Name: measuring granulocyte and monocyte phagocytosis and oxidative burst activity in human blood
Uploaded: 2016-09-12T13:00:00
Description: Watch this Scientific Journal Video about Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood at JoVE.com

The authors would like to acknowledge Zack Shue, Casey John, and Lynn Cialdella-Kam for their assistance during the optimization of this procedure.

NOTE: All blood collection procedures were conducted in accordance with the guidelines set forth by the Appalachian State University (ASU) Institutional Review Board (IRB).
1. Assay Preparation
<ol>
	<li>Gather, prepare, and organize the materials specified for the procedure (please refer to Table of Specific Materials/Equipment).
	<ol>
		<li>Label 12 x 75 mm tubes.
		<ol>
			<li>Label two tubes for each participant: one tube for the phagocytosis assay (label this tube with black ink and an ...

<ol>
	<li>Nieman, D. C., 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=Influence+of+pistachios+on+performance+and+exercise-induced+inflammation,+oxidative+stress,+immune+dysfunction,+and+metabolite+shifts+in+cyclists:+a+randomized,+crossover+trial.">Influence of pistachios on performance and exercise-induced inflammation, oxidative stress, immune dysfunction, and metabolite shifts in cyclists: a randomized, crossover trial.</a> PLoS One. 9, e113725 (2014).</li><li>Knab, A. M., 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=Effects+of+a+freeze-dried+juice+blend+powder+on+exercise-induced+inflammation,+oxidative+stress,+and+immune+function+in+cyclists.">Effects of a freeze-dried juice blend powder on exercise-induced inflammation, oxidative stress, and immune function in cyclists.</a> Appl Physiol Nutr Metab. 39, 381-385 (2014).</li><li>Konrad, M., 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=The+acute+effect+of+ingesting+a+quercetin-based+supplement+on+exercise-induced+inflammation+and+immune+changes+in+runners.">The acute effect of ingesting a quercetin-based supplement on exercise-induced inflammation and immune changes in runners.</a> Int J Sport Nutr Exerc Metab. 21, 338-346 (2011).</li><li>Nieman, D. C., 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=Immune+and+inflammation+responses+to+a+3-day+period+of+intensified+running+versus+cycling.">Immune and inflammation responses to a 3-day period of intensified running versus cycling.</a> Brain Behav Immun. 39, 180-185 (2014).</li><li>Nieman, D. C., 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=Carbohydrate+supplementation+affects+blood+granulocyte+and+monocyte+trafficking+but+not+function+after+2.5+h+of+running.">Carbohydrate supplementation affects blood granulocyte and monocyte trafficking but not function after 2.5 h of running.</a> Am J Clin Nutr. 66, 153-159 (1997).</li><li>Tidball, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammatory+cell+response+to+acute+muscle+injury.">Inflammatory cell response to acute muscle injury.</a> Med Sci Sports Exerc. 27, 1022-1032 (1995).</li><li>McFarlin, B. K., Williams, R. R., Venable, A. S., Dwyer, K. C., Haviland, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image-based+cytometry+reveals+three+distinct+subsets+of+activated+granulocytes+based+on+phagocytosis+and+oxidative+burst.">Image-based cytometry reveals three distinct subsets of activated granulocytes based on phagocytosis and oxidative burst.</a> Cytometry A. 83, 745-751 (2013).</li><li>Liao, Y., Liu, P., Guo, F., Zhang, Z. Y., Zhang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxidative+burst+of+circulating+neutrophils+following+traumatic+brain+injury+in+human.">Oxidative burst of circulating neutrophils following traumatic brain injury in human.</a> PLoS One. 8, e68963 (2013).</li><li>McFarlin, B. K., Venable, A. S., Prado, E. A., Henning, A. L., Williams, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image-based+flow+cytometry+technique+to+evaluate+changes+in+granulocyte+function+in+vitro.">Image-based flow cytometry technique to evaluate changes in granulocyte function in vitro.</a> J Vis Exp. , (2014).</li><li>Nieman, D. C., 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=Immune+response+to+obesity+and+moderate+weight+loss.">Immune response to obesity and moderate weight loss.</a> Int J Obes Relat Metab Disord. 20, 353-360 (1996).</li><li>Nieman, D. C., 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=Immune+function+in+female+elite+rowers+and+non-athletes.">Immune function in female elite rowers and non-athletes.</a> Br J Sports Med. 34, 181-187 (2000).</li><li>Ichii, H., 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=Iron+sucrose+impairs+phagocytic+function+and+promotes+apoptosis+in+polymorphonuclear+leukocytes.">Iron sucrose impairs phagocytic function and promotes apoptosis in polymorphonuclear leukocytes.</a> Am J Nephrol. 36, 50-57 (2012).</li><li>Gomes, A., Fernandes, E., Lima, 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=Fluorescence+probes+used+for+detection+of+reactive+oxygen+species.">Fluorescence probes used for detection of reactive oxygen species.</a> J Biochem Biophys Methods. 65, 45-80 (2005).</li><li>Gruger, 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=Negative+impact+of+linezolid+on+human+neutrophil+functions+in+vitro.">Negative impact of linezolid on human neutrophil functions in vitro.</a> Chemotherapy. 58, 206-211 (2012).</li><li>Dementhon, K., El-Kirat-Chatel, S., Noel, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+in+vitro+model+for+the+multi-parametric+quantification+of+the+cellular+interactions+between+Candida+yeasts+and+phagocytes.">Development of an in vitro model for the multi-parametric quantification of the cellular interactions between Candida yeasts and phagocytes.</a> PLoS One. 7, e32621 (2012).</li></ol>

This manuscript provides a step-by-step protocol for the determination of two indicators of granulocyte and monocyte function. We have identified a few steps that are critical to the success of this assay. One such step is the ice-bath incubation that occurs immediately prior to the addition of bacteria. Thorough cooling of all samples will minimize the effect of temperature on phagocytosis and OB activity. Another critical step is the addition of bacteria to each sample. A 8:1 bacteria:phagocyte ratio produces optimal r...

The granulocyte and monocyte phagocytosis/oxidative burst (OB) activity assay is a simple, straight-forward technique that is frequently used to gather information about innate immune function following prolonged and intensive exercise.1-4 When studying the response of the immune system to a stimulus, granulocytes and monocytes are of particular interest because they play a central role in host defense and are the first immune cells to accumulate at the site of infection.5 Neutrophils, a type of granulocyte, are the first cells to translocate into damaged muscle tissue following intensive exercise.6 Phagocytosis and OB activity are the most common measures of granulocyte and monocyte function,7 and are preferred as indicators of innate immune cell function because of their central role in both the infection process and the repair process.
The assay described in this manuscript utilizes a basic flow cytometer to provide a quantitative determination of granulocyte and monocyte phagocytosis and OB activity in whole blood. The use of whole blood in this technique is advantageous because it allows the assay to be conducted without a time-consuming purification procedure. This assay can easily be modified to accommodate a variety of research designs and lab capabilities. For example, Liao et al. utilized a similar technique to show that neutrophil OB activity is increased and neutrophil phagocytosis is decreased following traumatic brain injury.8 In another example, McFarlin et al. described an elegant modification of this technique that uses allophycocyanin (APC)-conjugated CD66b and high-performance tandem APC-conjugated CD45 labeling with image-based flow cytometry to classify granulocytes in terms of their phagocytosis and OB activities.7,9
Our research group has used various adaptations of this technique as an indicator of innate immune function in overweight, obese, and athletic populations for almost 20 years.10,11 The text below will provide the reader with step-by-step instructions for performing this assay and highlight areas that the reader may adapt to meet the needs of their research.

<table border="0" cellpadding="0" cellspacing="0" width="1070">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">12 x 75 mm tubes, with cap</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">20170-579</td>
			<td style="width:425px;">You will need 2 tubes per sample (&#34;H&#34; and &#34;F&#34;) plus 3 tubes per batch (for assay controls; &#34;F&#34;, &#34;H&#34;, and &#34;Blood only&#34;).</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">PBS (10X), pH 7.4</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">70011-044</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Bottle top 0.2 &#181;m cellulose acetate filter (500 ml capacity)</td>
			<td style="width:215px;">Fisher Scientific</td>
			<td style="width:215px;">09-741-07&#160;</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Glucose, powder</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">15023-021</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:215px;">Citric acid (anhydrous, cell culture tested, plant cell culture tested)</td>
			<td style="width:215px;">Sigma-Aldrich</td>
			<td style="width:215px;">C2404-100G</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Sodium citrate tribasic dihydrate</td>
			<td style="width:215px;">Sigma-Aldrich</td>
			<td style="width:215px;">S4641-25G</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Dihydroethidium (Hydroethidine) (HE)</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">D11347</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Dimethyl sulfoxide (DMSO) Anhydrous</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">D12345</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:215px;">Staphylococcus aureus&#160;(Wood strain without protein A) BioParticles, fluorescein conjugate (FITC)</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">S2851</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:215px;">Staphylococcus aureus&#160;(Wood strain without protein A) BioParticles, unlabeled</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">S-2859</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Trypan Blue Solution, 0.4%</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">15250-061</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:215px;">4.0 mL vacutainer containing 7.2 mg K2EDTA, spray-dried</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">BD367861</td>
			<td style="width:425px;">You will need 1 K2 EDTA blood collection tube per sample.</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:215px;">4.0 mL vacutainer containing 68 USP units Lithium Heparin, spray-coated</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">BD367884</td>
			<td style="width:425px;">You will need 1 Lithium Heparin blood collection tube per sample.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER Ac&#183;T 5diff CP</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">6605705</td>
			<td style="width:425px;">i.e., hematology analyzer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER Ac&#183;T 5diff Rinse</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547167</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER Ac&#183;T 5diff Fix</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547171</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER Ac&#183;T 5diff WBC Lyse</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547170</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER Ac&#183;T 5diff Hgb Lyse</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547168</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">COULTER Ac&#183;T 5diff Cal Calibrator</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">7547175</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">COULTER Ac&#183;T 5diff Control Plus</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">7547198</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">AcT 5diff Diluent</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547169</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">200 &#181;L extended-length pipette tips</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">37001-526</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Insulated ice pan</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">89198-980</td>
			<td style="width:425px;">For ice water bath.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Open metal tube rack</td>
			<td style="width:215px;">VWR</td>
			<td style="width:215px;">60916-702</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Fetal Bovine Serum</td>
			<td style="width:215px;">Life Technologies</td>
			<td style="width:215px;">26140-111</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">TQ-Prep Workstation</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">6605429</td>
			<td style="width:425px;">i.e., automated cell lyse preparation workstation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">ImmunoPrep Reagent System</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">7546999</td>
			<td style="width:425px;">i.e., RBC lyse/WBC fix reagent system</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:215px;">Cytomics FC500 MCL Flow Cytometry System with CXP Software</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">626553</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">IsoFlow Sheath Fluid</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8547008</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">COULTER CLENZ</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">8546929</td>
			<td style="width:425px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Flow-Check Fluorospheres&#160;</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:215px;">6605359</td>
			<td style="width:425px;">i.e., alignment and fluidics verification fluorospheres</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">FlowJo Software</td>
			<td style="width:215px;">FlowJo</td>
			<td style="width:215px;">FlowJo</td>
			<td style="width:425px;">i.e., flow cytometry analysis software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Excel Software</td>
			<td style="width:215px;">Microsoft</td>
			<td style="width:215px;">Microsoft Excel</td>
			<td style="width:425px;">i.e., electronic spreadsheet program</td>
		</tr>
	</tbody>
</table>

measuring granulocyte and monocyte phagocytosis and oxidative burst activity in human blood

The granulocyte and monocyte phagocytosis and oxidative burst (OB) activity assay can be used to study the innate immune system. This manuscript provides the necessary methodology to add this assay to an exercise immunology arsenal. The first step in this assay is to prepare two aliquots ("H" and "F") of whole blood (heparin). Then, dihydroethidium is added to the H aliquot, and both aliquots are incubated in a warm water bath followed by a cold water bath. Next, Staphylococcus aureus (S. aureus) is added to the H aliquot and fluorescein isothiocyanate-labeled S. aureus is added to the F aliquot (bacteria:phagocyte = 8:1), and both aliquots are incubated in a warm water bath followed by a cold water bath. Then, trypan blue is added to each aliquot to quench extracellular fluorescence, and the cells are washed with phosphate-buffered saline. Next, the red blood cells are lysed, and the white blood cells are fixed. Finally, a flow cytometer and appropriate analysis software are used to measure granulocyte and monocyte phagocytosis and OB activity. This assay has been used for over 20 years. After heavy and prolonged exertion, athletes experience a significant but transient increase in phagocytosis and an extended decrease in OB activity. The post-exercise increase in phagocytosis is correlated with inflammation. In contrast to normal weight individuals, granulocyte and monocyte phagocytosis is chronically elevated in overweight and obese participants, and is modestly correlated with C-reactive protein. In summary, this flow cytometry-based assay measures the phagocytosis and OB activity of phagocytes and can be used as an additional measure of exercise- and obesity-induced inflammation.

Prolonged and intensive exercise has profound effects on innate immune function, including natural killer cell function, macrophage cytokine-mediated response to viral infection, and granulocyte and monocyte phagocytosis and OB activity. Multiple studies indicate that granulocyte and monocyte phagocytosis increases significantly post-exercise, reflecting the inflammation induced by muscle damage and metabolic demands. In contrast, granulocyte and monocyte OB activity decreases post-exerci...

Watch this Scientific Journal Video about Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood at JoVE.com

Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood

The purpose of this manuscript is to present a method for measuring monocyte and granulocyte phagocytosis and oxidative burst activity in human blood samples.

The granulocyte and monocyte phagocytosis and oxidative burst (OB) activity assay can be used to study the innate immune system. This manuscript provides ...

Research

JoVE Journal

Immunology and Infection

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1&#946; in Human Monocyte-derived Dendritic Cells

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1ÃƒÅ½Ã‚Â² in Human Monocyte-derived Dendritic Cells

Inflammatory processes resulting from the secretion of Interleukin (IL)-1 family cytokines by immune cells lead to local or systemic inflammation, tissue ...

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

Mitochondrial dysfunction is known to play a significant role in a number of pathological conditions such as atherosclerosis, diabetes, septic shock, and ...

Culture of Macrophage Colony-stimulating Factor Differentiated Human Monocyte-derived Macrophages

A protocol is presented for cell culture of macrophage colony-stimulating factor (M-CSF) differentiated human monocyte-derived macrophages. For initiation ...

Flow Cytometric Analysis of Natural Killer Cell Lytic Activity in Human Whole Blood

NK cell cytotoxicity is a widely used measure to determine the effect of outside intervention on NK cell function. However, the accuracy and ...

Generation of Human Monocyte-derived Dendritic Cells from Whole Blood

Dendritic cells (DCs) recognize foreign structures of different pathogens, such as viruses, bacteria, and fungi, via a variety of pattern recognition ...

Use of a Monocyte Monolayer Assay to Evaluate Fc&#947; Receptor-mediated Phagocytosis

Use of a Monocyte Monolayer Assay to Evaluate FcÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â³ Receptor-mediated Phagocytosis

Although originally developed for predicting transfusion outcomes of serologically incompatible blood, the monocyte monolayer assay (MMA) is a highly ...

Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells

The efficient transduction of specific genes into natural killer (NK) cells has been a major challenge. Successful transductions are critical to defining ...

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Chemotaxis is migration along a specific chemical gradient1. Chemokines are chemotactic cytokines that promote cellular trafficking with anatomic and ...

Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis

Monocytes are key contributors in various inflammatory disorders and alterations to these cells, including their subset proportions and functions, can ...

Pattern-Triggered Oxidative Burst and Seedling Growth Inhibition Assays in Arabidopsis thaliana

Pattern-Triggered Oxidative Burst and Seedling Growth Inhibition Assays in Arabidopsis thaliana

Plants have evolved a robust immune system to perceive pathogens and protect against disease. This paper describes two assays that can be used to measure ...