Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved]
Background: Interferon-γ (IFN-γ) secretion by T cells is a key correlate of immune protection against many pathogens including tuberculosis and the neglected tropical disease melioidosis. Clinical studies in tropical regions of immune responses to pathogens and vaccine monitoring studies require the...
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| Format: | Article |
| Langue: | English |
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Wellcome
2023-04-01
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| Collection: | Wellcome Open Research |
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| Accès en ligne: | https://wellcomeopenresearch.org/articles/8-188/v1 |
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| author | Susanna J. Dunachie Barbara Kronsteiner Patpong Rongkard |
| author_facet | Susanna J. Dunachie Barbara Kronsteiner Patpong Rongkard |
| author_sort | Susanna J. Dunachie |
| collection | DOAJ |
| description | Background: Interferon-γ (IFN-γ) secretion by T cells is a key correlate of immune protection against many pathogens including tuberculosis and the neglected tropical disease melioidosis. Clinical studies in tropical regions of immune responses to pathogens and vaccine monitoring studies require the collection of samples in resource-limited rural areas and subsequent shipment to central laboratories for downstream assays and long-term storage. Here, we studied the impact of two different shipping temperatures on the viability, composition and function of peripheral blood mononuclear cells (PBMC) using multi-colour flow cytometry and IFN-g enzyme-linked immunospot assay (IFN-g ELISpot), in order to provide guidance on sample shipment conditions for future clinical studies. Methods: Paired peripheral blood mononuclear cell (PBMC) samples from recovered melioidosis patients were stored in liquid nitrogen (-196°C) and then shipped from Bangkok, Thailand to Oxford, UK at either -80°C (dry ice) or -196°C (dry shipper). After thawing, cell viability and composition were assessed by flow cytometry and antigen specific responses to Burkholderia pseudomallei (BP) were measured using IFN-g ELISpot. Results: We observed modest lowering of viability in the majority of samples and a reduction in IFN-g responses to BP which correlated to a decrease of monocytes and natural killer cells in samples shipped at -80°C compared to -196°C. Despite being lower in magnitude antigen-specific responses remained detectable in the majority of samples. Conclusions: Here we demonstrate that shipment of cryopreserved PBMC at -196°C has a benefit on cell viability, recovery and T cell responses to bacterial antigens, although useful information can still be obtained from samples shipped at -80°C, thus providing important guidance for sample management in future clinical trials. |
| first_indexed | 2025-03-21T15:12:37Z |
| format | Article |
| id | doaj.art-fae3b65b51cc4f76b3afd4a77295da38 |
| institution | Directory Open Access Journal |
| issn | 2398-502X |
| language | English |
| last_indexed | 2025-03-21T15:12:37Z |
| publishDate | 2023-04-01 |
| publisher | Wellcome |
| record_format | Article |
| series | Wellcome Open Research |
| spelling | doaj.art-fae3b65b51cc4f76b3afd4a77295da382024-06-20T01:00:00ZengWellcomeWellcome Open Research2398-502X2023-04-01820868Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved]Susanna J. Dunachie0https://orcid.org/0000-0001-5665-6293Barbara Kronsteiner1Patpong Rongkard2https://orcid.org/0000-0002-4094-5143Faculty of Tropical Medicine, Mahidol-Oxford Tropical Medicine Research Unit, Mahidol University, Bangkok, 10400, ThailandCentre for Tropical Medicine and Global Health, Nuffield Department of Medicine, University of Oxford, Oxford, OX3 7LG, UKFaculty of Tropical Medicine, Mahidol-Oxford Tropical Medicine Research Unit, Mahidol University, Bangkok, 10400, ThailandBackground: Interferon-γ (IFN-γ) secretion by T cells is a key correlate of immune protection against many pathogens including tuberculosis and the neglected tropical disease melioidosis. Clinical studies in tropical regions of immune responses to pathogens and vaccine monitoring studies require the collection of samples in resource-limited rural areas and subsequent shipment to central laboratories for downstream assays and long-term storage. Here, we studied the impact of two different shipping temperatures on the viability, composition and function of peripheral blood mononuclear cells (PBMC) using multi-colour flow cytometry and IFN-g enzyme-linked immunospot assay (IFN-g ELISpot), in order to provide guidance on sample shipment conditions for future clinical studies. Methods: Paired peripheral blood mononuclear cell (PBMC) samples from recovered melioidosis patients were stored in liquid nitrogen (-196°C) and then shipped from Bangkok, Thailand to Oxford, UK at either -80°C (dry ice) or -196°C (dry shipper). After thawing, cell viability and composition were assessed by flow cytometry and antigen specific responses to Burkholderia pseudomallei (BP) were measured using IFN-g ELISpot. Results: We observed modest lowering of viability in the majority of samples and a reduction in IFN-g responses to BP which correlated to a decrease of monocytes and natural killer cells in samples shipped at -80°C compared to -196°C. Despite being lower in magnitude antigen-specific responses remained detectable in the majority of samples. Conclusions: Here we demonstrate that shipment of cryopreserved PBMC at -196°C has a benefit on cell viability, recovery and T cell responses to bacterial antigens, although useful information can still be obtained from samples shipped at -80°C, thus providing important guidance for sample management in future clinical trials.https://wellcomeopenresearch.org/articles/8-188/v1IFN-gamma ELISpot PBMC cryopreservation shipping storageeng |
| spellingShingle | Susanna J. Dunachie Barbara Kronsteiner Patpong Rongkard Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] Wellcome Open Research IFN-gamma ELISpot PBMC cryopreservation shipping storage eng |
| title | Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] |
| title_full | Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] |
| title_fullStr | Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] |
| title_full_unstemmed | Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] |
| title_short | Impact of shipping temperature on cell viability and T cell responses to bacterial antigens [version 1; peer review: 2 approved] |
| title_sort | impact of shipping temperature on cell viability and t cell responses to bacterial antigens version 1 peer review 2 approved |
| topic | IFN-gamma ELISpot PBMC cryopreservation shipping storage eng |
| url | https://wellcomeopenresearch.org/articles/8-188/v1 |
| work_keys_str_mv | AT susannajdunachie impactofshippingtemperatureoncellviabilityandtcellresponsestobacterialantigensversion1peerreview2approved AT barbarakronsteiner impactofshippingtemperatureoncellviabilityandtcellresponsestobacterialantigensversion1peerreview2approved AT patpongrongkard impactofshippingtemperatureoncellviabilityandtcellresponsestobacterialantigensversion1peerreview2approved |