Hospital Coordination and Protocols Using Serum and Peripheral Blood Cells from Patients and Healthy Donors in a Longitudinal Study of Guillain-Barré Syndrome

Abstract

Background/Objectives: Guillain-Barré syndrome (GBS) is a rare autoimmune peripheral neuropathy that affects both the myelin sheaths and axons of the peripheral nervous system. It is the leading cause of acute neuromuscular paralysis worldwide, with an annual incidence of less than two cases per 100,000 people. Although most patients recover, a small proportion do not regain mobility and even remain dependent on mechanical ventilation. In this study, we refer to the analysis of samples collected from GBS patients at different defined time points during hospital recovery and performed by a medical or research group. Methods: The conditions for whole blood collection, peripheral blood mononuclear cell isolation, and serum collection from GBS patients and volunteer donors are explained. Aliquots of these human samples have been used for red blood cell phenotyping, transcriptomic and proteomic analyses, and serum biochemical parameter studies. Results: The initial sporadic preservation of human samples from GBS patients and control volunteers enabled the creation of a biobank collection for current and future studies related to the diagnosis and treatment of GBS. Conclusions: In this article, we describe the laboratory procedures and the integration of a GBS biobank collection, local medical services, and academic institutions collaborating in its respective field. The report establishes the intra-disciplinary and inter-institutional network to conduct long-term longitudinal studies on GBS.

Keywords: Guillain–Barre syndrome; biobank collection; biochemical parameters; biomarkers; diagnosis; phenotyping; proteomics; transcriptomics.

Figure 1

Patient and control individuals included in the current BioGBS study. In (A): composition of each group with demographic, clinical diagnosis, and inclusion/exclusion criteria for the BioGBS study; (*) number of healthy donors increases progressively until reaching the same number of GBS group for stratification by age and sex. In (B): current distribution of GBS Subtypes in GBS Biobank Collection: MFS, Miller Fisher Syndrome; AIDP, Acute Inflammatory Demyelinating Polyradiculoneuropathy; AMAN, Acute Motor Axonal Neuropathy; AMSAN, Acute Motor Sensory Axonal Neuropathy; and Undefined GBS subtype. N, number of individuals; (F/M) female/male; GBS, Guillain–Barre syndrome; tSCI, traumatic spinal cord injury.

Figure 2

Whole blood processing from volunteer donors and selected blood collection time points from Guillain–Barre syndrome onset to recovery. The different steps for obtaining serum, PBMCs, red blood cells (RBC) phenotyping, and final storage of samples in the GBS Biobank Collection (TOSGB) are shown (A). Monophasic behavior curve for GBS progression and recovery after infection; blood collections performed in general and rehabilitation hospitals at time points T0, T1, T2, and T3, corresponding to the early, plateau, onset recovery, and moderate recovery phases of GBS (B).

Figure 3

Examples of quantitative real-time PCR in PBMCs, serum proteomic analysis, and serum antibodies from samples in the TOSGB Biobank collection. The mRNA from PBMCs was used to determine gene expression levels of GBS-specific genetic markers (A). Serum from GBS patients and controls were processed for proteomic analysis (B), expression of GBS-specific proteins is shown in the left panel (a), and the corresponding affected biological function in the right panel (b,c). Specific anti-α-Gal antibody types in serum from GBS patients and GBS/COVID-19 patients stored in the GBS Biobank, compared with COVID-19 patients and healthy controls (C). Modified from [17,18,22] with permission.

Figure 4

Representative determination of serum biochemical parameters from a patient with GBS and phenotyping of red blood cells from a control donor. Serum analysis of various biochemical parameters at time points T1, T2, and T3 during the recovery of a male patient with GBS (A). Phenotyping of red blood cells obtained from the blood of a male volunteer donor (B). AST, serum glutamic–oxaloacetic transaminase or aspartate aminotransferase; ALT, serum glutamic–pyruvic transaminase or alanine aminotransferase; GGT, gamma–glutamyl transferase.

Figure 5

Central role of GBS Biobank Collection (TOSGB) for intra-hospital and inter-institutional interaction in BioGBS study.