Inhibition of activation induced CD154 on CD4+ CD25- cells: a valid surrogate for human Treg suppressor function.
Natural Regulatory T cells (Tregs) are defined by stable expression of the cell surface proteins CD4 and CD25, low surface expression of CD127 and expression of the transcription factor FOXP3. The contribution of Treg to the prevention of autoimmunity and the maintenance of immune homoestasis is the subject of ongoing interest, as alterations in Treg numbers and function are implicated in a wide range of diseases. The in vitro benchmark for determining Treg function is suppression of proliferation of unmatched effector T cells in a mixed lymphocyte reaction (MLR) over a 3-6-day time period. As an alternative to this assay, we show that a 7-h CD154 expression assay is rapid, simple and provides a reliable readout of suppressor function. Using multiple Treg-like cell types including natural (n)Treg, inducible (i)Treg and Treg cell lines, we show that suppression of CD154 expression is a surrogate for suppression of proliferation. We propose this as a suitable alternative to the MLR assay, as it is rapid and may be more amenable to high-throughput screening, analysing large cohorts of clinical samples or assaying transiently suppressive populations.
Molecular Immunology Laboratory, Women's and Children's Health Research Institute, Women's and Children's Hospital, North Adelaide, South Australia, Australia.
Danika
D
Hill
Nicola
N
Eastaff-Leung
Suzanne
S
Bresatz-Atkins
Noel
N
Warner
Joyce
J
Ruitenberg
Doreen
D
Krumbiegel
Steve
S
Pederson
Natasha
N
McInnes
Cheryl Y
CY
Brown
Timothy
T
Sadlon
Simon C
SC
Barry
D000328:Adult; D023201:CD40 Ligand / Q000276:immunology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D005312:Fetal Blood / Q000166:cytology; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D007118:Immunoassay; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D010641:Phenotype; D013194:Staining and Labeling; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology*
C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053645:Interleukin-2 Receptor alpha Subunit; D023201:CD40 Ligand