the haematocrit did not increase significantly. Posttransfusion shivering occurred in two patients (7%). Bacteriological cultures of the lost blood and of venous samples obtained after postoperative autotransfusion remained sterile. Postoperative autotransfusion is a simple technique with few side-effects, which might be safely associated with other methods used to decrease the rate of homologous blood transfusion.

Adult↗
PubMed1993

Maturation of human myelomonocytic leukemia cells following ligation of the low affinity receptor for IgE (Fc epsilon RII/CD23).

The regulation of the low affinity receptor for IgE (Fc epsilon RII/CD23) expression and its role were investigated in U937 cell line and in leukemic cells from a patient (Amb) with acute myeloblastic leukemia. Both cell populations were CD23- but could acquire CD23 expression following treatment with IL-4. CD23+ cells, however, remained blastic and did not show any significant phenotypical and functional modifications. Following ligation of the CD23 on U937 and Amb cells by anti-CD23 mAb, these leukemic cells differentiated into mature monocyte/macrophage-like cells. CD23 ligation promoted the expression of the monocyte marker, CD14, increased the expression of the common beta chain of the LFA-1 family (CD18), and down-regulated the expression of the promonocytic marker CD33. Morphological and phenotypical changes were associated with functional modifications as CD23 ligation allowed the acquisition of the oxidative metabolism in leukemic cells as revealed by luminol-dependent chemiluminescence. As in mature monocytes, CD23 ligation induced an accumulation of intracellular cAMP in leukemic cells. These data indicate that ligation of CD23 may induce the maturation of myelomonocytic cells into monocytic-like cells.

Cyclic AMP↗
PubMed1993

Interleukin 4 inhibits the proliferation and promotes the maturation of human leukemic early B cells.

The effects of interleukin 4 (IL-4) on human leukemic precursor B-cell lines were investigated. Recombinant IL-4 (rIL-4) was added to three acute lymphoblastic leukemia-derived pre B-cell lines: Reh, Km3 and Nalm-6. Our results show that rIL-4 significantly decreases continuous proliferation of Reh and Km3 cells while Nalm-6 cells have a limited response in this respect. This rIL-4 effect is dose-dependent and can be neutralized by anti-IL-4 monoclonal antibody (mAb). Furthermore, rIL-4 down-regulated IL-3-induced proliferation of Reh cells. Phenotypic analysis of rIL-4-treated cells points to significant induction of surface marker maturation of leukemic cells by this cytokine. Together, these in vitro data suggest that IL-4: 1) inhibits the proliferation and 2) promotes the differentiation of certain human leukemic B-cell precursors.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1993

Purification of normal human bone-marrow-derived basophils.

It is well established that basophils and eosinophils share a common differentiation pathway, although the factors regulating their terminal commitment (towards one or other lineage) are not yet fully defined. Interleukin-3 (IL-3) is a major differentiation factor for both human eosinophils and basophils, yielding a mixed population composed predominantly of eosinophilic cells (65 +/- 9%; n = 4), basophils at different stages of maturity (29 +/- 6%; n = 4) and monocytes/macrophages (6 +/- 3%; n = 4), after 3-4 weeks in culture. Using a relatively rapid and simple method involving a first step of gradient density centrifugation over a Percoll gradient (d = 1.063 g/ml) and a subsequent step of adhesion on tissue-culture-treated plastic, a cell population composed of 94 +/- 5% normal basophils and their precursors, with no demonstrable mast cells, was reproducibly obtained from human hematopoietic cells cultured for 3-4 weeks in the presence of recombinant IL-3. These cells contained high levels of histamine (1.39 +/- 0.14 pg/cell) and released this mediator upon stimulation with calcium ionophore A23187 and in a dose dependent manner upon stimulation with IgE-anti IgE, demonstrating their functional capacity. This relatively simple method therefore permits the production of large quantities of pure populations of normal and functional human bone-marrow-derived-basophils.

Basophils↗
PubMed1992

Cytokine effects of CD23 are mediated by an epitope distinct from the IgE binding site.

Human CD23 and its soluble forms (sCD23) display various biological activities, in addition to their IgE binding function (IgE/BF). The IgE binding domain was recently mapped to residues between Cys163 and Cys282 but its involvement in IgE-independent, CD23 functions remains unknown. In order to clarify this point, a series of N-terminal, C-terminal and internal deletion mutants of CD23 or sCD23 were expressed in CHO cells and tested for their ability (i) to bind to IgE, (ii) to induce colony formation by human myeloid precursor cells, (iii) to promote mature T cell marker expression by early prothymocytes, and (iv) to regulate IgE synthesis. The present study indicates that cytokine activities require the presence of Cys288, while this amino acid is not necessary for IgE/BF. Blocking experiments using various conformation-sensitive monoclonal antibodies further suggest that active epitope(s) of CD23 in cytokine assays is(are) distinct from those involved in IgE/BF.

Animals↗
PubMed1992

Epidermal keratinocyte-derived basophil promoting activity. Role of interleukin 3 and soluble CD23.

Human epidermal keratinocytes (EK) secrete factors able to sustain the proliferation of early myeloid cells and, in particular, the generation of basophils. This activity was previously attributed to IL-3, although no definitive in situ demonstration of this cytokine was provided. In regard to the possible physiological relevance of these data, we investigated herein the nature of EK-derived factors responsible for basophil promotion. Our data show that EK-derived supernatants (EK-sup) contain IL-3 as well as soluble CD23 (sCD23), both known for their colony stimulating activity. Messenger RNA for IL-3 and CD23 were also detected in EK. Blocking experiments using specific neutralizing monoclonal antibodies (mAb) further indicate that EK-derived basophil promoting activity is mainly due to the presence of IL-3 and sCD23 in EK-sup. Furthermore, by contrast to IL-3, sCD23 secretion by EK is cortisone sensitive and highly enhanced by IL-4, suggesting distinct regulatory mechanisms for their production.

Antibodies, Monoclonal↗
PubMed1992

Effect of CD23 on purified human hematopoietic cells.

We report herein the effect of soluble CD23 (sCD23) on the differentiation of lymphoid and myeloid precursors. CD23 is known as the low affinity receptor for IgE. In addition to it, our results indicate that sCD23 in synergy with IL1 is able to promote the maturation of normal and leukemic hematopoietic progenitor cells.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1992

[Demonstration of a protector effect of interleukin-1 against hematologic toxicity of azidothymidine (AZT)].

3'-azido-3'-deoxythymidine (Azidothymidine or AZT) has attained wide critical utility in the treatment of acquired immunodeficiency syndrome (AIDS). Unfortunately, treatment with AZT is associated with the development of severe hematopoietic toxicity. The AZT sensitivity of marrow progenitors was different with an IC 50 of 10(-8) M and 10(-6) M for respectively BFU-E and CFU-GM/GEMM. Data reported here show that recombinant human interleukin-1 alpha (IL-1 alpha), a pleiotropic cytokine, was demonstrated to be efficient to protect normal human as well as murine hematopoietic progenitors (CFU-GM, CFU-GEMM and BFU-E) from the toxic effect of AZT. The maximal effect was observed with 30 U/ml (Human cells) or 100 U/ml (murine cells) IL-1 alpha for BFU-E and CFU-GM/GEMM, with a marked effect at 1 U/ml. The results demonstrate that marrow progenitors respond differently to AZT and point out the potential efficacy of IL-1 alpha to enhance the proliferation of hematopoietic stem cells treated with growth factors (IL-3, erythropoietin) and to minimize the hematopoietic toxicity associated with AZT treatment.

Adult↗
PubMed1992

[Studying the mast cell. Recent data].

In the light of recently published data, the mast cell can now be viewed as a key cell, not only in allergic reactions such as immediate hypersensitivity responses, but also in a broad spectrum of other biologic responses including host-parasite interactions, nonspecific inflammatory reactions, fibrosis, angiogenesis, tissue reconstruction, and wound healing. Nevertheless the molecular basis for the intervention of mast cells in many of these biologic responses is still unclear. Very recent studies have demonstrated that mast cells are capable of producing a wide range of cytokines, a property which may influence various physiologic, immunologic and disease processes. Furthermore, although substantial differences have been reported between mast cells located in different tissues, the reasons and mechanisms underlying this heterogeneity long remained obscure. The recent development of two original experimental approaches, i.e., in vitro culture of mast cells, mainly derived from mouse bone marrow precursors, and replenishment of mast cell-deficient mice, has provided new insight into the mechanisms by which tissue microenvironment influence of regulation mast cell phenotype. Extrapolation to humans of data obtained in rodents is, however, hazardous. In the review presented here, the most recent data from the literature provide the basis for outlining avenues of research which can be expected, in the near or remote future, to solve what mast cell experts term "the riddle of the mast cells".

Animals↗
PubMed1991

Human thymic epithelial cells produce interleukin-3.

Interleukin-3 (IL-3) is a hematopoietic growth factor suggested to be produced by activated T lymphocytes. Meanwhile, supernatants from human thymic stroma could promote the proliferation of myeloid stem cells. Thus, we investigated whether IL-3 accounts for this activity. Therefore, human thymic epithelial cells (TEC), fibroblasts, and adherent cells were isolated, and their culture supernatants assayed for myeloid colony promotion. Only supernatants from thymic epithelial cells supported colony-forming unit growth in semisolid media. This effect decreased following anti-IL-3 monoclonal antibody addition to these cultures. Furthermore, in situ hybridization showed the presence of IL-3 mRNA in epithelial cells. Effect of TEC culture conditions on IL-3 production by these cells was also studied. Together, these data show that IL-3 production is not the exclusive property of human activated T lymphocytes.

Antibodies, Monoclonal↗
PubMed1991

Soluble CD23 increases IL-3 induction of histamine synthesis by human bone marrow cells.

Interleukin 3 (IL-3) and soluble CD23 (sCD23/IgERII) have similar capacities to induce basophil development from human bone marrow (BM) precursors. IL-3 also has the capacity to induce de novo histamine synthesis from human BM cells. In the present study, we examined the effect of sCD23 on histamine synthesis by human BM cells and its relationship with IL-3. Our data showed that recombinant 25 KDa sCD23, although on its own failed to induce histamine release, significantly enhanced IL-3-stimulated histamine synthesis by BM cells. These data provide further support for the cytokine-like role of sCD23 on hematopoietic cells.

Antigens, Differentiation, B-Lymphocyte↗