Name: in vitro methods for comparing target binding and cdc induction between therapeutic antibodies: applications in biosimilarity analysis
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Description: Watch this Scientific Journal Video about In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis at JoVE.com

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1. Evaluación de la unión de diana por citometría de flujo <ol><li> Preparación de materiales biológicos y reactivos <ol><li> Hacer 500 mL de medio de cultivo RPMI suplementado con 10% de suero fetal bovino inactivado por calor (H-IFBS). </li><li> Cultivo de linfoma Daudi Burkitt (Daudi) y células Daudi GFP + usando RPMI y frascos de cultivo de 75 cm2. Mantener los cultivos a 37 ° C en una atmósfera humidificada al 5% de CO2 hasta que alcancen 6 - 9 x 105 células / ml. </li><li> Ha...

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A., 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=Comparability+of+Antibody-Mediated+Cell+Killing+Activity+Between+a+Proposed+Biosimilar+RTXM83+and+the+Originator+Rituximab.">Comparability of Antibody-Mediated Cell Killing Activity Between a Proposed Biosimilar RTXM83 and the Originator Rituximab.</a> Bio Drugs. 30 (3), 225-231 (2016).</li><li>Iwamoto, N., 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=Validated+LC/MS+Bioanalysis+of+Rituximab+CDR+Peptides+Using+Nano-surface+and+Molecular-Orientation+Limited+(nSMOL)+Proteolysis.">Validated LC/MS Bioanalysis of Rituximab CDR Peptides Using Nano-surface and Molecular-Orientation Limited (nSMOL) Proteolysis.</a> Biol Pharm Bull. 39 (7), 1187-1194 (2016).</li><li>Chapman, K., 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=Waiving+in+vivo+studies+for+monoclonal+antibody+biosimilar+development:+National+and+global+challenges.">Waiving in vivo studies for monoclonal antibody biosimilar development: National and global challenges.</a> MAbs. 8 (3), 427-435 (2016).</li><li>Zembruski, N. 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P., 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=Assessment+of+physicochemical+properties+of+rituximab+related+to+its+immunomodulatory+activity.">Assessment of physicochemical properties of rituximab related to its immunomodulatory activity.</a> J Immunol Res. 2015, 910763 (2015).</li><li>Visser, 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=Physicochemical+and+functional+comparability+between+the+proposed+biosimilar+rituximab+GP2013+and+originator+rituximab.">Physicochemical and functional comparability between the proposed biosimilar rituximab GP2013 and originator rituximab.</a> BioDrugs. 27 (5), 495-507 (2013).</li><li>Ylera, F., 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=Off-rate+screening+for+selection+of+high-affinity+anti-drug+antibodies.">Off-rate screening for selection of high-affinity anti-drug antibodies.</a> Anal Biochem. 441 (2), 208-213 (2013).</li><li>Broyer, L., Goetsch, L., Broussas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+complement-dependent+cytotoxicity+using+ATP+measurement+and+C1q/C4b+binding.">Evaluation of complement-dependent cytotoxicity using ATP measurement and C1q/C4b binding.</a> Methods Mol Biol. 988, 319-329 (2013).</li><li>Herbst, R., 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=B-cell+depletion+in+vitro+and+in+vivo+with+an+afucosylated+anti-CD19+antibody.">B-cell depletion in vitro and in vivo with an afucosylated anti-CD19 antibody.</a> J Pharm Exp Ther. 335 (1), 213-222 (2010).</li><li>Lazar, G. A., 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=Engineered+antibody+Fc+variants+with+enhanced+effector+function.">Engineered antibody Fc variants with enhanced effector function.</a> Proc Natl Acad Sci U S A. 103 (11), 4005-4010 (2006).</li><li>Winiarska, 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=Statins+impair+antitumor+effects+of+rituximab+by+inducing+conformational+changes+of+CD20.">Statins impair antitumor effects of rituximab by inducing conformational changes of CD20.</a> PLoS medicine. 5 (3), e64 (2008).</li><li>Zhou, X., Hu, W., Qin, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+complement+in+the+mechanism+of+action+of+rituximab+for+B-cell+lymphoma:+implications+for+therapy.">The role of complement in the mechanism of action of rituximab for B-cell lymphoma: implications for therapy.</a> Oncologist. 13 (9), 954-966 (2008).</li><li>Hayashi, K., 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=Gemcitabine+enhances+rituximab-mediated+complement-dependent+cytotoxicity+to+B+cell+lymphoma+by+CD20+upregulation.">Gemcitabine enhances rituximab-mediated complement-dependent cytotoxicity to B cell lymphoma by CD20 upregulation.</a> Cancer Sci. 107 (5), 682-689 (2016).</li><li>Mossner, E., 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=Increasing+the+efficacy+of+CD20+antibody+therapy+through+the+engineering+of+a+new+type+II+anti-CD20+antibody+with+enhanced+direct+and+immune+effector+cell-mediated+B-cell+cytotoxicity.">Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell-mediated B-cell cytotoxicity.</a> Blood. 115 (22), 4393-4402 (2010).</li><li>Lapalombella, R., 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=A+novel+Raji-Burkitt's+lymphoma+model+for+preclinical+and+mechanistic+evaluation+of+CD52-targeted+immunotherapeutic+agents.">A novel Raji-Burkitt's lymphoma model for preclinical and mechanistic evaluation of CD52-targeted immunotherapeutic agents.</a> Clin Cancer Res. 14 (2), 569-578 (2008).</li><li>Mitoma, 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=Mechanisms+for+cytotoxic+effects+of+anti-tumor+necrosis+factor+agents+on+transmembrane+tumor+necrosis+factor+alpha-expressing+cells:+comparison+among+infliximab,+etanercept,+and+adalimumab.">Mechanisms for cytotoxic effects of anti-tumor necrosis factor agents on transmembrane tumor necrosis factor alpha-expressing cells: comparison among infliximab, etanercept, and adalimumab.</a> Arthritis Rheum. 58 (5), 1248-1257 (2008).</li><li>Kaymakcalan, Z., 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=Comparisons+of+affinities,+avidities,+and+complement+activation+of+adalimumab,+infliximab,+and+etanercept+in+binding+to+soluble+and+membrane+tumor+necrosis+factor.">Comparisons of affinities, avidities, and complement activation of adalimumab, infliximab, and etanercept in binding to soluble and membrane tumor necrosis factor.</a> Clin Immunol. 131 (2), 308-316 (2009).</li><li>Zent, C. 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=Direct+and+complement+dependent+cytotoxicity+in+CLL+cells+from+patients+with+high-risk+early-intermediate+stage+chronic+lymphocytic+leukemia+(CLL)+treated+with+alemtuzumab+and+rituximab.">Direct and complement dependent cytotoxicity in CLL cells from patients with high-risk early-intermediate stage chronic lymphocytic leukemia (CLL) treated with alemtuzumab and rituximab.</a> Leuk Res. 32 (12), 1849-1856 (2008).</li><li>Goswami, M. 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=Regulation+of+complement-dependent+cytotoxicity+by+TGF-beta-induced+epithelial-mesenchymal+transition.">Regulation of complement-dependent cytotoxicity by TGF-beta-induced epithelial-mesenchymal transition.</a> Oncogene. 35 (15), 1888-1898 (2016).</li><li>Wang, A., 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=Induction+of+anti-EGFR+immune+response+with+mimotopes+identified+from+a+phage+display+peptide+library+by+panitumumab.">Induction of anti-EGFR immune response with mimotopes identified from a phage display peptide library by panitumumab.</a> Oncotarget. , (2016).</li><li>Ueda, N., 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+cytotoxic+effects+of+certolizumab+pegol+and+golimumab+mediated+by+transmembrane+tumor+necrosis+factor+alpha.">The cytotoxic effects of certolizumab pegol and golimumab mediated by transmembrane tumor necrosis factor alpha.</a> Inflamm Bowel Dis. 19 (6), 1224-1231 (2013).</li><li>Nesbitt, A., 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=Mechanism+of+action+of+certolizumab+pegol+(CDP870):+in+vitro+comparison+with+other+anti-tumor+necrosis+factor+alpha+agents.">Mechanism of action of certolizumab pegol (CDP870): in vitro comparison with other anti-tumor necrosis factor alpha agents.</a> Inflamm Bowel Dis. 13 (11), 1323-1332 (2007).</li><li>Teeling, J. 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La expiración de la patente de un mAb terapéutico está promoviendo el desarrollo de los biosimilares. Por lo tanto, existe una necesidad de métodos simples que pueden identificar diferencias en las actividades de relevancia clínica de estos productos. Se emplearon células CD20 + cultivadas para la evaluación de dos características funcionales clave de rituximab: unión a la diana y la inducción CDC. La actividad anterior requiere el reconocimiento de CD20 por la región Fab del mAb, mientras que el ?...

Los anticuerpos terapéuticos son anticuerpos monoclonales recombinantes (mAbs) desarrollados para el tratamiento de diferentes patologías, incluyendo cánceres, enfermedades autoinmunes y crónicas, trastornos neurológicos y otros 1 . Actualmente, la FDA ha concedido la aprobación a más de 40 mAb terapéuticos, y se espera que lleguen más al mercado en los años siguientes. Rituximab es un anticuerpo IgG1 monoclonal quimérico de alta afinidad aprobado para el tratamiento del linfoma no Hodgkin de células B CD20 + , LNH folicular CD20 + , leucemia linfocítica crónica y artritis reumatoide 2 , 3 . El reconocimiento de CD20, que está sobreexpresado en células B, por rituximab induce apoptosis; activación del complemento; Y citotoxicidad mediada por células dependientes de anticuerpos (ADCC) 3 . Las patentes de este fármaco expiraron en Europa y en los EE.UU. en 2013 y 2016, Respectivamente. Así, las compañías farmacéuticas en todo el mundo están desarrollando biosimilares de rituximab. Como en cualquier otro medicamento para consumo humano, los biosimilares requieren la aprobación de las agencias reguladoras. Directrices internacionales indican que para los mAbs, la biosimilaridad debe demostrarse mediante la comparación de las características físico-químicas, farmacocinéticas, la eficacia y la seguridad de los productos nuevos y de referencia [ 4] . Por consiguiente, las metodologías utilizadas en tales comparaciones deben evaluar las características estructurales y funcionales de los mAb, especialmente aquellos con relevancia clínica. Para ello, los ensayos in vitro muestran varias ventajas sobre los experimentos in vivo (revisados ​​en Chapman et al. ) 5 : i) los estudios in vitro son más sensibles a las diferencias entre el biosimilar propuesto y el producto de referencia; Ii) deben realizarse estudios in vivo en especies relevantes, que para muchos mAbs sonPrimates no humanos; Y iii) ya que el mecanismo de acción, la toxicología preclínica y los efectos clínicos del producto de referencia son bien conocidos, los estudios in vivo con biosimilares pueden no proporcionar información útil adicional. En consecuencia, la Orientación de la Unión Europea para biosimilares permite a los candidatos a entrar en ensayos clínicos basados ​​en sólidos datos in vitro sólo [ 6] . Presentamos dos ensayos rápidos, económicos y sencillos que evalúan la actividad biológica de rituximab utilizando células cultivadas con CD20 + . Estos ensayos pueden incluirse como parte del ejercicio de comparabilidad para los candidatos biosimilares de rituximab.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>RPMI-1640 medium</td>
			<td>ATCC</td>
			<td>30-2001</td>
			<td>Modify the culture depending on the cell line</td>
		</tr>
		<tr>
			<td>Trypan Blue solution</td>
			<td>Sigma</td>
			<td>T8154</td>
			<td>0.4%, liquid, sterile-filtered, suitable for cell culture</td>
		</tr>
		<tr>
			<td>Daudi Burkitt&#39;s Lymphoma Cells</td>
			<td>ATCC</td>
			<td>CCL-213</td>
			<td>You can modify the cell line depending on the antibody of interest</td>
		</tr>
		<tr>
			<td>Fetal bovine serum(FBS)</td>
			<td>GIBCO</td>
			<td>16000-044</td>
			<td>You can modify the source of serum depending of requirements of the cell line</td>
		</tr>
		<tr>
			<td>Normal Human Serum Complement</td>
			<td>Quidel</td>
			<td>A113</td>
			<td>It is therefore appropriate for use in biocompatibility experiments including drug development, biomaterials testing and other applications</td>
		</tr>
		<tr>
			<td>7AA-D</td>
			<td>BDPharmigen</td>
			<td>559925</td>
			<td>You can use broad range of color options, compatible with most instrument configurations for to analyze viability.</td>
		</tr>
		<tr>
			<td>PECy5 Mouse Anti-human IgG</td>
			<td>BDPharmigen</td>
			<td>551497</td>
			<td>Change fluorochrome depending on the filter and laser of your flow cytometer.</td>
		</tr>
		<tr>
			<td>Human IgG Isotype Control</td>
			<td>ThermoFisher Scientific</td>
			<td>07-7102</td>
			<td>Change depending to mAb</td>
		</tr>
		<tr>
			<td>BDCytofix</td>
			<td>BDPharmigen</td>
			<td>554655</td>
			<td>Flow Cytometry Fixation Buffer (1-4% formaldehyde or paraformaldehyde )</td>
		</tr>
		<tr>
			<td>PBS pH 7.4 10X (Phosphate buffer saline)</td>
			<td>GIBCO</td>
			<td>70011-044</td>
			<td>Phosphatebuffer without Ca2+/Mg2+ [137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, 1.46 mM KH2PO4] and endotoxin free.</td>
		</tr>
		<tr>
			<td>Cell culture plates 96 well, V-bottom</td>
			<td>Corning</td>
			<td>29442-068</td>
			<td>12 x 75 mm round bottom test tubes or 96-well V- or U- bottom microtiter plates</td>
		</tr>
		<tr>
			<td>MabThera (Rituximab)</td>
			<td>Roche</td>
			<td>&#8212;</td>
			<td>Reference product</td>
		</tr>
		<tr>
			<td>Rituximab</td>
			<td>Indian</td>
			<td>&#8212;</td>
			<td>Biosimilar product</td>
		</tr>
		<tr>
			<td>15- or 50-mL conical centrifuge tubes</td>
			<td>Corning</td>
			<td>430290 or 430052</td>
			<td>&#8212;</td>
		</tr>
		<tr>
			<td>Pipette Tips</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Multiple volume configurations are necessary</td>
		</tr>
		<tr>
			<td>Pipettes</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Adjustable-volume pipettes are necessary</td>
		</tr>
		<tr>
			<td>Centrifuge 5430/ 5430R model</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Refrigerated variable-speed centrifuge (4 to 25 &#176; C) with speeds ranging from 10 to 30,130 &#215; g</td>
		</tr>
		<tr>
			<td>Flow cytometer</td>
			<td>BD Dickinson</td>
			<td>&#8212;</td>
			<td>BD FACSAria III or other flow cytometer</td>
		</tr>
		<tr>
			<td>Olympus optical and light microscope</td>
			<td>Olympus</td>
			<td>&#8212;</td>
			<td>To quantify and evaluate cell growth</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>SANYO</td>
			<td>&#8212;</td>
			<td>Incubatorfor temperature andCO2 control to culture cells</td>
		</tr>
		<tr>
			<td>Biological Safety Cabinet</td>
			<td>CHC BIOLUS</td>
			<td>&#8212;</td>
			<td>Biological safety cabinet that is used to protect the researcher, product and environment.</td>
		</tr>
	</tbody>
</table>

in vitro methods for comparing target binding and cdc induction between therapeutic antibodies: applications in biosimilarity analysis

anticuerpos monoclonales terapéuticos (mAbs) son relevantes para el tratamiento de diferentes patologías, incluyendo los cánceres. El desarrollo de anticuerpos monoclonales biosimilares por las compañías farmacéuticas es una oportunidad de mercado, pero también es una estrategia para aumentar la accesibilidad de drogas y reducir los costes asociados a la terapia. Los protocolos detallados aquí describen la evaluación de unión a la diana y la inducción CDC por rituximab en células Daudi. Estas dos funciones requieren diferentes regiones estructurales del anticuerpo y son relevantes para el efecto clínico inducida por rituximab. Los protocolos permiten la comparación de lado a lado de un rituximab referencia y un biosimilar rituximab comercializado. Los productos evaluados mostraron diferencias tanto en unión a la diana y la inducción CDC, lo que sugiere que hay diferencias fisicoquímicas subyacentes y poner de relieve la necesidad de analizar el impacto de esas diferencias en el entorno clínico. Los métodos aquí presentados constituyen simple y barato in vitro &lt;/ Em&gt; para la evaluación de la actividad de rituximab biosimilares. Por lo tanto, pueden ser útiles durante el desarrollo biosimilar, así como para el control de calidad en la producción biosimilar. Además, los métodos presentados pueden extrapolarse a otros mAbs terapéuticos.

Usando los protocolos descritos anteriormente, se comparó la unión de diana y la inducción de CDC de rituximab de referencia en paralelo con las de un rituximab biosimilar producido y comercialmente disponible en Asia. En las células de Daudi, ambos mAbs se unieron a CD20 de una manera dependiente de la concentración ( Figura 1D ). Las regresiones no lineales de datos de unión mostró un r 2 d...

Watch this Scientific Journal Video about In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis at JoVE.com

In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis

Este protocolo describe la comparación in vitro de dos características funcionales clave del rituximab: la unión al diana y la inducción de la citotoxicidad dependiente del complemento (CDC). Los métodos se emplearon para una comparación lado a lado entre el rituximab de referencia y un biosimilar de rituximab. Estos ensayos se pueden emplear durante el desarrollo biosimilar o como un control de calidad en su producción.

Métodos in vitro para comparar la unión de diana y la inducción de CDC entre anticuerpos terapéuticos: Aplicaciones en análisis de biosimilitud

Therapeutic monoclonal antibodies (mAbs) are relevant to the treatment of different pathologies, including cancers. The development of biosimilar mAbs by ...

Research

JoVE Journal

Immunology and Infection

In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

Aislamiento de linfocitos T humanos, cultura y análisis de la migración In Vitro</em

The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular ...

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Inmunología e Infección

Introducing Shear Stress in the Study of Bacterial Adhesion

La introducción de la tensión de cizallamiento en el Estudio de la adhesión bacteriana

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

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

El establecimiento de un cultivo líquido cubierto de las vías respiratorias humanas epiteliales polarizadas Calu-3 células para estudiar la respuesta del huésped celular a patógenos respiratorios<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 ...

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Extracción de proteínas totales y 2-D Métodos gel de electroforesis para<em&gt; Burkholderia</em&gt; Especies

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

Una Novela<em&gt; In vitro</em&gt; Modelo para el estudio de las interacciones entre Whole Sangre humana y del endotelio

The majority of all known diseases are accompanied by disorders of the cardiovascular system. Studies into the complexity of the interacting pathways ...

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

Aislamiento de neutrófilos y análisis para determinar su papel en el linfoma de células sensibles a los agentes terapéuticos

Neutrophils are the most abundant (40% to 75%) type of white blood cells and among the first inflammatory cells to migrate towards the site of ...

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

Un<em&gt; In Vitro</em&gt; Caseosa Ensayo de unión que permite predecir la penetración de fármacos en la tuberculosis lesiones

The eradication of tuberculosis disease requires drug regimens that can penetrate the multiple layers of complex pulmonary lesions. Drug distribution in ...

Induction of Paralysis and Visual System Injury in Mice by T Cells Specific for Neuromyelitis Optica Autoantigen Aquaporin-4

Inducción de parálisis y lesión del sistema Visual en ratones por T las células específicas para el Optica del Neuromyelitis Autoantigen Acuaporina-4

While it is recognized that aquaporin-4 (AQP4)-specific T cells and antibodies participate in the pathogenesis of neuromyelitis optica (NMO), a human ...

Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity

Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity

Métodos alternativos In Vitro para la determinación de la integridad estructural de la cápside Viral

Human norovirus exacts considerable public health and economic losses worldwide. Emerging in vitro cultivation advances are not yet applicable for routine ...

Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium

Monocapas de Colonoid humano para estudiar las interacciones entre patógenos, comensales y epitelio Intestinal Host

Human 3-dimensional (3D) enteroid or colonoid cultures derived from crypt base stem cells are currently the most advanced ex vivo model of the intestinal ...