ctl cryotm abc media kit  (Cellular Technology Ltd)

Journal: Frontiers in Immunology

Article Title: Apheresis for the treatment of relapses in MS and NMOSD: reduced antibody reactivities, gene expression changes and potential clinical response indicators

doi: 10.3389/fimmu.2025.1531447

Figure Lengend Snippet: Study design and experimental workflow. The analyses were based on two independent cohorts: the main study cohort ( n =33 patients) and the validation cohort ( n =30 patients). Patients with MS (solid lines) and patients with NMOSD (broken line) were recruited at the time of an acute relapse. The patients received either high-dose methylprednisolone (green line) or therapeutic apheresis (blue lines). Peripheral blood was taken immediately before and after the relapse treatment. The blood samples were used to separate plasma or serum as well as PBMC. The plasma and serum samples were used to measure antibody reactivities against potential linear epitopes by indirect ELISAs. The PBMC samples from the validation cohort were cryopreserved and later used for a basic immunophenotyping. CD19 + B cells and CD4 + T cells were isolated from a subset of PBMC samples, and their purity was assessed by flow cytometry. RNA from the isolated cells was employed for a transcriptome profiling using Clariom D arrays (main study) and for real-time PCR measurements (validation). The data were analyzed to explore changes in the humoral and cellular immune signature following apheresis and to identify markers of the patients’ clinical response to relapse therapy. We refer to our previous study regarding transcriptome data of MS patients receiving methylprednisolone . ELISA, enzyme-linked immunosorbent assay; MS, multiple sclerosis; NMOSD, neuromyelitis optica spectrum disorder; PBMC, peripheral blood mononuclear cells; PCR, polymerase chain reaction.

Article Snippet: The PBMC samples were cryopreserved in CTL-Cryo ABC medium (Immunospot) in Bochum and shipped in liquid nitrogen to Rostock for the analyses of the present study.

Techniques: Biomarker Discovery, Clinical Proteomics, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction