Assessment of a multisite standardized biospecimen collection protocol for immune phenotyping in neurodevelopmental disorders

Shane Cleary; Grace Teskey; Craig Mathews; Russell J Sachachar; Robert Nicolson; Rosanna Weksberg; Evdokia Anagnostou; Dawn M E Bowdish; Jane A Foster

doi:10.1038/s41598-023-33380-z

Assessment of a multisite standardized biospecimen collection protocol for immune phenotyping in neurodevelopmental disorders

Sci Rep. 2023 Apr 28;13(1):6971. doi: 10.1038/s41598-023-33380-z.

Authors

Shane Cleary^{1

2}, Grace Teskey³, Craig Mathews^{1

2}, Russell J Sachachar⁴, Robert Nicolson⁵, Rosanna Weksberg^{6

7

8

9}, Evdokia Anagnostou^{7

10}, Dawn M E Bowdish^{3

11}, Jane A Foster^{12

13}

Affiliations

¹ Department of Psychiatry and Behavioural Neurosciences, McMaster University, Hamilton, ON, Canada.
² The Research Institute at St. Joe's, Hamilton, ON, Canada.
³ Department of Medicine, McMaster University, Hamilton, ON, Canada.
⁴ Department of Psychiatry, University of Toronto, The Hospital for Sick Children, Toronto, ON, Canada.
⁵ Lawson Health Research Institute and Western University, London, ON, Canada.
⁶ Division of Clinical and Metabolic Genetics and Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto, ON, Canada.
⁷ Departments of Pediatrics, University of Toronto, Toronto, ON, Canada.
⁸ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
⁹ Institiute of Medical Sciences, University of Toronto, Toronto, ON, Canada.
¹⁰ Bloorview Research Institute, Holland Bloorview Kids Rehabilitation Hospital, Toronto, ON, Canada.
¹¹ McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.
¹² Department of Psychiatry and Behavioural Neurosciences, McMaster University, Hamilton, ON, Canada. [email protected].
¹³ The Research Institute at St. Joe's, Hamilton, ON, Canada. [email protected].

Abstract

Multisite collection and preservation of peripheral blood mononuclear cells (PBMCs) for centralized analysis is an indispensable strategy for large cohort immune phenotyping studies. However, the absence of cross-site standardized protocols introduces unnecessary sample variance. Here we describe the protocol implemented by the Province of Ontario Neurodevelopmental Disorders (POND) Network's immune platform for the multisite collection, processing, and cryopreservation of PBMCs. We outline quality control standards and evaluate the performance of our PBMC processing and storage protocol. We also describe the Child Immune History Questionnaire results, an assessment tool evaluating pre-existing immune conditions in children with neurodevelopmental disorders (NDDs). Cell viability was assessed in samples from 178 participants based on strict quality control criteria. Overall, 83.1% of samples passed quality control standards. Samples collected and processed at the same site had higher quality control pass rates than samples that were collected and subsequently shipped to another site for processing. We investigated if freezer time impacted sample viability and found no difference in mean freezer time between samples that passed and failed quality control. The Child Immune History Questionnaire had a response rate of 87.1%. The described protocol produces viable samples that may be used in future immune phenotyping experiments.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blood Preservation* / methods
Child
Cryopreservation
Humans
Leukocytes, Mononuclear*
Quality Control
Reference Standards