{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T22:28:23Z","timestamp":1775341703605,"version":"3.50.1"},"reference-count":33,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2020,9,22]],"date-time":"2020-09-22T00:00:00Z","timestamp":1600732800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["FIS PI16\/01605"],"award-info":[{"award-number":["FIS PI16\/01605"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2021,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Primary antibody deficiencies (PADs) are characterized by hypogammaglobulinemia and impaired B\u2010cell differentiation. Patients with common variable immunodeficiency (CVID) present severe reductions in at least 2 serum immunoglobulins and impaired terminal differentiation of B cells. Most patients with CVID do not appear to present monogenic defects. Activated phosphoinositide 3\u2010kinase delta syndrome (APDS), caused by gain\u2010of\u2010function mutations in the PIK3CD gene (p110\u03b4), can present in patients with a CVID\u2010like phenotype. Memory B\u2010cell differentiation requires the orchestrated activation of numerous intracellular signaling pathways, which promote transcriptional programs required for long\u2010term B\u2010cell survival. The aim of this study was to develop a flow cytometry assay to trace the PI3K\u2010Akt\u2010mTOR pathway, a critical component of B\u2010cell homeostasis, and analyze its status in PADs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We analyzed the intracellular expression of Akt and S6 by flow cytometry and their phosphorylation status in both baseline conditions and upon B\u2010cell receptor activation with anti\u2010IgM in various primary B\u2010cell subsets of patients with CVID and APDS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>B cells from CVID patients showed reduced phosphorylation in Akt and S6 proteins after anti\u2010IgM stimulation. Constitutive high baseline B\u2010cell levels of Akt and S6 phosphorylation in a patient with APDS were reduced once m\u2010TOR inhibition therapy was initiated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Intracellular flow cytometry can be routinely employed to explore alterations in the PI3K\u2010Akt\u2010mTOR pathway in B cells from patients with PADs. AKT and S6 phosphorylation levels are informative biomarkers that could be employed as mTOR inhibitors for monitoring therapies targeting this pathway.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.21956","type":"journal-article","created":{"date-parts":[[2020,9,23]],"date-time":"2020-09-23T00:50:59Z","timestamp":1600822259000},"page":"460-466","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Evaluation of B\u2010cell intracellular signaling by monitoring the PI3K\u2010Akt axis in patients with common variable immunodeficiency and activated phosphoinositide 3\u2010kinase delta syndrome"],"prefix":"10.1002","volume":"100","author":[{"given":"Luc\u00eda","family":"del Pino\u2010Molina","sequence":"first","affiliation":[{"name":"Clinical Immunology Department La Paz University Hospital and Lymphocyte Pathophysiology in Immunodeficiencies Group, La Paz Institute for Health Research (IdiPAZ) and Center for Biomedical Network Research on Rare Diseases (CIBERER U767)  Madrid Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juan M.","family":"Torres Canizales","sequence":"additional","affiliation":[{"name":"Clinical Immunology Department La Paz University Hospital and Lymphocyte Pathophysiology in Immunodeficiencies Group, La Paz Institute for Health Research (IdiPAZ) and Center for Biomedical Network Research on Rare Diseases (CIBERER U767)  Madrid Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebeca","family":"Rodr\u00edguez\u2010Pena","sequence":"additional","affiliation":[{"name":"Clinical Immunology Department La Paz University Hospital and Lymphocyte Pathophysiology in Immunodeficiencies Group, La Paz Institute for Health Research (IdiPAZ) and Center for Biomedical Network Research on Rare Diseases (CIBERER U767)  Madrid Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eduardo","family":"L\u00f3pez\u2010Granados","sequence":"additional","affiliation":[{"name":"Clinical Immunology Department La Paz University Hospital and Lymphocyte Pathophysiology in Immunodeficiencies Group, La Paz Institute for Health Research (IdiPAZ) and Center for Biomedical Network Research on Rare Diseases (CIBERER U767)  Madrid Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,9,22]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1243292"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2019.02.017"},{"key":"e_1_2_8_4_1","first-page":"38","article-title":"International consensus document (ICON): Common variable immunodeficiency disorders","volume":"4","author":"Bonilla F. 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