OMIP‐025: Evaluation of human T‐ and NK‐cell responses including memory and follicular helper phenotype by intracellular cytokine staining

Abstract

Purpose and Appropriate Sample Types This panel was developed to assess antigen-specific T cells using peptide pools to various antigens of interest, although other types of antigens such as recombinant proteins or whole pathogens could be considered using different stimulation times. In addition to multiple functional markers, the panel includes differentiation markers and markers to assess follicular helper T cells and NK cells (Table 1). It was optimized using cryopreserved peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV) uninfected and HIV infected adults with known cytomegalovirus (CMV) responses and it underwent assay qualification. The panel is being used to evaluate the responses to HIV and malaria vaccine candidates in adults and children from different geographic areas.

Safety and immune responses after a 12-month booster in healthy HIV-uninfected adults in HVTN 100 in South Africa: A randomized double-blind placebo-controlled trial of ALVAC-HIV (vCP2438) and bivalent subtype C gp120/MF59 vaccines

“…The neutralizing antibody (nAb) assay was performed with Env-pseudotyped viruses in TZM-bl cells as previously described [10]. Intracellular cytokine staining (ICS) was performed to measure HIV-specific T cells expressing IFN-γ, IL-2, or CD40L [15] using a 16-color assay. Details on the antigens used in laboratory assays are shown in S2 Table.…”

Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube

“…The diverse functional subsets of CD4+ T cells, particularly Th cells, were first identified based on their unique in vitro profiles of cytokine secretion (1,17,18). However, this requires in vitro culture for variable periods of time (19,20), which is time-consuming and very difficult to standardize for the clinical settings (20). To overcome these limitations, identification of the major subsets of CD4+ T cells has also been performed in the last decades based on their surrogate cell surface membrane phenotypes, by both multiparameter flow cytometry (MFC) (21-25) and mass cytometry (26)(27)(28)(29)(30).…”

OMIP‐056: Evaluation of Human Conventional T Cells, Donor‐Unrestricted T Cells, and NK Cells Including Memory Phenotype by Intracellular Cytokine Staining

“…This panel is unique in the combination of functional and phenotypic markers, but it can be considered an expansion of two of our prior ICS assays, OMIP‐014 , and OMIP‐025 .…”