title: Culturing at atmospheric oxygen levels impacts lymphocyte function.
abstract: 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.
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305-5318, USA. atkuri@stanford.edu
authors.forename: Kondala R
authors.initials: KR
authors.lastname: Atkuri
authors.forename: Leonard A
authors.initials: LA
authors.lastname: Herzenberg
authors.forename: Leonore A
authors.initials: LA
authors.lastname: Herzenberg
mesh_terms: D018106:CD28 Antigens / Q000502:physiology; D017252:CD3 Complex / Q000502:physiology; D018929:Cell Culture Techniques; D003208:Concanavalin A / Q000494:pharmacology; D006801:Humans; D008213:Lymphocyte Activation; D010100:Oxygen / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D018106:CD28 Antigens; D017252:CD3 Complex; D003208:Concanavalin A; D010100:Oxygen
references.citation: Biophys J. 2001 Aug;81(2):675-84
references.pmid: 11463616
references.citation: Clin Exp Immunol. 1996 Dec;106(3):560-7
references.pmid: 8973628
references.citation: Cytometry. 2001 Nov 1;45(3):194-205
references.pmid: 11746088
references.citation: Clin Lab Med. 2001 Dec;21(4):697-712, vii
references.pmid: 11770282
references.citation: Cytotherapy. 2001;3(6):435-47
references.pmid: 11953028
references.citation: Cytotherapy. 2000;2(1):25-37
references.pmid: 12042052
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4001-5
references.pmid: 12642656
references.citation: J Immunol Methods. 2003 Dec;283(1-2):173-83
references.pmid: 14659909
references.citation: Proc Natl Acad Sci U S A. 2003 Dec 9;100(25):15229-34
references.pmid: 14645704
references.citation: Biotechnol Bioeng. 2004 Aug 20;87(4):437-50
references.pmid: 15286980
references.citation: J Exp Med. 1967 Sep 1;126(3):423-42
references.pmid: 6034749
references.citation: Int J Immunopharmacol. 1996 Oct;18(10):545-52
references.pmid: 9080248
references.citation: J Immunol. 2001 Dec 1;167(11):6140-9
references.pmid: 11714773
references.citation: J Cell Physiol. 1968 Dec;72(3):149-52
references.pmid: 4177706
references.citation: Br J Cancer. 1992 Oct;66(4):619-22
references.pmid: 1419598
references.citation: Respiration. 1997;64(3):200-5
references.pmid: 9154671