PubMed Health⌕ Search

Biomedical subjects

F Patrone

Publications and source records attributed to F Patrone.

At least 37 records · Page 2Linked to original sources
PubMed1986

Expression of cytolytic functions in HL-60 leukaemic cells after induction of polymorphonuclear leukocyte differentiation.

Mature polymorphonuclear leukocytes (PMN) are capable of mediating phorbol myristate acetate (PMA)- and antibody (A)-dependent cellular cytotoxicity (DCC) against ox red blood cells (ORBC) by using oxidative means. The purpose of the present study was to investigate the acquirement of these cytotoxic functions during PMN ontogeny, using the promyelocytic HL-60 cell line as a model for PMN differentiation. HL-60 cells were induced to differentiate along the PMN pathway by exposure to dimethyl sulfoxide (DMSO). Uninduced HL-60 cells were found to be completely devoid of PMA-DCC and ADCC activity. DMSO-induced cells progressively acquired the capacity to kill ORBC and to undergo the activation of oxidative metabolic burst when triggered by PMA. Despite approximately 40% of them also were capable of binding IgG-sensitized ORBC, no ADCC activity and respiratory burst activation was observed: this finding indicates that maturing HL-60 cells require a more complete maturation than that induced by DMSO to actually exert ADCC. Together the results suggest that: a. the acquirement of both PMA-DCC and ADCC potential is a post-promyelocytic event; b. the cytotoxicity activating stimuli, PMA and IgG-coated targets, follow different post-receptor transductional pathways to trigger the effector cell lytic systems: only the PMA receptor-linked pathway develops during DMSO-driven differentiation of HL-60 cells.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1986

Inhibition of neutrophil cytolysin production by target cells.

Neutrophils, triggered by heat-aggregated human IgG (Agg.IgG), were found to lyse chicken red blood cells (CRBC) as determined by a 51Cr release method. The lysis was inhibited by azide, catalase, chloride-free medium and amino acids, suggesting the requirement for myeloperoxidase (MPO), hydrogen peroxide (H2O2), chloride ions (Cl-), and hypochlorous acid (HOC1), respectively. These results indicate that neutrophils lyse CRBC through an HOCl-(ie, MPO-H2O2-Cl-) dependent process. Although HOCl can react with neutrophil-derived nitrogenous (N-) compounds to yield chloramines, the main and well-characterized chloramines did not play a direct role in the lysis of CRBC in our model system. Thus, it appears that lysis is due either to HOCl or to an unknown compound derived from and with characteristics similar to HOCl. When CRBC were replaced with HRBC targets, no lysis could be observed. Treatment of HRBC with carmustine, to inhibit the glutathione cycle, did not affect the cell resistance to lysis by neutrophils. Conversely, the inhibition of HRBC catalase activity with aminotriazole (AT) made the cells susceptible to neutrophil-mediated HOCl-dependent lysis: this suggests that HRBC escape lysis by neutrophils through an AT-inhibitable, ie catalase-dependent, process. Through an identical catalase-dependent process, HRBC were capable of efficiently preventing the H2O2 and HOCl recovery from Agg.IgG-triggered neutrophils, tested under experimental conditions similar to those used for cytolytic assays. Together, these data suggest that HRBC targets, endowed with high catalase activity, escape neutrophil-mediated lysis by consuming (by catalase) neutrophil-derived H2O2, so that HOCl cannot be produced in amounts sufficient to promote lysis. Parallel experiments, performed with AT-treated CRBC, showed that these cells, endowed with a relatively low catalase content, only partially limit neutrophil cytolytic efficiency by a process qualitatively similar to that observed with HRBC targets. The results provide evidence that target cells can restrain neutrophil cytolytic efficiency by interfering with the MPO-H2O2-Cl system through their catalase activity.

Amitrole↗
PubMed1986

Role of hypochlorous acid and chloramines in the extracellular cytolysis by neutrophil polymorphonuclear leukocytes.

The lysis of human red blood cells (HRBC) by neutrophil polymorphonuclear leukocytes (PMN), triggered with opsonized zymosan (OPZ) particles, was inhibited by azide, catalase, Cl- -free medium and amino acids indicating the involvement of myeloperoxidase (MPO), hydrogen peroxide (H2O2), Cl- ions and hypochlorous acid (HOCl) respectively. Thus, the cytolytic process depends on the following reaction: (Formula: see text). Because the oxidizing agent HOCl is also the precursor of the chloramines, a group of oxidants formed by the reaction between HOCl and PMN-derived ammonia (NH4+) or amines (R-NH2), the observed HRBC lysis can be theoretically due to HOCl and/or chloramines. Nevertheless, we found that PMN-mediated cytotoxicity occurs as an unidirectional process, being HRBC targets lysed and PMN unaffected. This finding indicates that the cytotoxin must be relatively more efficient against HRBC as compared with PMN. In fact, reagent HOCl (used at concentrations comparable to those generated by PMN) but not chloramines displayed such a type of property. Taken together, the data suggest that HRBC are killed by PMN-derived HOCl without the requirement for chloramines: this implies that NH4+ and R-NH2, released by PMN, act as down-modulators of the cytotoxic process, serving as HOCl trapping agents.

Animals↗
PubMed1986

Interference of target cell catalase with an early step of the neutrophil cytolytic pathway.

The hypochlorous acid (HOCL)-dependent lysis of human red blood cells (HRBC) targets by neutrophils, activated with opsonized zymosan particles (OPZ), was increased by inhibiting HRBC catalatic activity with aminotriazole (AT; HRBCAT). The inhibition of HRBC glutathione cycle activity with carmustine (BCNU; HRBCBCNU) had no effect. In addition, the recovery of hydrogen peroxide (H2O2) and HOCL from neutrophils, activated under conditions similar to those used for cytotoxicity assay, was reduced by the presence of HRBC and restored by replacing HRBC with HRBCAT, but not with HRBCBCNU. Linear relationships were found between the increments in the neutrophil-mediated lysis, observed by using HRBCAT instead of HRBC, and the increments in the H2O2 or HOCL recovery, detected by replacing HRBC with HRBCAT. Together these data, coupled with the results obtained by probing neutrophil cytolysis with chemical agents, suggest that the increased cytolytic efficiency displayed by neutrophils against HRBCAT, inhibited in their catalatic activity, is due to an enhanced availability of neutrophil-derived H2O2, with a consequent enhancement in the HOCL production (according to the following reaction: (formula; see text). Thus it appears that HRBC catalase restrains the neutrophil cytolytic activity, by interfering with an early step of the pathway through which neutrophils generate cytotoxins.

Amitrole↗
PubMed1985

Restoration of defective EAG-rosetting capacity of cancer patient neutrophils by levamisole.

Levamisole, used in vitro at therapeutic concentrations, was found able to restore the defective Fc-receptor activity of cancer patient neutrophils. In addition, the drug prevented the inhibition of normal neutrophils Fc-receptor function by cancer patient sera. The neutrophil Fc-receptor function was also restored in six of seven cancer patients after in vivo administration of levamisole (2.5 mg/Kg/day for 3 days). Due to the central role played by the Fc-receptor function in host defence mechanisms, including phagocytosis as well as antibody dependent cellular cytotoxicity, the capacity of levamisole to restore defective neutrophil Fc-receptor function in cancer patients could contribute to the "immunomodulating" effect of the drug.

Cell Membrane↗
PubMed1985

Cellular cytotoxicity mediated by granule-depleted neutrophil cytoplasts.

Neutrophil-derived nucleus-and granule-free cytoplasts, consisting of cytosol enclosed by an intact plasma membrane, were able to destroy 51Cr-labelled ox red blood cells (ORBC) in the presence of phorbol myristate acetate (PMA). The slope of the target cell lysis vs the log of the cytoplast number was similar to that observed with neutrophils as effector cells. Nevertheless, a number of cytoplasts 60-80 times higher than that of neutrophils was required to obtain a common level of cytotoxicity. The ability of cytoplasts and neutrophils to lyse ORBC was completely abolished by catalase and unaffected by superoxide dismutase and mannitol, suggesting the involvement of hydrogen peroxide in the target cell damage. Addition of myeloperoxidase (MPO) to cytoplasts increased lysis. The MPO lysis by cytoplasts, except when experiments were carried out in the presence of MPO. The results indicate that neutrophil cytosol and plasma membrane represent the basic requirement for the PMA-dependent cytolytic process, whereas MPO behaves as a device to amplify lysis.

Catalase↗
PubMed1985

Extracellular cytotoxicity by phagocytosing polymorphonuclear neutrophilic leukocytes: enhancement by a chemotactic stimulus.

This study investigated the influence of a chemotactic stimulus on the extracellular cytotoxicity mediated by phagocytosing polymorphonuclear neutrophilic leukocytes (PMN). We used N-formyl-methionyl-leucyl-phenylalanine (FMLP) as chemotactic peptide, opsonized zymosan as phagocytosable particle, and ox red blood cells (ORBC) as extracellular bystander targets. Phagocytosing PMN were found to kill ORBC efficiently, as determined by the 51Cr-release assay. FMLP, at the concentration of 100 nM, significantly enhanced the target cell lysis. PMN from two patients with chronic granulomatous disease and normal PMN plus catalase or free radical scavengers (mannitol, benzoate, histidine) were completely devoid of cytolytic activity both in the presence and in the absence of FMLP. The results indicate that the target cell lysis by phagocytosing PMN as well as the chemotactic peptide-related amplification of the lysis itself depend on the expression of the PMN oxidative cytotoxic potential. A similar response to a chemotactic stimulus in vivo could provide a mechanism for regulating PMN-dependent cytotoxic and inflammatory processes.

Catalase↗
PubMed1985

Erythrocyte lysis by PMA-triggered neutrophil polymorphonuclears: evidence for an hypochlorous acid-dependent process.

Human red blood cells (HRBC) were efficiently lysed when incubated with neutrophil polymorphonuclears (PMN) in the presence of phorbol-myristate-acetate (PMA), as detected by a 4-hr 51Cr release assay. The lysis was virtually absent in the presence of catalase, azide or cyanide and in the absence of chloride ions. These findings indicate the involvement of the myeloperoxidase (MPO)-hydrogen peroxide (H2O2)-chloride (Cl-) system in the cytolytic process. As the MPO-H2O2-Cl- system is capable of generating the powerful oxidant hypochlorous acid (HOCl), cytotoxicity assays were performed in the presence of taurine, glycine, serine and valine to scavenge this potentially lytic agent. Each of these compounds efficiently inhibited the HRBC lysis by PMA-triggered PMN, as well as the lysis caused by HOCl in a cell-free system. Thus, the results suggest that HOCl, or an agent with similar reactivity, plays a key role in the PMA-dependent PMN-mediated cytotoxicity against HRBC targets.

Chlorine↗
PubMed1985

Down-regulation of K cell activity by neutrophils.

Human neutrophils, activated by phorbol-myristate acetate (PMA), (A-neutrophils), were found to suppress lymphocytic killer (K) cell-mediated antibody-dependent cellular cytotoxicity (ADCC). Resting (R) neutrophils, ie, PMA-untreated cells, were completely ineffective. Suppression was optimal when A-neutrophils were added at the beginning of the ADCC assay. Furthermore, A-neutrophils were found to cause an approximately 80% reduction in the number of Raji target cell-bound lymphocytes. These data indicate that A-neutrophils inhibit K cell activity by interfering with the target cell recognition. A-neutrophils were capable of reducing the percentage of Fc receptor (FcR)-bearing lymphocytes with a half-time of 7.2 minutes, through a process preventable by the serine-protease inhibitors tosyl-lysine-chloromethyl ketone (TLCK) and lima bean trypsin inhibitor (LBTI). Conversely, A-neutrophils caused a very slow decrease in the amount of Raji cell-bound antibodies, as detected by the complement-mediated lytic assay. Thus, only lymphocyte FcR structures seem to be highly susceptible to neutrophil-derived TLCK- and LBTI-inhibitable proteases. Furthermore, supernatants from A-neutrophils were found to inhibit K cell ADCC and lymphocyte binding to Raji target cells. In addition, LBTI prevented the A-neutrophil-dependent and the supernatant-dependent inhibition of both K cell ADCC activity and lymphocyte-target cell conjugate formation. Together these data suggest that A-neutrophils suppress K cell function through a protease-mediated impairment of the FcR binding capacity. The results provide evidence that human neutrophils are endowed with mechanisms to regulate K cell ADCC activity.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1979

Defective neutrophil chemotaxis and bactericidal power in a child with hyperimmunoglobulinemia E.

An 11-year-old boy with a life-long history of atopic-like dermatitis and recurrent staphylococcal abscesses was found to have defective neutrophil chemotaxis, impaired-T-lymphocyte functions, hyperimmunoglobulinemia E, and delayed neutrophil bactericidal power. This latter defect has never been found in such patients. The patient's mother revealed a panhypogammaglobulinemia, while his sister and maternal grandmother who had repeated infections were immunologically normal.

Chemotaxis, Leukocyte↗
PubMed1979

In vitro function of chronic myelocytic leukemia granulocytes. Effects of irradiation and storage.

Granulocyte function was studied in 9 patients with untreated, Ph1-positive chronic myelocytic leukemia (CML). The nitroblue tetrazolium reduction by stimulated granulocytes was impaired in all patients; 4 patients also had diminished phagocytosis and 2 others defective chemotaxis. In spite of this variety of polymorphonuclear (PMN) functional impairments, there is little evidence of increased susceptibility to infections in CML patients. This suggests that CML-PMN leucocytes (PMNs) may be successfully used for transfusion into neutropenic recipients, as previously reported. To evaluate the effects of irradiation and liquid storage on CML-PMNs, 5 of our patients were subjected to leukapheresis by continuous-flow centrifugation in the Aminco Celltrifuge, and granulocyte functional capacities were also evaluated on the cell-rich plasma immediately after collection and after short-term storage at 4 degrees C with or without irradiation (1500 rads). As evaluated by in vitro studies, granulocytes maintained, even after irradiation, functional activities similar to those found immediately after collection up to 24 h of storage at 4 degrees C and presented a moderate loss of function after 48 h. Chemotaxis appeared to be the most sensitive detector for cellular damage of stored leucocytes, irradiated and non-irradiated, so that it might be used for assessment of leucocyte function before transfusion.

Adult↗
PubMed1979

Lazy leukocyte syndrome.

A-35-year-old woman with a long-lasting history of neutropenia and recurrent infections was found to have defective neutrophil chemotaxis, random motility, and in vivo migration. Although the bone marrow granulocyte reserve was normal, the patient failed to release an appropriate amount of granulocytes after injection of etiocholanolone. These features are characteristic of the so-called "Lazy leukocyte syndrome". The clinical presentation of the five cases of this syndrome so far reported and its pathophysiological aspects are discussed.

Adult↗
PubMed1979

Neutrophil functions in acne conglobata.

Chemotaxis, random migration, phagocytosis, spontaneous and stimulated NBT reduction were evaluated in neutrophils from the peripheral blood of 8 patients with acne conglobata and were found to be normal. Only 2 patients had an increased spontaneous NBT reduction, and another exhibited a defect in neutrophil adhesiveness. On this basis, intrinsic defects of the major neutrophil functions, as evaluated in vitro, can be excluded in acne conglobata.

Acne Vulgaris↗
PubMed1979

[Vitamin C and phagocytic system: present status and perspectives (author's transl)].

Host defence mechanisms against microbial invasion requires an adequate number and function of phagocytic cells. There is evidence that vitamin C, which has been claimed to play a role in several aspects of host defence mechanisms, is involved significantly at least in phagocytic function. Vitamin C has been shown to interfere with oxidative metabolism, bactericidal power and chemotaxis of neutrophil granulocytes. The authors present evidence that, in vitro, vitamin C stimulates the true chemotactic response of normal human granulocytes. In addition the results of clinical trials in patients with recurrent infections and primary deficiencies of neutrophil function are presented. In such patients vitamin C therapy induced the restoration of impaired functions. A significant clinical improvement was also seen in the majority of cases. Vitamin C represents the specific therapy for primary defects of phagocytic function in patients with recurrent infections.

Ascorbic Acid↗
PubMed1979

Effects of irradiation and storage on granulocytes harvested by continuous-flow centrifugation.

Five normal subjects were subjected to leukapheresis by continuous-flow-centrifugation (CFC) in the Aminco Celltrifuge. Granulocyte functional capacities were evaluated on the venous blood samples drawn before apheresis and on the cell-rich plasma collected by CFC, immediately after collection and after short-term storage at 4 degrees C with or without previous irradiation (1500 rad, 50 rad/min). The CFC technique has been shown to provide cells without functional damage. Irradiation did not appear to influence granulocyte function, as evaluated by in vitro studies. The data demonstrate that granulocytes maintain, even after irradiation, functional activities similar to those found immediately after collection for up to 24 hours of storage at 4 degrees C and exhibit only a moderate loss of function after 48 h. Chemotaxis appears to be the most sensitive detector of cellular damage of stored granulocytes, either irradiated or non-irradiated; this technique may be the most useful for assessment of granulocyte function before transfusion.

Blood Preservation↗
PubMed1978

Identification of mouse bone marrow colony growth in agar-cultures by the nitroblue tetrazolium dye.

Identification of mouse bone marrow colony growth in soft agar cultures has been realized by layering of the plates with nitroblue tetrazolium at the concentrations of 1 mg/ml. Granulocyte-macrophage colonies appear dark-blue coloured on the yellow surrounding medium and can be fixed, stored at 4 degrees C and counted few days later. This method allows the cultures to be assessed macroscopically and makes a better enumeration of small clusters even if spread and consisting of only 30-40 cells.

Animals↗