Journal of immunological methods最新文献

Immediate allergic responses, orchestrated by basophils and mast cells, are pivotal in severe allergic reactions. The flow cytometry-based Basophil Activation Test (BAT) is a clinically important assay for testing allergic reactions using CD63 and CD203c as endpoints. The test measures the concentration dependent response to the allergens providing a functional readout of the patients' allergies. BAT is presently in clinical use within the Unites States as well as several other countries for the diagnosis and monitoring of allergies, most commonly against food allergens. This article details assay validation, both analytical and clinical with reference to existing regulations/recommendations through CAP, CLIA, NYS and CLSI on issues including accuracy, precision, linearity, reportable range, reference range, analytical sensitivity & specificity, pre-analytical considerations, the utility of the assay in diagnosis or monitoring and interpretation of the results; and the assay's limitations. The BAT plays a crucial role in assessing patient suitability for food challenges and therapies such as oral immunotherapy, sublingual immunotherapy or omalizumab and can aid in predicting treatment outcomes. We further review the current research on advancing the test, focused on improvements in its clinical utility. Continuous efforts are warranted for enhanced regulatory oversight and comprehensive clinical validation, ensuring BAT's seamless integration into diverse clinical settings.

Despite great advancements in the discovery of genetic variants underlying inborn errors of immunity, functional assessment of the immunological profile of patients in routine clinical practice remains challenging. The lymphocyte proliferation assay using ³H-thymidine incorporation has been the gold standard for decades for functional evaluation of T cells in the clinical laboratory, however, recently developed flow cytometric methods allowing for single cell analysis provide non-radioactive alternatives. Understanding the technical and analytical challenges of test development, validation and maintenance is essential for correct interpretation and test utilization, to assure appropriate and timely patient care. This review will discuss the technological aspects, validation and clinical utilization of in vitro lymphocyte proliferation assays performed in the clinical immunology laboratory, providing a diagnostic readout for T lymphocyte function, an essential hallmark of a functional cellular immune response, and allowing for the detection of impaired responses such as in patients with functional T cell defects.

Hemophagocytic lymphohistiocytosis (HLH) is a rare clinical syndrome caused by severe systemic hyperinflammation. HLH can be rapidly fatal if unrecognized or inadequately treated. It is important that clinicians are able to utilize diagnostic testing to assess for HLH and determine the underlying causes including possible inborn errors of immunity (IEI). This article summarizes many of the tools available to aid with the diagnostic evaluation of patients with possible HLH and underlying IEI.

Detection of changes in phosphorylation of cell signaling molecules using flow cytometry is termed "phosphoflow" or "phospho-flow cytometry". Phosphoflow has wide application for basic research into the mechanics of cell signaling, for evaluating aberrant signaling in cancerous cells and tissues, for studying efficacy or off-target effects during drug and vaccine development, and for functional assessment of pathogenic variants of genes that are known to play a role in development or function of the immune system. Phosphoflow has not been widely adopted in clinical laboratories owing to the challenges with developing and validating robust assays consistent with clinical laboratory regulatory standards. This review provides a brief overview of the utility of phosphoflow and points of consideration for development and validation of phosphoflow assays for diagnostic use, with a focus on inborn errors of immunity.