Work

349: Higher-order cytokine flow cytometry reveals distinct functional signatures of maturation subsets of CD4+ and CD8+ T cells

Year: 2007

Type: article

Abstract: Our clinical studies have suggested that impaired immune recovery after SCT is associated with greater proportions of circulating memory and late memory cells, as defined by phenotyping. To characterize the relationship between immunophenotypic markers of T cell maturation and functional attributes of T cells, we optimized an 8-color, 10-parameter cytokine flow cytometry (CFC) approach and examined PBMC in 3 healthy donors. PBMC were exposed to stimuli that both bypass and signal through the T cell receptor (PMA/Ionomycin—P:I and staph enterotoxin B—SEB, respectively) and stained with CD45RA and CD27 to demarcate naı̈ve (N, CD45RA+CD27+), memory (M, CD45RA-CD27-) and late-memory (LM, CD45RA+CD27-) CD4+ and CD8+ T cells. We assessed the production of effector cytokines (IL-2, IFNγ, TNFα and MIP1β) within the three maturation subsets. Following PMA and SEB stimulation, all maturation subsets (N, M and LM) of CD4+ and CD8+ T cells produced IFNγ. TNFα production was similarly distributed after P:I stimulation. However, after SEB stimulation, LM CD4+ and CD8+ cells produced only 2.5% and 8.5% of total TNFα, respectively. Production of IL-2 and MIP1-β was much more skewed, with LM cells contributing little to total IL-2 production after stimulation with P:I (0.2% of total, CD4; 0%, CD8) or SEB (CD4, 2.1%; CD8, 2.8%). While LM cells produced little or no IL-2, they produced the bulk of MIP1-β following stimulation with P:I (96.5%, CD4; 84.8%, CD8) or SEB (97.5%, CD4; 70.7%, CD8). Co-production of IL-2 and IFNγ in T cells has recently been postulated to be important for protective immunity. Importantly, no IL-2+IFNγ+ cells were seen in the LM compartment following P:I stimulation (CD4+ Distribution: 8.9% N, 91.1% M, 0% LM; CD8: 15.7% N, 84.3% M, 0% LM). Results following SEB stimulation were similar (CD4: 3.3% N, 94.9% M, 1.8% LM; CD8: 12.2% N, 87.8% M, 0% LM). Overall, these results suggest that: 1) T cell maturation stage does correlate strongly with functional capacity; 2) Nearly all IL-2 production occurs in N and M cells; 3) LM cells produce little or no IL-2; but most of total MIP-1β; 4) IL-2/IFNγ co-production is absent in LM cells, perhaps suggesting why late-stage skewing after SCT is associated with functional T cell impairment; and 5) Maturation stage appears to be more closely associated with functional status than CD4 or CD8 lineage. These results may have important implications for clinical studies of post-SCT immune recovery and GVHD.

Source: Transplantation and Cellular Therapy

DOI: 10.1016/j.bbmt.2006.12.354

Authors: T.K. Kim, L.S. St. John, E.D. Wieder, J.J. Molldrem, K.V. Komanduri

Institutions: The University of Texas MD Anderson Cancer Center, University of Houston

Language: English

Topic: T-cell and B-cell Immunology

Subfield: Immunology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,650