Future perspectives on in-vitro diagnosis of drug allergy by the lymphocyte transformation test

Amol Fatangare,Andreas Glässner,Bernhardt Sachs,Albert Sickmann

doi:10.1016/j.jim.2021.113072

Abstract

This article aims to envisage future perspectives of the lymphocyte transformation test (LTT). We describe the select innovative techniques, which can be integrated at different stages of the LTT to potentially improve the sensitivity, specificity, or practicability of the LTT. We first focus upon the cell sorting techniques comprising immunomagnetic cell separation and flow cytometry, which can be implemented prior and after the LTT culturing step to concentrate and quantify specific immune cell types. Further, we elaborate upon three important omics techniques such as transcriptomics, proteomics, and metabolomics, which can be integrated downstream of the LTT to analyze molecular changes in specific immune cells following drug induced activation and proliferation. We also develop visions, how state of the art techniques used in other scientific fields, can be transferred and applied in the context of in-vitro detection of drug allergy.

Highlights

The classical lymphocyte transformation test (LTT) offers valuable information on the diagnosis of hypersensitivity caused by the specific drug
Modern day omics methods generate quantitative information on an entire complement of expressed genes, proteins, or metabolites. The integration of this infor mation in a systems biology approach is a promising way to enhance our understanding of drug allergy. We discuss these three important omics technologies, what potential they carry if applied in the context of in-vitro drug allergy detection, and how they could be implemented in the context of the classical LTT platform (Fig. 1), thereby furthering our understanding of the molecular changes upon T cell activation and proliferation
The successful implementation of quantitative MS-proteomics in the LTT platform could potentially extend our understanding of the proteins involved in the T cell activation and proliferation process, and identify inducible proteins which could serve as potential read-out parameters or markers for the in-vitro detection of drug allergy

Summary

Introduction

The classical lymphocyte transformation test (LTT) offers valuable information on the diagnosis of hypersensitivity caused by the specific drug. The immunological basis of T cell mediated drug allergy is addressed in the separate manuscript “Lymphocyte transformation test: history and current approaches” by Sachs et al in this “Special Issue on the vitro detection of drug allergy” (Fig. 1) Multiple mechanisms such as haptenation, pharmacological interaction with immune re ceptors, or the altered peptide repertoire, have been proposed to explain the observed T cell activation and formation of drug-specific memory T cells and are detailed elsewhere (Pichler, 2019; Weltzien et al, 1996; Yun et al, 2016). We will look at the future perspectives offered by selected innovative technologies to analyze cellular and molecular changes in the drug allergic reactions

Cell isolation and sorting: immunomagnetic cell separation and flow cytometry

Transcriptomics

Proteomics

Metabolomics

Conclusion

Funding source