Measurement of Cellular Immune Response to Viral Infection and Vaccination

Bouwman, Wilbert; Verhaegh, Wim; Holtzer, Laurent; Stolpe, Anja van de

doi:10.3389/fimmu.2020.575074

Cited by 26 publications

(49 citation statements)

References 62 publications

(77 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Subsequently, the comparison was made with CD4+ T cell samples from breast cancer patients to identify the CD4+ T cell subset responsible for immunotolerance in TIL, and to investigate whether the immunotolerant CD4+ T cell pathway profile can also be detected in blood samples from breast cancer patients. The signal transduction pathway assays used in the current study have been biologically validated on multiple cell types, including immune cell types (12)(13)(14)16). Affymetrix expression microarray data were used for the signaling pathway analysis, however, it is good to keep in mind that with this assay technology Affymetrix microarrays are only used as a method to measure a carefully preselected set of mRNA levels, and not to discover a new gene profile.…”

Section: Discussionmentioning

confidence: 99%

“…Increased PI3K (inferred from a decrease in FOXO activity), NFκB, JAK-STAT1/2, and JAK-STAT3 pathway activity reflects T cell activation, and is of crucial importance for clonal proliferation and execution of specific immune response functions (6)(7)(8)(27)(28)(29)(30)(31)(32). Differentiation of activated CD4+ T cells to Th1, Th2 and Treg cells appeared to be associated with characteristic alterations in the pathway activity profile, reflecting specific cell functions determined by signaling pathway activity (13,14,16,19). For example, In Th1 compared to Th2 cells, a higher JAK-STAT1/2 pathway activity is necessary for the Th1 role to activate CD8+ T cells, for example in viral infections (7), while a high PI3K pathway activity is required for expression of the TBET transcription factor, essential for Th1 function (6).…”

Section: Discussionmentioning

confidence: 99%

“…Tests to quantitatively measure functional activity of PI3K, NFκB, TGFβ, JAK-STAT, and Notch signal transduction pathways on Affymetrix Human Genome U133 Plus 2.0 expression microarrays data have been described before (13,14,16,18,19). In brief, the pathway assays are based on the concept of a Bayesian network computational model which calculates from mRNA levels of a selected set, usually between 20 and 30, target genes of the pathway-associated transcription factor a probability score for pathway activity, which is translated to a log2 value of the transcription factor odds, i.e.…”

Section: Signaling Pathway Activity Analysis Of Preclinical and Clinical Studiesmentioning

confidence: 99%

“…the PI3K, JAK-STAT1/2, JAK-STAT3, NFκB, TGFβ, and Notch pathways (6)(7)(8)(9)(10)(11). Recently, assays have been developed which enable quantitative measurement of the activity of these signal transduction pathways in tissue and blood samples (12)(13)(14)(15)(16).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Characterization of immunoactive and immunotolerant CD4+ T cells in breast cancer by measuring activity of signaling pathways that determine immune cell function

Wesseling-Rozendaal

Doorn

Willard-Gallo

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Cancer immunotolerance can be reversed by checkpoint blockade immunotherapy in some patients, but response prediction remains a challenge. CD4+ T cells play an important role in activating adaptive immune responses against cancer. Conversion to an immune suppressive state impairs the anti-cancer immune response and is mainly effected by CD4+ Treg cells. A number of signal transduction pathways activate and control functions of CD4+ T cell subsets. As previously described, assays have been developed which enable quantitative measurement of the activity of signal transduction pathways (e.g. TGFβ, NFκB, PI3K-FOXO, JAK-STAT1/2, JAK-STAT3, Notch) in a cell or tissue sample. Using these assays, pathway activity profiles for various CD4+ T cell subsets were defined and cellular mechanisms underlying breast cancer-induced immunotolerance investigated in vitro. Results were used to measure the immune response state in a clinical breast cancer study.MethodsSignal transduction pathway activity scores were measured on Affymetrix expression microarray data of resting, immune-activated, and immune-activated CD4+ T cells incubated with breast cancer tissue supernatants, and of CD4+ Th1, Th2, and Treg cells, and in a clinical study in which CD4+ T cells were derived from blood, lymph node and cancer tissue from primary breast cancer patients (n=10).ResultsIn vitro CD4+ T cell activation induced PI3K, NFκB, JAK-STAT1/2, and JAK-STAT3 pathway activity. Simultaneous incubation with primary cancer supernatant reduced PI3K and NFκB, and partly reduced JAK-STAT3, pathway activity, while simultaneously increasing TGFβ pathway activity; characteristic of an immune tolerant state. CD4+ Th1, Th2, and Treg cells all had a specific pathway activity profile, with activated immune suppressive Treg cells characterized by NFκB, JAK-STAT3, TGFβ, and Notch pathway activity. An immune tolerant pathway profile was identified in CD4+ T cells from tumor infiltrate of a subset of primary breast cancer patients which could be contributed to activated Treg cells. A Treg pathway profile was also identified in blood samples.ConclusionSignaling pathway assays can be used to quantitatively measure the functional immune response state of lymphocyte subsets in vitro and in vivo. Clinical results suggest that in primary breast cancer the adaptive immune response of CD4+ T cells has frequently been replaced by immunosuppressive Treg cells, potentially causing resistance to checkpoint inhibition. In vitro study results suggest that this effect is mediated by soluble factors from cancer tissue (e.g. TGFβ). Signaling pathway activity analysis on TIL and/or blood samples is expected to improve predicting and monitoring response to checkpoint inhibitor immunotherapy.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Signaling Pathway Activity Analysis Of Preclinical and Clinical Studiesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of immunoactive and immunotolerant CD4+ T cells in breast cancer by measuring activity of signaling pathways that determine immune cell function

Wesseling-Rozendaal

Doorn

Willard-Gallo

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…The measurement of the functional activity of these pathways in tumor biopsies from individual patients is expected to improve the prediction of therapy response. We have previously described a novel approach to quantitatively measure the activity levels of individual signal transduction pathways in various cell and tissue types [22][23][24][25]. In addition to the development of assays to measure the activity of the estrogen and androgen receptor pathways, the PI3K, JAK-STAT3, Wnt, Hedgehog, TGFβ, NFκB and JAK-STAT1/2 pathways, we now report the development and biological validation of a quantitative Notch pathway activity assay.…”

Section: Introductionmentioning

confidence: 99%

A Molecular Test for Quantifying Functional Notch Signaling Pathway Activity in Human Cancer

Canté-Barrett

Holtzer

Ooijen

et al. 2020

Cancers

Self Cite

View full text Add to dashboard Cite

Background: The Notch signal transduction pathway is pivotal for various physiological processes, including immune responses, and has been implicated in the pathogenesis of many diseases. The effectiveness of various targeted Notch pathway inhibitors may vary due to variabilities in Notch pathway activity among individual patients. The quantitative measurement of Notch pathway activity is therefore essential to identify patients who could benefit from targeted treatment. Methods: We here describe a new assay that infers a quantitative Notch pathway activity score from the mRNA levels of generally conserved direct NOTCH target genes. Following the calibration and biological validation of our Notch pathway activity model over a wide spectrum of human cancer types, we assessed Notch pathway activity in a cohort of T-ALL patient samples and related it to biological and clinical parameters, including outcome. Results: We developed an assay using 18 select direct target genes and high-grade serous ovarian cancer for calibration. For validation, seven independent human datasets (mostly cancer series) were used to quantify Notch activity in agreement with expectations. For T-ALL, the median Notch pathway activity was highest for samples with strong NOTCH1-activating mutations, and T-ALL patients of the TLX subtype generally had the highest levels of Notch pathway activity. We observed a significant relationship between ICN1 levels and the absence/presence of NOTCH1-activating mutations with Notch pathway activity scores. Patients with the lowest Notch activity scores had the shortest event-free survival compared to other patients. Conclusions: High Notch pathway activity was not limited to T-ALL samples harboring strong NOTCH1 mutations, including juxtamembrane domain mutations or hetero-dimerization combined with PEST-domain or FBXW7 mutations, indicating that additional mechanisms may activate Notch signaling. The measured Notch pathway activity was related to intracellular NOTCH levels, indicating that the pathway activity score more accurately reflects Notch pathway activity than when it is predicted on the basis of NOTCH1 mutations. Importantly, patients with low Notch pathway activity had a significantly shorter event-free survival compared to patients showing higher activity.

show abstract