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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

Differentiation between active and latent Mtb-infected subjects. (a) ROC curve (plotting sensitivity versus 1 − specificity) to discriminate active TB from LTBI patients. The area under curve (AUC) was 0.8444. (b) Analysis of triple-positive IFN-γ+ IL-2+ TNF-α+ CD4+ T cells, using a cut-off to score responses as either positive or negative. The subjects were considered as positive (black) whether the frequency of CD4+ T cells was >0.182% and negative (white) when the frequency was <0.182%. Horizontal bars represent the median values and horizontal dashed line indicates the cut-off of 0.182%.
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fig4: Differentiation between active and latent Mtb-infected subjects. (a) ROC curve (plotting sensitivity versus 1 − specificity) to discriminate active TB from LTBI patients. The area under curve (AUC) was 0.8444. (b) Analysis of triple-positive IFN-γ+ IL-2+ TNF-α+ CD4+ T cells, using a cut-off to score responses as either positive or negative. The subjects were considered as positive (black) whether the frequency of CD4+ T cells was >0.182% and negative (white) when the frequency was <0.182%. Horizontal bars represent the median values and horizontal dashed line indicates the cut-off of 0.182%.

Mentions: Based on these differences, we performed a ROC analysis (Figure 4(a)) and cut-off < 0.182% for polyfunctional CD4+ T cells allowed the best combination of sensitivity (77.78%, 95% CI: 52.36–93.59) and specificity (70%, 95% CI: 34.75–93.33%; AUC 0.8444; 95% CI: 0.7021–0.9868%, P = 0.0002) to differentiate between active TB and LTBI subjects. Using this cut-off to score ICCFC responses as either positive or negative, we observed a positive response (>0.182%) in 4 out of 18 (22%) active TB patients and in 7 out of 10 (70%) LTBI patients (Figure 4(b)). In our hands, the frequency of polyfunctional CD4+ T cells which simultaneously produced IFN-γ, IL-2, and TNF-α may be indicative of LTBI status.


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)

Differentiation between active and latent Mtb-infected subjects. (a) ROC curve (plotting sensitivity versus 1 − specificity) to discriminate active TB from LTBI patients. The area under curve (AUC) was 0.8444. (b) Analysis of triple-positive IFN-γ+ IL-2+ TNF-α+ CD4+ T cells, using a cut-off to score responses as either positive or negative. The subjects were considered as positive (black) whether the frequency of CD4+ T cells was >0.182% and negative (white) when the frequency was <0.182%. Horizontal bars represent the median values and horizontal dashed line indicates the cut-off of 0.182%.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Differentiation between active and latent Mtb-infected subjects. (a) ROC curve (plotting sensitivity versus 1 − specificity) to discriminate active TB from LTBI patients. The area under curve (AUC) was 0.8444. (b) Analysis of triple-positive IFN-γ+ IL-2+ TNF-α+ CD4+ T cells, using a cut-off to score responses as either positive or negative. The subjects were considered as positive (black) whether the frequency of CD4+ T cells was >0.182% and negative (white) when the frequency was <0.182%. Horizontal bars represent the median values and horizontal dashed line indicates the cut-off of 0.182%.
Mentions: Based on these differences, we performed a ROC analysis (Figure 4(a)) and cut-off < 0.182% for polyfunctional CD4+ T cells allowed the best combination of sensitivity (77.78%, 95% CI: 52.36–93.59) and specificity (70%, 95% CI: 34.75–93.33%; AUC 0.8444; 95% CI: 0.7021–0.9868%, P = 0.0002) to differentiate between active TB and LTBI subjects. Using this cut-off to score ICCFC responses as either positive or negative, we observed a positive response (>0.182%) in 4 out of 18 (22%) active TB patients and in 7 out of 10 (70%) LTBI patients (Figure 4(b)). In our hands, the frequency of polyfunctional CD4+ T cells which simultaneously produced IFN-γ, IL-2, and TNF-α may be indicative of LTBI status.

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