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CD161+ MAIT cells are severely reduced in peripheral blood and lymph nodes of HIV-infected individuals independently of disease progression.

Eberhard JM, Hartjen P, Kummer S, Schmidt RE, Bockhorn M, Lehmann C, Balagopal A, Hauber J, van Lunzen J, Schulze zur Wiesch J - PLoS ONE (2014)

Bottom Line: Mononuclear cells from blood and lymph node samples as well as plasma from 63 patients and 26 healthy donors were analyzed by multicolor flow cytometry and ELISA for IL-18, sCD14 and sCD163.CD161+ MAIT cell numbers did not recover even after successful antiretroviral treatment.In vitro stimulation of MAIT cells with IL-18 and IL-12, IL-7 and fixed E.coli also resulted in a rapid and additive reduction of the MAIT cell frequency defined by CD161, IL-18R and CCR6.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases Unit, Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.

ABSTRACT
Mucosal-associated invariant T (MAIT) cells are characterized by the combined expression of the semi-invariant T cell receptor (TCR) Vα7.2, the lectin receptor CD161, as well as IL-18R, and play an important role in antibacterial host defense of the gut. The current study characterized CD161(+) MAIT and CD161-TCRVα7.2(+) T cell subsets within a large cohort of HIV patients with emphasis on patients with slow disease progression and elite controllers. Mononuclear cells from blood and lymph node samples as well as plasma from 63 patients and 26 healthy donors were analyzed by multicolor flow cytometry and ELISA for IL-18, sCD14 and sCD163. Additionally, MAIT cells were analyzed after in vitro stimulation with different cytokines and/or fixed E.coli. Reduced numbers of CD161(+) MAIT cells during HIV infection were detectable in the blood and lymph nodes of all patient groups, including elite controllers. CD161+ MAIT cell numbers did not recover even after successful antiretroviral treatment. The loss of CD161(+) MAIT cells was correlated with higher levels of MAIT cell activation; an increased frequency of the CD161-TCRVα7.2(+)T cell subset in HIV infection was observed. In vitro stimulation of MAIT cells with IL-18 and IL-12, IL-7 and fixed E.coli also resulted in a rapid and additive reduction of the MAIT cell frequency defined by CD161, IL-18R and CCR6. In summary, the irreversible reduction of the CD161(+) MAIT cell subset seems to be an early event in HIV infection that is independent of later stages of the disease. This loss appears to be at least partially due to the distinctive vulnerability of MAIT cells to the pronounced stimulation by microbial products and cytokines during HIV-infection.

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Frequencies of CD161+ MAIT cells are reduced upon in vitro stimulation with PFA fixed E.coli, IL-7 and combined IL-12 and IL-18.A) Representative FACS plots depicting frequencies of CD161–TCRVα7.2+ cells, CD161+ MAIT cells and all TCRVα7.2+ cells as frequencies of CD3+ T cells after 28 hours of stimulation with IL-12 (100 µg/ml), IL-18 (100 µg/ml) and their combination, IL-7 (100 µg/ml) and PFA fixed E. coli (bacteria per cell ratio of 100∶1 PBMC). PBMCs were healthy donor-derived and seeded in 1×106 cells/well. B) Cumulative display of normalized frequencies of CD161+ MAIT cells and CD161–TCRVα7.2+ cells from 3 healthy donors in duplicate measurements for each stimulation setting (apart from E.coli in combination with cytokines, which was tested once) measured after 28 hours of stimulation (left panel). Activation levels were assessed by determining the percentage of CD69+ cells within the CD161+ MAIT cell and the CD161–TCRVα7.2+ population.
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pone-0111323-g004: Frequencies of CD161+ MAIT cells are reduced upon in vitro stimulation with PFA fixed E.coli, IL-7 and combined IL-12 and IL-18.A) Representative FACS plots depicting frequencies of CD161–TCRVα7.2+ cells, CD161+ MAIT cells and all TCRVα7.2+ cells as frequencies of CD3+ T cells after 28 hours of stimulation with IL-12 (100 µg/ml), IL-18 (100 µg/ml) and their combination, IL-7 (100 µg/ml) and PFA fixed E. coli (bacteria per cell ratio of 100∶1 PBMC). PBMCs were healthy donor-derived and seeded in 1×106 cells/well. B) Cumulative display of normalized frequencies of CD161+ MAIT cells and CD161–TCRVα7.2+ cells from 3 healthy donors in duplicate measurements for each stimulation setting (apart from E.coli in combination with cytokines, which was tested once) measured after 28 hours of stimulation (left panel). Activation levels were assessed by determining the percentage of CD69+ cells within the CD161+ MAIT cell and the CD161–TCRVα7.2+ population.

Mentions: To determine possible factors that might contribute to the reduction of the CD161+ MAIT cell population in HIV-infection on the one hand or possibly could lead to proliferation on the other hand, we tested the effect of cytokines (i.e. cytokines for which MAIT cells highly express the respective receptor) on CD161+ MAIT cells. Moreover, the hypothesis of microbial translocation as a driving force for MAIT cell reduction in HIV was tested by adding fixed E.coli in a bacteria per cell ratio of 100∶1 to the in vitro culture. In an in vitro experiment, PBMCs from healthy donors were stimulated with IL-12, IL-18, IL-7, fixed E.coli alone or in combination to elucidate the effect on MAIT cell activation (Fig. 4A, B). No significant alteration in the level of activation or frequency of the CD161+ MAIT cell population was observed with addition of IL-12 or IL-18 alone (Fig. 4A, B). Combined stimulation with IL-12 and IL-18 in contrast led to potent activation of the MAIT cell subset as measured by CD69 expression (unstimulated cells mean = 2.98±1.16% vs. IL-12+IL-18 mean = 53.88±27.04%) accompanied by a slight reduction of the MAIT cell frequency among CD3+ T cells, reflecting synergistic effects of IL-18 and IL-12 (Fig. 4A, B). The reduction of the MAIT cell frequency was confirmed using IL18R or CCR6 expression in addition to CD161 expression as alternative MAIT cell characterization (Fig. S4).


CD161+ MAIT cells are severely reduced in peripheral blood and lymph nodes of HIV-infected individuals independently of disease progression.

Eberhard JM, Hartjen P, Kummer S, Schmidt RE, Bockhorn M, Lehmann C, Balagopal A, Hauber J, van Lunzen J, Schulze zur Wiesch J - PLoS ONE (2014)

Frequencies of CD161+ MAIT cells are reduced upon in vitro stimulation with PFA fixed E.coli, IL-7 and combined IL-12 and IL-18.A) Representative FACS plots depicting frequencies of CD161–TCRVα7.2+ cells, CD161+ MAIT cells and all TCRVα7.2+ cells as frequencies of CD3+ T cells after 28 hours of stimulation with IL-12 (100 µg/ml), IL-18 (100 µg/ml) and their combination, IL-7 (100 µg/ml) and PFA fixed E. coli (bacteria per cell ratio of 100∶1 PBMC). PBMCs were healthy donor-derived and seeded in 1×106 cells/well. B) Cumulative display of normalized frequencies of CD161+ MAIT cells and CD161–TCRVα7.2+ cells from 3 healthy donors in duplicate measurements for each stimulation setting (apart from E.coli in combination with cytokines, which was tested once) measured after 28 hours of stimulation (left panel). Activation levels were assessed by determining the percentage of CD69+ cells within the CD161+ MAIT cell and the CD161–TCRVα7.2+ population.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4219715&req=5

pone-0111323-g004: Frequencies of CD161+ MAIT cells are reduced upon in vitro stimulation with PFA fixed E.coli, IL-7 and combined IL-12 and IL-18.A) Representative FACS plots depicting frequencies of CD161–TCRVα7.2+ cells, CD161+ MAIT cells and all TCRVα7.2+ cells as frequencies of CD3+ T cells after 28 hours of stimulation with IL-12 (100 µg/ml), IL-18 (100 µg/ml) and their combination, IL-7 (100 µg/ml) and PFA fixed E. coli (bacteria per cell ratio of 100∶1 PBMC). PBMCs were healthy donor-derived and seeded in 1×106 cells/well. B) Cumulative display of normalized frequencies of CD161+ MAIT cells and CD161–TCRVα7.2+ cells from 3 healthy donors in duplicate measurements for each stimulation setting (apart from E.coli in combination with cytokines, which was tested once) measured after 28 hours of stimulation (left panel). Activation levels were assessed by determining the percentage of CD69+ cells within the CD161+ MAIT cell and the CD161–TCRVα7.2+ population.
Mentions: To determine possible factors that might contribute to the reduction of the CD161+ MAIT cell population in HIV-infection on the one hand or possibly could lead to proliferation on the other hand, we tested the effect of cytokines (i.e. cytokines for which MAIT cells highly express the respective receptor) on CD161+ MAIT cells. Moreover, the hypothesis of microbial translocation as a driving force for MAIT cell reduction in HIV was tested by adding fixed E.coli in a bacteria per cell ratio of 100∶1 to the in vitro culture. In an in vitro experiment, PBMCs from healthy donors were stimulated with IL-12, IL-18, IL-7, fixed E.coli alone or in combination to elucidate the effect on MAIT cell activation (Fig. 4A, B). No significant alteration in the level of activation or frequency of the CD161+ MAIT cell population was observed with addition of IL-12 or IL-18 alone (Fig. 4A, B). Combined stimulation with IL-12 and IL-18 in contrast led to potent activation of the MAIT cell subset as measured by CD69 expression (unstimulated cells mean = 2.98±1.16% vs. IL-12+IL-18 mean = 53.88±27.04%) accompanied by a slight reduction of the MAIT cell frequency among CD3+ T cells, reflecting synergistic effects of IL-18 and IL-12 (Fig. 4A, B). The reduction of the MAIT cell frequency was confirmed using IL18R or CCR6 expression in addition to CD161 expression as alternative MAIT cell characterization (Fig. S4).

Bottom Line: Mononuclear cells from blood and lymph node samples as well as plasma from 63 patients and 26 healthy donors were analyzed by multicolor flow cytometry and ELISA for IL-18, sCD14 and sCD163.CD161+ MAIT cell numbers did not recover even after successful antiretroviral treatment.In vitro stimulation of MAIT cells with IL-18 and IL-12, IL-7 and fixed E.coli also resulted in a rapid and additive reduction of the MAIT cell frequency defined by CD161, IL-18R and CCR6.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases Unit, Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.

ABSTRACT
Mucosal-associated invariant T (MAIT) cells are characterized by the combined expression of the semi-invariant T cell receptor (TCR) Vα7.2, the lectin receptor CD161, as well as IL-18R, and play an important role in antibacterial host defense of the gut. The current study characterized CD161(+) MAIT and CD161-TCRVα7.2(+) T cell subsets within a large cohort of HIV patients with emphasis on patients with slow disease progression and elite controllers. Mononuclear cells from blood and lymph node samples as well as plasma from 63 patients and 26 healthy donors were analyzed by multicolor flow cytometry and ELISA for IL-18, sCD14 and sCD163. Additionally, MAIT cells were analyzed after in vitro stimulation with different cytokines and/or fixed E.coli. Reduced numbers of CD161(+) MAIT cells during HIV infection were detectable in the blood and lymph nodes of all patient groups, including elite controllers. CD161+ MAIT cell numbers did not recover even after successful antiretroviral treatment. The loss of CD161(+) MAIT cells was correlated with higher levels of MAIT cell activation; an increased frequency of the CD161-TCRVα7.2(+)T cell subset in HIV infection was observed. In vitro stimulation of MAIT cells with IL-18 and IL-12, IL-7 and fixed E.coli also resulted in a rapid and additive reduction of the MAIT cell frequency defined by CD161, IL-18R and CCR6. In summary, the irreversible reduction of the CD161(+) MAIT cell subset seems to be an early event in HIV infection that is independent of later stages of the disease. This loss appears to be at least partially due to the distinctive vulnerability of MAIT cells to the pronounced stimulation by microbial products and cytokines during HIV-infection.

Show MeSH
Related in: MedlinePlus