Cryopreservation-related loss of antigen-specific IFNγ producing CD4 + T-cells can skew immunogenicity data in vaccine trials: Lessons from a malaria vaccine trial substudy
Autor: | Katie J. Ewer, Tom Ford, Jill Gilmour, Nicola Winstone, Len Dally, Claire Wenden, Josephine H. Cox, Alison Mbekeani, Merribeth Morin, Adrian V. S. Hill |
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Přispěvatelé: | Bill & Melinda Gates Foundation, PATH |
Rok vydání: | 2017 |
Předmět: |
CD4-Positive T-Lymphocytes
0301 basic medicine Enzyme-Linked Immunospot Assay MULTICENTER Drug Evaluation Preclinical Research & Experimental Medicine BLOOD MONONUCLEAR-CELLS TRAP 0302 clinical medicine INFECTION PROGRAM IMMUNE-RESPONSES Malaria vaccine ELISPOT Immunogenicity 11 Medical And Health Sciences MVA Flow Cytometry 3. Good health medicine.anatomical_structure Infectious Diseases Medicine Research & Experimental Molecular Medicine Life Sciences & Biomedicine CLINICAL-TRIALS T cell Immunology EFFECTOR ChAd63 Biology Article Specimen Handling Interferon-gamma 03 medical and health sciences Clinical Trials Phase II as Topic CSP Virology Immunology and Microbiology(all) Malaria Vaccines medicine Humans ASSAYS Cryopreservation Science & Technology Staining and Labeling General Veterinary General Immunology and Microbiology MEMORY PBMC Vaccine trial Public Health Environmental and Occupational Health 06 Biological Sciences medicine.disease veterinary(all) Malaria Clinical trial 030104 developmental biology HIV/AIDS VACCINE 07 Agricultural And Veterinary Sciences CD8 030215 immunology |
Zdroj: | Vaccine |
ISSN: | 0264-410X |
DOI: | 10.1016/j.vaccine.2017.02.038 |
Popis: | Ex vivo functional immunoassays such as ELISpot and intracellular cytokine staining (ICS) by flow cytometry are crucial tools in vaccine development both in the identification of novel immunogenic targets and in the immunological assessment of samples from clinical trials. Cryopreservation and subsequent thawing of PBMCs via validated processes has become a mainstay of clinical trials due to processing restrictions inherent in the disparate location and capacity of trial centres, and also in the need to standardize biological assays at central testing facilities. Logistical and financial requirement to batch process samples from multiple study timepoints are also key. We used ELISpot and ICS assays to assess antigen-specific immunogenicity in blood samples taken from subjects enrolled in a phase II malaria heterologous prime-boost vaccine trial and showed that the freeze thaw process can result in a 3 to 5-fold reduction of malaria antigen-specific IFNγ-producing CD3+CD4+ effector populations from PBMC samples taken post vaccination. We have also demonstrated that peptide responsive CD8+ T cells are relatively unaffected, as well as CD4+ T cell populations that do not produce IFNγ. These findings contribute to a growing body of data that could be consolidated and synthesised as guidelines for clinical trials with the aim of increasing the efficiency of vaccine development pipelines. |
Databáze: | OpenAIRE |
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