Limits...
Multifunctional Analysis of CD4+ T-Cell Response as Immune-Based Model for Tuberculosis Detection.

Lichtner M, Mascia C, Sauzullo I, Mengoni F, Vita S, Marocco R, Belvisi V, Russo G, Vullo V, Mastroianni CM - J Immunol Res (2015)

Bottom Line: Among TB-infected subjects, the frequencies of multifunctional CD4(+) T cells, simultaneously producing all 3 cytokines, are lower in active TB than LTBI subjects (P = 0.003).Thus, assigning to triple-positive CD4(+) T cells a cut-off <0.182%, TB-infected individuals could be classified as active TB subjects (<0.182%) or LTBI subjects (>0.182%).The magnitude of CD8(+) T-cell responses showed no differences between active TB and LTBI.

View Article: PubMed Central - PubMed

Affiliation: Department of Public Health and Infectious Diseases, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University, Piazzale Aldo Moro 5, 00185 Rome, Italy ; Infectious Diseases Unit, Sapienza University, Corso della Repubblica 79, 04100 Latina, Italy.

ABSTRACT
Mono- and multifunctional specific CD4(+) and CD8(+) T-cell responses were evaluated to improve the immune-based detection of active tuberculosis (TB) and latent infection (LTBI). We applied flow cytometry to investigate cytokines profile (IFN-γ, TNF-α, and IL-2) of T cells after stimulation with TB antigens in 28 TB-infected subjects (18 active TB and 10 LTBI) and 10 uninfected controls. Cytokines production by CD4(+) T cells at single-cell levels was higher in TB-infected subjects than uninfected controls (P < 0.0001). Assigning to activated CD4(+) T cells, producing any of the three cytokines, a cut-off >0.45%, it was possible to differentiate TB-infected (>0.45%) by uninfected subjects (<0.45%). Among TB-infected subjects, the frequencies of multifunctional CD4(+) T cells, simultaneously producing all 3 cytokines, are lower in active TB than LTBI subjects (P = 0.003). Thus, assigning to triple-positive CD4(+) T cells a cut-off <0.182%, TB-infected individuals could be classified as active TB subjects (<0.182%) or LTBI subjects (>0.182%). The magnitude of CD8(+) T-cell responses showed no differences between active TB and LTBI. Multifunctional CD4(+) T-cell responses could have the potential to identify at single time point subjects without TB infection and patients having active or latent TB.

No MeSH data available.


Related in: MedlinePlus

Analysis of cytokine production by CD8+ T cells at the single-cell level. (a) Frequencies of activated Mtb-specific CD8+T cells producing any of the 3 cytokines (IFN-γ, IL-2, or TNF-α) cells in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) are show. ((b), (c), and (d)) Frequency of “total IFN-γ+,” “total IL-2+,” and “total TNF-α+” Mtb-specific CD8+T in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) is shown. Horizontal bars represent the median values. Statistical analysis was performed using Kruskal-Wallis ANOVA with Dunn's posttest comparison and significant differences are indicated by asterisks (**P < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4539126&req=5

fig5: Analysis of cytokine production by CD8+ T cells at the single-cell level. (a) Frequencies of activated Mtb-specific CD8+T cells producing any of the 3 cytokines (IFN-γ, IL-2, or TNF-α) cells in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) are show. ((b), (c), and (d)) Frequency of “total IFN-γ+,” “total IL-2+,” and “total TNF-α+” Mtb-specific CD8+T in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) is shown. Horizontal bars represent the median values. Statistical analysis was performed using Kruskal-Wallis ANOVA with Dunn's posttest comparison and significant differences are indicated by asterisks (**P < 0.01).

Mentions: The analysis of activated CD8+ T cells, producing any of the 3 cytokines (IFN-γ or IL-2 or TNF-α) showed similar results to CD4+ T cells, revealing a significant greater frequency of activated CD8+ T cells in both active TB (median 0.599%, range 0–4.55%) and LTBI patients (0.489%, 0–1.796%) compared to healthy controls (0%, range 0–0.249%, P < 0.0020 by Kruskal-Wallis test), but no difference was found between two groups of infected individuals (Figure 5(a)). Likewise, the frequency of “total IFN-γ+ CD8+ T cells” (Figure 5(b)), “total IL-2+ CD8+ T cells” (Figure 5(c)), and “total TNF-α+ CD8+ T cells” (Figure 5(d)) was significantly higher in active TB patients compared to the other 2 groups of patients. These differences were only statistically significant between active TB subjects and healthy controls (P = 0.0045 for IFN-γ+; P = 0.0033 for IL-2; P = 0.0078 for TNF-α by Kruskal-Wallis test).


Multifunctional Analysis of CD4+ T-Cell Response as Immune-Based Model for Tuberculosis Detection.

Lichtner M, Mascia C, Sauzullo I, Mengoni F, Vita S, Marocco R, Belvisi V, Russo G, Vullo V, Mastroianni CM - J Immunol Res (2015)

Analysis of cytokine production by CD8+ T cells at the single-cell level. (a) Frequencies of activated Mtb-specific CD8+T cells producing any of the 3 cytokines (IFN-γ, IL-2, or TNF-α) cells in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) are show. ((b), (c), and (d)) Frequency of “total IFN-γ+,” “total IL-2+,” and “total TNF-α+” Mtb-specific CD8+T in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) is shown. Horizontal bars represent the median values. Statistical analysis was performed using Kruskal-Wallis ANOVA with Dunn's posttest comparison and significant differences are indicated by asterisks (**P < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4539126&req=5

fig5: Analysis of cytokine production by CD8+ T cells at the single-cell level. (a) Frequencies of activated Mtb-specific CD8+T cells producing any of the 3 cytokines (IFN-γ, IL-2, or TNF-α) cells in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) are show. ((b), (c), and (d)) Frequency of “total IFN-γ+,” “total IL-2+,” and “total TNF-α+” Mtb-specific CD8+T in active TB patients (n = 18), in LTBI patients (n = 10), and in healthy controls (n = 10) is shown. Horizontal bars represent the median values. Statistical analysis was performed using Kruskal-Wallis ANOVA with Dunn's posttest comparison and significant differences are indicated by asterisks (**P < 0.01).
Mentions: The analysis of activated CD8+ T cells, producing any of the 3 cytokines (IFN-γ or IL-2 or TNF-α) showed similar results to CD4+ T cells, revealing a significant greater frequency of activated CD8+ T cells in both active TB (median 0.599%, range 0–4.55%) and LTBI patients (0.489%, 0–1.796%) compared to healthy controls (0%, range 0–0.249%, P < 0.0020 by Kruskal-Wallis test), but no difference was found between two groups of infected individuals (Figure 5(a)). Likewise, the frequency of “total IFN-γ+ CD8+ T cells” (Figure 5(b)), “total IL-2+ CD8+ T cells” (Figure 5(c)), and “total TNF-α+ CD8+ T cells” (Figure 5(d)) was significantly higher in active TB patients compared to the other 2 groups of patients. These differences were only statistically significant between active TB subjects and healthy controls (P = 0.0045 for IFN-γ+; P = 0.0033 for IL-2; P = 0.0078 for TNF-α by Kruskal-Wallis test).

Bottom Line: Among TB-infected subjects, the frequencies of multifunctional CD4(+) T cells, simultaneously producing all 3 cytokines, are lower in active TB than LTBI subjects (P = 0.003).Thus, assigning to triple-positive CD4(+) T cells a cut-off <0.182%, TB-infected individuals could be classified as active TB subjects (<0.182%) or LTBI subjects (>0.182%).The magnitude of CD8(+) T-cell responses showed no differences between active TB and LTBI.

View Article: PubMed Central - PubMed

Affiliation: Department of Public Health and Infectious Diseases, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University, Piazzale Aldo Moro 5, 00185 Rome, Italy ; Infectious Diseases Unit, Sapienza University, Corso della Repubblica 79, 04100 Latina, Italy.

ABSTRACT
Mono- and multifunctional specific CD4(+) and CD8(+) T-cell responses were evaluated to improve the immune-based detection of active tuberculosis (TB) and latent infection (LTBI). We applied flow cytometry to investigate cytokines profile (IFN-γ, TNF-α, and IL-2) of T cells after stimulation with TB antigens in 28 TB-infected subjects (18 active TB and 10 LTBI) and 10 uninfected controls. Cytokines production by CD4(+) T cells at single-cell levels was higher in TB-infected subjects than uninfected controls (P < 0.0001). Assigning to activated CD4(+) T cells, producing any of the three cytokines, a cut-off >0.45%, it was possible to differentiate TB-infected (>0.45%) by uninfected subjects (<0.45%). Among TB-infected subjects, the frequencies of multifunctional CD4(+) T cells, simultaneously producing all 3 cytokines, are lower in active TB than LTBI subjects (P = 0.003). Thus, assigning to triple-positive CD4(+) T cells a cut-off <0.182%, TB-infected individuals could be classified as active TB subjects (<0.182%) or LTBI subjects (>0.182%). The magnitude of CD8(+) T-cell responses showed no differences between active TB and LTBI. Multifunctional CD4(+) T-cell responses could have the potential to identify at single time point subjects without TB infection and patients having active or latent TB.

No MeSH data available.


Related in: MedlinePlus