Name: an in vitro model to study immune responses of human peripheral blood mononuclear cells to human respiratory syncytial virus infection
Uploaded: 2013-12-10T21:00:00
Description: Watch this Scientific Journal Video about An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection at JoVE.com

RSV A2 was kindly provided by Dr. R. de Swart (Erasmus MC, Rotterdam, The Netherlands). MV and GF are supported by the Virgo consortium, funded by the Dutch government project number FES0908, and by the Netherlands Genomics Initiative (NGI) project number 050-060-452. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Note:
Experiments with HRSV must be performed in a biological safety cabinet in a biosafety level 2 laboratory. Collect all viral waste and material that has been in contact with the virus for proper disposal due to biological hazards.
Special precaution must be taken when using potentially infected human blood.
It is crucial to use endotoxin-free reagents throughout all protocols.
1. Culturing of HRSV A2
<ol>
	...

<ol>
	<li>Nair, H., Nokes, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+burden+of+acute+lower+respiratory+infections+due+to+respiratory+syncytial+virus+in+young+children:+a+systematic+review+and+meta-analysis.">Global burden of acute lower respiratory infections due to respiratory syncytial virus in young children: a systematic review and meta-analysis.</a> Lancet. 375, 1545-1555 (2010).</li><li>Falsey, A. R., Hennessey, P. A., Formica, M. A., Cox, C., Walsh, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+infection+in+elderly+and+high-risk+adults.">Respiratory syncytial virus infection in elderly and high-risk adults.</a> N. Eng. J. Med. , 352-1749 (2005).</li><li>Openshaw, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antiviral+Immune+Responses+and+Lung+Inflammation+after+Respiratory+Syncytial+Virus+Infection.">Antiviral Immune Responses and Lung Inflammation after Respiratory Syncytial Virus Infection.</a> Proc. Am. Thorac. Soc. 2 (2), 121-125 (2005).</li><li>Openshaw, P. J., Tregoning, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+responses+and+disease+enhancement+during+respiratory+syncytial+virus+infection.">Immune responses and disease enhancement during respiratory syncytial virus infection.</a> Clin. Microbio. Rev. 18 (3), 541-555 (2005).</li><li>Villenave, R., Thavagnanam, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+modeling+of+respiratory+syncytial+virus+infection+of+pediatric+bronchial+epithelium,+the+primary+target+of+infection+in+vivo.">In vitro modeling of respiratory syncytial virus infection of pediatric bronchial epithelium, the primary target of infection in vivo.</a> Proc. Natl. Acad. Sci. U.S.A. 109 (13), 5040-5045 (2012).</li><li>Tulic, M. K., Hurrelbrink, R. J., Pr&#234;le, C. M., Laing, I. A., Upham, J. W., Le Souef, P., Sly, P. D., Holt, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TLR4+polymorphisms+mediate+impaired+responses+to+respiratory+syncytial+virus+and+lipopolysaccharide.">TLR4 polymorphisms mediate impaired responses to respiratory syncytial virus and lipopolysaccharide.</a> J. Immunol. 179 (1), 132-140 (2007).</li><li>Vissers, M., Remijn, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+infection+augments+NOD2+signaling+in+an+IFN-&#946;-dependent+manner+in+human+primary+cells.">Respiratory syncytial virus infection augments NOD2 signaling in an IFN-&#946;-dependent manner in human primary cells.</a> Eur. J. Immunol. 42 (10), 2727-2735 (2012).</li><li>Cherukuri, A., Patton, K., Gasser, R. A., Zuo, F., Woo, J., Esser, M. T., Tang, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adults+65+years+old+and+older+have+reduced+number+of+functional+memory+T+cells+to+respiratory+syncytial+virus+fusion+protein.">Adults 65 years old and older have reduced number of functional memory T cells to respiratory syncytial virus fusion protein.</a> Clin. Vacc. Immunol. 20 (2), 239-247 (2013).</li><li>Shafique, M., Wilschut, J., de Haan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+mucosal+and+systemic+immunity+against+respiratory+syncytial+virus+by+inactivated+virus+supplemented+with+TLR9+and+NOD2+ligands.">Induction of mucosal and systemic immunity against respiratory syncytial virus by inactivated virus supplemented with TLR9 and NOD2 ligands.</a> Vaccine. 30, 597-606 (2012).</li><li>Kwilas, S., Liesman, R. M., Zhang, L., Walsh, E., Pickles, R. J., Peeples, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+grown+in+Vero+cells+contains+a+truncated+attachment+protein+that+alters+its+infectivity+and+dependence+on+glycosaminoglycans.">Respiratory syncytial virus grown in Vero cells contains a truncated attachment protein that alters its infectivity and dependence on glycosaminoglycans.</a> J. Virol. 83 (20), 10710-10718 (2009).</li><li>Treuhaft, M. W., Beem, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defective+interfering+particles+of+respiratory+syncytial+virus.">Defective interfering particles of respiratory syncytial virus.</a> Infect. Immun. 37 (2), 439-444 (1982).</li><li>Korns Johnson, D., Homann, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+and+improved+quantification+of+lymphocytic+choriomeningitis+virus+(LCMV)+titers+by+flow+cytometry.">Accelerated and improved quantification of lymphocytic choriomeningitis virus (LCMV) titers by flow cytometry.</a> PLoS One. 7 (5), e37337 (2012).</li><li>Wurfel, M. M., Park, W. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+high+and+low+responders+to+lipopolysaccharide+in+normal+subjects:+an+unbiased+approach+to+identify+modulators+of+innate+immunity.">Identification of high and low responders to lipopolysaccharide in normal subjects: an unbiased approach to identify modulators of innate immunity.</a> J. Immunol. 175 (4), 2570-2578 (2005).</li><li>Kimpen, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+and+asthma.+The+role+of+monocytes.">Respiratory syncytial virus and asthma. The role of monocytes.</a> Am. J. Respir. Crit. Care Med. 163, 7-9 (2001).</li><li>Wang, S. Z., Forsyth, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+interaction+of+neutrophils+with+respiratory+epithelial+cells+in+viral+infection.">The interaction of neutrophils with respiratory epithelial cells in viral infection.</a> Respirology. 5 (1), 1-10 (2000).</li><li>San-Juan-Vergara, H., Sampayo-Escobar, V., Reyes, N., Cha, B., Pacheco-Lugo, L., Wong, T., Peeples, M. E., Collins, P. L., Castano, M. E., Mohapatra, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholesterol-rich+microdomains+as+docking+platforms+for+respiratory+syncytial+virus+in+normal+human+bronchial+epithelial+cells.">Cholesterol-rich microdomains as docking platforms for respiratory syncytial virus in normal human bronchial epithelial cells.</a> J. Virol. 86 (3), 1832-1843 (2012).</li><li>Davidson, D., Zaytseva, A., Miskolci, V., Castro-Alcaraz, S., Vancurova, I., Patel, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+expression+profile+of+endotoxin-stimulated+leukocytes+of+term+new+born:+control+of+cytokine+gene+expression+by+interleukin-10.">Gene expression profile of endotoxin-stimulated leukocytes of term new born: control of cytokine gene expression by interleukin-10.</a> Plos One. 8 (1), e53641 (2013).</li></ol>

In this study, we demonstrate that infection of PBMC with HRSV is a fast and reliable model system in which inflammatory pathways can be studied. In order to stimulate PBMCs an HRSV stock has to be prepared and quantified.
Multiple cell lines are susceptible to HRSV infection. Cell lines most used for HRSV culturing are HEp-2, HeLa,&#160;and Vero cells. However, we would not recommend the usage of Vero cells for culturing of HRSV because it has been shown that culturing of HRSV in Vero cells r...

Human Respiratory Syncytial Virus (HRSV) is the most common cause of lower respiratory tract infections in children. Each year, over 33 million children under the age of five are infected with HRSV, leading to over three million hospitalizations and almost 200,000 deaths1. A growing body of evidence suggests that HRSV also poses a significant threat to the elderly and adults with underlying chronic illnesses2.
The majority of HRSV infections in young children presents with mild symptoms, comparable to the common cold, and do not require clinical intervention. However, a small proportion of patients require hospitalization and mechanical ventilation due to severe bronchiolitis.
Part of the pathogenesis of HRSV disease is the host&#39;s overexuberant and inadequate immune response to infection3,4. This is illustrated by several observations. The period of maximal illness is often preceded by the peak of viral infection and coincides better with cellular infiltration of infected tissues and the release of inflammatory cytokines3. Another line of evidence comes from the formalin inactivated HRSV (FI-RSV) trials in the 1960s. Instead of inducing protection in young infants, the vaccine resulted in an exaggerated immune response resulting in enhanced respiratory disease and higher morbidity and mortality.
Several models have been developed to study the pathogenesis of HRSV infections. Continuous cell lines, like HEp-2 and A549, have been used extensively to study HRSV infection in vitro. Epithelial cells are the primary targets of HRSV infection5, therefore a lot of focus has been on these cell types. However, the in vivo situation is much more complex and not limited to one cell type. In order to examine these complex interactions, several animal infection models have been developed to study HRSV pathogenesis. Thus far, two different strategies have been employed, focusing either on heterologous (nonhuman) models as well as cognate host-pneumovirus models. Examples of heterologous models for HRSV include chimpanzees, sheep, cotton rats and mice. RSV infection in its natural host has been studied in cattle and in mice, using bovine RSV and pneumovirus, respectively.
Whilst each of these models has provided important insights into disease pathogenesis, HRSV is highly adapted to humans. Therefore, as an addition to the cell lines and animal models currently used, we propose to use human primary PBMCs as a model for stimulation with HRSV. Human PBMCs can be obtained from a range of individuals to address specific research questions, including young children6, immunocompromised individuals as well as healthy adults7 or elderly individuals8. PBMCs can be used to study innate aspects of infection as well as the adaptive response to the virus. Activated inflammatory pathways important for the pathogenesis of HRSV disease can be studied at the mRNA and protein level. Further, viral infection of (subsets of) immune cells can be determined by flow cytometry. We have used this model previously to identify the synergistic effects of HRSV infection together with the bacterial ligand muramyl dipeptide (MDP) on the induction of proinflammatory cytokines7. We propose to use HRSV stimulation of human primary PBMCs as a robust, easy and fast in vitro model to study inflammatory pathways involved in the pathogenesis of HRSV disease.

<table border="1">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Reagent</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Lonza</td>
			<td>BE17-516F</td>
			<td>PBS without Ca2+ Mg2+ or phenol red</td>
		</tr>
		<tr>
			<td>HBSS</td>
			<td>Invitrogen</td>
			<td>14025-100</td>
			<td>HBSS, Calcium, Magnesium, no Phenol Red</td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Invitrogen</td>
			<td>31966-047</td>
			<td>DMEM, High Glucose, GlutaMAX, Pyruvate</td>
		</tr>
		<tr>
			<td>RPMI</td>
			<td>Invitrogen</td>
			<td>72400054</td>
			<td>RPMI 1640 Medium, GlutaMAX, HEPES</td>
		</tr>
		<tr>
			<td>Reduced Serum Medium (Opti-MEM)</td>
			<td>Invitrogen</td>
			<td>51985-026</td>
			<td>Opti-MEM I Reduced Serum Medium, GlutaMAX</td>
		</tr>
		<tr>
			<td>FCS</td>
			<td>Greiner Bio-one</td>
			<td></td>
			<td>FBS (Fetal Bovine Serum)</td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Sigma Aldrich</td>
			<td>A7030</td>
			<td>Albumin from bovine serum</td>
		</tr>
		<tr>
			<td>P/S</td>
			<td>Invitrogen</td>
			<td>15140-122</td>
			<td>Penicillin-Streptomycin, liquid</td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Invitrogen</td>
			<td>25300-054</td>
			<td>0.05% Trypsin-EDTA (1x), Phenol Red</td>
		</tr>
		<tr>
			<td>HeLa cells</td>
			<td>ATCC</td>
			<td>CCL-2</td>
			<td></td>
		</tr>
		<tr>
			<td>A549 cells</td>
			<td>ATCC</td>
			<td>CCL-185</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma Aldrich</td>
			<td>S7903</td>
			<td>Sucrose BioXtra, &#8805;99.5%</td>
		</tr>
		<tr>
			<td>IgG2 anti-mouse-FITC</td>
			<td>BD Pharmingen</td>
			<td>555057</td>
			<td>Ms IgG2b K FITC isotype control</td>
		</tr>
		<tr>
			<td>Anti-NP-RSV-FITC</td>
			<td>Abcam</td>
			<td>ab25849</td>
			<td>Anti-Respiratory Syncytial Virus antibody [671] (FITC)</td>
		</tr>
		<tr>
			<td>Density gradient medium (Lymphoprep)</td>
			<td>Axis-Shield</td>
			<td>1114545</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA tubes</td>
			<td>BD Biosciences</td>
			<td>367525</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>Merck Millipore</td>
			<td>1000141000</td>
			<td>Acetone for analysis</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>[header]</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Material </td>
		</tr>
		<tr>
			<td>Centrifuge Tubes</td>
			<td>Beckman Coulter</td>
			<td>326823</td>
			<td>Thinwall, Polyallomer, 38.5 ml, 25 mm x 89 mm</td>
		</tr>
		<tr>
			<td>SureSpin 630 Rotor with 36 ml buckets</td>
			<td>Sorvall</td>
			<td>79368</td>
			<td>Swinging bucket titanium rotor</td>
		</tr>
		<tr>
			<td>Sorvall WX80 Ultracentrifuge</td>
			<td>Sorvall</td>
			<td>46900</td>
			<td></td>
		</tr>
		<tr>
			<td>CB 150l CO2 Incubator</td>
			<td>Binder</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

an in vitro model to study immune responses of human peripheral blood mononuclear cells to human respiratory syncytial virus infection

O vírus sincicial respiratório humano (HRSV) infecções apresentar um largo espectro de gravidade da doença, variando desde as infecções suaves a bronquiolite com risco de vida. Uma parte importante da patogênese da doença grave é uma resposta imune melhorada levando a imunopatologia. Aqui, nós descrevemos um protocolo usado para investigar a resposta imune de células do sistema imunológico humano a uma infecção HRSV. Em primeiro lugar, descrevemos métodos utilizados para a cultura, purificação e quantificação de HRSV. Subsequentemente, foi descrito um modelo humano in vitro, em que as células mononucleares do sangue periférico (PBMC) são estimuladas com HRSV vivo. Este sistema modelo pode ser usado para estudar vários parâmetros que podem contribuir para a gravidade da doença, incluindo a resposta imune inata e adaptativa. Estas respostas podem ser medido ao nível da transcrição e da tradução. Além disso, a infecção virai de células pode ser facilmente medida utilizando citometria de fluxo. Tomadojuntos, a estimulação de PBMC com HRSV vivo fornece um sistema modelo rápido e reprodutível para examinar os mecanismos envolvidos na doença induzida por HRSV.

The HRSV A2 cultivation and purification method described here will give a yield between 2 x&#160;107 and 2 x&#160;108 FITC-50 quantified infectious viral particles/ml. Results of microscopic analysis, where FITC-stained infected cells were counted (results not shown), were compared to results of quantification by FACS (Figure 1). Both methods resulted in similar titers, although the FACS method had a much lower standard deviation between different experiments and was less laborious...

Watch this Scientific Journal Video about An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection at JoVE.com

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

Human respiratory syncytial virus (HRSV) can cause severe bronchiolitis in young infants. Part of the pathogenesis of severe HRSV disease is caused by the host immune response. Stimulation of primary human immune cells with HRSV provides a fast and reproducible model system to study activation of inflammatory pathways and infection.

Uma<em&gt; In vitro</em&gt; Modelo para estudar respostas imunes de sangue periférico humano células mononucleares para Vírus Respiratório Sincicial Humano Infecção

Human respiratory syncytial  virus (HRSV) infections present a broad spectrum of disease severity, ranging  from mild infections to life-threatening ...

Research

JoVE Journal

Immunology and Infection

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

Um<em&gt; In vitro</em&gt; Modelo de Co-infecção para Estudar<em&gt; Plasmodium falciparum</em&gt;-HIV-1 Interacções em monócitos humanos primários derivados de células imunes

Plasmodium falciparum, the causative agent  of the deadliest form of malaria, and human immunodeficiency virus type-1 (HIV-1) are among the most important ...

Imunologia e Infecção

Establishing a Liquid-covered Culture of Polarized Human Airway Epithelial Calu-3 Cells to Study Host Cell Response to Respiratory Pathogens In vitro

Establishing a Liquid-covered Culture of Polarized Human Airway Epithelial Calu-3 Cells to Study Host Cell Response to Respiratory Pathogens In vitro

Estabelecer uma cultura líquida, coberta de vias respiratórias humanas epiteliais Polarized Calu-3 células para estudar a resposta Cell Host para patógenos respiratórios<em&gt; In vitro</em

The  apical and basolateral surfaces of airway epithelial cells demonstrate  directional responses to pathogen exposure in  vivo. Thus, ideal in vitro ...

An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization

An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization

Um<em&gt; In vitro</em&gt; Modelo para estudar a heterogeneidade de diferenciação de macrófagos humanos e Polarização

Monocyte-derived macrophages represent an important cell type of the innate immune system. Mouse models studying macrophage biology suffer from the ...

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Um<em&gt; In Vitro</em&gt; Modelo para medição de respostas imunes a Malária no Contexto do HIV co-infecção

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. ...

A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood

Uma abordagem automatizada-Semi Preparando Antibody Cocktails para Análise imunofenotípica de Sangue Periférico Humano

Immunophenotyping of peripheral blood by flow cytometry determines changes in the frequency and activation status of peripheral leukocytes during disease ...

A Galleria mellonella Oral Administration Model to Study Commensal-Induced Innate Immune Responses

A Galleria mellonella Oral Administration Model to Study Commensal-Induced Innate Immune Responses

Um modelo de administração Oral Galleria mellonella para respostas imunes Study Commensal-Induced inata

The investigation of the immunogenic potential of commensal bacteria on the host immune system is one essential component when studying intestinal ...

Isolation of Tonsillar Mononuclear Cells to Study Ex Vivo Innate Immune Responses in a Human Mucosal Lymphoid Tissue

Isolamento de células mononucleares tonsiláres para estudar respostas imunes inatas ex vivo em um tecido linfoide mucosal humano

Studying isolated cells from mucosa-associated lymphoid tissues (MALT) allows understanding of immune cells response in pathologies involving mucosal ...

Assessing Respiratory Immune Responses to Haemophilus Influenzae

Assessing Respiratory Immune Responses to Haemophilus Influenzae

Avaliando as respostas imunes respiratórias ao Haemophilus influenzae

Haemophilus influenzae (Hi) is a prevalent bacterium found in a range of respiratory conditions. A variety of different assays/techniques may be used to ...

Xenograft Skin Model to Manipulate Human Immune Responses In Vivo

Xenograft Skin Model to Manipulate Human Immune Responses In Vivo

Modelo de pele de xenoenxerto para manipular respostas imunes humanas in vivo

The human skin xenograft model, in which human donor skin is transplanted onto an immunodeficient mouse host, is an important option for translational ...

Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

Separação de subpopulações de células imunes em amostras de sangue periférico de crianças com mononucleose infecciosa

Infectious mononucleosis (IM) is an acute syndrome mostly associated with primary Epstein&#8211;Barr virus (EBV) infection. The main clinical symptoms ...