Diagnosis of DOCK8 deficiency using Flow cytometry Biomarkers: an Egyptian Center experience

Meshaal, Safa; Hawary, Rabab El; Eldash, Alia; Grimbacher, Bodo; Camacho-Ordóñez, Nadezhda; Elaziz, Dalia Abd; Galal, Nermeen; Boutros, Jeannette; Shawky, Shereen; Elmarsafy, Aisha

doi:10.1016/j.clim.2018.07.011

Cited by 10 publications

(11 citation statements)

References 25 publications

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“…FCM was proved to be a powerful diagnostic tool in many IEI disorders while in others it was useful for screening and narrowing the possibilities of diagnoses [ 16 , 18 ]. Protein expression assays increased the spectrum of the diagnoses achieved and helped reach definitive diagnosis in several disorders like, DOCK8 deficiency, LRBA deficiency, XLA, CGD, Perforin deficiency, ADA deficiency, IL7RA deficiency, LADI and MHC-II deficiency [ 12 , 21 , 22 , 27 ]. Also, sending out for genetic testing has been cut down based on optimizing the molecular diagnosis strategy adapted in our center guided by the FCM [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

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Flow cytometry optimizing the diagnostic approach in inborn errors of immunity: experience from Egypt

Meshaal

Hawary

Eldash

et al. 2022

Allergy Asthma Clin Immunol

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Background Human inborn errors of immunity (IEI) are a group of inherited genetic disorders of the immune system. IEI Patients suffer from severe repeated infections, autoimmunity, lymphadenopathy and/or increased susceptibility to malignancies. IEI are due to absence, disproportion, or loss of function of immune cells; mostly inherited in autosomal recessive manner, hence are more common in countries with high rate of consanguinity. Definite diagnosis of IEI is achieved by genetic analysis, however it is not always available. Aim: to report on different IEI categories and impact of expanding the use of flow cytometry (FCM) in diagnosis, categorization and follow up of IEI patients in a highly consanguineous population. Methods Retrospective chart review on different IEI categories diagnosed at the primary immunodeficiency center in Cairo University Specialized Pediatric hospital from 2011 to 2021 based on expanding the use of FCM. Results 1510 IEI patients were diagnosed; 480 were diagnosed genetically with FMF, 11 with cystic fibrosis and 1019 patients were diagnosed with other IEI disorders. Phagocytic defects were the commonest (30%) followed by severe combined immunodeficiency (22%) and combined immunodeficiency (18.3%). FCM testing properly diagnosed and categorized 73% of the cases. Conclusion Using multi-color FCM to evaluate immune cells populations, subpopulations, functions, and intracellular proteins expression is proved a useful cost-effective method for screening, categorization and follow up of IEI patients. FCM can improve the diagnosis of IEI significantly when tests are properly targeted and well designed. This study presents a 10-year experience in diagnosis of IEI using FCM at a tertiary referral center in a setting of limited resources and yet high prevalence of IEI.

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Section: Discussionmentioning

confidence: 99%

“…We used some purified monoclonal antibodies that were not previously tested on FCM. In these experiments, careful titration was done, and test results were confirmed by targeted genetic analysis, until optimization of the FCM test was achieved [ 12 , 21 , 22 ].…”

Section: Methodsmentioning

confidence: 99%

Flow cytometry optimizing the diagnostic approach in inborn errors of immunity: experience from Egypt

Meshaal

Hawary

Eldash

et al. 2022

Allergy Asthma Clin Immunol

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“…DOCK8 deficiency causes reduced humoral immunity and self-tolerance. The germinal centers, located in secondary lymphoid organs, facilitate the selection and maturation of antigen-activated B-cell clones and provide an optimal immunological response to infections or immunization ( Meshaal et al, 2018 ). However, in absence of DOCK8, the migration of Tfh cells into the germinal center is impaired ( Zhang et al, 2016 ).…”

Section: The Role Of Dock8mentioning

confidence: 99%

CRISPR/Cas-Based Gene Editing Strategies for DOCK8 Immunodeficiency Syndrome

et al. 2022

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Defects in the DOCK8 gene causes combined immunodeficiency termed DOCK8 immunodeficiency syndrome (DIDS). DIDS previously belonged to the disease category of autosomal recessive hyper IgE syndrome (AR-HIES) but is now classified as a combined immunodeficiency (CID). This genetic disorder induces early onset of susceptibility to severe recurrent viral and bacterial infections, atopic diseases and malignancy resulting in high morbidity and mortality. This pathological state arises from impairment of actin polymerization and cytoskeletal rearrangement, which induces improper immune cell migration-, survival-, and effector functions. Owing to the severity of the disease, early allogenic hematopoietic stem cell transplantation is recommended even though it is associated with risk of unintended adverse effects, the need for compatible donors, and high expenses. So far, no alternative therapies have been developed, but the monogenic recessive nature of the disease suggests that gene therapy may be applied. The advent of the CRISPR/Cas gene editing system heralds a new era of possibilities in precision gene therapy, and positive results from clinical trials have already suggested that the tool may provide definitive cures for several genetic disorders. Here, we discuss the potential application of different CRISPR/Cas-mediated genetic therapies to correct the DOCK8 gene. Our findings encourage the pursuit of CRISPR/Cas-based gene editing approaches, which may constitute more precise, affordable, and low-risk definitive treatment options for DOCK8 deficiency.

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“…Different conditions are needed, such as: not stimulated, stimulated, and both with STAT3 antibodies or with an isotype control to have internal controls. Phosphorylation is clearly decreased in a patient with STAT3 deficiency compared to a healthy control [45,46]. See STAT phosphorylation staining protocol in Section 4.2.…”

Section: Stat3 Phosphorylation For the Diagnosis Of Hiper Ige Syndrommentioning

confidence: 99%

Flow Cytometry Applied to the Diagnosis of Primary Immunodeficiencies

Martínez-Gallo¹,

García-Prat²

2020

Innovations in Cell Research and Therapy

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Primary immunodeficiencies are the result of biological defects associated with functional immune abnormalities. It consists of a group of disorders showing a higher incidence and severity of infections, expression of immunological dysregulation such as inflammation and lymphoproliferation. The immunophenotyping and in vitro functional characterization of immunodeficient patients contribute, together with the clinical aspects, to define the underlying immune defect particularities. Flow cytometry applications in primary immunodeficiency assessment are multiple and include the study of a wide range of specific cell lymphocyte subpopulations. This chapter describes the main techniques used in the diagnosis of a wide variety of primary immunodeficiencies, in which intracellular proteins or activation markers involved in immunity are evaluated, as well as functional proliferation, cytokine production, phosphorylation of transcription factors, cytotoxic and degranulation capacity. Flow cytometry is a tool that allows rapid and accurate evaluation of multiple lymphocyte populations and immunological function, and this information is essential for the diagnosis and evaluation of patients with primary immunodeficiencies.

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Diagnosis of DOCK8 deficiency using Flow cytometry Biomarkers: an Egyptian Center experience

Cited by 10 publications

References 25 publications

Flow cytometry optimizing the diagnostic approach in inborn errors of immunity: experience from Egypt

Flow cytometry optimizing the diagnostic approach in inborn errors of immunity: experience from Egypt

CRISPR/Cas-Based Gene Editing Strategies for DOCK8 Immunodeficiency Syndrome

Flow Cytometry Applied to the Diagnosis of Primary Immunodeficiencies

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