CD4 T-helper cell cytokine phenotypes and antibody response following tetanus toxoid booster immunization

Kimberly A. Livingston, Xiaowen Jiang, Charles B. Stephensen

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Routine methods for enumerating antigen-specific T-helper cells may not identify low-frequency phenotypes such as Th2 cells. We compared methods of evaluating such responses to identify tetanus toxoid- (TT) specific Th1, Th2, Th17 and IL10+ cells. Eight healthy subjects were given a TT booster vaccination. Blood was drawn before, 3, 7, 14, and 28days after vaccination and peripheral blood mononuclear cells (PBMC) were cultured for 7days with TT, negative control (diluent), and a positive control (Staphylococcus enterotoxin B [SEB]). Activation markers (CD25 and CD69) were measured after 44h (n=8), cytokines in supernatant after 3 and 7days, and intracellular cytokine staining (ICS) of proliferated cells (identified by dye dilution) after 7days (n=6). Vaccination increased TT-specific expression of CD25 and CD69 on CD3+CD4+ lymphocytes, and TT-specific proliferation at 7, 14 and 28days post vaccination. Vaccination induced TT-specific Th1 (IFN-γ, TNF-α, and IL-2) Th2 (IL-13, IL-5, and IL-4), Th17 (IL-17A) and IL-10+ cells as measured by ICS. TT-specific Th1 cells were the most abundant (12-15% of all TT-specific CD4+ T-cells) while IL10+ (1.8%) Th17 (1.1%) and Th2 cells (0.2-0.6%) were less abundant. TT-specific cytokine concentrations in PBMC supernatants followed the same pattern where a TT-specific IL-9 response was also seen. In conclusion, TT booster vaccination induced a broad T-helper cell response. This method of evaluating cytokine phenotypes may be useful in examining the impact of nutrition and environmental conditions on the plasticity of T-helper cell memory responses.

Original languageEnglish (US)
Pages (from-to)18-29
Number of pages12
JournalJournal of Immunological Methods
Volume390
Issue number1-2
DOIs
StatePublished - Apr 30 2013

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Secondary Immunization
Tetanus Toxoid
Helper-Inducer T-Lymphocytes
Antibody Formation
Cytokines
Phenotype
Vaccination
Interleukin-10
Th17 Cells
Th2 Cells
Blood Cells
Interleukin-9
Staining and Labeling
Th1 Cells
Interleukin-13
Interleukin-17
Interleukin-5
Interleukin-4
Interleukin-2
Healthy Volunteers

Keywords

  • Intracellular cytokine staining
  • T-cell proliferation
  • Tetanus

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

CD4 T-helper cell cytokine phenotypes and antibody response following tetanus toxoid booster immunization. / Livingston, Kimberly A.; Jiang, Xiaowen; Stephensen, Charles B.

In: Journal of Immunological Methods, Vol. 390, No. 1-2, 30.04.2013, p. 18-29.

Research output: Contribution to journalArticle

Livingston, Kimberly A. ; Jiang, Xiaowen ; Stephensen, Charles B. / CD4 T-helper cell cytokine phenotypes and antibody response following tetanus toxoid booster immunization. In: Journal of Immunological Methods. 2013 ; Vol. 390, No. 1-2. pp. 18-29.
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abstract = "Routine methods for enumerating antigen-specific T-helper cells may not identify low-frequency phenotypes such as Th2 cells. We compared methods of evaluating such responses to identify tetanus toxoid- (TT) specific Th1, Th2, Th17 and IL10+ cells. Eight healthy subjects were given a TT booster vaccination. Blood was drawn before, 3, 7, 14, and 28days after vaccination and peripheral blood mononuclear cells (PBMC) were cultured for 7days with TT, negative control (diluent), and a positive control (Staphylococcus enterotoxin B [SEB]). Activation markers (CD25 and CD69) were measured after 44h (n=8), cytokines in supernatant after 3 and 7days, and intracellular cytokine staining (ICS) of proliferated cells (identified by dye dilution) after 7days (n=6). Vaccination increased TT-specific expression of CD25 and CD69 on CD3+CD4+ lymphocytes, and TT-specific proliferation at 7, 14 and 28days post vaccination. Vaccination induced TT-specific Th1 (IFN-γ, TNF-α, and IL-2) Th2 (IL-13, IL-5, and IL-4), Th17 (IL-17A) and IL-10+ cells as measured by ICS. TT-specific Th1 cells were the most abundant (12-15{\%} of all TT-specific CD4+ T-cells) while IL10+ (1.8{\%}) Th17 (1.1{\%}) and Th2 cells (0.2-0.6{\%}) were less abundant. TT-specific cytokine concentrations in PBMC supernatants followed the same pattern where a TT-specific IL-9 response was also seen. In conclusion, TT booster vaccination induced a broad T-helper cell response. This method of evaluating cytokine phenotypes may be useful in examining the impact of nutrition and environmental conditions on the plasticity of T-helper cell memory responses.",
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