T-cell proliferation by surface molecules expression on polymorphonuclear neutrophils stimulated with IL-4 in superantigen presence

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Quezada, S. Ramos, M. Garcia, X. Norambuena, D. Pavon" "autores" => array:5 [ 0 => array:2 [ "nombre" => "A." "apellidos" => "Quezada" ] 1 => array:2 [ "nombre" => "S." "apellidos" => "Ramos" ] 2 => array:2 [ "nombre" => "M." "apellidos" => "Garcia" ] 3 => array:2 [ "nombre" => "X." "apellidos" => "Norambuena" ] 4 => array:2 [ "nombre" => "D." "apellidos" => "Pavon" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054611001510?idApp=UINPBA00004N" "url" => "/03010546/0000004000000002/v1_201304101104/S0301054611001510/v1_201304101104/en/main.assets" ] "itemAnterior" => array:18 [ "pii" => "S0301054611001534" "issn" => "03010546" "doi" => "10.1016/j.aller.2011.02.010" "estado" => "S300" "fechaPublicacion" => "2012-03-01" "aid" => "286" "copyright" => "SEICAP" "documento" => "article" "crossmark" => 0 "subdocumento" => "fla" "cita" => "Allergol Immunopathol (Madr). 2012;40:75-80" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:2 [ "total" => 2128 "formatos" => array:3 [ "EPUB" => 7 "HTML" => 1656 "PDF" => 465 ] ] "en" => array:12 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Beneficial effects of erythropoietin on airway histology in a murine model of chronic asthma" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => "en" "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "75" "paginaFinal" => "80" ] ] "contieneResumen" => array:1 [ "en" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 764 "Ancho" => 2354 "Tamanyo" => 100628 ] ] "descripcion" => array:1 [ "en" => "Murine model of airway inflammation and treatment with EPO." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "M. Karaman, F. Firinci, M. Kiray, T. Tuncel, A. Bagriyanik, O. Yilmaz, N. Uzuner, O. Karaman" "autores" => array:8 [ 0 => array:2 [ "nombre" => "M." "apellidos" => "Karaman" ] 1 => array:2 [ "nombre" => "F." "apellidos" => "Firinci" ] 2 => array:2 [ "nombre" => "M." "apellidos" => "Kiray" ] 3 => array:2 [ "nombre" => "T." "apellidos" => "Tuncel" ] 4 => array:2 [ "nombre" => "A." "apellidos" => "Bagriyanik" ] 5 => array:2 [ "nombre" => "O." "apellidos" => "Yilmaz" ] 6 => array:2 [ "nombre" => "N." "apellidos" => "Uzuner" ] 7 => array:2 [ "nombre" => "O." "apellidos" => "Karaman" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054611001534?idApp=UINPBA00004N" "url" => "/03010546/0000004000000002/v1_201304101104/S0301054611001534/v1_201304101104/en/main.assets" ] "en" => array:19 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "T-cell proliferation by surface molecules expression on polymorphonuclear neutrophils stimulated with IL-4 in superantigen presence" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "81" "paginaFinal" => "87" ] ] "autores" => array:1 [ 0 => array:3 [ "autoresLista" => "B.K.A. Abdel-Salam" "autores" => array:1 [ 0 => array:3 [ "nombre" => "B.K.A." "apellidos" => "Abdel-Salam" "email" => array:1 [ 0 => "Manar_Muhamad@yahoo.com" ] ] ] "afiliaciones" => array:1 [ 0 => array:1 [ "entidad" => "Department of Zoology, Faculty of Science, 61519 Minia University, El-Minia, Egypt" ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 2062 "Ancho" => 3243 "Tamanyo" => 138547 ] ] "descripcion" => array:1 [ "en" => "Changes in HLA class II (A), CD80 (B) and CD86 (C) expression on unstimulated and IL-4 stimulated PMN. Statistical analysis showed that, there is a significant difference between IL-4 stimulated cells and unstimulated cells, P<0.05." ] ] ] "textoCompleto" => "IntroductionPolymorphonuclear neutrophils (PMNs) possess a very short half-life in the circulation because they constitutively undergo apoptosis.<a class="elsevierStyleCrossRefs" href="#bib0005">1,2</a> Under certain conditions PMNs play an important role in the effectors arm of host immune defence through the clearance of immune complexes, the phagocytosis of opsonised particles, and the release of inflammatory mediators.<a class="elsevierStyleCrossRefs" href="#bib0015">3–5</a> During the last years the image of PMNs has changed considerably. Traditionally considered to be the first line of defense against bacterial infection, it became increasingly clear that PMNs also participate in chronic inflammation disease and regulation of the immune response when appropriately activated.<a class="elsevierStyleCrossRef" href="#bib0030">6</a> For surface molecules, there are several reports that PMNs from a variety of species can express human leukocyte antigen (HLA) class II and costimulatory molecules (CD80 and CD86).<a class="elsevierStyleCrossRefs" href="#bib0035">7–10</a> Under certain stimulation murine neutrophils present HLA class II restricted antigen.<a class="elsevierStyleCrossRef" href="#bib0055">11</a>PMNs function and recruitment to the site of inflammation have been shown to be upregulated by various cytokines, including interleukin (IL)-1, IL-8, tumour necrosis factor (TNF), interferon-γ (IFN-γ), and granulocyte macrophage-colony stimulating factor (GM-CSF).<a class="elsevierStyleCrossRefs" href="#bib0025">5,12</a> In addition, IL-15 stimulated PMNs acquire HLA-DR.<a class="elsevierStyleCrossRef" href="#bib0065">13</a>IL-4 production has been found to occur in thymocytes, mature T-cells, certain malignant T-cells, mast-cells and basophiles and occasionally, in transformed B-cells.<a class="elsevierStyleCrossRef" href="#bib0070">14</a> It has an effect on B-cells, T-cells, monocytes, mast-cells, endothelial cells, and fibroblasts.<a class="elsevierStyleCrossRef" href="#bib0075">15</a> Directly and/or indirectly, IL-2 has a prominent role in the regulation of IL-4 producing cells.<a class="elsevierStyleCrossRef" href="#bib0070">14</a> IL-4 binds to a high-affinity cell-surface receptor (IL-4R) to exert its effects.<a class="elsevierStyleCrossRef" href="#bib0080">16</a> It promotes the growth and differentiation of activated human B-lymphocytes and shares many biological functions with IL-13.<a class="elsevierStyleCrossRef" href="#bib0085">17</a>Following this approach, the present study was aimed to estimate the expression of the antigen presenting molecule HLA class II and the co-stimulatory molecules CD80 and CD86 on PMNs stimulated with IL-4. The expression of these surface molecules prompted us to test the T-cell proliferation by their co-culture with IL-4 stimulated PMNs in the presence Staphylococcus aureus enterotoxin (A) as a superantigen.Materials and methodsBlood was taken from 10 healthy Egyptian people by venous puncture using 7.5ml heparin-coated tubes (Sarstedt, Nümbrecht, Germany) and was analysed within 2h. Recombinant human (rh) IL-4 was purchased from Sigma (St Louis, MO, USA). For fluorescence-activated cell sorter (FACS) analysis of whole blood, erythrocyte FACS lysing solution was obtained from Becton Dichinson (Heidelberg, Germany) and diluted 1:10 in bidistilled water. For cytofluorometry fluorescein isothiocyanate (FITC) and phycoerythrin (PE)-labelled murine MoAbs were used. Mouse IgG1 FITC, IgG2a PE, CD66b-FITC, HLA-DP+DQ+DR:PE, CD80:PE and CD86:PE were obtained from Coulter Immunotech (Marseille, France).For double labelling, anti-CD66b-FITC as a PMNs marker and the respective PE-labelled antibody were used in equal protein concentrations. Cells in whole blood were analysed by FACSCalibur and CellQuest software (Becton Dickinson, SanDiego, CA, USA). Results were expressed as the percentage of positive cells in the respective gate or quadrant. In FACS plots, there are different populations. So, gates were made around population with high CD66b-FITC.In all FACS experiments, PMNs in heparinised blood were cultured in 24-well plate, 2ml/well and incubated in the presence or absence of IL-4 (6ng/ml) for about 24h at 37°C with 5% CO2.For the co-culture of T-cells and PMNs, cells were isolated by Polymorph Prep® (Nycomed; Oslo, Norway). PMNs and T-cells fraction were further purified by adsorption to CD15 and CD3 beads (Miltenyi Biotec; Bergisch Gladbach, Germany), respectively, by magnetic cell separation using the devices supplied by Milteny Biotech (Bergisch-Gladbach, Germany).Highly purified PMNs (1×106/ml) were cultivated in AIM V (Gibco BRL; Paisley, Scotland)) with 2.5% autologous normal human serum, NHS (inactivated at 56°C for 30min.). T-cells were cultured in RPMI 1640 (Gibco BRL; Paisley, Scotland) supplemented with 10% fetal calf serum (FCS) (PAN Biotech GmbH; Aidenbach, Germany), 100U/ml penicillin/streptomycin (Gibco BRL; Paisley, Scotland), 2mM l-glutamine (Gibco BRL; Paisley, Scotland)), and 10mM HEPES (Gibco BRL; Paisley, Scotland). All cells were incubated at 37°C and 5% CO2 for the times indicated.Unstimulated and stimulated PMNs (1×103) in 100μl were added per well of a 96-well concave-bottom plate (Greiner; Nuertingen, Germany). Then, 1×104 T-cells (100μl) were added to each well together with 25ng S. aureus enterotoxins A (Sigma; Munich, Germany). After coincubation for four days at 37°C with 5% CO2, proliferation was tested by adding 1mCi of 3H-thymidine (Amer-sham Life Science; Braunschweig, Germany) for 6–8h [3H] TdR incorporation into DNA was measured and expressed as counts per minute (cpm). The values represent the mean±SE of 6–12 parallel wells.Statistical analysis of the obtained data was performed using one way analysis of variance (ANOVA) test followed by least square differences (LSD) analysis for comparison between means. Results were expressed as mean±standard error (SE), and differences were considered to be significant at P<0.05.ResultsResults were expressed as percentage of positive cells in the respective gate or quadrant. Each set of experiments was repeated in vitro ten times.In vitro expression of HLA class II on PMNsThe majority of healthy donors PMNs expressed CD66b (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>a–c). In the first set of experiments the effect of IL-4 on the expression of HLA class II was tested, where we found that PMNs on whole blood showed expression of HLA class II recording 7.77% (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>c) by using IL-4, as stimulators. In contrast, fresh and unstimulated cells cultured with medium only counted 1.05% (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>a) and 2.71% (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>b), respectively.<elsevierMultimedia ident="fig0005"></elsevierMultimedia>In vitro induction of CD80 on PMNsAs shown for HLA class II, surface expression of CD80 was most impressive following cultivation of whole blood with the stimuli for 24h. The proportion of double-positive cells (right upper quadrant) was estimated, where CD80 positive cells was slightly higher in stimulated cells recording 4.06% by using IL-4 (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>c) than unstimulated PMNs cultured with medium for 24h (1.12%) and fresh cells, 0.05% (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>a and b, respectively).<elsevierMultimedia ident="fig0010"></elsevierMultimedia>In vitro induction of CD86 on PMNsFor CD86, high expression was recorded on the surface of IL-4 stimulated PMNs in whole blood recorded 20.48% (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>c), while percentage of CD86 molecules on the surface of unstimulated PMNs measured 6.97 (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>b). Fresh PMNs in while blood had 1.55% (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>a).<elsevierMultimedia ident="fig0015"></elsevierMultimedia>Statistical analysis of HLA class II, CD80 and CD86 experimental sets showed that there is a significant difference between IL-4 stimulated cells and unstimulated cells, P<0.05 (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>).<elsevierMultimedia ident="fig0020"></elsevierMultimedia>Interaction of IL-4 stimulated PMNs with peripheral T-cellsFor these experiments highly purified PMNs were cultivated with IL-4 for 24h and then co-cultivated with highly purified isolated T-cells in the absence or presence of SEA. The differences between the groups in the sets of experiments related to the interaction of HLA class II positive PMNs with peripheral T-cells showed a significant difference, where P<0.05 (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>).<elsevierMultimedia ident="fig0025"></elsevierMultimedia>DiscussionPMNs are considered short-lived cells undergoing spontaneous apoptosis in vivo as well as in culture.<a class="elsevierStyleCrossRef" href="#bib0090">18</a> Previous studies have demonstrated that PMNs can be induced in vitro to synthesize and release various cytokines, suggesting that these cells can contribute significantly to the initiation and amplification of cellular and humoral immune responses.<a class="elsevierStyleCrossRef" href="#bib0025">5</a> The detection of these molecules, therefore, provides strong support for the hypothesis that human PMNs can actively synthesize immunoregulatory molecules<a class="elsevierStyleCrossRef" href="#bib0095">19</a> and have the potential to act as antigen presenting cells.<a class="elsevierStyleCrossRef" href="#bib0040">8</a>Recently, it has become increasingly evident that culturing PMNs in the presence of cytokines extends their life span.<a class="elsevierStyleCrossRefs" href="#bib0100">20–23</a> Cultured PMNs synthesize and release immunomodulatory cytokines by which they may participate in the afferent limb of the immune response.<a class="elsevierStyleCrossRef" href="#bib0025">5</a>This study has clearly shown that PMNs could be induced to express HLA class II, CD80 and CD86 after activation with IL-4. These data are in accordance with other results where HLA class II, CD80 and CD86 were expressed on the PMNs surface after exposure to GM-CSF and/or INF-γ.<a class="elsevierStyleCrossRef" href="#bib0030">6</a>The antigen presenting molecule HLA class II and the co-stimulatory molecules CD80 and CD86 play an important role in T-cell proliferation, where HLA class II presents the engulfed antigen to T-cells.<a class="elsevierStyleCrossRef" href="#bib0030">6</a> CD80 and CD86 act as second signal molecules involving in the stimulation of T-cells to produce the autocrine growth factor IL-2 without which T-cells are thus unable to proliferate.<a class="elsevierStyleCrossRef" href="#bib0120">24</a>The activation and recruitment of PMNs were also regulated by IL-15,<a class="elsevierStyleCrossRef" href="#bib0065">13</a> IFN-γ,<a class="elsevierStyleCrossRef" href="#bib0125">25</a> CSF-CSF<a class="elsevierStyleCrossRefs" href="#bib0130">26–28</a> and IL-8.<a class="elsevierStyleCrossRef" href="#bib0145">29</a> In addition, PMNs posses IL-2Rβ chain<a class="elsevierStyleCrossRefs" href="#bib0150">30,31</a> and IL-2Rγ chain<a class="elsevierStyleCrossRef" href="#bib0160">32</a> that have the ability to bind with IL-4.<a class="elsevierStyleCrossRef" href="#bib0165">33</a> These published data prompted us to study the effect of IL-4 on PMNs functions.When comparing the number of monocytes and PMNs required to induce T-cell proliferation, it was observed that 10 times more PMNs than monocytes were necessary to yield the same extent of T-cell proliferation. However, the cell preparation used in this work never contained more than 1% of contaminating cells and certainly not the 10% of monocytes that would be required to affect the results.<a class="elsevierStyleCrossRef" href="#bib0150">30</a> The observation that ten times more PMNs than monocytes were required to induce a similar extent of T-cell proliferation has to be considered together with the observation that only a proportion of PMNs acquired HLA class II. Thus, when only fully equipped PMNs are considered, the ability to process and to present antigen is similar to that of monocytes. Whether HLA class II-positive PMNs participate in the immune defence or play a role in pathophysiological events, is a matter of speculation. The fact that only a minor proportion of PMNs acquire HLA class II might lead to the conclusion that a possible accessory function of PMNs would be rather weak. One has; however, to bear in mind that PMNs are numerous in the peripheral blood, and that even a low percentage of PMNs expressing HLA class II would exceed both circulating monocytes and dendritic cells in number.Due to the notion that the presence of Staphylococcus entrotoxin coincides with relapses of Wegener's granulomatosis,<a class="elsevierStyleCrossRef" href="#bib0170">34</a> we tested whether PMNs was able to also present Staphylococcus enterotoxin A as a superantigen, so-called because it binds outside of the peptide-binding groove of the HLA class II and the antigen-specific domain of the T-cell receptor, and consequently activates a large portion of T-cells, preferentially those with a V beta 2 domain.<a class="elsevierStyleCrossRef" href="#bib0175">35</a> In accordance with previous studies,<a class="elsevierStyleCrossRefs" href="#bib0040">8,36</a> co-culture of PMNs with T-cells and SEA resulted in T-cell proliferation. Taken together, our data demonstrate that by synthesizing and expressing of HLA class II antigens, CD80 and CD86; PMNs acquire the capacity to present superantigens to T-cells.Data showed that there is a clear increase in T-lymphocyte proliferation in the co-culture of stimulated PMNs+T-cells. Because there is no antigen present, this means that it is not caused by the co-stimulatory molecules on the surface of the PMNs but probably due to mediators secreted by the PMNs. Furthermore, unstimulated PMNs+T-cells+SEA give almost a 1000-fold increase of T-cell proliferation while only a few percent of PMNs express co-stimulatory molecules in a naive state. Probably the results obtained are both from released mediators and the co-stimulatory molecules. Mediators secreted by IL-4 stimulated PMNs will be investigated in our future research.In conclusion, stimulated PMNs with IL-4 lead to expression of the antigen presenting molecules (HLA class II) and the co-stimulatory molecules (CD80 and CD86). These molecules play an important role in antigen presentation and consequently T-cell proliferation in the presence of SEA. This means that IL-4 stimulated PMNs might be involved indirectly in acquired immune response in addition to their role in innate immunity.Conflict of interestThe authors have no conflict of interest to declare." "textoCompletoSecciones" => array:1 [ "secciones" => array:10 [ 0 => array:2 [ "identificador" => "xres86088" "titulo" => array:6 [ 0 => "Abstract" 1 => "Background" 2 => "Aim of the study" 3 => "Methods" 4 => "Results" 5 => "Conclusions" ] ] 1 => array:2 [ "identificador" => "xpalclavsec74252" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "xpalclavsec74251" "titulo" => "Abbreviations" ] 3 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 4 => array:2 [ "identificador" => "sec0010" "titulo" => "Materials and methods" ] 5 => array:3 [ "identificador" => "sec0015" "titulo" => "Results" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0020" "titulo" => "In vitro expression of HLA class II on PMNs" ] 1 => array:2 [ "identificador" => "sec0025" "titulo" => "In vitro induction of CD80 on PMNs" ] 2 => array:2 [ "identificador" => "sec0030" "titulo" => "In vitro induction of CD86 on PMNs" ] 3 => array:2 [ "identificador" => "sec0035" "titulo" => "Interaction of IL-4 stimulated PMNs with peripheral T-cells" ] ] ] 6 => array:2 [ "identificador" => "sec0040" "titulo" => "Discussion" ] 7 => array:2 [ "identificador" => "sec0045" "titulo" => "Conflict of interest" ] 8 => array:2 [ "identificador" => "xack31869" "titulo" => "Acknowledgements" ] 9 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2010-12-28" "fechaAceptado" => "2011-02-14" "PalabrasClave" => array:1 [ "en" => array:2 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec74252" "palabras" => array:5 [ 0 => "IL-4" 1 => "HLA class II" 2 => "PMNs" 3 => "Staphylococcus aureus A" 4 => "T-cells" ] ] 1 => array:4 [ "clase" => "abr" "titulo" => "Abbreviations" "identificador" => "xpalclavsec74251" "palabras" => array:3 [ 0 => "PMNs" 1 => "HLA" 2 => "IL" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "BackgroundPolymorphonuclear neutrophils (PMNs) were originally described as short lived and terminally differentiated phagocytes that contribute only to the innate immune response. Some studies of PMNs cytokine production and expression of numerous cell surface proteins has suggested that PMNs are likely to influence adaptive responses and may satisfy the criteria of antigen presenting cells. Aim of the studyThis work aimed to study the effect of IL-4 in the function of PMNs as antigen presenting cells. MethodsFlow cytometry was used in the present study for the detection of cell surface human leukocyte antigen (HLA) class II, CD80 and CD86 required for antigen presentation and subsequent T-cell activation in the presence of Staphylococcus aureus enterotoxin (A). Human peripheral blood neutrophils were used for this purpose. ResultsThis study has shown that IL-4 stimulated PMNs for 24h expressed HLA class II, CD80 and CD86 that involved in antigen presentation. It also indicated that co-cultivation of IL-4 stimulated PMNs with autologous T-cells and in the presence of S. aureus enterotoxin (A) induced T-cell proliferation. ConclusionsIn vitro stimulation of PMNs with IL-4 showed expression of surface molecules involved in antigen presentation. In addition, the co-culture of T-Cells and stimulated PMNs showed high T-Cells proliferation in the presence of superantigens." ] ] "multimedia" => array:5 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2764 "Ancho" => 2674 "Tamanyo" => 291075 ] ] "descripcion" => array:1 [ "en" => "Direct flow cytometry of the HLA class II induction in whole blood PMN. (A) Unstimulated PMN (0h). (B) Unstimulated PMN (24h). (C) Stimulated PMN with IL-4 (24h)." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 2597 "Ancho" => 2424 "Tamanyo" => 254580 ] ] "descripcion" => array:1 [ "en" => "Direct flow cytometry of the CD80 induction in whole blood PMN. (A) Unstimulated PMN (0h). (B) Unstimulated PMN (24h). (C) Stimulated PMN with IL-4 (24h)." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 2674 "Ancho" => 2417 "Tamanyo" => 270841 ] ] "descripcion" => array:1 [ "en" => "FACS for CD86 surface expression in whole blood PMN. (A) Unstimulated PMN (0h). (B) Unstimulated PMN (24h). (C) Stimulated PMN with IL-4 (24h)." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 2062 "Ancho" => 3243 "Tamanyo" => 138547 ] ] "descripcion" => array:1 [ "en" => "Changes in HLA class II (A), CD80 (B) and CD86 (C) expression on unstimulated and IL-4 stimulated PMN. Statistical analysis showed that, there is a significant difference between IL-4 stimulated cells and unstimulated cells, P<0.05." ] ] 4 => array:7 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 986 "Ancho" => 1611 "Tamanyo" => 58762 ] ] "descripcion" => array:1 [ "en" => "Changes in the proliferation of T-Cells co-cultured with either unstimulated or IL-4 stimulated PMN in the presence and absence of superantigen. 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(Zoology Department, Faculty of Science, El-Minia University, El-Minia, Egypt) for his efforts in data analysis." ] ] ] "idiomaDefecto" => "en" "url" => "/03010546/0000004000000002/v1_201304101104/S030105461100156X/v1_201304101104/en/main.assets" "Apartado" => array:4 [ "identificador" => "5554" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Original articles" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/03010546/0000004000000002/v1_201304101104/S030105461100156X/v1_201304101104/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S030105461100156X?idApp=UINPBA00004N"]

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B.K.A. Abdel-Salam

Department of Zoology, Faculty of Science, 61519 Minia University, El-Minia, Egypt

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there are several reports that PMNs from a variety of species can express human leukocyte antigen &#40;HLA&#41; class II and costimulatory molecules &#40;CD80 and CD86&#41;&#46;<a class="elsevierStyleCrossRefs" href="#bib0035"><span class="elsevierStyleSup">7&#8211;10</span></a> Under certain stimulation murine neutrophils present HLA class II restricted antigen&#46;<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">PMNs function and recruitment to the site of inflammation have been shown to be upregulated by various cytokines&#44; including interleukin &#40;IL&#41;-1&#44; IL-8&#44; tumour necrosis factor &#40;TNF&#41;&#44; interferon-&#947; &#40;IFN-&#947;&#41;&#44; and granulocyte macrophage-colony stimulating factor &#40;GM-CSF&#41;&#46;<a class="elsevierStyleCrossRefs" href="#bib0025"><span class="elsevierStyleSup">5&#44;12</span></a> In addition&#44; IL-15 stimulated PMNs acquire HLA-DR&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">IL-4 production has been found to occur in thymocytes&#44; mature T-cells&#44; certain malignant T-cells&#44; mast-cells and basophiles and occasionally&#44; in transformed B-cells&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> It has an effect on B-cells&#44; T-cells&#44; monocytes&#44; mast-cells&#44; endothelial cells&#44; and fibroblasts&#46;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">15</span></a> Directly and&#47;or indirectly&#44; IL-2 has a prominent role in the regulation of IL-4 producing cells&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> IL-4 binds to a high-affinity cell-surface receptor &#40;IL-4R&#41; to exert its effects&#46;<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">16</span></a> It promotes the growth and differentiation of activated human B-lymphocytes and shares many biological functions with IL-13&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Following this approach&#44; the present study was aimed to estimate the expression of the antigen presenting molecule HLA class II and the co-stimulatory molecules CD80 and CD86 on PMNs stimulated with IL-4&#46; The expression of these surface molecules prompted us to test the T-cell proliferation by their co-culture with IL-4 stimulated PMNs in the presence <span class="elsevierStyleItalic">Staphylococcus aureus</span> enterotoxin &#40;A&#41; as a superantigen&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Materials and methods</span><p id="par0025" class="elsevierStylePara elsevierViewall">Blood was taken from 10 healthy Egyptian people by venous puncture using 7&#46;5<span class="elsevierStyleHsp" style=""></span>ml heparin-coated tubes &#40;Sarstedt&#44; N&#252;mbrecht&#44; Germany&#41; and was analysed within 2<span class="elsevierStyleHsp" style=""></span>h&#46; Recombinant human &#40;rh&#41; IL-4 was purchased from Sigma &#40;St Louis&#44; MO&#44; USA&#41;&#46; For fluorescence-activated cell sorter &#40;FACS&#41; analysis of whole blood&#44; erythrocyte FACS lysing solution was obtained from Becton Dichinson &#40;Heidelberg&#44; Germany&#41; and diluted 1&#58;10 in bidistilled water&#46; For cytofluorometry fluorescein isothiocyanate &#40;FITC&#41; and phycoerythrin &#40;PE&#41;-labelled murine MoAbs were used&#46; Mouse IgG1 FITC&#44; IgG2a PE&#44; CD66b-FITC&#44; HLA-DP&#43;DQ&#43;DR&#58;PE&#44; CD80&#58;PE and CD86&#58;PE were obtained from Coulter Immunotech &#40;Marseille&#44; France&#41;&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">For double labelling&#44; anti-CD66b-FITC as a PMNs marker and the respective PE-labelled antibody were used in equal protein concentrations&#46; Cells in whole blood were analysed by FACSCalibur and CellQuest software &#40;Becton Dickinson&#44; SanDiego&#44; CA&#44; USA&#41;&#46; Results were expressed as the percentage of positive cells in the respective gate or quadrant&#46; In FACS plots&#44; there are different populations&#46; So&#44; gates were made around population with high CD66b-FITC&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">In all FACS experiments&#44; PMNs in heparinised blood were cultured in 24-well plate&#44; 2<span class="elsevierStyleHsp" style=""></span>ml&#47;well and incubated in the presence or absence of IL-4 &#40;6<span class="elsevierStyleHsp" style=""></span>ng&#47;ml&#41; for about 24<span class="elsevierStyleHsp" style=""></span>h at 37<span class="elsevierStyleHsp" style=""></span>&#176;C with 5&#37; CO<span class="elsevierStyleInf">2</span>&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">For the co-culture of T-cells and PMNs&#44; cells were isolated by Polymorph Prep<span class="elsevierStyleSup">&#174;</span> &#40;Nycomed&#59; Oslo&#44; Norway&#41;&#46; PMNs and T-cells fraction were further purified by adsorption to CD15 and CD3 beads &#40;Miltenyi Biotec&#59; Bergisch Gladbach&#44; Germany&#41;&#44; respectively&#44; by magnetic cell separation using the devices supplied by Milteny Biotech &#40;Bergisch-Gladbach&#44; Germany&#41;&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Highly purified PMNs &#40;1<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">6</span>&#47;ml&#41; were cultivated in AIM V &#40;Gibco BRL&#59; Paisley&#44; Scotland&#41;&#41; with 2&#46;5&#37; autologous normal human serum&#44; NHS &#40;inactivated at 56<span class="elsevierStyleHsp" style=""></span>&#176;C for 30<span class="elsevierStyleHsp" style=""></span>min&#46;&#41;&#46; T-cells were cultured in RPMI 1640 &#40;Gibco BRL&#59; Paisley&#44; Scotland&#41; supplemented with 10&#37; fetal calf serum &#40;FCS&#41; &#40;PAN Biotech GmbH&#59; Aidenbach&#44; Germany&#41;&#44; 100<span class="elsevierStyleHsp" style=""></span>U&#47;ml penicillin&#47;streptomycin &#40;Gibco BRL&#59; Paisley&#44; Scotland&#41;&#44; 2<span class="elsevierStyleHsp" style=""></span>mM <span class="elsevierStyleSmallCaps">l</span>-glutamine &#40;Gibco BRL&#59; Paisley&#44; Scotland&#41;&#41;&#44; and 10<span class="elsevierStyleHsp" style=""></span>mM HEPES &#40;Gibco BRL&#59; Paisley&#44; Scotland&#41;&#46; All cells were incubated at 37<span class="elsevierStyleHsp" style=""></span>&#176;C and 5&#37; CO<span class="elsevierStyleInf">2</span> for the times indicated&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Unstimulated and stimulated PMNs &#40;1<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">3</span>&#41; in 100<span class="elsevierStyleHsp" style=""></span>&#956;l were added per well of a 96-well concave-bottom plate &#40;Greiner&#59; Nuertingen&#44; Germany&#41;&#46; Then&#44; 1<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">4</span> T-cells &#40;100<span class="elsevierStyleHsp" style=""></span>&#956;l&#41; were added to each well together with 25<span class="elsevierStyleHsp" style=""></span>ng <span class="elsevierStyleItalic">S&#46; aureus</span> enterotoxins A &#40;Sigma&#59; Munich&#44; Germany&#41;&#46; After coincubation for four days at 37<span class="elsevierStyleHsp" style=""></span>&#176;C with 5&#37; CO<span class="elsevierStyleInf">2</span>&#44; proliferation was tested by adding 1<span class="elsevierStyleHsp" style=""></span>mCi of <span class="elsevierStyleSup">3</span>H-thymidine &#40;Amer-sham Life Science&#59; Braunschweig&#44; Germany&#41; for 6&#8211;8<span class="elsevierStyleHsp" style=""></span>h &#91;<span class="elsevierStyleSup">3</span>H&#93; TdR incorporation into DNA was measured and expressed as counts per minute &#40;cpm&#41;&#46; The values represent the mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>SE of 6&#8211;12 parallel wells&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">Statistical analysis of the obtained data was performed using one way analysis of variance &#40;ANOVA&#41; test followed by least square differences &#40;LSD&#41; analysis for comparison between means&#46; Results were expressed as mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard error &#40;SE&#41;&#44; and differences were considered to be significant at <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Results</span><p id="par0060" class="elsevierStylePara elsevierViewall">Results were expressed as percentage of positive cells in the respective gate or quadrant&#46; Each set of experiments was repeated in vitro ten times&#46;</p><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">In vitro expression of HLA class II on PMNs</span><p id="par0065" class="elsevierStylePara elsevierViewall">The majority of healthy donors PMNs expressed CD66b &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>a&#8211;c&#41;&#46; In the first set of experiments the effect of IL-4 on the expression of HLA class II was tested&#44; where we found that PMNs on whole blood showed expression of HLA class II recording 7&#46;77&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>c&#41; by using IL-4&#44; as stimulators&#46; In contrast&#44; fresh and unstimulated cells cultured with medium only counted 1&#46;05&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>a&#41; and 2&#46;71&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>b&#41;&#44; respectively&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">In vitro induction of CD80 on PMNs</span><p id="par0070" class="elsevierStylePara elsevierViewall">As shown for HLA class II&#44; surface expression of CD80 was most impressive following cultivation of whole blood with the stimuli for 24<span class="elsevierStyleHsp" style=""></span>h&#46; The proportion of double-positive cells &#40;right upper quadrant&#41; was estimated&#44; where CD80 positive cells was slightly higher in stimulated cells recording 4&#46;06&#37; by using IL-4 &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>c&#41; than unstimulated PMNs cultured with medium for 24<span class="elsevierStyleHsp" style=""></span>h &#40;1&#46;12&#37;&#41; and fresh cells&#44; 0&#46;05&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>a and b&#44; respectively&#41;&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">In vitro induction of CD86 on PMNs</span><p id="par0075" class="elsevierStylePara elsevierViewall">For CD86&#44; high expression was recorded on the surface of IL-4 stimulated PMNs in whole blood recorded 20&#46;48&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>c&#41;&#44; while percentage of CD86 molecules on the surface of unstimulated PMNs measured 6&#46;97 &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>b&#41;&#46; Fresh PMNs in while blood had 1&#46;55&#37; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>a&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia><p id="par0080" class="elsevierStylePara elsevierViewall">Statistical analysis of HLA class II&#44; CD80 and CD86 experimental sets showed that there is a significant difference between IL-4 stimulated cells and unstimulated cells&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>&#41;&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Interaction of IL-4 stimulated PMNs with peripheral T-cells</span><p id="par0085" class="elsevierStylePara elsevierViewall">For these experiments highly purified PMNs were cultivated with IL-4 for 24<span class="elsevierStyleHsp" style=""></span>h and then co-cultivated with highly purified isolated T-cells in the absence or presence of SEA&#46; The differences between the groups in the sets of experiments related to the interaction of HLA class II positive PMNs with peripheral T-cells showed a significant difference&#44; where <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>&#41;&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia></span></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Discussion</span><p id="par0090" class="elsevierStylePara elsevierViewall">PMNs are considered short-lived cells undergoing spontaneous apoptosis in vivo as well as in culture&#46;<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">18</span></a> Previous studies have demonstrated that PMNs can be induced in vitro to synthesize and release various cytokines&#44; suggesting that these cells can contribute significantly to the initiation and amplification of cellular and humoral immune responses&#46;<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a> The detection of these molecules&#44; therefore&#44; provides strong support for the hypothesis that human PMNs can actively synthesize immunoregulatory molecules<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">19</span></a> and have the potential to act as antigen presenting cells&#46;<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a></p><p id="par0095" class="elsevierStylePara elsevierViewall">Recently&#44; it has become increasingly evident that culturing PMNs in the presence of cytokines extends their life span&#46;<a class="elsevierStyleCrossRefs" href="#bib0100"><span class="elsevierStyleSup">20&#8211;23</span></a> Cultured PMNs synthesize and release immunomodulatory cytokines by which they may participate in the afferent limb of the immune response&#46;<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a></p><p id="par0100" class="elsevierStylePara elsevierViewall">This study has clearly shown that PMNs could be induced to express HLA class II&#44; CD80 and CD86 after activation with IL-4&#46; These data are in accordance with other results where HLA class II&#44; CD80 and CD86 were expressed on the PMNs surface after exposure to GM-CSF and&#47;or INF-&#947;&#46;<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a></p><p id="par0105" class="elsevierStylePara elsevierViewall">The antigen presenting molecule HLA class II and the co-stimulatory molecules CD80 and CD86 play an important role in T-cell proliferation&#44; where HLA class II presents the engulfed antigen to T-cells&#46;<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a> CD80 and CD86 act as second signal molecules involving in the stimulation of T-cells to produce the autocrine growth factor IL-2 without which T-cells are thus unable to proliferate&#46;<a class="elsevierStyleCrossRef" href="#bib0120"><span class="elsevierStyleSup">24</span></a></p><p id="par0110" class="elsevierStylePara elsevierViewall">The activation and recruitment of PMNs were also regulated by IL-15&#44;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a> IFN-&#947;&#44;<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">25</span></a> CSF-CSF<a class="elsevierStyleCrossRefs" href="#bib0130"><span class="elsevierStyleSup">26&#8211;28</span></a> and IL-8&#46;<a class="elsevierStyleCrossRef" href="#bib0145"><span class="elsevierStyleSup">29</span></a> In addition&#44; PMNs posses IL-2R&#946; chain<a class="elsevierStyleCrossRefs" href="#bib0150"><span class="elsevierStyleSup">30&#44;31</span></a> and IL-2R&#947; chain<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">32</span></a> that have the ability to bind with IL-4&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">33</span></a> These published data prompted us to study the effect of IL-4 on PMNs functions&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">When comparing the number of monocytes and PMNs required to induce T-cell proliferation&#44; it was observed that 10 times more PMNs than monocytes were necessary to yield the same extent of T-cell proliferation&#46; However&#44; the cell preparation used in this work never contained more than 1&#37; of contaminating cells and certainly not the 10&#37; of monocytes that would be required to affect the results&#46;<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">30</span></a> The observation that ten times more PMNs than monocytes were required to induce a similar extent of T-cell proliferation has to be considered together with the observation that only a proportion of PMNs acquired HLA class II&#46; Thus&#44; when only fully equipped PMNs are considered&#44; the ability to process and to present antigen is similar to that of monocytes&#46; Whether HLA class II-positive PMNs participate in the immune defence or play a role in pathophysiological events&#44; is a matter of speculation&#46; The fact that only a minor proportion of PMNs acquire HLA class II might lead to the conclusion that a possible accessory function of PMNs would be rather weak&#46; One has&#59; however&#44; to bear in mind that PMNs are numerous in the peripheral blood&#44; and that even a low percentage of PMNs expressing HLA class II would exceed both circulating monocytes and dendritic cells in number&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">Due to the notion that the presence of <span class="elsevierStyleItalic">Staphylococcus entrotoxin</span> coincides with relapses of Wegener&#39;s granulomatosis&#44;<a class="elsevierStyleCrossRef" href="#bib0170"><span class="elsevierStyleSup">34</span></a> we tested whether PMNs was able to also present <span class="elsevierStyleItalic">Staphylococcus enterotoxin A</span> as a superantigen&#44; so-called because it binds outside of the peptide-binding groove of the HLA class II and the antigen-specific domain of the T-cell receptor&#44; and consequently activates a large portion of T-cells&#44; preferentially those with a V beta 2 domain&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">35</span></a> In accordance with previous studies&#44;<a class="elsevierStyleCrossRefs" href="#bib0040"><span class="elsevierStyleSup">8&#44;36</span></a> co-culture of PMNs with T-cells and SEA resulted in T-cell proliferation&#46; Taken together&#44; our data demonstrate that by synthesizing and expressing of HLA class II antigens&#44; CD80 and CD86&#59; PMNs acquire the capacity to present superantigens to T-cells&#46;</p><p id="par0125" class="elsevierStylePara elsevierViewall">Data showed that there is a clear increase in T-lymphocyte proliferation in the co-culture of stimulated PMNs<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>T-cells&#46; Because there is no antigen present&#44; this means that it is not caused by the co-stimulatory molecules on the surface of the PMNs but probably due to mediators secreted by the PMNs&#46; Furthermore&#44; unstimulated PMNs<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>T-cells<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>SEA give almost a 1000-fold increase of T-cell proliferation while only a few percent of PMNs express co-stimulatory molecules in a naive state&#46; Probably the results obtained are both from released mediators and the co-stimulatory molecules&#46; Mediators secreted by IL-4 stimulated PMNs will be investigated in our future research&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">In conclusion&#44; stimulated PMNs with IL-4 lead to expression of the antigen presenting molecules &#40;HLA class II&#41; and the co-stimulatory molecules &#40;CD80 and CD86&#41;&#46; These molecules play an important role in antigen presentation and consequently T-cell proliferation in the presence of SEA&#46; This means that IL-4 stimulated PMNs might be involved indirectly in acquired immune response in addition to their role in innate immunity&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Conflict of interest</span><p id="par0135" class="elsevierStylePara elsevierViewall">The authors have no conflict of interest to declare&#46;</p></span></span>"
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          "titulo" => "Keywords"
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          "titulo" => "Abbreviations"
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          "titulo" => "Introduction"
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          "titulo" => "Materials and methods"
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          "titulo" => "Results"
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            0 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "In vitro expression of HLA class II on PMNs"
            ]
            1 => array:2 [
              "identificador" => "sec0025"
              "titulo" => "In vitro induction of CD80 on PMNs"
            ]
            2 => array:2 [
              "identificador" => "sec0030"
              "titulo" => "In vitro induction of CD86 on PMNs"
            ]
            3 => array:2 [
              "identificador" => "sec0035"
              "titulo" => "Interaction of IL-4 stimulated PMNs with peripheral T-cells"
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        6 => array:2 [
          "identificador" => "sec0040"
          "titulo" => "Discussion"
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        7 => array:2 [
          "identificador" => "sec0045"
          "titulo" => "Conflict of interest"
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          "identificador" => "xack31869"
          "titulo" => "Acknowledgements"
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          "titulo" => "References"
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    "fechaRecibido" => "2010-12-28"
    "fechaAceptado" => "2011-02-14"
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            0 => "IL-4"
            1 => "HLA class II"
            2 => "PMNs"
            3 => "<span class="elsevierStyleItalic">Staphylococcus aureus A</span>"
            4 => "T-cells"
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        1 => array:4 [
          "clase" => "abr"
          "titulo" => "Abbreviations"
          "identificador" => "xpalclavsec74251"
          "palabras" => array:3 [
            0 => "PMNs"
            1 => "HLA"
            2 => "IL"
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        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Polymorphonuclear neutrophils &#40;PMNs&#41; were originally described as short lived and terminally differentiated phagocytes that contribute only to the innate immune response&#46; Some studies of PMNs cytokine production and expression of numerous cell surface proteins has suggested that PMNs are likely to influence adaptive responses and may satisfy the criteria of antigen presenting cells&#46;</p> <span class="elsevierStyleSectionTitle">Aim of the study</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">This work aimed to study the effect of IL-4 in the function of PMNs as antigen presenting cells&#46;</p> <span class="elsevierStyleSectionTitle">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Flow cytometry was used in the present study for the detection of cell surface human leukocyte antigen &#40;HLA&#41; class II&#44; CD80 and CD86 required for antigen presentation and subsequent T-cell activation in the presence of <span class="elsevierStyleItalic">Staphylococcus aureus</span> enterotoxin &#40;A&#41;&#46; Human peripheral blood neutrophils were used for this purpose&#46;</p> <span class="elsevierStyleSectionTitle">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">This study has shown that IL-4 stimulated PMNs for 24<span class="elsevierStyleHsp" style=""></span>h expressed HLA class II&#44; CD80 and CD86 that involved in antigen presentation&#46; It also indicated that co-cultivation of IL-4 stimulated PMNs with autologous T-cells and in the presence of <span class="elsevierStyleItalic">S&#46; aureus</span> enterotoxin &#40;A&#41; induced T-cell proliferation&#46;</p> <span class="elsevierStyleSectionTitle">Conclusions</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">In vitro stimulation of PMNs with IL-4 showed expression of surface molecules involved in antigen presentation&#46; In addition&#44; the co-culture of T-Cells and stimulated PMNs showed high T-Cells proliferation in the presence of superantigens&#46;</p>"
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          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Direct flow cytometry of the HLA class II induction in whole blood PMN&#46; &#40;A&#41; Unstimulated PMN &#40;0<span class="elsevierStyleHsp" style=""></span>h&#41;&#46; &#40;B&#41; Unstimulated PMN &#40;24<span class="elsevierStyleHsp" style=""></span>h&#41;&#46; &#40;C&#41; Stimulated PMN with IL-4 &#40;24<span class="elsevierStyleHsp" style=""></span>h&#41;&#46;</p>"
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          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Changes in the proliferation of T-Cells co-cultured with either unstimulated or IL-4 stimulated PMN in the presence and absence of superantigen&#46; Statistical analysis showed that&#44; there is a significant difference in T-Cells proliferation between T-Cells co-cultured with IL-4 stimulated and unstimulated PMN in superantigen presence&#44; where <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p>"
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Article information

ISSN: 03010546
Original language: English

DOI:10.1016/j.aller.2011.02.012

The statistics are updated each day

Year/Month	Html	Pdf	Total

2024 November	3	2	5
2024 October	10	5	15
2024 September	22	9	31
2024 August	10	3	13
2024 July	16	10	26
2024 June	7	7	14
2024 May	20	5	25
2024 April	20	9	29
2024 March	27	7	34
2024 February	14	11	25
2024 January	22	7	29
2023 December	24	9	33
2023 November	49	6	55
2023 October	30	9	39
2023 September	22	6	28
2023 August	20	9	29
2023 July	18	4	22
2023 June	16	5	21
2023 May	14	4	18
2023 April	23	3	26
2023 March	21	4	25
2023 February	20	10	30
2023 January	33	6	39
2022 December	21	10	31
2022 November	25	8	33
2022 October	37	7	44
2022 September	36	9	45
2022 August	15	9	24
2022 July	12	13	25
2022 June	18	9	27
2022 May	24	9	33
2022 April	16	5	21
2022 March	25	13	38
2022 February	17	7	24
2022 January	32	8	40
2021 December	32	11	43
2021 November	20	9	29
2021 October	22	18	40
2021 September	20	6	26
2021 August	19	21	40
2021 July	13	10	23
2021 June	8	5	13
2021 May	16	10	26
2021 April	29	30	59
2021 March	18	16	34
2021 February	6	6	12
2021 January	7	8	15
2020 December	2	0	2
2020 November	0	1	1
2020 October	0	1	1
2020 September	0	1	1
2020 August	0	1	1
2020 July	0	2	2
2020 May	0	2	2
2020 March	0	2	2
2020 January	0	2	2
2019 November	0	1	1
2019 September	0	3	3
2019 July	0	1	1
2019 June	0	5	5
2019 May	0	19	19
2018 August	0	1	1
2018 February	3	1	4
2018 January	6	1	7
2017 December	8	0	8
2017 November	7	6	13
2017 October	8	8	16
2017 September	10	6	16
2017 August	16	4	20
2017 July	8	3	11
2017 June	8	5	13
2017 May	5	4	9
2017 April	17	13	30
2017 March	12	52	64
2017 February	9	4	13
2017 January	20	0	20
2016 December	15	5	20
2016 November	19	1	20
2016 October	42	8	50
2016 September	28	1	29
2016 August	18	4	22
2016 July	17	5	22
2016 June	22	5	27
2016 May	11	12	23
2016 April	10	10	20
2016 March	17	15	32
2016 February	21	16	37
2016 January	21	15	36
2015 December	14	9	23
2015 November	15	11	26
2015 October	23	12	35
2015 September	19	7	26
2015 August	34	5	39
2015 July	31	5	36
2015 June	15	5	20
2015 May	22	2	24
2015 April	23	9	32
2015 March	20	6	26
2015 February	15	6	21
2015 January	27	4	31
2014 December	32	3	35
2014 November	16	2	18
2014 October	29	4	33
2014 September	33	4	37
2014 August	22	0	22
2014 July	11	3	14
2014 June	18	2	20
2014 May	8	1	9
2014 April	8	7	15
2014 March	24	8	32
2014 February	32	2	34
2014 January	28	10	38
2013 December	29	8	37
2013 November	29	2	31
2013 October	34	10	44
2013 September	45	5	50
2013 August	59	10	69
2013 July	41	9	50
2013 June	17	4	21
2013 May	44	5	49
2013 April	34	8	42
2013 March	23	4	27
2013 February	23	6	29
2013 January	25	3	28
2012 December	9	4	13
2012 November	1	2	3
2012 October	2	1	3
2012 September	1	1	2
2012 March	454	0	454

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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