Procedures.
cough, vomiting, fever, diarrhea, proteinuria, asthma, runny nose
INS, ADVERSE EVENT, RECURRENCE, ASTHMA
Following successful screening, a BM sample (50 mL of BM aspirated in an heparinized syringe) was harvested from iliac crests under sedation from each patient and processed within a timeframe of maximum 4–6 hours from harvesting. Both children and adults were hospitalized briefly for this procedure. BM cells were directly seeded in cell-stack flasks up to first harvesting (P1) and subsequently cells were expanded in multiple (up to 6) 5-layer flasks until final harvesting (P2) following identical procedures in the GMP-approved cell factory facilities in both participating centers, as previously reported (One month after the first BM-MSC infusion, steroid and/or concomitant immunosuppressive therapy was tapered and, whenever possible, progressively withdrawn. Immunosuppressive drugs were withdrawn in the following order: first glucocorticoids by reducing the prednisone dose by 2.5–5 mg every 1–2 weeks, then calcineurin inhibitors (cyclosporine A or tacrolimus), and finally antiproliferative agents (azathioprine, mycophenolate mofetil, or cyclophosphamide) by a quarter of the original dose every 2–3 weeks, in order to achieve complete withdrawal within 6 to 9 months. These guidelines were aimed at standardizing immunosuppressive therapy tapering/withdrawal, to limit the potential confounding effect of different approaches on data interpretation, but the principal investigator had the possibility to delay progressive dose reductions according to patient-specific conditions, reporting, and justifying this on the patient’s case report form.INS recurrence was defined as 3 or more positive Albustix dipsticks for proteinuria on 3 consecutive days or by positive dipsticks (1+ to 3+) for 7 consecutive days. Patients with a relapse of INS were treated with high-dose glucocorticoids as per center practice. If the reintroduction of a second-line steroid-sparing immunosuppressive agent was deemed necessary, the best approach was chosen as per center practice and recorded in the patient’s case report form.On follow-up, urine was tested every other day at home by Albustix. Routine laboratory exams, 24-hour proteinuria (or protein/creatinine ratio), and “immunomonitoring” were performed at baseline and after 1, 3, 6, 9, and 12 months from the first MSC infusion, according to the time schedule defined by the study protocol (The methods for classifying/recording AEs, the definitions used to describe an AE, an SAE, an unexpected AE together with the criteria used to assess severity of AE, relationship of AE to study drug(s), and outcome are detailed in the Study Protocol (Supplemental Material). Briefly, all AEs of any severity were recorded in the patient records and in trial case report forms for the duration of the study, from enrolment to study completion. No distinction was made between treatment-emergent AEs and others in terms of detection and reporting. Grading of severity was performed according to GCP, and relation to the study drug was at the investigator’s judgment. Safety reports were compiled every 6 months throughout the study and sent to the Safety Committee for evaluation. An interim analysis was performed as per protocol when more than 10 patients had been treated. Common Terminology Criteria for Adverse Events were employed for grading of all AEs.To record clinical parameters (including urinary protein dipstick, prednisone dose, antibiotic treatment) and tightly monitor for potential side effects of treatment (fever, runny nose, asthma, cough, vomiting, diarrhea), patients or parents of underage patients were asked to complete and submit each month a monthly spreadsheet with daily information. This was particularly useful to maintain a reliable monitoring of all main relevant clinical parameters in the emergency setting of the pandemic, during which for some months in-person visits were not possible in Bergamo.We assessed quality of life with age-appropriate questionnaires — the Short Form Health Survey 36 (SF-36) for adolescents and adults (