Proliferation assay amplification by IL-2 in model primary and recall antigen systems

Abstract

BackgroundIt can be difficult to register a weak proliferative response of T lymphocytes to an antigen, particularly in a simple culture system of peripheral blood mononuclear cells (PBMC). Here we assess the usefulness of the cytokine IL-2 in amplifying such a response.MethodsPBMC from healthy donors were cultured in the presence or absence of keyhole limpet haemocyanin (KLH), an antigen to which people have not been previously exposed. IL-2 was added from the beginning or on the fifth day of culture. Proliferation was determined by incorporation of tritiated thymidine at eight days. The recall antigen, tuberculin PPD, provided a positive control.ResultsIL-2 added at the beginning of culture can induce extremely high levels of proliferation even in the absence of antigen. However, when added on the fifth day it allowed the clear observation of a proliferative response to KLH that was barely detectable in its absence. Added late it was similarly able to boost low responses to PPD and to the mitogens lipopolysaccharide and poly(I:C), but it had no such effect with pokeweed mitogen.ConclusionsIL-2 added late in culture is highly effective in increasing the sensitivity of T lymphocyte proliferative assays.

RETRACTED ARTICLE: A biomimetic assay platform for the interrogation of antigen-dependent anti-tumor T-cell function

“…1b ). PBMC cultures were then expanded in the presence of low dose interleukin-2 (IL-2) for a further 3 days to increase the proliferation of CD8 + CD127 + memory T-cells 18 . PBMC cultures, which consisted mainly of CD4 + and CD8 + T-cells, were then cultured in a “resting” medium for a further 3 days in the presence of IL-2, IL-4, and IL-7 (Fig.…”

Characterisation of the Immunomodulatory Effects of Meningococcal Opa Proteins on Human Peripheral Blood Mononuclear Cells and CD4+ T Cells

“…with PHA or anti-CD3) is a standard method for assessing antigen-specific responses following a vaccination, proliferative responses measured in vitro in the absence of such a recent stimulus can be more difficult to detect [ 33 ]. Further experiments were therefore performed with additional IL-2 to amplify proliferative responses [ 34 ].…”

Comparative Analysis of T-Cell Responses to Aquaporin-4 and Myelin Oligodendrocyte Glycoprotein in Inflammatory Demyelinating Central Nervous System Diseases

“…However, the CFSE dilution assay used here is a powerful and sensitive method for directly detecting proliferation of rare autoantigen-specific human T-cells (31,81). Moreover, the late addition of IL-2 during a re-stimulation step (82)(83)(84) offers high sensitivity and specificity. This strategy effectively increases the sensitivity for rare antigen-specific Tcells by selectively facilitating the proliferation of T lymphocytes that express the IL-2 receptor alpha-chain CD25 following antigen recognition (82,85).…”