Delayed Activation Kinetics of Th2- and Th17 Cells Compared to Th1 Cells

Duechting, Andrea; Przybyla, Anna; Küerten, Stefanie; Lehmann, Paul

doi:10.3390/cells6030029

Cited by 20 publications

(18 citation statements)

References 34 publications

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“…Using this ICS-based approach, only 2.1% of the seronegative donors were found to have HCMV-specific memory T cells [ 10 ], apparently contradicting our finding reported here that such memory cells are frequently present in HCMV-seronegative donors. The ELISPOT assays we performed to generate the data for Table 1 , including a 24 h antigen-stimulation culture, as has been established to be ideal for detecting IFN-γ-secreting T cells in general [ 25 ]. By studying the kinetics of the I-HCMV-induced IFN-γ response, we addressed the hypothesis that the underestimation of HCMV-specific T cells by ICS might have resulted from an insufficiently lengthy antigen stimulation period.…”

Section: Resultsmentioning

confidence: 99%

Direct Detection of T- and B-Memory Lymphocytes by ImmunoSpot® Assays Reveals HCMV Exposure that Serum Antibodies Fail to Identify

Terlutter¹,

Caspell²,

Nowacki

et al. 2018

Cells

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It is essential to identify donors who have not been infected with human cytomegalovirus (HCMV) in order to avoid transmission of HCMV to recipients of blood transfusions or organ transplants. In the present study, we tested the reliability of seronegativity as an indicator for the lack of HCMV exposure in healthy human blood donors. Eighty-two HCMV seronegative individuals were identified, and their peripheral blood mononuclear cells (PBMC) were tested in ImmunoSpot® assays for the presence of HCMV-specific T- and B-memory lymphocytes. Eighty-two percent (67 of 82) of these HCMV seronegative individuals featured at least one memory cell that was lineage specific for HCMV, with the majority of these subjects possessing CD4+ and CD8+ T cells, as well as B cells, providing three independent lines of evidence for having developed immunity to HCMV. Only 15 of these 82 donors (18%) showed neither T- nor B-cell memory to HCMV, consistent with immunological naïveté to the virus. The data suggest that measurements of serum antibodies frequently fail to reveal HCMV exposure in humans, which may be better identified by direct detection of HCMV-specific memory lymphocytes.

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Section: Resultsmentioning

confidence: 99%

Direct Detection of T- and B-Memory Lymphocytes by ImmunoSpot® Assays Reveals HCMV Exposure that Serum Antibodies Fail to Identify

Terlutter¹,

Caspell²,

Nowacki

et al. 2018

Cells

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“…ELISPOT assays detecting IL-2, IL-4, IL-5, and IL-17 were performed in the same way using the respective kits for IL-2 (cat.# CTL-HIL2-1/5), IL-4 (cat.# HIL4-1/5), IL-5 (cat.# HIL5-1/5) and IL-17 (cat.# CTL- HIL17-1/5). Due to the delayed cytokine secretion kinetics of IL-4, IL-5 and IL-17-producing CD4 cells [ 14 ], the antigen stimulation culture for the IL-4 assays was 48 h, and for IL-5 and IL-17 assays, 72 h. The plates were air-dried in a laminar flow hood prior to analysis. The ELISPOT plates were scanned and analyzed using an ImmunoSpot ® S6 Ultimate Reader from CTL.…”

Section: Methodsmentioning

confidence: 99%

A Positive Control for Detection of Functional CD4 T Cells in PBMC: The CPI Pool

Schiller¹,

Zhang²,

Li³

et al. 2017

Cells

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Testing of peripheral blood mononuclear cells (PBMC) for immune monitoring purposes requires verification of their functionality. This is of particular concern when the PBMC have been shipped or stored for prolonged periods of time. While the CEF (Cytomegalo-, Epstein-Barr and Flu-virus) peptide pool has become the gold standard for testing CD8 cell functionality, a positive control for CD4 cells is so far lacking. The latter ideally consists of proteins so as to control for the functionality of the antigen processing and presentation compartments, as well. Aiming to generate a positive control for CD4 cells, we first selected 12 protein antigens from infectious/environmental organisms that are ubiquitous: Varicella, Influenza, Parainfluenza, Mumps, Cytomegalovirus, Streptococcus, Mycoplasma, Lactobacillus, Neisseria, Candida, Rubella, and Measles. Of these antigens, three were found to elicited interferon (IFN)-γ-producing CD4 cells in the majority of human test subjects: inactivated cytomegalo-, parainfluenza-, and influenza virions (CPI). While individually none of these three antigens triggered a recall response in all donors, the pool of the three (the ‘CPI pool’), did. One hundred percent of 245 human donors tested were found to be CPI positive, including Caucasians, Asians, and African-Americans. Therefore, the CPI pool appears to be suitable to serve as universal positive control for verifying the functionality of CD4 and of antigen presenting cells.

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“…[56][57][58] The low detection of vaccineinduced Th17 responses in our experiments cannot be linked to the stimulation protocol; however, since Th17 responses typically require a time-frame of ≤5 days of stimulation. 39,59 Thus, our data suggest that both AT and ClfA are ineffective activators of Th17 responses in the vaccine context.…”

Section: Discussionmentioning

confidence: 73%

Role and plasticity of Th1 and Th17 responses in immunity to Staphylococcus aureus

Ferraro

Buonocore

Auquier

et al. 2019

Human Vaccines & Immunotherapeutics

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The human commensal Staphylococcus aureus (SA) is a leading cause of skin/soft tissue and surgical-site infections, and bacteremia. Functional antibodies and T-cell-mediated immunity, particularly Th1/Th17 responses, are thought to mediate protection. Vaccine development may be hindered by modulation of vaccine-induced T cells by pathogen-activated immunoregulatory responses, e.g., via IL-10.We screened SA proteins for CD4+ T-cell-activating and IL-10/IL-17-inducing capacities using healthy donor-derived PBMCs. Responses were characterized (Th1/Th17/Th22/immunosuppressive IL-10-producing cells) using intracellular cytokine staining and flow cytometry. Phenotypic plasticity of Th1/Th17 cells was evaluated under pro- or anti-inflammatory conditions using modulatory cytokines. The impact of vaccination on SA-specific memory responses was assessed using samples from a clinical trial evaluating AS03-adjuvanted and non-adjuvanted multicomponent (CPS5/CPS8/α-toxin/ClfA) vaccines (NCT01160172).The donors exhibited SA-specific memory T-cell responses, indicative of pre-existing immunity to SA. We identified effective activators of Th1 responses (EbhA/IsaA/SdrE/MntC/Aaa/α-toxin), and Th17 and Th1/Th17 responses (EbhA/IsaA/SdrE and, to a lesser extent, α-toxin), but not of Th22 responses or IL-10 production. MRPII, IsdA, and ClfA were inefficient CD4+ T-cell activators in our assays. IL-10, likely produced by innate immune cells, influenced mainly Th1 cells by suppressing IFN-γ production. The memory CD4+ T-cells observed after long-term stimulation with α-toxin and ClfA indicated that vaccination with these proteins had induced expansion of pre-existing Th1 but not Th17 responses, without apparent adjuvant effect, confirming the trial data. The Th1/Th17-driving proteins (EbhA/IsaA/SdrE) shared low IL-10-promoting abilities and restricted phenotypic plasticity under pro- and anti-inflammatory conditions.Given the complex immunopathology and multiple virulence factors, identification of Th1/Th17-driving antigens, adjuvants and administration routes, and delineation of the role of memory responses, may advance vaccine development.

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Delayed Activation Kinetics of Th2- and Th17 Cells Compared to Th1 Cells

Cited by 20 publications

References 34 publications

Direct Detection of T- and B-Memory Lymphocytes by ImmunoSpot® Assays Reveals HCMV Exposure that Serum Antibodies Fail to Identify

Direct Detection of T- and B-Memory Lymphocytes by ImmunoSpot® Assays Reveals HCMV Exposure that Serum Antibodies Fail to Identify

A Positive Control for Detection of Functional CD4 T Cells in PBMC: The CPI Pool

Role and plasticity of Th1 and Th17 responses in immunity to Staphylococcus aureus

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