Scientific letter
CD64 expression on monocytes and granulocytes in pediatric acute appendicitis scenario: A pilot study in pediatric critical care
Expresión de CD64 en monocitos y granulocitos en apendicitis aguda: estudio piloto en cuidados intensivos pediátricos
A. García-Salidoa,
, M. De Lucio-Rodríguezb, J.L. Alonso Calderónb, G. De Lama Caro-Patóna, M. Ramírez-Orellanac,d, A. Serrano-Gonzáleza
Autor para correspondencia
a Pediatric Critical Care Unit, Hospital Infantil Universitario Niño Jesús, Madrid, Spain
b Pediatric Surgery Unit, Hospital Infantil Universitario Niño Jesús, Madrid, Spain
c Pediatric Hematology and Oncology, Hospital Infantil Universitario Niño Jesús, Madrid, Spain
d Instituto de Investigaciones Sanitarias La Princesa, Madrid, Spain
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García-Salido, M. De Lucio-Rodríguez, J.L. Alonso Calderón, G. De Lama Caro-Patón, M. Ramírez-Orellana, A. Serrano-González" "autores" => array:6 [ 0 => array:4 [ "nombre" => "A." "apellidos" => "García-Salido" "email" => array:1 [ 0 => "citopensis@yahoo.es" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">*</span>" "identificador" => "cor0005" ] ] ] 1 => array:3 [ "nombre" => "M." "apellidos" => "De Lucio-Rodríguez" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 2 => array:3 [ "nombre" => "J.L." "apellidos" => "Alonso Calderón" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 3 => array:3 [ "nombre" => "G." 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"apellidos" => "Serrano-González" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] ] "afiliaciones" => array:4 [ 0 => array:3 [ "entidad" => "Pediatric Critical Care Unit, Hospital Infantil Universitario Niño Jesús, Madrid, Spain" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Pediatric Surgery Unit, Hospital Infantil Universitario Niño Jesús, Madrid, Spain" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Pediatric Hematology and Oncology, Hospital Infantil Universitario Niño Jesús, Madrid, Spain" "etiqueta" => "c" "identificador" => "aff0015" ] 3 => array:3 [ "entidad" => "Instituto de Investigaciones Sanitarias La Princesa, Madrid, Spain" "etiqueta" => "d" "identificador" => "aff0020" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Expresión de CD64 en monocitos y granulocitos en apendicitis aguda: estudio piloto en cuidados intensivos pediátricos" ] ] "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" => 695 "Ancho" => 2342 "Tamanyo" => 79459 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Spearman correlation between CD64 expression and days of PICU stay. %gCD64 is the percentage of CD64-positive cells, gCD64 is the CD64 degree of expression in granulocytes by mean fluorescence intensity and mCD64 is the CD64 degree of expression in monocytes by mean fluorescence intensity.</p>" ] ] ] "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">Acute appendicitis (AA) is one of the most common childhood surgical entities.<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">1</span></a> Anamnesis, physical examination, blood tests and imaging provide an accurate diagnosis in 98% of patients. However, there is a lack of reliable instruments to predict evolution and anticipate complications.<a class="elsevierStyleCrossRefs" href="#bib0050"><span class="elsevierStyleSup">2,3</span></a> To this aim, recent studies using flow cytometry (FC) have been done to evaluate expression profile in patients with acute abdomen. The FC is a laboratory technique that allows to evaluate immune status or response of an individual based on marking leukocyte populations with monoclonal antibodies. One of the biomarkers that have demonstrated clinical applicability is CD64. Increasing of its density on surface is directly related to the intensity of stimulation received by inflammatory cytokines. It could be quantified by flow cytometry as mean fluorescence intensity (MFI).<a class="elsevierStyleCrossRef" href="#bib0060"><span class="elsevierStyleSup">4</span></a> The main objective of this preliminary study was to describe the CD64 expression in pediatric patients after appendectomy and pediatric critical care unit (PICU) admission. For this purpose, the percentage of CD64-positive cells and the degree of expression (MFI) of CD-64 on monocytes (mCD64) and granulocytes (gCD64) were determined. Afterwards, it was analyzed its association with common biomarkers, the therapeutic care, the hospitalization time and complications.</p><p id="par0010" class="elsevierStylePara elsevierViewall">The study was prospectively conducted from November 2016 to January 2017. Peripheral blood samples were extracted after parents or legal guardians consent at PICU admission. The PICU admission was done by surgeon criteria and not protocoled. Samples were collected at first 24<span class="elsevierStyleHsp" style=""></span>h after surgery in sterile EDTA at room temperature or refrigerated at 4<span class="elsevierStyleHsp" style=""></span>°C, used for CD45+ cells marking and analyzed by flow cytometry in a time period shorter than 24<span class="elsevierStyleHsp" style=""></span>h. Fifteen patients (6 female and 9 male) were recruited with a median age 7.3<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>4.2 years. Median PICU stay was 1<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>1.4 days with a median hospitalization of 5.5<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>4.4 days. The CD64 expression on neutrophils and monocytes were mCD64 median MFI of 16,366<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>6762 and gCD64 median MFI of 2071<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>2791.8 with a percentage of CD64+ granulocytes (%gCD64+) 75<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>24.4% and a mCD64/gCD64 ratio of 5<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>1.7. Non-parametric test were applied. Time of stay in the PICU correlated with gCD64 (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.56; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.03), % gCD64+ (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.6; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.02), and mCD64/gCD64 ratio (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>−0.6; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.02) (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>). Also the hospitalization time showed correlation with mCD64/gCD64 ratio (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.5; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.04). The use of nasogastric tube, as an expression of bowel movement, was correlated with gCD64 (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.5; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.03) and % gCD64+ (<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.5; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.05). Finally, Granulocytes of patients with parenteral nutrition necessity (PNT) showed greater positivity for CD64 (% gCD64+ – TPN 90<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>14 vs not TPN 68<span class="elsevierStyleHsp" style=""></span>±<span class="elsevierStyleHsp" style=""></span>27, <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>0.045). Among the classical parameters, none of them showed correlation or statistically significant association with the clinical data except for the C-reactive protein (CRP). At PICU admission leukocytes, C reactive protein and procalcitonin were obtained and there was observed a positive correlation with no signification with CD64 expression.</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0015" class="elsevierStylePara elsevierViewall">This pilot work describes, for the first time, the CD64 expression on peripheral blood monocytes and granulocytes from pediatric patients after appendectomy.<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">5</span></a> Also, this is the first study that tries to correlate clinical and evolution data from acute appendicitis pediatric patients with CD64 values obtained by FC.</p><p id="par0020" class="elsevierStylePara elsevierViewall">As said the FC interpretation may provide unique information letting know immunological changes in real-time. These properties would help to substantiate diagnostic suspicion, anticipate the evolution and modify therapeutic attitudes. In this study, CD64 expression in terms on granulocyte of MFI is lower, as our group has previously seen,<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">6</span></a> than in other inflammatory and infectious contexts.<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">7</span></a> These results could have two explanation. On the one hand, the AA is, with exceptions, a localized infection in which the systemic expression through CD64 quantification on granulocytes and monocytes in peripheral blood, may be underestimated. In this process, participating granulocytes would not be predominant in the bloodstream. At the same time, in this study, determination by FC was performed after surgery, so after having removed the inflammatory insult and having received, at least, one dose of antibiotic therapy. It is well known that CD64 reduces its expression once received appropriate antibiotic therapy. In the same way, it is known that the half-life of a human granulocyte in peripheral blood is 6<span class="elsevierStyleHsp" style=""></span>h.<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">8</span></a> Thus, in our experiment, leukocyte populations studied might not be under the same inflammatory context than those present at the onset of AA underestimating the value of MFI.<a class="elsevierStyleCrossRefs" href="#bib0065"><span class="elsevierStyleSup">5,6</span></a></p><p id="par0025" class="elsevierStylePara elsevierViewall">In order to minimize the potential effect of this confusion factor, CD64 expression ratio (mCD64/gCD64) was studied knowing that the presence of CD64 on granulocytes should be null in case of no inflammation (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>). Changes in this ratio could be used to quantify upregulation of this molecule in granulocytes. Our study showed that mCD64/gCD64 ratio inversely correlated with total hospital stay. Also, and with the same objective, CD64+ granulocytes percentage was quantified. Since baseline percentage of positive CD64 granulocytes must be zero, any higher value would indicate activation. In the studied population CD64 positivity existed in all cases, being always superior to 45% in granulocytes. As said, the percentage of CD64+ granulocytes was higher in patients with prolonged PICU stay, days of hospital admission and need of PNT. This correlation with the need of PNT, is of great interest due to its novelty.<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">2</span></a></p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0030" class="elsevierStylePara elsevierViewall">This study presents limitations. It was done in a single center, with a small number of patients and without strictly defined PICU admission criteria. At the same time, it cannot be assumed that CD64 is the only responsible for the clinical course in the case of AA patients. Their values should be compared with traditional biomarkers in larger samples. This aspect could also be attenuated with the inclusion of other cell surface markers in further analysis. Finally, just a single blood sample was obtained after the surgery. These values should be compared to previous surgery data and/or later samples. The study about CD64 dynamic variation should be included in future studies.</p><p id="par0035" class="elsevierStylePara elsevierViewall">As conclusion, in our preliminary work, the expression of CD64 on granulocytes at PICU admission correlated with the hospitalization time and need of nasogastric tube and non-enteral nutrition. Based on these findings, it seems of interest to continue the study of CD64 in order to provide dynamic and reliable information about the actual utility of the flow cytometry in these patients.</p></span>" "pdfFichero" => "main.pdf" "tienePdf" => true "multimedia" => array:2 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 695 "Ancho" => 2342 "Tamanyo" => 79459 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Spearman correlation between CD64 expression and days of PICU stay. %gCD64 is the percentage of CD64-positive cells, gCD64 is the CD64 degree of expression in granulocytes by mean fluorescence intensity and mCD64 is the CD64 degree of expression in monocytes by mean fluorescence intensity.</p>" ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1391 "Ancho" => 2334 "Tamanyo" => 317914 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">(A) CD64 staining on granulocytes, monocytes and lymphocytes in a healthy child. From left to right the expression on them can be observed. CD64 is expressed in monocytes but not in lymphocytes and neutrophils. (B) CD64 staining in a child with bacterial infection, the granulocytes are CD64 positive (displaced from left to right) as a consequence on CD64 upregulation. The lymphocytes act as an internal negative control.</p>" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0015" "bibliografiaReferencia" => array:8 [ 0 => array:3 [ "identificador" => "bib0045" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "The magic of an appendicitis score" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "R.E. 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| año/Mes | Html | Total | |
|---|---|---|---|
| 2024 Noviembre | 11 | 10 | 21 |
| 2024 Octubre | 79 | 60 | 139 |
| 2024 Septiembre | 85 | 54 | 139 |
| 2024 Agosto | 100 | 67 | 167 |
| 2024 Julio | 78 | 53 | 131 |
| 2024 Junio | 87 | 59 | 146 |
| 2024 Mayo | 70 | 38 | 108 |
| 2024 Abril | 81 | 31 | 112 |
| 2024 Marzo | 73 | 39 | 112 |
| 2024 Febrero | 91 | 38 | 129 |
| 2024 Enero | 118 | 29 | 147 |
| 2023 Diciembre | 130 | 43 | 173 |
| 2023 Noviembre | 120 | 48 | 168 |
| 2023 Octubre | 110 | 27 | 137 |
| 2023 Septiembre | 83 | 36 | 119 |
| 2023 Agosto | 73 | 21 | 94 |
| 2023 Julio | 79 | 40 | 119 |
| 2023 Junio | 100 | 33 | 133 |
| 2023 Mayo | 99 | 42 | 141 |
| 2023 Abril | 77 | 37 | 114 |
| 2023 Marzo | 123 | 50 | 173 |
| 2023 Febrero | 115 | 41 | 156 |
| 2023 Enero | 129 | 36 | 165 |
| 2022 Diciembre | 134 | 36 | 170 |
| 2022 Noviembre | 145 | 58 | 203 |
| 2022 Octubre | 133 | 34 | 167 |
| 2022 Septiembre | 102 | 55 | 157 |
| 2022 Agosto | 104 | 52 | 156 |
| 2022 Julio | 108 | 50 | 158 |
| 2022 Junio | 120 | 26 | 146 |
| 2022 Mayo | 114 | 42 | 156 |
| 2022 Abril | 81 | 47 | 128 |
| 2022 Marzo | 101 | 54 | 155 |
| 2022 Febrero | 84 | 17 | 101 |
| 2022 Enero | 117 | 39 | 156 |
| 2021 Diciembre | 101 | 45 | 146 |
| 2021 Noviembre | 134 | 38 | 172 |
| 2021 Octubre | 130 | 65 | 195 |
| 2021 Septiembre | 98 | 29 | 127 |
| 2021 Agosto | 73 | 35 | 108 |
| 2021 Julio | 50 | 28 | 78 |
| 2021 Junio | 54 | 37 | 91 |
| 2021 Mayo | 84 | 66 | 150 |
| 2021 Abril | 140 | 76 | 216 |
| 2021 Marzo | 194 | 36 | 230 |
| 2021 Febrero | 91 | 25 | 116 |
| 2021 Enero | 81 | 24 | 105 |
| 2020 Diciembre | 64 | 27 | 91 |
| 2020 Noviembre | 50 | 22 | 72 |
| 2020 Octubre | 26 | 12 | 38 |
| 2020 Septiembre | 2 | 0 | 2 |
| 2020 Agosto | 2 | 0 | 2 |
| 2020 Julio | 50 | 21 | 71 |
| 2020 Junio | 58 | 41 | 99 |
| 2020 Mayo | 34 | 27 | 61 |
| 2020 Abril | 19 | 15 | 34 |
| 2020 Marzo | 3 | 0 | 3 |
| 2019 Mayo | 2 | 0 | 2 |
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