Culturing at atmospheric oxygen levels impacts lymphocyte function
Open accessAbstract
To determine whether culturing peripheral blood mononuclear cells at atmospheric oxygen levels skews responses in comparison with culturing lymphocytes at physiologic oxygen levels, we cultured peripheral blood mononuclear cells at 5%, 10%, and atmospheric (20%) gas-phase oxygen for 5 days. We found that incubator oxygen levels influenced lymphocyte proliferation stimulated by two commonly used stimuli: Con A and antibodies that crosslink surface CD3 and CD28 to mimic antigen presentation. In both cases, proliferation increased as gas-phase oxygen levels increased. In contrast, oxygen levels did not influence proliferation stimulated by phytohemagglutinin, another commonly used mitogen. Similarly, oxygen levels did not impact cell viability in unstimulated cultures. Thus, we conclude that the influence of oxygen levels on proliferation depends on the stimulus, and, most importantly from the standpoint of immune responses, culturing cells at atmospheric rather than physiologic oxygen levels results in significantly increased proliferation responses to the CD3/CD28 crosslinking, a proliferation stimulus commonly used to mimic T cell antigen receptor signaling.
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T cells activated at physiologic O2 tension (2-5% O2) have reduced proliferation and expansion following TCR stimulation.
“when T cells are activated in conditions of physiologic oxygen tension (2–5 % O2, ~2–5 kPa, or ~15–38 mm Hg), there are a number of changes in T cell dynamics. First, T cells activated at physiologic O2 tension have reduced proliferation and expansion following TCR stimulation.”
From Hypoxia and hypoxia-inducible factors as regulators of T cell development, differentiation, and function (2013) · cites this paper as [74]
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- Authors
- Atkuri, K. R., Herzenberg, L. A., & Herzenberg, L. A.
- Affiliations
- Stanford Medicine (US)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2005-02-28
- Volume
- 102
- Issue
- 10
- Pages
- 3756-3759
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC553335
- OpenAlex
- W2055563043
- Cited by
- 114
- References
- 16
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 2 · 2023: 6 · 2022: 7 · 2021: 2 · 2020: 6 · 2019: 3 · 2018: 3 · 2017: 9 · 2016: 8 · 2015: 7 · 2014: 7 · 2013: 10 · 2012: 7
- FWCI
- 1.89
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (AI566223)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
- Keywords
- oxygen concentration, lymphocyte proliferation, peripheral blood mononuclear cells, atmospheric oxygen, T cell activation, phytohemagglutinin, concanavalin A, cell viability, culture conditions
- MeSH terms
- Concanavalin A, Humans, Lymphocyte Activation, Oxygen, T-Lymphocytes, CD3 Complex, CD28 Antigens, Cell Culture Techniques
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