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Clin Chim Acta. 2018 Nov 5;488:135-142. doi: 10.1016/j.cca.2018.11.008. [Epub ahead of print]

Multi-centre validation of a flow cytometry method to identify optimal responders to interferon-beta in multiple sclerosis.

Author information

1
Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: noelia.villarrubia@salud.madrid.org.
2
Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: eulalia.rodriguez@salud.madrid.org.
3
Immunology and Neurology Dpt., Universitat Pompeu Fabra, Institut Hospital del Mar d'Investigacions Mèdiques (IMIM), Carrer del Dr. Aiguader 88, 08003 Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: ealari@imim.es.
4
Immunology and Neurology Dpt., Hospital Universitario de Donostia, Biodonostia, P° Dr. Beguiristain 107-111, 20014 San Sebastián, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: larraitz.aragonirusquieta@osakidetza.eus.
5
Immunology and Neurology Dpt., Hospital Universitario de Donostia, Biodonostia, P° Dr. Beguiristain 107-111, 20014 San Sebastián, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: tamaracastillo.tri@gmail.co.
6
Servei de Neurologia-Neuroimmunologia, Centre d'Esclerosi Múltiple de Catalunya, Cemcat, Vall d'Hebron Institut de Recerca, Hospital Universitari Vall d'Hebron, Passeig Vall d'Hebron, 119-129, 08035 Barcelona, Spain. Universitat Autònoma de Barcelona, 08193 Bellaterra (Cerdanyola del Vallès), Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: herena.eixarch@vhir.org.
7
Immunology and Neurology Dpt., Hospital Universitari Son Espases, Instituto de Investigación Sanitaria Illes Balears, IdISBa, Ctra. Valldemossa 79, Palma 07010, Spain.
8
Immunology and Neurology Dpt., Universitat Pompeu Fabra, Institut Hospital del Mar d'Investigacions Mèdiques (IMIM), Carrer del Dr. Aiguader 88, 08003 Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: jmartinezr@parcdesalutmar.cat.
9
Immunology and Neurology Dpt., Hospital Universitari Son Espases, Instituto de Investigación Sanitaria Illes Balears, IdISBa, Ctra. Valldemossa 79, Palma 07010, Spain. Electronic address: margaritam.massot@ssib.es.
10
Clinical Management Unit of Neurosciences, Instituto de Biomedicina de Málaga (IBIMA), Hospital Regional Universitario de Málaga, Universidad de Málaga, Plaza del Hospital Civil s/n., Málaga, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: mjpmedel@ibima.eu.
11
Immunology and Neurology Dpt., Hospital Universitario de Donostia, Biodonostia, P° Dr. Beguiristain 107-111, 20014 San Sebastián, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: alvarojose.pradainurrategui@osakidetza.eu.
12
Servei de Neurologia-Neuroimmunologia, Centre d'Esclerosi Múltiple de Catalunya, Cemcat, Vall d'Hebron Institut de Recerca, Hospital Universitari Vall d'Hebron, Passeig Vall d'Hebron, 119-129, 08035 Barcelona, Spain. Universitat Autònoma de Barcelona, 08193 Bellaterra (Cerdanyola del Vallès), Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: brodriguez@cem-cat.org.
13
Clinical Management Unit of Neurosciences, Instituto de Biomedicina de Málaga (IBIMA), Hospital Regional Universitario de Málaga, Universidad de Málaga, Plaza del Hospital Civil s/n., Málaga, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
14
Neuroimmunology Unit, Hospital Clínic Universitari de València, Av. de Blasco Ibáñez 17, 46010 València, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
15
Flow Cytometry Unit, UCIM, INCLIVA-Universidad de Valencia, Avda Blasco Ibañez 13, 46010 València, Spain.
16
Immunology and Neurology Dpt., Hospital Universitario Virgen de la Arrixaca, Ctra. Madrid-Cartagena, s/n, 30120 El Palmar, Murcia, Spain.
17
Immunology and Neurology Dpt., IDIVAL, Hospital Universitario Marqués de Valdecilla, Avda. Valdecilla 25, 39008 Santander, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
18
Immunology and Neurology Dpt., IDIVAL, Hospital Universitario Marqués de Valdecilla, Avda. Valdecilla 25, 39008 Santander, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: david.sansegundo@scsalud.es.
19
Immunology and Neurology Dpt., Hospital General Universitario and Instituto de Investigación Sanitaria "Gregorio Marañón", C/ Dr Esquerdo 46, 28007 Madrid, Spain. Electronic address: juanpablo.cuello@salud.madrid.org.
20
Immunology and Neurology Dpt., Hospital General Universitario and Instituto de Investigación Sanitaria "Gregorio Marañón", C/ Dr Esquerdo 46, 28007 Madrid, Spain. Electronic address: juana.gil@salud.madrid.org.
21
Immunology and Neurology Dpt., Hospital San Pedro de Alcántara, Avda. Pablo Naranjo s/n, 10003 Cáceres, Spain. Electronic address: ccamarah@salud.madrid.org.
22
Immunology and Neurology Dpt., Hospital San Pedro de Alcántara, Avda. Pablo Naranjo s/n, 10003 Cáceres, Spain.
23
Immunology and Neurology Dpt., Hospital Clínic de Barcelona, Carrer de Villarroel, 170, 08036 Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: emmartinez@clinic.ub.es.
24
Immunology and Neurology Dpt., Instituto de Investigación Sanitaria La Princesa, IIS-IP, Demyelinating Diseases Unit, Hospital Universitario de La Princesa, C/ Diego de León 62, 28006 Madrid, Spain.
25
Immunology and Neurology Dpt., Hospital de la Santa Creu i Sant Pau, Universitat Autònoma de Barcelona, C/ Sant Antoni Maria Claret 167, 08025 Barcelona, Ciberer, Madrid, Spain. Electronic address: mmoga@santpau.cat.
26
Immunology and Neurology Dpt., Instituto de Investigación Sanitaria La Princesa, IIS-IP, Demyelinating Diseases Unit, Hospital Universitario de La Princesa, C/ Diego de León 62, 28006 Madrid, Spain. Electronic address: cmunozc@salud.madrid.org.
27
Immunology and Neurology Dpt., Hospital de la Santa Creu i Sant Pau, Universitat Autònoma de Barcelona, C/ Sant Antoni Maria Claret 167, 08025 Barcelona, Ciberer, Madrid, Spain. Electronic address: lquerol@santpau.cat.
28
Immunology and Neurology Dpt., Multiple Sclerosis Unit, Hospital Universitario e Instituto de Investigación Germans Trias i Pujol, Campus Can Ruti, 08916 Badalona, Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
29
Immunology and Neurology Dpt., Multiple Sclerosis Unit, Hospital Universitario e Instituto de Investigación Germans Trias i Pujol, Campus Can Ruti, 08916 Badalona, Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: ateniente.germanstrias@gencat.cat.
30
Immunology and Neurology Dpt., Hospital Clínic de Barcelona, Carrer de Villarroel, 170, 08036 Barcelona, Spain. Electronic address: vlagea@clinic.cat.
31
Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain. Electronic address: alfonso.muriel@hrc.es.
32
Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: ernesto.roldan@salud.madrid.org.
33
Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain; Red Española de Esclerosis Múltiple (REEM), Spain. Electronic address: luisamaria.villar@salud.madrid.org.

Abstract

BACKGROUND AND OBJECTIVES:

Percentages of blood CD19+CD5+ B cells and CD8+perforin+ T lymphocytes can predict response to Interferon (IFN)-beta treatment in relapsing-remitting multiple sclerosis (RRMS) patients. We aimed to standardize their detection in a multicenter study, prior to their implementation in clinical practice.

METHODS:

Fourteen hospitals participated in the study. A reference centre was established for comparison studies. Peripheral blood cells of 105 untreated RRMS patients were studied. Every sample was analyzed in duplicate in the participating centre and in the reference one by flow cytometry. When needed, participating centres corrected fluorescence compensations and negative cut-off position following reference centre suggestions. Concordance between results obtained by participating centres and by reference one was evaluated by intraclass correlation coefficients (ICC) and Spearman correlation test. Centre performance was measured by using z-scores values.

RESULTS:

After results review and corrective actions implementation, overall ICC was 0.86 (CI: 0.81-0.91) for CD19+CD5+ B cell and 0.89 (CI: 0.85-0.93) for CD8+ perforin+ T cell quantification; Spearman r was 0.92 (0.89-0.95; p <0.0001) and 0.92 (0.88-0.95; p <0.0001) respectively. All centres obtained z-scores≤0.5 for both biomarkers.

CONCLUSION:

Homogenous percentages of CD19+CD5+ B cells and CD8 perforin+ T lymphocytes can be obtained if suitable compensation values and negative cut-off are pre-established.

KEYWORDS:

Biomarkers; Flow cytometry; Immunology; Multiple sclerosis

PMID:
30408481
DOI:
10.1016/j.cca.2018.11.008

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