Dissection of the Pre-Germinal Center B-Cell Maturation Pathway
in Common Variable Immunodeficiency Based on Standardized Flow
Cytometric EuroFlow Tools

J 2021

Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools

DEL PINO-MOLINA, L., E. LOPEZ-GRANADOS, Q. LECREVISSE, J. CANIZALES, M. PEREZ-ANDRES et. al.

Základní údaje

Originální název

Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools

Autoři

DEL PINO-MOLINA, L., E. LOPEZ-GRANADOS (garant), Q. LECREVISSE, J. CANIZALES, M. PEREZ-ANDRES, E. BLANCO, M. WENTINK, C. BONROY, Jana NECHVÁTALOVÁ (203 Česká republika, domácí), Tomas MILOTA (203 Česká republika), A. K. KIENZLER, J. PHILIPPE, A. E. SOUSA, M. VAN DER BURG, Tomas KALINA (203 Česká republika), J. J. M. VAN DONGEN a A. ORFAO

Vydání

Frontiers in Immunology, LAUSANNE, FRONTIERS MEDIA SA, 2021, 1664-3224

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30102 Immunology

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 8.786

Kód RIV

RIV/00216224:14110/21:00121774

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.3389/fimmu.2020.603972

UT WoS

000624523100001

Klíčová slova anglicky

CVID; Pre-GC B-cell tube; pre-GC maturation pathway; expression markers; EuroFlow standardization

Štítky

14110114, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 15. 6. 2021 09:26, Mgr. Tereza Miškechová

Anotace

V originále

Introduction Common Variable Immunodeficiency (CVID) is characterized by defective antibody production and hypogammaglobulinemia. Flow cytometry immunophenotyping of blood lymphocytes has become of great relevance for the diagnosis and classification of CVID, due to an impaired differentiation of mature post-germinal-center (GC) class-switched memory B-cells (MBC) and severely decreased plasmablast/plasma cell (Pb) counts. Here, we investigated in detail the pre-GC B-cell maturation compartment in blood of CVID patients. Methods In this collaborative multicentric study the EuroFlow PID 8-color Pre-GC B-cell tube, standardized sample preparation procedures (SOPs) and innovative data analysis tools, were used to characterize the maturation profile of pre-GC B-cells in 100 CVID patients, vs 62 age-matched healthy donors (HD). Results The Pre-GC B-cell tube allowed identification within pre-GC B-cells of three subsets of maturation associated immature B-cells and three subpopulations of mature naive B-lymphocytes. CVID patients showed overall reduced median absolute counts (vs HD) of the two more advanced stages of maturation of both CD5(+) CD38(+/++) CD21(het) CD24(++) (2.7 vs 5.6 cells/mu l, p=0.0004) and CD5(+) CD38(het) CD21(+) CD24(+) (6.5 vs 17 cells/mu l, p<0.0001) immature B cells (below normal HD levels in 22% and 37% of CVID patients). This was associated with an expansion of CD21(-)CD24(-) (6.1 vs 0.74 cells/mu l, p<0.0001) and CD21(-)CD24(++) (1.8 vs 0.4 cells/mu l, p<0.0001) naive B-cell counts above normal values in 73% and 94% cases, respectively. Additionally, reduced IgMD(+) (21 vs 32 cells/mu l, p=0.03) and IgMD(-) (4 vs 35 cells/mu l, p<0.0001) MBC counts were found to be below normal values in 25% and 77% of CVID patients, respectively, always together with severely reduced/undetectable circulating blood pb. Comparison of the maturation pathway profile of pre-GC B cells in blood of CVID patients vs HD using EuroFlow software tools showed systematically altered patterns in CVID. These consisted of: i) a normally-appearing maturation pathway with altered levels of expression of >1 (CD38, CD5, CD19, CD21, CD24, and/or smIgM) phenotypic marker (57/88 patients; 65%) for a total of 3 distinct CVID patient profiles (group 1: 42/88 patients, 48%; group 2: 8/88, 9%; and group 3: 7/88, 8%) and ii) CVID patients with a clearly altered pre-GC B cell maturation pathway in blood (group 4: 31/88 cases, 35%). Conclusion Our results show that maturation of pre-GC B-cells in blood of CVID is systematically altered with up to four distinctly altered maturation profiles. Further studies, are necessary to better understand the impact of such alterations on the post-GC defects and the clinical heterogeneity of CVID.

Citovat

DEL PINO-MOLINA, L., E. LOPEZ-GRANADOS, Q. LECREVISSE, J. CANIZALES, M. PEREZ-ANDRES, E. BLANCO, M. WENTINK, C. BONROY, Jana NECHVÁTALOVÁ, Tomas MILOTA, A. K. KIENZLER, J. PHILIPPE, A. E. SOUSA, M. VAN DER BURG, Tomas KALINA, J. J. M. VAN DONGEN a A. ORFAO. Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools. Frontiers in Immunology. LAUSANNE: FRONTIERS MEDIA SA, 2021, roč. 11, FEB 2021, s. 1-14. ISSN 1664-3224. Dostupné z: https://dx.doi.org/10.3389/fimmu.2020.603972.

@article{1776207,
   author = {del PinoandMolina, L. and LopezandGranados, E. and Lecrevisse, Q. and Canizales, J. and PerezandAndres, M. and Blanco, E. and Wentink, M. and Bonroy, C. and Nechvátalová, Jana and Milota, Tomas and Kienzler, A. K. and Philippe, J. and Sousa, A. E. and van der Burg, M. and Kalina, Tomas and van Dongen, J. J. M. and Orfao, A.},
   article_location = {LAUSANNE},
   article_number = {FEB 2021},
   doi = {http://dx.doi.org/10.3389/fimmu.2020.603972},
   keywords = {CVID; Pre-GC B-cell tube; pre-GC maturation pathway; expression markers; EuroFlow standardization},
   language = {eng},
   issn = {1664-3224},
   journal = {Frontiers in Immunology},
   title = {Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools},
   url = {https://www.frontiersin.org/articles/10.3389/fimmu.2020.603972/full},
   volume = {11},
   year = {2021}
}

TY  - JOUR
ID  - 1776207
AU  - del Pino-Molina, L. - Lopez-Granados, E. - Lecrevisse, Q. - Canizales, J. - Perez-Andres, M. - Blanco, E. - Wentink, M. - Bonroy, C. - Nechvátalová, Jana - Milota, Tomas - Kienzler, A. K. - Philippe, J. - Sousa, A. E. - van der Burg, M. - Kalina, Tomas - van Dongen, J. J. M. - Orfao, A.
PY  - 2021
TI  - Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools
JF  - Frontiers in Immunology
VL  - 11
IS  - FEB 2021
SP  - 1-14
EP  - 1-14
PB  - FRONTIERS MEDIA SA
SN  - 16643224
KW  - CVID
KW  - Pre-GC B-cell tube
KW  - pre-GC maturation pathway
KW  - expression markers
KW  - EuroFlow standardization
UR  - https://www.frontiersin.org/articles/10.3389/fimmu.2020.603972/full
N2  - Introduction Common Variable Immunodeficiency (CVID) is characterized by defective antibody production and hypogammaglobulinemia. Flow cytometry immunophenotyping of blood lymphocytes has become of great relevance for the diagnosis and classification of CVID, due to an impaired differentiation of mature post-germinal-center (GC) class-switched memory B-cells (MBC) and severely decreased plasmablast/plasma cell (Pb) counts. Here, we investigated in detail the pre-GC B-cell maturation compartment in blood of CVID patients. Methods In this collaborative multicentric study the EuroFlow PID 8-color Pre-GC B-cell tube, standardized sample preparation procedures (SOPs) and innovative data analysis tools, were used to characterize the maturation profile of pre-GC B-cells in 100 CVID patients, vs 62 age-matched healthy donors (HD). Results The Pre-GC B-cell tube allowed identification within pre-GC B-cells of three subsets of maturation associated immature B-cells and three subpopulations of mature naive B-lymphocytes. CVID patients showed overall reduced median absolute counts (vs HD) of the two more advanced stages of maturation of both CD5(+) CD38(+/++) CD21(het) CD24(++) (2.7 vs 5.6 cells/mu l, p=0.0004) and CD5(+) CD38(het) CD21(+) CD24(+) (6.5 vs 17 cells/mu l, p<0.0001) immature B cells (below normal HD levels in 22% and 37% of CVID patients). This was associated with an expansion of CD21(-)CD24(-) (6.1 vs 0.74 cells/mu l, p<0.0001) and CD21(-)CD24(++) (1.8 vs 0.4 cells/mu l, p<0.0001) naive B-cell counts above normal values in 73% and 94% cases, respectively. Additionally, reduced IgMD(+) (21 vs 32 cells/mu l, p=0.03) and IgMD(-) (4 vs 35 cells/mu l, p<0.0001) MBC counts were found to be below normal values in 25% and 77% of CVID patients, respectively, always together with severely reduced/undetectable circulating blood pb. Comparison of the maturation pathway profile of pre-GC B cells in blood of CVID patients vs HD using EuroFlow software tools showed systematically altered patterns in CVID. These consisted of: i) a normally-appearing maturation pathway with altered levels of expression of >1 (CD38, CD5, CD19, CD21, CD24, and/or smIgM) phenotypic marker (57/88 patients; 65%) for a total of 3 distinct CVID patient profiles (group 1: 42/88 patients, 48%; group 2: 8/88, 9%; and group 3: 7/88, 8%) and ii) CVID patients with a clearly altered pre-GC B cell maturation pathway in blood (group 4: 31/88 cases, 35%). Conclusion Our results show that maturation of pre-GC B-cells in blood of CVID is systematically altered with up to four distinctly altered maturation profiles. Further studies, are necessary to better understand the impact of such alterations on the post-GC defects and the clinical heterogeneity of CVID.
ER  -

DEL PINO-MOLINA, L., E. LOPEZ-GRANADOS, Q. LECREVISSE, J. CANIZALES, M. PEREZ-ANDRES, E. BLANCO, M. WENTINK, C. BONROY, Jana NECHVÁTALOVÁ, Tomas MILOTA, A. K. KIENZLER, J. PHILIPPE, A. E. SOUSA, M. VAN DER BURG, Tomas KALINA, J. J. M. VAN DONGEN a A. ORFAO. Dissection of the Pre-Germinal Center B-Cell Maturation Pathway in Common Variable Immunodeficiency Based on Standardized Flow Cytometric EuroFlow Tools. \textit{Frontiers in Immunology}. LAUSANNE: FRONTIERS MEDIA SA, 2021, roč.~11, FEB 2021, s.~1-14. ISSN~1664-3224. Dostupné z: https://dx.doi.org/10.3389/fimmu.2020.603972.

Podrobný výpis o publikaci