PubMed Health⌕ Search

Biomedical subjects

C Galatiuc

Publications and source records attributed to C Galatiuc.

17 recordsLinked to original sources
PubMed2004

Interaction of human immunoglobulin G with CD16 on natural killer cells: ligand clearance, FcgammaRIIIA turnover and effects of metalloproteinases on FcgammaRIIIA-mediated binding, signal transduction and killing.

Human natural killer (NK) cells express low-affinity Fc immunoglobulin G (IgG) receptor (FcgammaRIIIA/CD16). The binding of monomeric IgG (mIgG) and F(ab')(2) fragments of 3G8 anti-CD16 monoclonal antibody (mAb) to FcgammaRIIIA was investigated by flow cytometry. Over 90% of NK cells bound endogenous IgG, and during incubation at 37 degrees C, the FcgammaRIIIA occupancy decreased slowly. Approximately 90% of NK cells bind mIgG or F(ab')(2) fragments of 3G8 anti-CD16 mAb. The calculated half-time (T(1/2)) of in vitro mIgG dissociation from FcgammaRIIIA was 130 min. By cross-linking the mIgG ligand with F(ab')(2) fragments of anti-human IgG antibody, the T(1/2) decreases to 85 min. In kinetics study, it has been shown that (125)I-mIgG bound to FcgammaRIIIA is slowly released in the culture supernatant, maybe eluted at acid pH, or partially internalized and degraded. The binding of IgG to FcgammaRIIIA was increased by 53.8% on cells cultured in the presence of RU36156, a matrix metalloproteinase (MMP) inhibitor. Furthermore, an increase in phosphorylation of Lyn tyrosine kinase, after cross-linking of mIgG-FcgammaRIIIA complex, was observed on NK cells treated with RU36156. When the FcgammaRIIIA was occupied by mIgG, the capacity of NK cells to kill K562 target cells was decreased by RU36156, because the MMP inhibitor protects CD16 from proteolysis. Our data demonstrate that binding of mIgG to human NK cells is followed by ligand dissociation and/or internalization, enzymatic degradation and exocytosis. The RU36156 MMP inhibitor protects FcgammaRIIIA from cleavage, augments NK-cell activation and may interfere in their killing capacity.

Cytotoxicity Tests, Immunologic↗
PubMed1995

Natural killer (NK) activity in human responders and nonresponders to stimulation by anti-CD16 antibodies.

Various anti-Fc gamma RIII (CD16) monoclonal antibodies (mAbs) are shown here to have positive or negative modulatory effects on human NK cells. Thus, 3G8 mAb (IgG1) triggered a dose-dependent augmentation of NK activity in 67% (23/34) of individuals tested, who were designated as responders. All four IgG1 anti-CD16 mAb tested (BL-LGL/1, B73.1, Leu11c, and 3G8) were stimulatory for NK cells isolated from responders, whereas six non-IgG1 anti-CD16 mAbs were either inhibitory or had no significant effects on NK activity. The upregulation of NK activity in responders was not attributable to an increase in either the conjugate formation or the delivery of the lethal hit to target cells. This mAb-mediated up-regulation of NK activity was shown to be associated with a recycling capacity higher than that of controls and with enhanced release of cytokines by activated NK cells. Anti-CD16 mAb inhibited binding of either monomeric or polymeric IgG to Fc gamma RIIIA on NK cells. Also, mAb 3G8 or its F(ab')2 fragments decreased or reversed inhibition of NK activity induced by monomeric IgG (mIgG). Our data indicate that regulation of NK activity via the Fc gamma RIIIA is influenced by dose-dependent interactions between cytophilic mIgG and anti-CD16 mAb of IgG1 isotype.

Antibodies, Monoclonal↗
PubMed1993

Regulation of human natural cytotoxicity by IgG. IV. Association between binding of monomeric IgG to the Fc receptors on large granular lymphocytes and inhibition of natural killer (NK) cell activity.

The regulation of human natural killer (NK) activity by IgG described previously by us depends on the ability of cytophilic molecules of monomeric IgG (mIgG) to inhibit the subsequent killing of NK-sensitive targets. Highly purified NK cells obtained from human peripheral blood are able to directly bind mIgG as well as antigen-complexed IgG through its Fc region. The demonstration that NK cells bear labile cytophilic IgG, a property which usually has been attributed to L cells, indicates that mIgG-induced inhibition of NK activity is mediated by direct interactions between the inhibitory ligand and cytotoxic effector cells. The Fc receptor (FcR) mediating downregulation of NK cytotoxicity appeared to be FcR gamma III, previously found to be selectively expressed on NK cells and granulocytes. In studies of unidirectional cross-inhibition of mIgG binding to NK cells by various anti-CD16 monoclonal antibodies, binding characteristics of mIgG or complexed IgG were similar. Thus, the FcR gamma III for mIgG appear to be indistinguishable from receptors responsible for binding of polymeric IgG on human NK cells. The negative regulation of NK activity by mIgG was not attributable to inhibition of conjugate formation between effector cells and K532 targets, but rather to inhibition of a post-binding event involved in killing of conjugated targets. The data presented suggest that the Fc gamma RIII on human NK cells can either mediate killing against IgG antibody-coated target cells or, upon interaction with cytophilic monomeric ligand in soluble form, induce inhibition of NK activity.

Antigen-Antibody Complex↗
PubMed1991

Expression of Fc mu receptors on human natural killer cells.

Fc receptors for IgG (CD16) have been described as the only type of immunoglobulin receptor on large granular lymphocytes (LGL). However, the ability of natural killer (NK) cells to mediate antibody-dependent cellular cytotoxicity (ADCC) in the presence of monoclonal or polyclonal IgM and the inhibition of NK activity by highly purified IgM could not be explained on the basis of FcR for IgG. In order to directly assess the expression of Fc receptors for IgM (Fc mu R), NK cells were treated with human polyclonal IgM, and its binding was visualized by a direct anti-globulin rosette assay with identification of rosette-forming LGL on Giemsa-stained smears. The data indicated that a high proportion of LGL (up to 68%) were Fc mu R-positive cells. However, this percentage varied depending on the IgM preparation (polyclonal or monoclonal), the indicator reagent used for the rosette assays, and the cell preparations studied. Two-color flow cytometry of human nonadherent lymphocyte preparations confirmed the presence of CD56+IgM+ cells, which represented from 43 to 78% of CD56+ cells. Flow cytometry was also performed using highly enriched preparations of human NK cells (the mean percentage of CD3-CD56+ cells was 84%). Up to 88% of purified NK cells bound FITC-labeled monoclonal IgM at a saturating concentration. By indirect immunofluorescence, from 34 to 62% of NK cells purified from the peripheral blood of normal donors were able to bind polyclonal IgM. Similar results were obtained with LGL from a patient with NK lymphoproliferative disease. Thus the presence of Fc mu R on a majority of human NK cells was demonstrated by different techniques, using unseparated peripheral blood mononuclear leukocytes, purified normal NK cells, and also LGL from a patient with NK lymphoproliferative disease.

Animals↗
PubMed1989

Regulation of human natural cytotoxicity by IgG. III. Interaction between negative regulation by monomeric IgG and positive regulation by interferons of different types.

Our prior reported results have demonstrated the dose-dependent inhibition of human natural killer (NK) cell activity upon treatment of peripheral blood mononuclear cells (PBMC) with monomeric IgG (mIgG) prior to the cytotoxic assay. In the present study, the combined effects on NK activity of human interferon (IFN) of each of the three types and mIgG, respectively, were determined. NK cells incubated with IFN alpha or IFN beta had augmented cytotoxicity against K562 target cells but remained responsive to negative regulation by mIgG. PBMC treated with human recombinant IFN gamma had unchanged cytotoxic activity but became partially resistant to suppression by mIgG. This ability of IFN gamma to interfere with the negative regulation of NK activity by cytophilic mIgG was seen when the cytokine was preincubated with effector cells prior to, simultaneously with, or after their exposure to inhibitor protein. These data provide some clues regarding the possible biological significance of the mIgG-induced down-regulation of NK cells which, when required for host protection, might be appreciably reversed or blocked by IFN gamma produced by NK cells or T cells in response to various agents.

Cytotoxicity Tests, Immunologic↗
PubMed1983

Herpes simplex virus (HSV) antigens on HSV-transformed rat cells can be demonstrated by rosette formation with Staphylococcus aureus protein-A-coated sheep red blood cells.

Two rat embryo cell lines transformed by herpes simplex virus (HSV) and four tumor cell lines derived from inoculation of HSV-transformed cells formed rosettes after treatment with rabbit or human anti-HSV IgG and sheep red blood cells coated with protein A of Staphylococcus aureus, Cowan I strain. The frequency of rosette-forming cells after treatment with rabbit anti-HSV IgG at a concentration of 0.01 mg/10(6) cells varied between 41.6 and 73% depending on the cell line. Pretreatment of HSV-transformed cells with rabbit anti-HSV IgG Fab prevented rosette formation after treatment with rabbit or human anti-HSV IgG. Trypsin digestion of transformed cells reduced significantly the proportion of rosettes formed with human or rabbit anti-HSV IgG. The microscopic visualization of rosette formation on HSV-transformed cells is consistent with the expression of HSV-specific antigens on the cell surface.

Animals↗
PubMed1982

Receptors for IgA on human thymus cells in myasthenia gravis.

Specific surface receptors for the Fc region of human immunoglobulin A (Fc alpha R) were identified on the thymus cells of all 15 patients affected by myasthenia gravis. A large variation of the percentage of Fc alpha R bearing thymus cells was recorded (9 . 5-66 . 0%; mean value 27 . 9 +/- 17 . 8) the high values being associated in most cases with the thymus hyperplasia and the low values with a lipomatosis of the organ. Even the lowest percentage of Fc alpha R bearing cells recorded in myasthenia gravis are considerably higher than those found in normal subjects (0 . 4-4 . 1; mean value 1 . 8) demonstrating that the expression of Fc alpha R on the thymus cells is the consequence of the disease. No attempts were made to identify the cell population bearing Fc alpha R but indirect results indicate that the cells belong to E-rosette forming population, and, in some cases, to a null cell population devoid of the ability to form E and EAC rosettes, Fc alpha R can bind various IgA ligands and the binding is confined to a small fraction of IgA (not more than 5%) considered as cytophilic IgA.

Absorption↗
PubMed1982

Inhibition of human natural killer cell activity by cytophilic immunoglobulin G.

The natural killer (NK) cell activity of human peripheral blood mononuclear cells (PBL) was found to be increased after incubation at 37 degrees C for 2 hr. The observed increase was shown to be associated with release from inhibition by human serum factors, because incubation in autologous serum interfered with augmentation. The serum-mediated effect appeared attributable to the degree of binding of labile IgG to PBL and could be reduced by selective depletion of IgG from the serum. Human monomeric IgG was found to efficiently inhibit the culture-induced augmentation of NK activity; the inhibitory IgG had properties consistent with those described for cytophilic IgG and was mediated through the Fc region of IgG. The inhibition by monomeric IgG occurred at 0 degrees C as well as at 37 degrees C and this could be induced even after culture-induced augmentation of NK activity. Thus, binding of monomeric IgG to human PBL appears to reversibly inhibit their NK activity. These results provide evidence for a novel mechanism for negative regulation of NK activity.

Animals↗
PubMedDate not supplied

A non-dissociable rabbit IgG-protein a complex.

Glutaraldehyde treatment of rabbit IgG antibody complexed with protein A of Staphylococcus aureus (SpA) enabled the complex (IgG2-SpA1)2 to maintain its molecular weight unchanged and to become non-dissociable at low pH and in excess of IgG. The glutaraldehyde-treated (IgG2-SpA1)2 complex had the same half-life, complement-activating capacity and ability to interact with the Fc receptor as the non-treated complex. Moreover, the glutaraldehyde-treated complex had a stronger immunosuppressive effect on the synthesis of anti-sheep red blood cell antibody than either the non-complexed antibody or the IgG antibody complexed with SpA but untreated with glutaraldehyde.

Animals↗
PubMedDate not supplied

Phospholipase A2 modulates respiratory burst developed by neutrophils in patients with rheumatoid arthritis.

Activated by bacterial peptides, phorbol esters, calcium ionophores and other agonists, neutrophils (PMNs) release the proinflammatory mediator, arachidonic acid (AA) via the intervention of phospholipase A(2) (PLA(2)). AA may play an essential role in activation of NADPH-oxidase, which is involved in the generation of superoxide anion by neutrophils. The present study is focused on the involvement of PLA(2) in the respiratory burst developed by PMNs isolated from patients with rheumatoid arthritis (RA). PLA(2) exists in very high levels in diseases such as rheumatoid arthritis and may cause acute inflammatory and proliferative changes in synovial structures. The respiratory burst was evaluated as superoxide anion release, using an amplified chemiluminescence method. The assays were performed using PMNs untreated or treated with different doses of stimulatory reagents (phorbol 12-myristate-13-acetate (PMA), calcium ionophore (A23187)). Our data suggested that PMA stimulated the production of superoxide anion in a dose-response manner, as compared with A23187, which did not induce a significant release of superoxide anion in PMNs-RA. The exogenous addition of AA significantly amplified the superoxide anion release by PMNs-RA stimulated with PMA and to a lesser extent, by PMNs stimulated with A23187. AA has also reversed the inhibitory effect of arachidonyl-trifluorometylketone and E-6-(bromomethylene)tetrahydro-3-(1-naphthalenyl)2H-pyran-2-one (BEL) on the superoxide anion release by PMNs-RA. In conclusion, the differential responses to these two agents suggested that different isoforms of PLA(2) were activated by A23187 or PMA, and support the idea that activation of these different PLA(2) served distinct functions of PMNs. Therefore, the inhibition of PLA(2) enzymes might be of great importance in the immunotherapy of rheumatoid arthritis.

Arachidonic Acid↗
PubMedDate not supplied

IgA monoclonal and polyclonal proteins as regulatory factors of the NK cytotoxic activity.

The presence of receptors for IgA (IgAR) on natural killer (NK) cells was only indirectly suggested yet. To elucidate the presence of IgAR on NK cells and its possible role in modulation of the NK activity, was initiated a preliminary study carried out in a homologous system, using human non adherent lymphocytes (NAL) and human seric or secretory IgA. A proportion of over 20% NK cells (CD16+ CD56+) was determined by flow-cytometry in the NAL-cells population. Dose-dependent IgA-binding to NAL cells was determined, showing a limitation of the IgA+ NAL proportion for a cell population of 32% and a ligand concentration of 4 mg/ml/10(7) cells. The binding parameters of the IgA/IgAR system, calculated by Scatchard procedure, revealed values of the affinity constant (K) and of the maximum number of ligand molecules bound/cell (n) depending on the aggregation degree of the ligands: K-values of 1.0 and 0.4 x 10(7) M-1 for dimeric and respectively, monomeric IgA, and n-values of 0.8 and 1.7 molecules/cell, respectively. A proportion of 3% IgA molecules endowed with cytophilic property was also calculated. The turnover rate of secretory IgA (sIgA) on the NAL cells surface showed values of the ligand half time (T 1/2) of 1 h. The effect of polyclonal IgA, (sIgA and normal seric IgA) and of monoclonal IgA myeloma (monomeric and dimeric IgA) on the NK cytotoxicity target K562 cell line was of inhibition, depended on the ligand doses and varied with the IgA type. The possible relevance of immunoglobulins A and of the activity of natural killer cells in processing of some mechanisms of the antiviral protection was discussed.

Cytotoxicity, Immunologic↗
PubMedDate not supplied

Expression and function of Fc gamma RII on human natural killer cells.

In this report, we present data on the expression and function of Fc gamma RII (CD32) by natural killer (NK) cells. Highly enriched NK cell populations were isolated from peripheral blood lymphocytes by negative selection and consisted of > or = 95% CD3-/CD56+ cells. Flow cytometric analyses with anti-CD32 monoclonal antibodies (mAbs) demonstrated that a small proportion of NK cells were recognized by mAbs IV.3 and 41H16. Two-color flow cytometric analysis indicated coexpression of the epitope on NK cells recognized by both these mAbs. Verification of expression of CD32 on NK cells was obtained by demonstrating coexpression of CD32 on either CD16+ or CD56+ cells. The CD32+/CD16+ and CD32+/CD56+ cells represented approximately 7 and 3% of the total, respectively. CD32 transcripts were identified from highly purified NK cells using reverse transcription-polymerase chain reaction with CD32-specific primers, followed by Southern blotting. Enhanced chemiluminescence-Western blot (ECL-WB) analysis of lysates of purified NK cells indicated that mAb IV.3 recognized a molecule of approximately 40 kD. The Fc gamma RII on NK cells was able to transduce intracellular signals in several types of assay. Cross-linking of anti-CD32 resulted in a mobilization of intracellular Ca2+, although to a lesser extent than that induced by cross-linking CD16. Both mAbs IV.3 and 41H16 were found to be capable of inducing reverse antibody-dependent cellular cytotoxicity against FcR+ target cells (e.g. P815). These data represent the first direct description of the expression and function of Fc gamma RII on human NK cells.

Antibodies, Monoclonal↗