Abstract
Flow cytometry has emerged as a useful technology that has facilitated our understanding of the human immune system. Primary immune deficiency disorders (PIDDs) are a heterogeneous group of inherited disorders affecting the immune system. More than 350 genes causing various PIDDs have been identified. While the initial suspicion and recognition of PIDDs is clinical, laboratory tools such as flow cytometry and genetic sequencing are essential for confirmation and categorization. Genetic sequencing, however, are prohibitively expensive and not readily available in resource constrained settings. Flow cytometry remains a simple, yet powerful, tool for multi-parametric analysis of cells. While it is confirmatory of diagnosis in certain conditions, in others it helps in narrowing the list of putative genes to be analyzed. The utility of flow cytometry in diagnosis of PIDDs can be divided into four major categories: (a) Enumeration of lymphocyte subsets in peripheral blood. (b) Detection of intracellular signaling molecules, transcription factors, and cytokines. (c) Functional assessment of adaptive and innate immune cells (e.g., T cell function in severe combined immune deficiency and natural killer cell function in familial hemophagocytic lymphohistiocytosis). (d) Evaluation of normal biological processes (e.g., class switching in B cells by B cell immunophenotyping). This review focuses on use of flow cytometry in disease-specific diagnosis of PIDDs in the context of a developing country.
Highlights
Amit Rawat †, Kanika Arora †, Jitendra Shandilya, Pandiarajan Vignesh, Deepti Suri, Gurjit Kaur, Rashmi Rikhi, Vibhu Joshi, Jhumki Das, Babu Mathew and Surjit Singh*
Flow cytometry has emerged as a useful technology that has facilitated our understanding of the human immune system
The utility of flow cytometry in diagnosis of Primary immune deficiency disorders (PIDDs) can be divided into four major categories: (a) Enumeration of lymphocyte subsets in peripheral blood. (b) Detection of intracellular signaling molecules, transcription factors, and cytokines. (c) Functional assessment of adaptive and innate immune cells (e.g., T cell function in severe combined immune deficiency and natural killer cell function in familial hemophagocytic lymphohistiocytosis). (d) Evaluation of normal biological processes
Summary
Amit Rawat †, Kanika Arora †, Jitendra Shandilya, Pandiarajan Vignesh, Deepti Suri, Gurjit Kaur, Rashmi Rikhi, Vibhu Joshi, Jhumki Das, Babu Mathew and Surjit Singh*. Laboratory Work Flow for SCID at Our Center In a suspected case of SCID or other CID, we first perform lymphocyte subset analysis using a limited panel of four antibodies, i.e., CD45, CD3, CD19, and CD56. Laboratory Work Flow for DOCK8 at Our Center In a suspected case of DOCK8 deficiency, we perform this assay on lymphocytes and neutrophils.
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