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• W2889076804 abstract "Flow cytometry is a routinely available laboratory method to study cells in suspension from a variety of human sources. Application of this technology as a clinical laboratory method has evolved from the identification of cell-surface proteins to characterizing intracellular proteins and providing multiple different techniques to assess specific features of adaptive and innate immune function. This expanded menu of flow cytometric testing approaches has increased the utility of this platform in characterizing and diagnosing disorders of immune function. Flow cytometry is a routinely available laboratory method to study cells in suspension from a variety of human sources. Application of this technology as a clinical laboratory method has evolved from the identification of cell-surface proteins to characterizing intracellular proteins and providing multiple different techniques to assess specific features of adaptive and innate immune function. This expanded menu of flow cytometric testing approaches has increased the utility of this platform in characterizing and diagnosing disorders of immune function. Flow cytometry is a routinely available laboratory method to study cells in suspension, including peripheral blood, bone marrow, cerebrospinal fluid, and other body fluids or tissue suspensions. The clinical application of flow cytometry evolved as a tool for enumeration of CD4+ T cells in the blood of patients with HIV infection and to characterize hematologic malignancies. More recently, the role of flow cytometry has broadened to include the study of disorders of the immune system, including primary immunodeficiency disorders (PID). This review describes the use of flow cytometry in identifying cell-surface markers to characterize cells of the adaptive and innate immune system and then focuses on alternative applications of flow cytometry in clinical immunology, including evaluation of intracellular characteristics and immune cell function. Evaluation for the presence or absence of multiple cell-surface markers by using polychromatic flow cytometry constitutes the basis of clinical immunophenotyping. This allows for the characterization of cell populations and subpopulations, identification of the status of cell differentiation, and quantification of surface proteins associated with specific cellular functions. Determination of immunophenotypic characteristics of T, B, and natural killer (NK) cell subsets by using flow cytometry allows classification of patients with severe combined immunodeficiency (SCID) into different categories that have historically narrowed the search for possible underlying genetic defects.1Oliveira J.B. Notarangelo L.D. Fleisher T.A. Applications of flow cytometry for the study of primary immune deficiencies.Curr Opin Allergy Clin Immunol. 2008; 8: 499-509Crossref PubMed Scopus (38) Google Scholar, 2Villa A. Sobacchi C. Notarangelo L.D. Bozzi F. Abinun M. Abrahamsen T.G. et al.V(D)J recombination defects in lymphocytes due to RAG mutations: severe immunodeficiency with a spectrum of clinical presentations.Blood. 2001; 97: 81-88Crossref PubMed Scopus (281) Google Scholar T-cell subpopulation characterization might also be useful in the setting of abnormal results on newborn screening for severe T-cell immunodeficiency.1Oliveira J.B. Notarangelo L.D. Fleisher T.A. Applications of flow cytometry for the study of primary immune deficiencies.Curr Opin Allergy Clin Immunol. 2008; 8: 499-509Crossref PubMed Scopus (38) Google Scholar When low or absent numbers of T-cell receptor (TCR) excision circles are detected at birth, immunophenotyping to evaluate for numbers of naive T cells based on CD45RA in combination with CD31, CD127, and/or CD62 ligand (CD62L) is typically a part of the follow-up evaluation. In addition, both hypomorphic SCID mutations and typical SCID associated with maternal engraftment result in circulating nonnaive T cells that have upregulated activation markers, such as HLA-DR and CD69.2Villa A. Sobacchi C. Notarangelo L.D. Bozzi F. Abinun M. Abrahamsen T.G. et al.V(D)J recombination defects in lymphocytes due to RAG mutations: severe immunodeficiency with a spectrum of clinical presentations.Blood. 2001; 97: 81-88Crossref PubMed Scopus (281) Google Scholar Other surface antigens that are found only on T-activated cells are receptors for specific growth factors, such as CD25; receptors for critical elements required for cell growth, such as the transferrin receptor (CD71); and ligands for cell-to-cell communication after cell activation, including CD40 ligand (CD40L) on activated CD4+ T cells. Flow cytometry is important in the diagnosis of autoimmune lymphoproliferative syndrome (ALPS) because increased numbers of double-negative T (DNT) cells expressing the α/β TCR present in peripheral blood and lymphoid tissues is a diagnostic marker of the disease. These patients usually have mutations in genes that regulate the extrinsic Fas-mediated cell death pathway (FAS, FASL, and CASP10), and DNT cells homogeneously express multiple surface markers, including CD45RA, CD57, CD27, CD28, perforin, and HLA-DR, but lack CD45RO and CD56. This finding contrasts with DNT cells in peripheral blood of healthy control subjects, which predominantly express the γ/δ TCR receptor.3Oliveira J.B. Bleesing J.J. Dianzani U. Fleisher T.A. Jaffe E.S. Lenardo M.J. et al.Revised diagnostic criteria and classification for the autoimmune lymphoproliferative syndrome (ALPS): report from the 2009 NIH International Workshop.Blood. 2010; 116: e35-e40Crossref PubMed Scopus (346) Google Scholar Flow cytometry is a crucial diagnostic tool for MHC class II deficiency. This is a rare autosomal recessive form of PID characterized by the deficiency of different MHC class II molecules.4Aluri J. Gupta M. Dalvi A. Mhatre S. Kulkarni M. Hule G. et al.Clinical, immunological, and molecular findings in five patients with major histocompatibility complex class II deficiency from India.Front Immunol. 2018; 9: 188Crossref PubMed Scopus (19) Google Scholar These defects affect the cellular and humoral immune responses by impairing the development of CD4+ TH cells and TH cell–dependent antibody production by B cells.5Hanna S. Etzioni A. MHC class I and II deficiencies.J Allergy Clin Immunol. 2014; 134: 269-275Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar Affected children typically present with severe respiratory and gastrointestinal tract infections, and hematopoietic stem cell transplantation is the only curative therapy available.4Aluri J. Gupta M. Dalvi A. Mhatre S. Kulkarni M. Hule G. et al.Clinical, immunological, and molecular findings in five patients with major histocompatibility complex class II deficiency from India.Front Immunol. 2018; 9: 188Crossref PubMed Scopus (19) Google Scholar Complete absence of HLA-DR expression on B cells and monocytes by using flow cytometry is diagnostic of this disorder.4Aluri J. Gupta M. Dalvi A. Mhatre S. Kulkarni M. Hule G. et al.Clinical, immunological, and molecular findings in five patients with major histocompatibility complex class II deficiency from India.Front Immunol. 2018; 9: 188Crossref PubMed Scopus (19) Google Scholar, 5Hanna S. Etzioni A. MHC class I and II deficiencies.J Allergy Clin Immunol. 2014; 134: 269-275Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar Application of B-cell immunophenotyping has emerged as a useful tool in the evaluation of common variable immunodeficiency (CVID), a heterogenous group of disorders characterized by hypogammaglobulinemia, impaired antibody production, increased susceptibility to sinopulmonary infections, and immune dysregulation and cancer.6Cunningham-Rundles C. Bodian C. Common variable immunodeficiency: clinical and immunological features of 248 patients.Clin Immunol. 1999; 92: 34-48Crossref PubMed Scopus (1282) Google Scholar Patients with CVID often have normal or low numbers of B cells. However, evaluating B cells based on the distribution of naive (CD27−IgD+IgM+), nonswitched memory (CD27+IgD+IgM+), and switched memory (CD27+IgD−IgM−) cells has proved to be a useful tool in categorizing patients with CVID.7Warnatz K. Denz A. Drager R. Braun M. Groth C. Wolff-Vorbeck G. et al.Severe deficiency of switched memory B cells (CD27(+)IgM(-)IgD(-)) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease.Blood. 2002; 99: 1544-1551Crossref PubMed Scopus (503) Google Scholar, 8Rosel A.L. Scheibenbogen C. Schliesser U. Sollwedel A. Hoffmeister B. Hanitsch L. et al.Classification of common variable immunodeficiencies using flow cytometry and a memory B-cell functionality assay.J Allergy Clin Immunol. 2015; 135: 198-208Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar, 9Wehr C. Kivioja T. Schmitt C. Ferry B. Witte T. Eren E. et al.The EUROclass trial: defining subgroups in common variable immunodeficiency.Blood. 2008; 111: 77-85Crossref PubMed Scopus (611) Google Scholar, 10Bonilla F.A. Barlan I. Chapel H. Costa-Carvalho B.T. Cunningham-Rundles C. de la Morena M.T. et al.International Consensus Document (ICON): common variable immunodeficiency disorders.J Allergy Clin Immunol Pract. 2016; 4: 38-59Abstract Full Text Full Text PDF PubMed Scopus (481) Google Scholar Furthermore, immunophenotyping of these patients can be extended to various B-cell subsets beyond naive and memory cells to include marginal zone B cells (CD19+CD27+IgM+IgD+), transitional B cells (CD19+CD27−CD24hiCD38hiIgMhiCD10+), plasmablasts (CD19+CD20−IgM−CD38intCD27+), and B cells with different levels of CD21 expression.10Bonilla F.A. Barlan I. Chapel H. Costa-Carvalho B.T. Cunningham-Rundles C. de la Morena M.T. et al.International Consensus Document (ICON): common variable immunodeficiency disorders.J Allergy Clin Immunol Pract. 2016; 4: 38-59Abstract Full Text Full Text PDF PubMed Scopus (481) Google Scholar A subgroup of CVID has been associated with abnormalities in the inducible T-cell costimulator, CD19, and B-cell activating factor receptor (TNFRSF13C) molecules. Screening for these defects is performed by using flow cytometry to detect reduced upregulation of inducible T-cell costimulator on activated T cells, or reduced expression of B-cell activating factor receptor or CD19 on B cells.11Grimbacher B. Hutloff A. Schlesier M. Glocker E. Warnatz K. Drager R. et al.Homozygous loss of ICOS is associated with adult-onset common variable immunodeficiency.Nat Immunol. 2003; 4: 261-268Crossref PubMed Scopus (606) Google Scholar, 12van Zelm M.C. Reisli I. van der Burg M. Castano D. van Noesel C.J. van Tol M.J. et al.An antibody-deficiency syndrome due to mutations in the CD19 gene.N Engl J Med. 2006; 354: 1901-1912Crossref PubMed Scopus (432) Google Scholar, 13Warnatz K. Salzer U. Rizzi M. Fischer B. Gutenberger S. Bohm J. et al.B-cell activating factor receptor deficiency is associated with an adult-onset antibody deficiency syndrome in humans.Proc Natl Acad Sci U S A. 2009; 106: 13945-13950Crossref PubMed Scopus (289) Google Scholar Decreased numbers of switched memory B cells are also observed in patients with hyper-IgM (HIGM) syndromes.14Agematsu K. Nagumo H. Shinozaki K. Hokibara S. Yasui K. Terada K. et al.Absence of IgD-CD27(+) memory B cell population in X-linked hyper-IgM syndrome.J Clin Invest. 1998; 102: 853-860Crossref PubMed Scopus (169) Google Scholar These syndromes represent a group of genetic disorders affecting molecules involved in B-cell class-switch recombination and somatic hypermutation.15Gulino A.V. Notarangelo L.D. Hyper IgM syndromes.Curr Opin Rheumatol. 2003; 15: 422-429Crossref PubMed Scopus (56) Google Scholar, 16Lee W.I. Torgerson T.R. Schumacher M.J. Yel L. Zhu Q. Ochs H.D. Molecular analysis of a large cohort of patients with the hyper immunoglobulin M (IgM) syndrome.Blood. 2005; 105: 1881-1890Crossref PubMed Scopus (171) Google Scholar Affected patients present with normal or increased serum IgM levels and low levels of IgG and IgA. Although mutations in several genes have been associated with HIGM, the most frequently affected gene is the X-linked CD40L gene. Because CD40L (CD154) is important for normal T-cell function, deficient patients have not only bacterial but also opportunistic infections and malignancies.17Seyama K. Nonoyama S. Gangsaas I. Hollenbaugh D. Pabst H.F. Aruffo A. et al.Mutations of the CD40 ligand gene and its effect on CD40 ligand expression in patients with X-linked hyper IgM syndrome.Blood. 1998; 92: 2421-2434Crossref PubMed Google Scholar CD40L expression by activated CD4+ T cells is absent or reduced in approximately 80% to 90% of the patients when assessed with anti-CD40L–specific mAbs, whereas the remaining patients have mutations that result in expression of nonfunctional protein that is still detected by using the mAbs.16Lee W.I. Torgerson T.R. Schumacher M.J. Yel L. Zhu Q. Ochs H.D. Molecular analysis of a large cohort of patients with the hyper immunoglobulin M (IgM) syndrome.Blood. 2005; 105: 1881-1890Crossref PubMed Scopus (171) Google Scholar Flow cytometric detection can be further improved by using a biotinylated CD40-immunoglobulin fusion protein to detect functional CD40L, identifying more than 90% of the patients with confirmed CD40L mutations.17Seyama K. Nonoyama S. Gangsaas I. Hollenbaugh D. Pabst H.F. Aruffo A. et al.Mutations of the CD40 ligand gene and its effect on CD40 ligand expression in patients with X-linked hyper IgM syndrome.Blood. 1998; 92: 2421-2434Crossref PubMed Google Scholar CD40 deficiency, one of several autosomal recessive HIGM syndromes, is a clinical phenocopy of CD40L deficiency that can be identified by assessing for CD40 expression on B cells, monocytes, and/or dendritic cells.18Ferrari S. Giliani S. Insalaco A. Al-Ghonaium A. Soresina A.R. Loubser M. et al.Mutations of CD40 gene cause an autosomal recessive form of immunodeficiency with hyper IgM.Proc Natl Acad Sci U S A. 2001; 98: 12614-12619Crossref PubMed Scopus (304) Google Scholar The remaining classic HIGM syndromes involve B-cell proteins that currently cannot be detected by using flow cytometry. Genetic defects affecting Toll-like receptor (TLR) pathways have been described in immunodeficient patients with unique patterns of infection.19Picard C. Casanova J.L. Puel A. Infectious diseases in patients with IRAK-4, MyD88, NEMO, or IkappaBalpha deficiency.Clin Microbiol Rev. 2011; 24: 490-497Crossref PubMed Scopus (289) Google Scholar These include IRAK4 mutations in patients with recurrent and/or severe pneumococcal infections, NEMO and IKBA defects in patients with atypical mycobacteriosis and other bacterial infections, and UNC93B and TLR3 mutations in children with Herpes simplex encephalitis.19Picard C. Casanova J.L. Puel A. Infectious diseases in patients with IRAK-4, MyD88, NEMO, or IkappaBalpha deficiency.Clin Microbiol Rev. 2011; 24: 490-497Crossref PubMed Scopus (289) Google Scholar A reliable screening method for some of these diseases is a flow cytometric assay based on L-selectin (CD62L) shedding after TLR stimulation. In this method incubation of whole fresh blood with various TLR ligands induces rapid shedding of CD62L from the surfaces of granulocytes. The absence of shedding can identify patients with IRAK4 and UNC93B mutations. Importantly, this procedure is not useful to identify patients with TLR3, NEMO, and IKBA mutations.20von Bernuth H. Ku C.L. Rodriguez-Gallego C. Zhang S. Garty B.Z. Marodi L. et al.A fast procedure for the detection of defects in Toll-like receptor signaling.Pediatrics. 2006; 118: 2498-2503Crossref PubMed Scopus (51) Google Scholar Leukocyte adhesion deficiency (LAD) type 1 is associated with recurrent skin and deep-seated bacterial infections. It is caused by a defect in β2-integrin expression that can be diagnosed by using flow cytometry, evaluating for the expression of cell-surface CD18 and its partner proteins, CD11a, CD11b, and CD11c.21van de Vijver E. van den Berg T.K. Kuijpers T.W. Leukocyte adhesion deficiencies.Hematol Oncol Clin North Am. 2013; 27 (viii): 101-116Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar In patients with LAD type 1, CD18 is usually less than 5% to 10% of normal values, and unlike control subjects, the level of expression is not upregulated after neutrophil activation. By contrast, in patients with LAD type 2 associated with defective fucosylation, the diagnosis can be suggested by demonstrating failure of CD15s (sialyl-Lewis X antigen) expression together with red blood cell typing that demonstrates the rare Bombay blood type.21van de Vijver E. van den Berg T.K. Kuijpers T.W. Leukocyte adhesion deficiencies.Hematol Oncol Clin North Am. 2013; 27 (viii): 101-116Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar, 22Etzioni A. Defects in the leukocyte adhesion cascade.Clin Rev Allergy Immunol. 2010; 38: 54-60Crossref PubMed Scopus (75) Google Scholar Cell-surface staining can also allow for investigation of lymphocyte clonality. T-cell clonality studies are particularly useful in evaluation of PIDs associated with a restricted T-cell repertoire. Omenn syndrome, a form of leaky SCID, is associated with multiple different genetic defects that impair but do not completely abrogate T-cell generation.23Villa A. Santagata S. Bozzi F. Giliani S. Frattini A. Imberti L. et al.Partial V(D)J recombination activity leads to Omenn syndrome.Cell. 1998; 93: 885-896Abstract Full Text Full Text PDF PubMed Scopus (374) Google Scholar, 24Villa A. Notarangelo L.D. Roifman C.M. Omenn syndrome: inflammation in leaky severe combined immunodeficiency.J Allergy Clin Immunol. 2008; 122: 1082-1086Abstract Full Text Full Text PDF PubMed Scopus (182) Google Scholar This disorder is characterized by oligoclonal T-cell expansion that can be detected in peripheral blood, as well as in T cells infiltrating the skin, liver, spleen, and lymph nodes.24Villa A. Notarangelo L.D. Roifman C.M. Omenn syndrome: inflammation in leaky severe combined immunodeficiency.J Allergy Clin Immunol. 2008; 122: 1082-1086Abstract Full Text Full Text PDF PubMed Scopus (182) Google Scholar Similarly, a restricted T-cell repertoire is seen in patients with SCID with maternal engraftment.25Sottini A. Quiros-Roldan E. Notarangelo L.D. Malagoli A. Primi D. Imberti L. Engrafted maternal T cells in a severe combined immunodeficiency patient express T-cell receptor variable beta segments characterized by a restricted V-D-J junctional diversity.Blood. 1995; 85: 2105-2113PubMed Google Scholar The method uses mAbs directed at specific T-cell antigen receptor variable β (Vβ) chains (Fig 1). This approach identifies significant underrepresentation or overrepresentation of a specific Vβ chain family expressing T cells and is complementary to PCR–based TCR spectratyping.26Pilch H. Hohn H. Freitag K. Neukirch C. Necker A. Haddad P. et al.Improved assessment of T-cell receptor (TCR) VB repertoire in clinical specimens: combination of TCR-CDR3 spectratyping with flow cytometry-based TCR VB frequency analysis.Clin Diagn Lab Immunol. 2002; 9: 257-266PubMed Google Scholar In the setting of hematologic malignancies, issues of monoclonality can be fully or partially addressed by using flow cytometry when analyzing B-cell malignancies and, in some circumstances, when studying T-cell disease. Normally, B cells are a heterogeneous mixture of mutually exclusive κ or λ light chain–expressing cells. Measuring the distribution of κ or λ light chain–expressing B cells or plasmacytes can be informative with respect to the presence or absence of monoclonality.27Berliner N. Ault K.A. Martin P. Weinberg D.S. Detection of clonal excess in lymphoproliferative disease by kappa/lambda analysis: correlation with immunoglobulin gene DNA rearrangement.Blood. 1986; 67: 80-85PubMed Google Scholar The capacity to evaluate T-cell monoclonality by using flow cytometry is less definitive and consists of using the method described previously to identify evidence of significant overrepresentation of one Vβ family. This is an indirect measure that suggests possible clonality. Traditional flow cytometry testing for Vβ chain expression requires 8 tubes (24 Vβ-specific mAbs), but a more recently described method combines all TCR Vβ mAbs into 1 tube.28Wu D. Anderson M.M. Othus M. Wood B.L. Clinical experience with modified, single-tube T-cell receptor vbeta flow cytometry analysis for T-cell clonality.Am J Clin Pathol. 2016; 145: 467-485Crossref PubMed Google Scholar This latter technique enables linking the aberrant immunophenotype of a neoplastic T-cell clone to its apparent clonal TCR Vβ restriction. A false-positive rate for identifying a clonal population with this approach was estimated to be less than 6.2%.28Wu D. Anderson M.M. Othus M. Wood B.L. Clinical experience with modified, single-tube T-cell receptor vbeta flow cytometry analysis for T-cell clonality.Am J Clin Pathol. 2016; 145: 467-485Crossref PubMed Google Scholar Despite these results, molecular approaches for T-cell clonality by using RNA sequencing have increased sensitivity and diagnostic utility compared with flow cytometry.29Brown S.D. Hapgood G. Steidl C. Weng A.P. Savage K.J. Holt R.A. Defining the clonality of peripheral T cell lymphomas using RNA-seq.Bioinformatics. 2017; 33: 1111-1115PubMed Google Scholar Additionally, cell surface–based flow cytometry is able to detect rare events, including detecting CD34+ hematopoietic stem cells in peripheral blood under resting conditions30Sutherland D.R. Keating A. Nayar R. Anania S. Stewart A.K. Sensitive detection and enumeration of CD34+ cells in peripheral and cord blood by flow cytometry.Exp Hematol. 1994; 22: 1003-1010PubMed Google Scholar and evaluating for minimal residual disease in patients with hematologic malignancy, such as hairy cell leukemia.31Garnache Ottou F. Chandesris M.O. Lhermitte L. Callens C. Beldjord K. Garrido M. et al.Peripheral blood 8 colour flow cytometry monitoring of hairy cell leukaemia allows detection of high-risk patients.Br J Haematol. 2014; 166: 50-59Crossref PubMed Scopus (29) Google Scholar Despite advances in the evaluation of cell-surface markers (Table I), this approach has limitations because it does not assess the functional status of cells. For example, in patients with CVID, the presence of normal B-cell numbers does not correlate with immunoglobulin production and an antigen-specific antibody response.32Warnatz K. Schlesier M. Flowcytometric phenotyping of common variable immunodeficiency.Cytometry B Clin Cytom. 2008; 74: 261-271Crossref PubMed Scopus (115) Google Scholar Likewise, in patients with SCID caused by X-linked common γ chain defects, normal surface protein expression of this protein does not rule out a disease-causing mutation, resulting in expression of a defective protein incapable of initiating a signal, a consistent problem with immunoassay-based protein detection, such as flow cytometry.Table IEvaluation of PIDs by using flow cytometryCell-surface protein stainingLow or absent TRECs on NBS: naive T cells and recent thymic emigrantsMHC class II deficiency: absent HLA-DRALPS: increased α/β TCR double-negative (CD4−CD8−) T-cell countsOmenn syndrome and SCID: restricted T-cell repertoireCVID: CD19 (B cells), BAFF-R (B cells), ICOS (activated T cells)CVID: decreased switched memory B-cell countsX-linked HIGM: CD40L (activated T cells)Autosomal recessive HIGM: CD40 (B cells)LAD type 1: CD18 (granulocytes)LAD type 2: CD15sIRAK4 and UNC93B deficiency: CD62L (granulocytes)Intracellular protein stainingXLA: BTK (monocytes, platelets)WAS, WIP deficiency: WASp (lymphocytes, myeloid cells)XLP1/2: SAP (CD8 T cells, NK cells)/XIAP (lymphocytes)FHL: perforin (CD8 T cells, NK cells),IPEX: FoxP3 (regulatory T cells)CTLA4 haploinsufficiency: CTLA4 expression (regulatory T cells)LRBA deficiency: CTLA4 expression (regulatory T cells), LRBA expression (PBMCs)DOCK8 deficiency: DOCK8 expressionALPS, Autoimmune lymphoproliferative syndrome; BTK, Bruton tyrosine kinase; FHL, familial hemophagocytic lymphohistiocytosis; FoxP3, forkhead box P3; ICOS, inducible T-cell costimulator; IPEX, immune dysregulation, polyendocrinopathy, enteropathy, X-linked inheritance syndrome; IRAK4, IL-1 receptor–associated kinase 4; LRBA, LPS-responsive beige-like anchor; NBS, Newborn screening; SAP, Signaling lymphocyte activation molecule-associated protein; TREC, T-cell excision circle; WIP, WASP-interacting protein; XIAP, X-linked inhibitor of apoptosis; XLA, X-linked agammaglobulinemia; XLP, X-linked lymphoproliferative syndrome. Open table in a new tab ALPS, Autoimmune lymphoproliferative syndrome; BTK, Bruton tyrosine kinase; FHL, familial hemophagocytic lymphohistiocytosis; FoxP3, forkhead box P3; ICOS, inducible T-cell costimulator; IPEX, immune dysregulation, polyendocrinopathy, enteropathy, X-linked inheritance syndrome; IRAK4, IL-1 receptor–associated kinase 4; LRBA, LPS-responsive beige-like anchor; NBS, Newborn screening; SAP, Signaling lymphocyte activation molecule-associated protein; TREC, T-cell excision circle; WIP, WASP-interacting protein; XIAP, X-linked inhibitor of apoptosis; XLA, X-linked agammaglobulinemia; XLP, X-linked lymphoproliferative syndrome. Clinical flow cytometry now includes the capacity to identify and quantify intracellular proteins associated with immune function (Table I). This method requires fixation and permeabilization to allow the mAb to pass through the cell membrane. A clinical application of this approach is demonstrated by screening of patients for Wiskott-Aldrich syndrome (WAS).33Nakajima M. Yamada M. Yamaguchi K. Sakiyama Y. Oda A. Nelson D.L. et al.Possible application of flow cytometry for evaluation of the structure and functional status of WASP in peripheral blood mononuclear cells.Eur J Haematol. 2009; 82: 223-230Crossref PubMed Scopus (7) Google Scholar Either the absence or decreased intracellular expression of the Wiskott-Aldrich syndrome protein (WASP) can confirm the diagnosis of WAS and/or X-linked thrombocytopenia (XLT; Fig 2).34Kawai S. Minegishi M. Ohashi Y. Sasahara Y. Kumaki S. Konno T. et al.Flow cytometric determination of intracytoplasmic Wiskott-Aldrich syndrome protein in peripheral blood lymphocyte subpopulations.J Immunol Methods. 2002; 260: 195-205Crossref PubMed Scopus (42) Google Scholar Additionally, this testing can also identify carriers of WAS and XLT.35Notarangelo L.D. Miao C.H. Ochs H.D. Wiskott-Aldrich syndrome.Curr Opin Hematol. 2008; 15: 30-36Crossref PubMed Scopus (159) Google Scholar However, the presence of normal intracellular WASP expression, as determined by using flow cytometry, does not rule out WAS/XLT because some patients express a dysfunctional protein detectable at levels comparable with those of control subjects.33Nakajima M. Yamada M. Yamaguchi K. Sakiyama Y. Oda A. Nelson D.L. et al.Possible application of flow cytometry for evaluation of the structure and functional status of WASP in peripheral blood mononuclear cells.Eur J Haematol. 2009; 82: 223-230Crossref PubMed Scopus (7) Google Scholar Evaluation of WASP is also an effective test for WASP-interacting protein deficiency syndrome, an autosomal recessive disease with clinical features similar to WAS caused by impaired expression of WASP-interacting protein, resulting in WASP degradation.36Lanzi G. Moratto D. Vairo D. Masneri S. Delmonte O. Paganini T. et al.A novel primary human immunodeficiency due to deficiency in the WASP-interacting protein WIP.J Exp Med. 2012; 209: 29-34Crossref PubMed Scopus (127) Google Scholar This method has also proved useful in monitoring the presence of somatic reversion of the WAS gene37Wada T. Schurman S.H. Otsu M. Garabedian E.K. Ochs H.D. Nelson D.L. et al.Somatic mosaicism in Wiskott-Aldrich syndrome suggests in vivo reversion by a DNA slippage mechanism.Proc Natl Acad Sci U S A. 2001; 98: 8697-8702Crossref PubMed Scopus (126) Google Scholar and evaluation of donor chimerism after hematopoietic stem cell transplantation.38Yamaguchi K. Ariga T. Yamada M. Nelson D.L. Kobayashi R. Kobayashi C. et al.Mixed chimera status of 12 patients with Wiskott-Aldrich syndrome (WAS) after hematopoietic stem cell transplantation: evaluation by flow cytometric analysis of intracellular WAS protein expression.Blood. 2002; 100: 1208-1214Crossref PubMed Scopus (35) Google Scholar Post-transplantation WASP-positive cells of lymphoid or myeloid lineage in protein-negative patients with WAS represent donor cells, allowing for accurate assessment of the degree of chimerism among these various cell types.38Yamaguchi K. Ariga T. Yamada M. Nelson D.L. Kobayashi R. Kobayashi C. et al.Mixed chimera status of 12 patients with Wiskott-Aldrich syndrome (WAS) after hematopoietic stem cell transplantation: evaluation by flow cytometric analysis of intracellular WAS protein expression.Blood. 2002; 100: 1208-1214Crossref PubMed Scopus (35) Google Scholar A further example of intracellular protein testing by using flow cytometry in patients with PIDs involves screening for patients with possible immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome. This diagnosis is confirmed in male patients whose CD4+CD25+ regulatory T cells demonstrate an absence of forkhead box P3 protein expression.39d'Hennezel E. Bin Dhuban K. Torgerson T. Piccirillo C.A. The immunogenetics of immune dysregulation, polyendocrinopathy, enteropathy, X linked (IPEX) syndrome.J Med Genet. 2012; 49: 291-302Crossref PubMed Scopus (113) Google Scholar Cytotoxic T lymphocyte–associated protein 4 (CTLA4 [CD152]) is another intracellular marker that can be routinely assessed by using flow cytometry in selected patients with CVID-like features, as well as severe enteropathy, brain lesions, and autoimmune cytopenias.40Schubert D. Bode C. Kenefeck R. Hou T.Z. Wing J.B. Kennedy A. et al.Autosom" @default.
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• W2889076804 date "2019-02-01" @default.
• W2889076804 modified "2023-09-30" @default.
• W2889076804 title "Flow cytometry: Surface markers and beyond" @default.
• W2889076804 cites W1208069797 @default.
• W2889076804 cites W1557066644 @default.
• W2889076804 cites W1574766328 @default.
• W2889076804 cites W1965284880 @default.
• W2889076804 cites W1967067685 @default.
• W2889076804 cites W1971734644 @default.
• W2889076804 cites W1982569850 @default.
• W2889076804 cites W1983244640 @default.
• W2889076804 cites W1995924318 @default.
• W2889076804 cites W1996059130 @default.
• W2889076804 cites W1996073092 @default.
• W2889076804 cites W2003530076 @default.
• W2889076804 cites W2015803971 @default.
• W2889076804 cites W2024593848 @default.
• W2889076804 cites W2031778332 @default.
• W2889076804 cites W2035478089 @default.
• W2889076804 cites W2047210789 @default.
• W2889076804 cites W2052632028 @default.
• W2889076804 cites W2057936936 @default.
• W2889076804 cites W2060273459 @default.
• W2889076804 cites W2061416534 @default.
• W2889076804 cites W2061729626 @default.
• W2889076804 cites W2065293336 @default.
• W2889076804 cites W2067517995 @default.
• W2889076804 cites W2074267158 @default.
• W2889076804 cites W2079178172 @default.
• W2889076804 cites W2081238546 @default.
• W2889076804 cites W2081255498 @default.
• W2889076804 cites W2085614469 @default.
• W2889076804 cites W2089543039 @default.
• W2889076804 cites W2089996985 @default.
• W2889076804 cites W2091712495 @default.
• W2889076804 cites W2100462578 @default.
• W2889076804 cites W2101934322 @default.
• W2889076804 cites W2105191357 @default.
• W2889076804 cites W2106134589 @default.
• W2889076804 cites W2108473200 @default.
• W2889076804 cites W2110297391 @default.
• W2889076804 cites W2114667045 @default.
• W2889076804 cites W2114833837 @default.
• W2889076804 cites W2122907933 @default.
• W2889076804 cites W2122970793 @default.
• W2889076804 cites W2123783417 @default.
• W2889076804 cites W2125573161 @default.
• W2889076804 cites W2129942992 @default.
• W2889076804 cites W2130651020 @default.
• W2889076804 cites W2132968076 @default.
• W2889076804 cites W2133488585 @default.
• W2889076804 cites W2138231253 @default.
• W2889076804 cites W2147982220 @default.
• W2889076804 cites W2150234903 @default.
• W2889076804 cites W2150744739 @default.
• W2889076804 cites W2151513117 @default.
• W2889076804 cites W2151863179 @default.
• W2889076804 cites W2154696313 @default.
• W2889076804 cites W2154951784 @default.
• W2889076804 cites W2161151902 @default.
• W2889076804 cites W2235970093 @default.
• W2889076804 cites W2328207062 @default.
• W2889076804 cites W2343832057 @default.
• W2889076804 cites W2346039638 @default.
• W2889076804 cites W2401862332 @default.
• W2889076804 cites W2413715398 @default.
• W2889076804 cites W2512972788 @default.
• W2889076804 cites W2529004588 @default.
• W2889076804 cites W2562042122 @default.
• W2889076804 cites W2593544935 @default.
• W2889076804 cites W2614661137 @default.
• W2889076804 cites W2726820748 @default.
• W2889076804 cites W2790279588 @default.
• W2889076804 cites W2799466648 @default.
• W2889076804 cites W2801254160 @default.
• W2889076804 cites W4248036783 @default.
• W2889076804 cites W4252663695 @default.
• W2889076804 doi "https://doi.org/10.1016/j.jaci.2018.08.011" @default.
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