QUANTITATION OF IMMUNOSUPPRESSION BY FLOW CYTOMETRIC MEASUREMENT OF THE CAPACITY OF T CELLS FOR INTERLEUKIN-2 PRODUCTION1

Abstract We examined the effect of interleukin-3 (IL-3) on human CD4+ cloned T cells, P607 and 1C2. By flow cytometric analysis, we found that IL-3 downregulated the surface expression of IL-2 receptor (R) beta chain in a dose-dependent manner but had little effect on that of IL-2R alpha chain. A simultaneous 125I-labeled IL-2 binding assay showed a decrease in the number of high-affinity, but not of low-affinity, IL-2Rs by IL- 3. The downregulation of the IL-2R beta chain began 3 hours after culture initiation, increased further thereafter, and was completely inhibited by anti-IL-3 antibodies. Expression of mRNA for either alpha or beta chain was not reduced by IL-3, and this suggests that the reduction of surface beta chain expression was not caused by the reduction of beta chain mRNA. IL-3-accelerating internalization of IL- 2R beta chain appeared to be one of the mechanisms for IL-3-induced downregulation of surface IL-2R beta chain expression. IL-3 alone increased the proliferation of T-cell clones but decreased the existing increment of their proliferation by IL-2. Accordingly, IL-3 may be one of the factors acting as a liaison between the hematopoietic and immune systems.

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Flow cytometric measurement of STAT5 phosphorylation in cytomegalovirus‐stimulated T cells

Cytometry Part A ◽

10.1002/cyto.a.24286 ◽

2020 ◽

Author(s):

Michael Bitar ◽

Marcus Boettcher ◽

Andreas Boldt ◽

Fabian Hauck ◽

Ulrike Köhl ◽

...

Keyword(s):

T Cells ◽

Flow Cytometric ◽

Flow Cytometric Measurement

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Quantitation of Immunosuppression by Tacrolimus Using Flow Cytometric Analysis of Interleukin-2 and Interferon-γ Inhibition in CD8− and CD8+ Peripheral Blood T Cells

Therapeutic Drug Monitoring ◽

10.1097/00007691-200108000-00006 ◽

2001 ◽

Vol 23 (4) ◽

pp. 354-362 ◽

Cited By ~ 35

Author(s):

Mamun Ahmed ◽

Raman Venkataraman ◽

Alison J. Logar ◽

Abdul S. Rao ◽

Griffith P. Bartley ◽

...

Keyword(s):

T Cells ◽

Peripheral Blood ◽

Interleukin 2 ◽

Flow Cytometric Analysis ◽

Interferon Γ ◽

Flow Cytometric

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Interleukin-3-induced downregulation of the expression of interleukin-2 receptor beta chain in human T cells

Blood ◽

10.1182/blood.v78.11.2908.bloodjournal78112908 ◽

1991 ◽

Vol 78 (11) ◽

pp. 2908-2917

Author(s):

R Onishi ◽

T Ishikawa ◽

T Kodaka ◽

M Okuma ◽

T Uchiyama

Keyword(s):

T Cells ◽

Interleukin 2 ◽

Flow Cytometric Analysis ◽

Surface Expression ◽

Dependent Manner ◽

Beta Chain ◽

Interleukin 3 ◽

Flow Cytometric ◽

Cell Clones ◽

Human T Cells

We examined the effect of interleukin-3 (IL-3) on human CD4+ cloned T cells, P607 and 1C2. By flow cytometric analysis, we found that IL-3 downregulated the surface expression of IL-2 receptor (R) beta chain in a dose-dependent manner but had little effect on that of IL-2R alpha chain. A simultaneous 125I-labeled IL-2 binding assay showed a decrease in the number of high-affinity, but not of low-affinity, IL-2Rs by IL- 3. The downregulation of the IL-2R beta chain began 3 hours after culture initiation, increased further thereafter, and was completely inhibited by anti-IL-3 antibodies. Expression of mRNA for either alpha or beta chain was not reduced by IL-3, and this suggests that the reduction of surface beta chain expression was not caused by the reduction of beta chain mRNA. IL-3-accelerating internalization of IL- 2R beta chain appeared to be one of the mechanisms for IL-3-induced downregulation of surface IL-2R beta chain expression. IL-3 alone increased the proliferation of T-cell clones but decreased the existing increment of their proliferation by IL-2. Accordingly, IL-3 may be one of the factors acting as a liaison between the hematopoietic and immune systems.

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Flow cytometric measurement of STAT1 and STAT3 phosphorylation in CD4 + and CD8 + T cells—clinical applications in primary immunodeficiency diagnostics

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2017.05.017 ◽

2017 ◽

Vol 140 (5) ◽

pp. 1439-1441.e9 ◽

Cited By ~ 7

Author(s):

Michael Bitar ◽

Andreas Boldt ◽

Stefanie Binder ◽

Michael Borte ◽

Karim Kentouche ◽

...

Keyword(s):

T Cells ◽

Primary Immunodeficiency ◽

Cd8 T Cells ◽

Clinical Applications ◽

Flow Cytometric ◽

Stat3 Phosphorylation ◽

Flow Cytometric Measurement

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Double Negative Alpha Beta T Cells in Pediatrics Patients with Hemophagocytic Syndromes

QJM ◽

10.1093/qjmed/hcaa063.014 ◽

2020 ◽

Vol 113 (Supplement_1) ◽

Author(s):

E M Hossny ◽

R H Elowaidy ◽

H M Afifi ◽

H R Ahmed

Keyword(s):

T Cells ◽

Controlled Study ◽

Induction Treatment ◽

Double Negative ◽

Cell Counts ◽

Flow Cytometric ◽

Flow Cytometric Measurement ◽

Lymphoproliferative Syndrome ◽

Alpha Beta

Abstract Introduction Autoimmune lymphoproliferative syndrome (ALPS) and hemophagocytic lymphohistiocytosis (HLH) share clinical and laboratory features including lymphandenpathy, splenomegaly and pancytopenia. We sought to measure αβ double negative T cells (αβ DN T cells) among patients with established diagnosis of HLH and study its relation to disease activity and severity. Methods we conducted a follow-up controlled study, comprising 25 patients fulfilling diagnosis of HLH according to criteria set forward by the pediatric HLH study Group of the Histiocyte Society, in addition to 25 healthy matched controls. Patients were subjected to clinical evaluation and flow cytometric measurement of αβ DN T Cells percentages at presentation and 9 weeks after start of HLH induction treatment. Results In 17 (68%) patients, infection was the trigger of HLH while the trigger was malignancy in three (12%), and rheumatological disorders in two patients (8%). At enrollment, 15 patients (60%) has elvated αβ DN T cells (>2%), with median (IQR) counts of 1.71 (1.25-2.12) that were significantly higher that the control values (median (IQR): 0.7 (0.4-0.8) (p < 0.001). Initial αβ DN T cells counts of patients were also higher at enrollment as compared to results at the end of week 9 (median (IQR): 0.76 (0.45–1.17), p = 0.018). Survivors (n = 17 [68%] )and non-survivors (n = 8 [32%]) had comparable levels of αβ DN T cells at enrollment (p = 0.861).αβ DN T cells correlated positively with ALT (p = 0.019) and negatively with CD4/CD8 ratio (p = 0.023). Conclusion Elevated αβ DN T cell counts may not be specific for ALPS and their mild elevation might be related to the HLH process. Wider scale studies with longer periods of follow up are needed to validate the results and properly anticipate the correlation between both medical conditions.

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