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Phenotypic and functional analyses of NK and NKT-like populations during the early stages of chikungunya infection.

Thanapati S, Das R, Tripathy AS - Front Microbiol (2015)

Bottom Line: Higher percentages of perforin(+)CD3(-)CD56(+) and perforin(+)CD3(+)CD56(+) cells were observed in acute and convalescent patients, respectively.IFN-γ expression on NK cells of convalescent patients and on NKT-like cells of both patient groups was indicative of the regulatory role of NK and NKT-like cells.Collectively, these data showed that higher expression of activating receptors on NK/NKT-like cells and perforin(+) NK cells in acute patients could be responsible for increased cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Hepatitis Group, National Institute of Virology Pune, India.

ABSTRACT
The aim of this study was to characterize NK (CD56(+)CD3(-)) and NKT-like cell (CD56(+)CD3(+)) responses early after chikungunya infection. Expression profiling and functional analysis of T/NK/NKT-like cells were performed on samples from 56 acute and 31 convalescent chikungunya patients and 56 control individuals. The percentages of NK cells were high in both patient groups, whereas NKT-like cell percentages were high only in the convalescent group. The percentages of NKp30(+)CD3(-)CD56(+), NKp30(+)CD3(+)CD56(+), CD244(+)CD3(-)CD56(+), and CD244(+)CD3(+)CD56(+)cells were high, whereas the percentages of NKG2D(+)CD3(-)CD56(+) and NKG2D(+)CD3(+)CD56(+)cells were low in both patient groups. The percentages of NKp44(+)CD3(-)CD56(+) cells were high in both patient groups, whereas the percentages of NKp44(+)CD3(+)CD56(+) cells were higher in the acute group than in convalescent and control groups. The percentages of NKp46(+)CD3(-)CD56(+) cells were high in both patient groups. Higher percentages of perforin(+)CD3(-)CD56(+) and perforin(+)CD3(+)CD56(+) cells were observed in acute and convalescent patients, respectively. Higher cytotoxic activity was observed in acute patients than in controls. IFN-γ expression on NK cells of convalescent patients and on NKT-like cells of both patient groups was indicative of the regulatory role of NK and NKT-like cells. Collectively, these data showed that higher expression of activating receptors on NK/NKT-like cells and perforin(+) NK cells in acute patients could be responsible for increased cytotoxicity. The observed expression of perforin(+) NK cells in the acute phase and IFN-γ(+) NKT-like cells in the subsequent convalescent stage showed that NK/NKT-like cells mount an early and efficient response to chikungunya virus. Further study of the molecular mechanisms that limit viral dissemination/establishment of chronic disease will aid in understanding how NK/NKT-like cells control chikungunya infection.

No MeSH data available.


Related in: MedlinePlus

Patterns of NK receptors (NKRs) on NK and NKT-like cells. Frequencies of NKRs were enumerated from the whole blood of 30 control subjects, 30 acute, and 31 convalescent patients. (A) Box plots (i-vi) show percentages of NKRs+ NK (CD3−CD56+) cells as (i) NKp30, (ii) NKp44, (iii) NKp46, (iv) NKG2D, (v) CD244, (vi) CD161. Box plots (vii–xii) shows FACS analysis of percentages of NKRs+ NKT-like (CD3+CD56+) cells as (vii) NKp30, (viii) NKp44, (ix) NKp46, (x) NKG2D, (xi) CD244, (xii) CD161. CD161 percentage was enumerated in acute patients and controls. (B) Box plots show mean fluorescence intensity (MFI) of NKRs+ NK cells (i–vi) and NKRs+ NKT-like cells (vii–xii). Kolmogorov–Smirnov test was used for intergroup comparison. p < 0.05 is considered significant.
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Figure 3: Patterns of NK receptors (NKRs) on NK and NKT-like cells. Frequencies of NKRs were enumerated from the whole blood of 30 control subjects, 30 acute, and 31 convalescent patients. (A) Box plots (i-vi) show percentages of NKRs+ NK (CD3−CD56+) cells as (i) NKp30, (ii) NKp44, (iii) NKp46, (iv) NKG2D, (v) CD244, (vi) CD161. Box plots (vii–xii) shows FACS analysis of percentages of NKRs+ NKT-like (CD3+CD56+) cells as (vii) NKp30, (viii) NKp44, (ix) NKp46, (x) NKG2D, (xi) CD244, (xii) CD161. CD161 percentage was enumerated in acute patients and controls. (B) Box plots show mean fluorescence intensity (MFI) of NKRs+ NK cells (i–vi) and NKRs+ NKT-like cells (vii–xii). Kolmogorov–Smirnov test was used for intergroup comparison. p < 0.05 is considered significant.

Mentions: The relationship between NKT-like cells and changes in expression of activation and inhibitory receptors was investigated. Higher percentages of NKT-like cells bearing NKp30+ receptors were observed in acute and convalescent patients [acute: 33.8 (2.3−100), convalescent: 42.8 (1.8−100) vs. controls: 13.5 (1.6−88.1), p = 0.014 for each] (Figure 3Avii). The MFI of NKp30+ NKT-like cells were high in acute patients than in controls, however, comparable among convalescent patients and controls (Figure 3Bvii). The percentages of NKp44+ NKT-like cells were higher in acute patients than in controls [acute: 35.2 (0−96.9) vs. controls: 19.5 (1.6−33.1), p = 0.001]. The percentage of NKp44+ NKT-like cells was higher in acute than in convalescent patients [acute: 35.2 (0−96.9) vs. convalescent: 19.6 (0.1−96), p = 0.035] (Figure 3Aviii). Higher percentages of NKT-like cells bearing CD244+ receptors were observed in both patient categories [acute: 40.7 (0−100), convalescent: 25.9 (0−90.5) vs. controls: 0.9 (0−3.3), p < 0.001 for each] (Figure 3Axi). The MFI of NKp44+ and CD244+ NKT-like cells were high in patient categories than in controls (Figures 3Bviii,xi). Among the patient categories, MFI of NKp44+ NKT-like cells was high in acute than in controls (Figure 3Bviii). Percentage of NKp46+ NKT-like cells was comparable between studied categories (Figure 3Aix) (data not shown); however, lower MFI was observed in the patient categories than in controls (Figure 3Bix). Lower percentages of NKG2D+ NKT-like cells were observed in both patient categories compared to controls [acute: 39.2 (4.9−93.5), convalescent: 36.2 (9.6−95.8) vs. controls: 81.3 (33.8−98.3), p < 0.001 for each] (Figure 3Ax). In consistent with the percentage of NKG2D+ NK cells, MFI was also lower in patient categories than in controls (Figure 3Bx). The percentages of NKT-like cells bearing CD161+ receptors were higher in acute patients than in controls [acute: 45.6 (19.7−68.6) vs. controls: 31.6 (12–72.2), p = 0.018]. CD161+ NK/NKT-like cells were not assessed in the convalescent group (Figures 3Avi,xii). The MFI of CD161+ NKT-like cells was higher in acute patients, whereas MFI of CD161+ NK cells was comparable (Figures 3Bvi,xii). The percentages of CD94+ and NKG2A+ NK/NKT-like cells in the acute and control groups were comparable (data not shown). The gating strategy used for NKR enumeration is shown in Figure 2iv.


Phenotypic and functional analyses of NK and NKT-like populations during the early stages of chikungunya infection.

Thanapati S, Das R, Tripathy AS - Front Microbiol (2015)

Patterns of NK receptors (NKRs) on NK and NKT-like cells. Frequencies of NKRs were enumerated from the whole blood of 30 control subjects, 30 acute, and 31 convalescent patients. (A) Box plots (i-vi) show percentages of NKRs+ NK (CD3−CD56+) cells as (i) NKp30, (ii) NKp44, (iii) NKp46, (iv) NKG2D, (v) CD244, (vi) CD161. Box plots (vii–xii) shows FACS analysis of percentages of NKRs+ NKT-like (CD3+CD56+) cells as (vii) NKp30, (viii) NKp44, (ix) NKp46, (x) NKG2D, (xi) CD244, (xii) CD161. CD161 percentage was enumerated in acute patients and controls. (B) Box plots show mean fluorescence intensity (MFI) of NKRs+ NK cells (i–vi) and NKRs+ NKT-like cells (vii–xii). Kolmogorov–Smirnov test was used for intergroup comparison. p < 0.05 is considered significant.
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Related In: Results  -  Collection

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Figure 3: Patterns of NK receptors (NKRs) on NK and NKT-like cells. Frequencies of NKRs were enumerated from the whole blood of 30 control subjects, 30 acute, and 31 convalescent patients. (A) Box plots (i-vi) show percentages of NKRs+ NK (CD3−CD56+) cells as (i) NKp30, (ii) NKp44, (iii) NKp46, (iv) NKG2D, (v) CD244, (vi) CD161. Box plots (vii–xii) shows FACS analysis of percentages of NKRs+ NKT-like (CD3+CD56+) cells as (vii) NKp30, (viii) NKp44, (ix) NKp46, (x) NKG2D, (xi) CD244, (xii) CD161. CD161 percentage was enumerated in acute patients and controls. (B) Box plots show mean fluorescence intensity (MFI) of NKRs+ NK cells (i–vi) and NKRs+ NKT-like cells (vii–xii). Kolmogorov–Smirnov test was used for intergroup comparison. p < 0.05 is considered significant.
Mentions: The relationship between NKT-like cells and changes in expression of activation and inhibitory receptors was investigated. Higher percentages of NKT-like cells bearing NKp30+ receptors were observed in acute and convalescent patients [acute: 33.8 (2.3−100), convalescent: 42.8 (1.8−100) vs. controls: 13.5 (1.6−88.1), p = 0.014 for each] (Figure 3Avii). The MFI of NKp30+ NKT-like cells were high in acute patients than in controls, however, comparable among convalescent patients and controls (Figure 3Bvii). The percentages of NKp44+ NKT-like cells were higher in acute patients than in controls [acute: 35.2 (0−96.9) vs. controls: 19.5 (1.6−33.1), p = 0.001]. The percentage of NKp44+ NKT-like cells was higher in acute than in convalescent patients [acute: 35.2 (0−96.9) vs. convalescent: 19.6 (0.1−96), p = 0.035] (Figure 3Aviii). Higher percentages of NKT-like cells bearing CD244+ receptors were observed in both patient categories [acute: 40.7 (0−100), convalescent: 25.9 (0−90.5) vs. controls: 0.9 (0−3.3), p < 0.001 for each] (Figure 3Axi). The MFI of NKp44+ and CD244+ NKT-like cells were high in patient categories than in controls (Figures 3Bviii,xi). Among the patient categories, MFI of NKp44+ NKT-like cells was high in acute than in controls (Figure 3Bviii). Percentage of NKp46+ NKT-like cells was comparable between studied categories (Figure 3Aix) (data not shown); however, lower MFI was observed in the patient categories than in controls (Figure 3Bix). Lower percentages of NKG2D+ NKT-like cells were observed in both patient categories compared to controls [acute: 39.2 (4.9−93.5), convalescent: 36.2 (9.6−95.8) vs. controls: 81.3 (33.8−98.3), p < 0.001 for each] (Figure 3Ax). In consistent with the percentage of NKG2D+ NK cells, MFI was also lower in patient categories than in controls (Figure 3Bx). The percentages of NKT-like cells bearing CD161+ receptors were higher in acute patients than in controls [acute: 45.6 (19.7−68.6) vs. controls: 31.6 (12–72.2), p = 0.018]. CD161+ NK/NKT-like cells were not assessed in the convalescent group (Figures 3Avi,xii). The MFI of CD161+ NKT-like cells was higher in acute patients, whereas MFI of CD161+ NK cells was comparable (Figures 3Bvi,xii). The percentages of CD94+ and NKG2A+ NK/NKT-like cells in the acute and control groups were comparable (data not shown). The gating strategy used for NKR enumeration is shown in Figure 2iv.

Bottom Line: Higher percentages of perforin(+)CD3(-)CD56(+) and perforin(+)CD3(+)CD56(+) cells were observed in acute and convalescent patients, respectively.IFN-γ expression on NK cells of convalescent patients and on NKT-like cells of both patient groups was indicative of the regulatory role of NK and NKT-like cells.Collectively, these data showed that higher expression of activating receptors on NK/NKT-like cells and perforin(+) NK cells in acute patients could be responsible for increased cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Hepatitis Group, National Institute of Virology Pune, India.

ABSTRACT
The aim of this study was to characterize NK (CD56(+)CD3(-)) and NKT-like cell (CD56(+)CD3(+)) responses early after chikungunya infection. Expression profiling and functional analysis of T/NK/NKT-like cells were performed on samples from 56 acute and 31 convalescent chikungunya patients and 56 control individuals. The percentages of NK cells were high in both patient groups, whereas NKT-like cell percentages were high only in the convalescent group. The percentages of NKp30(+)CD3(-)CD56(+), NKp30(+)CD3(+)CD56(+), CD244(+)CD3(-)CD56(+), and CD244(+)CD3(+)CD56(+)cells were high, whereas the percentages of NKG2D(+)CD3(-)CD56(+) and NKG2D(+)CD3(+)CD56(+)cells were low in both patient groups. The percentages of NKp44(+)CD3(-)CD56(+) cells were high in both patient groups, whereas the percentages of NKp44(+)CD3(+)CD56(+) cells were higher in the acute group than in convalescent and control groups. The percentages of NKp46(+)CD3(-)CD56(+) cells were high in both patient groups. Higher percentages of perforin(+)CD3(-)CD56(+) and perforin(+)CD3(+)CD56(+) cells were observed in acute and convalescent patients, respectively. Higher cytotoxic activity was observed in acute patients than in controls. IFN-γ expression on NK cells of convalescent patients and on NKT-like cells of both patient groups was indicative of the regulatory role of NK and NKT-like cells. Collectively, these data showed that higher expression of activating receptors on NK/NKT-like cells and perforin(+) NK cells in acute patients could be responsible for increased cytotoxicity. The observed expression of perforin(+) NK cells in the acute phase and IFN-γ(+) NKT-like cells in the subsequent convalescent stage showed that NK/NKT-like cells mount an early and efficient response to chikungunya virus. Further study of the molecular mechanisms that limit viral dissemination/establishment of chronic disease will aid in understanding how NK/NKT-like cells control chikungunya infection.

No MeSH data available.


Related in: MedlinePlus