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Mucosal-Associated Invariant T Cells in the Human Gastric Mucosa and Blood: Role in Helicobacter pylori Infection.

Booth JS, Salerno-Goncalves R, Blanchard TG, Patil SA, Kader HA, Safta AM, Morningstar LM, Czinn SJ, Greenwald BD, Sztein MB - Front Immunol (2015)

Bottom Line: We found that CD8(+) and CD4(-)CD8(-) (double negative) MAIT cell subsets respond to H. pylori-infected macrophages stimulation in a MR-1 restrictive manner by producing cytokines (IFN-γ, TNF-α, IL-17A) and exhibiting cytotoxic activity.Interestingly, we observed that blood MAIT cell frequency in Hp(+ve) individuals was significantly lower than in Hp(-ve) individuals.However, gastric MAIT cell frequency was not significantly different between Hp(+ve) and Hp(-ve) individuals, demonstrating a dichotomy between blood and gastric tissues.

View Article: PubMed Central - PubMed

Affiliation: Center for Vaccine Development, University of Maryland School of Medicine , Baltimore, MD , USA ; Department of Pediatrics, University of Maryland School of Medicine , Baltimore, MD , USA.

ABSTRACT
Mucosal-associated invariant T (MAIT) cells represent a class of antimicrobial innate-like T cells that have been characterized in human blood, liver, lungs, and intestine. Here, we investigated, for the first time, the presence of MAIT cells in the stomach of children, adults, and the elderly undergoing routine endoscopy and assessed their reactivity to Helicobacter pylori (H. pylori - Hp), a major gastric pathogen. We observed that MAIT cells are present in the lamina propria compartment of the stomach and display a similar memory phenotype to blood MAIT cells. We then demonstrated that gastric and blood MAIT cells are able to recognize H. pylori. We found that CD8(+) and CD4(-)CD8(-) (double negative) MAIT cell subsets respond to H. pylori-infected macrophages stimulation in a MR-1 restrictive manner by producing cytokines (IFN-γ, TNF-α, IL-17A) and exhibiting cytotoxic activity. Interestingly, we observed that blood MAIT cell frequency in Hp(+ve) individuals was significantly lower than in Hp(-ve) individuals. However, gastric MAIT cell frequency was not significantly different between Hp(+ve) and Hp(-ve) individuals, demonstrating a dichotomy between blood and gastric tissues. Further, we observed that the majority of gastric MAIT cells (>80%) expressed tissue-resident markers (CD69(+) CD103(+)), which were only marginally present on PBMC MAIT cells (<3%), suggesting that gastric MAIT cells are readily available to respond quickly to pathogens. These results contribute important new information to the understanding of MAIT cells function on peripheral and mucosal tissues and its possible implications in the host response to H. pylori.

No MeSH data available.


Related in: MedlinePlus

Blood MAIT cells from healthy adults exhibit multifunctional abilities and are cytotoxic to H. pylori-infected macrophages. (A) Multifunctional activities of MAIT cells were determined by simultaneous detection of three intracellular cytokines (IFN-γ, TNF-α, and IL-17A) and expression of CD107a by CD8+ MAIT cells following stimulation with H. pylori-infected THP-1 Mϕ (E:T – 50:1). Scatter plot shows single-cytokine-producing and CD107-expressing cells and the six predominant multi-cytokine-producing/CD107-expressing patterns using PBMCs from healthy adult volunteers. Horizontal red lines indicate the median responses. Multifunctionality was analyzed using the FCOM feature of WinList. (B)H. pylori CTL responses by MAIT cells were measured at various effector:target ratios (50:1, 25:1, 12.5:1, 6.25:1) in non-infected (NI) and H. pylori-infected (INF) THP-1 Mϕ. Blocking of CTL responses was performed using anti-human MR-1 antibody (goat, polyclonal) (10 μg/ml) or goat IgG control (immunoglobulin control; IgC; 10 μg/ml). (C)H. pylori CTL responses by MAIT cells were measured at two effector:target ratios (50:1 and 25:1) in NI and INF THP-1 Mϕ. Blocking of CTL responses were performed using the anti-human MR-1 mAb (clone 26.5) (10 μg/ml) or a matched isotype control (IC, 10 μg/ml). Lines show the mean percentages of cytotoxicity at different E/T ratios from triplicate wells. The data are representative of four separate experiments.
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Figure 5: Blood MAIT cells from healthy adults exhibit multifunctional abilities and are cytotoxic to H. pylori-infected macrophages. (A) Multifunctional activities of MAIT cells were determined by simultaneous detection of three intracellular cytokines (IFN-γ, TNF-α, and IL-17A) and expression of CD107a by CD8+ MAIT cells following stimulation with H. pylori-infected THP-1 Mϕ (E:T – 50:1). Scatter plot shows single-cytokine-producing and CD107-expressing cells and the six predominant multi-cytokine-producing/CD107-expressing patterns using PBMCs from healthy adult volunteers. Horizontal red lines indicate the median responses. Multifunctionality was analyzed using the FCOM feature of WinList. (B)H. pylori CTL responses by MAIT cells were measured at various effector:target ratios (50:1, 25:1, 12.5:1, 6.25:1) in non-infected (NI) and H. pylori-infected (INF) THP-1 Mϕ. Blocking of CTL responses was performed using anti-human MR-1 antibody (goat, polyclonal) (10 μg/ml) or goat IgG control (immunoglobulin control; IgC; 10 μg/ml). (C)H. pylori CTL responses by MAIT cells were measured at two effector:target ratios (50:1 and 25:1) in NI and INF THP-1 Mϕ. Blocking of CTL responses were performed using the anti-human MR-1 mAb (clone 26.5) (10 μg/ml) or a matched isotype control (IC, 10 μg/ml). Lines show the mean percentages of cytotoxicity at different E/T ratios from triplicate wells. The data are representative of four separate experiments.

Mentions: Our group has previously shown that MAIT cells can respond to S. typhi by secreting multiple cytokines simultaneously (14). Thus, we next investigated the multifunctionality of the responses exhibited by blood CD8+ MAIT cells against H. pylori-infected differentiated THP-1 macrophages. Analysis of multiple cytokines (IFN-γ, TNF-α, and IL-17A) and/or CD107 expression patterns (16 possible combinations) using the Winlist FCOM function revealed that MAIT cell responses were characterized by single, double, or triple cytokine producers/CD107 expressors, albeit at different percentages, following stimulation with H. pylori-infected THP-1 Mϕ (Figure 5A). Interestingly, CD8+ MAIT cells produced IL-17A mostly as single-cytokine-producing cell following stimulation with H. pylori-infected Mϕ. In contrast, CD8+ MAIT cells response to H. pylori-infected Mϕ produced IFN-γ mostly as multifunctional [triple-positive cells (IFN-γ+ TNF-α+ CD107a+)] (Figure 5A). We also noted that the cytotoxic marker, CD107a, was prominent in both multifunctional and single-positive cell subsets, suggesting that MAIT cells are highly cytotoxic to H. pylori-infected THP-1 Mϕ.


Mucosal-Associated Invariant T Cells in the Human Gastric Mucosa and Blood: Role in Helicobacter pylori Infection.

Booth JS, Salerno-Goncalves R, Blanchard TG, Patil SA, Kader HA, Safta AM, Morningstar LM, Czinn SJ, Greenwald BD, Sztein MB - Front Immunol (2015)

Blood MAIT cells from healthy adults exhibit multifunctional abilities and are cytotoxic to H. pylori-infected macrophages. (A) Multifunctional activities of MAIT cells were determined by simultaneous detection of three intracellular cytokines (IFN-γ, TNF-α, and IL-17A) and expression of CD107a by CD8+ MAIT cells following stimulation with H. pylori-infected THP-1 Mϕ (E:T – 50:1). Scatter plot shows single-cytokine-producing and CD107-expressing cells and the six predominant multi-cytokine-producing/CD107-expressing patterns using PBMCs from healthy adult volunteers. Horizontal red lines indicate the median responses. Multifunctionality was analyzed using the FCOM feature of WinList. (B)H. pylori CTL responses by MAIT cells were measured at various effector:target ratios (50:1, 25:1, 12.5:1, 6.25:1) in non-infected (NI) and H. pylori-infected (INF) THP-1 Mϕ. Blocking of CTL responses was performed using anti-human MR-1 antibody (goat, polyclonal) (10 μg/ml) or goat IgG control (immunoglobulin control; IgC; 10 μg/ml). (C)H. pylori CTL responses by MAIT cells were measured at two effector:target ratios (50:1 and 25:1) in NI and INF THP-1 Mϕ. Blocking of CTL responses were performed using the anti-human MR-1 mAb (clone 26.5) (10 μg/ml) or a matched isotype control (IC, 10 μg/ml). Lines show the mean percentages of cytotoxicity at different E/T ratios from triplicate wells. The data are representative of four separate experiments.
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Related In: Results  -  Collection

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Figure 5: Blood MAIT cells from healthy adults exhibit multifunctional abilities and are cytotoxic to H. pylori-infected macrophages. (A) Multifunctional activities of MAIT cells were determined by simultaneous detection of three intracellular cytokines (IFN-γ, TNF-α, and IL-17A) and expression of CD107a by CD8+ MAIT cells following stimulation with H. pylori-infected THP-1 Mϕ (E:T – 50:1). Scatter plot shows single-cytokine-producing and CD107-expressing cells and the six predominant multi-cytokine-producing/CD107-expressing patterns using PBMCs from healthy adult volunteers. Horizontal red lines indicate the median responses. Multifunctionality was analyzed using the FCOM feature of WinList. (B)H. pylori CTL responses by MAIT cells were measured at various effector:target ratios (50:1, 25:1, 12.5:1, 6.25:1) in non-infected (NI) and H. pylori-infected (INF) THP-1 Mϕ. Blocking of CTL responses was performed using anti-human MR-1 antibody (goat, polyclonal) (10 μg/ml) or goat IgG control (immunoglobulin control; IgC; 10 μg/ml). (C)H. pylori CTL responses by MAIT cells were measured at two effector:target ratios (50:1 and 25:1) in NI and INF THP-1 Mϕ. Blocking of CTL responses were performed using the anti-human MR-1 mAb (clone 26.5) (10 μg/ml) or a matched isotype control (IC, 10 μg/ml). Lines show the mean percentages of cytotoxicity at different E/T ratios from triplicate wells. The data are representative of four separate experiments.
Mentions: Our group has previously shown that MAIT cells can respond to S. typhi by secreting multiple cytokines simultaneously (14). Thus, we next investigated the multifunctionality of the responses exhibited by blood CD8+ MAIT cells against H. pylori-infected differentiated THP-1 macrophages. Analysis of multiple cytokines (IFN-γ, TNF-α, and IL-17A) and/or CD107 expression patterns (16 possible combinations) using the Winlist FCOM function revealed that MAIT cell responses were characterized by single, double, or triple cytokine producers/CD107 expressors, albeit at different percentages, following stimulation with H. pylori-infected THP-1 Mϕ (Figure 5A). Interestingly, CD8+ MAIT cells produced IL-17A mostly as single-cytokine-producing cell following stimulation with H. pylori-infected Mϕ. In contrast, CD8+ MAIT cells response to H. pylori-infected Mϕ produced IFN-γ mostly as multifunctional [triple-positive cells (IFN-γ+ TNF-α+ CD107a+)] (Figure 5A). We also noted that the cytotoxic marker, CD107a, was prominent in both multifunctional and single-positive cell subsets, suggesting that MAIT cells are highly cytotoxic to H. pylori-infected THP-1 Mϕ.

Bottom Line: We found that CD8(+) and CD4(-)CD8(-) (double negative) MAIT cell subsets respond to H. pylori-infected macrophages stimulation in a MR-1 restrictive manner by producing cytokines (IFN-γ, TNF-α, IL-17A) and exhibiting cytotoxic activity.Interestingly, we observed that blood MAIT cell frequency in Hp(+ve) individuals was significantly lower than in Hp(-ve) individuals.However, gastric MAIT cell frequency was not significantly different between Hp(+ve) and Hp(-ve) individuals, demonstrating a dichotomy between blood and gastric tissues.

View Article: PubMed Central - PubMed

Affiliation: Center for Vaccine Development, University of Maryland School of Medicine , Baltimore, MD , USA ; Department of Pediatrics, University of Maryland School of Medicine , Baltimore, MD , USA.

ABSTRACT
Mucosal-associated invariant T (MAIT) cells represent a class of antimicrobial innate-like T cells that have been characterized in human blood, liver, lungs, and intestine. Here, we investigated, for the first time, the presence of MAIT cells in the stomach of children, adults, and the elderly undergoing routine endoscopy and assessed their reactivity to Helicobacter pylori (H. pylori - Hp), a major gastric pathogen. We observed that MAIT cells are present in the lamina propria compartment of the stomach and display a similar memory phenotype to blood MAIT cells. We then demonstrated that gastric and blood MAIT cells are able to recognize H. pylori. We found that CD8(+) and CD4(-)CD8(-) (double negative) MAIT cell subsets respond to H. pylori-infected macrophages stimulation in a MR-1 restrictive manner by producing cytokines (IFN-γ, TNF-α, IL-17A) and exhibiting cytotoxic activity. Interestingly, we observed that blood MAIT cell frequency in Hp(+ve) individuals was significantly lower than in Hp(-ve) individuals. However, gastric MAIT cell frequency was not significantly different between Hp(+ve) and Hp(-ve) individuals, demonstrating a dichotomy between blood and gastric tissues. Further, we observed that the majority of gastric MAIT cells (>80%) expressed tissue-resident markers (CD69(+) CD103(+)), which were only marginally present on PBMC MAIT cells (<3%), suggesting that gastric MAIT cells are readily available to respond quickly to pathogens. These results contribute important new information to the understanding of MAIT cells function on peripheral and mucosal tissues and its possible implications in the host response to H. pylori.

No MeSH data available.


Related in: MedlinePlus