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Critical role for the chemokine receptor CXCR6 in NK cell-mediated antigen-specific memory of haptens and viruses.

Paust S, Gill HS, Wang BZ, Flynn MP, Moseman EA, Senman B, Szczepanik M, Telenti A, Askenase PW, Compans RW, von Andrian UH - Nat. Immunol. (2010)

Bottom Line: We report here that hepatic NK cells, but not splenic or naive NK cells, also developed specific memory of vaccines containing antigens from influenza, vesicular stomatitis virus (VSV) or human immunodeficiency virus type 1 (HIV-1).NK cell memory of haptens and viruses depended on CXCR6, a chemokine receptor on hepatic NK cells that was required for the persistence of memory NK cells but not for antigen recognition.Thus, hepatic NK cells can develop adaptive immunity to structurally diverse antigens, an activity that requires NK cell-expressed CXCR6.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
Hepatic natural killer (NK) cells mediate antigen-specific contact hypersensitivity (CHS) in mice deficient in T cells and B cells. We report here that hepatic NK cells, but not splenic or naive NK cells, also developed specific memory of vaccines containing antigens from influenza, vesicular stomatitis virus (VSV) or human immunodeficiency virus type 1 (HIV-1). Adoptive transfer of virus-sensitized NK cells into naive recipient mice enhanced the survival of the mice after lethal challenge with the sensitizing virus but not after lethal challenge with a different virus. NK cell memory of haptens and viruses depended on CXCR6, a chemokine receptor on hepatic NK cells that was required for the persistence of memory NK cells but not for antigen recognition. Thus, hepatic NK cells can develop adaptive immunity to structurally diverse antigens, an activity that requires NK cell-expressed CXCR6.

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Liver NK-cells develop specific memory to haptens. (a) Hapten-specific CHS responses in naïve Rag2−/−Il2rg−/− recipients of hepatic CD45+NK1.1+Thy1+ NK-cells (1×105) from naïve (vehicle-exposed), DNFB or OXA sensitized Rag1−/− donors. Recipients were challenged 24h or four months post transfer. Hapten-specific ear swelling was determined after 24hrs by subtracting background swelling in naïve mice from that in NK recipients. n=10–15 recipients/group. *p<10−2; **p<10−3 (b) Survival and expansion of adoptively transferred NK-cells is not altered by prior sensitization. Two weeks after the 4 month challenge, recipient mice shown in a were analyzed by FACS and the number of liver-resident CD45+ NK1.1+ cells was determined. Results were similar for DNFB or OXA challenged mice, so data were pooled. No NK1.1+ cells were detected in mock recipient Rag2−/−Il2rg−/− mice (not shown). (c) Liver-restricted memory NK-cells arise in the presence of T and B-cells. Sorted CD45+NK1.1+CD3−Thy1+ NK-cells (105) from actin-GFP transgenic donors were transferred to naïve C57BL/6 mice; recipients were challenged six weeks later and analyzed as in a. n=10–15 recipients/group. *p<10−3; **p<10−4 (d) Recruitment of memory NK-cells to sites of challenge is Ag-specific. Hepatic CD45+NK1.1+Thy1+ NK-cells from naïve CD45.1+ WT donors (C57BL/6) and from CD45.2+ DNFB or OXA sensitized WT or actin-GFP transgenic donors were mixed (105 each) and adoptively transferred into naïve Rag2−/−Il2rg−/− recipients. One month post adoptive transfer, recipient ears were challenged with either DNFB or OXA. Livers and ears were harvested at 24, 48 and 72 h, and analyzed for the presence of NK-cells whose origin was distinguished by congenic/fluorescent markers using FACS. No NK-cells were found in acetone challenged control ears (not shown). The mean of all mice analyzed at 24, 48 and 72 h is shown (n=6–7 recipients/group). *p<10−11.
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Figure 1: Liver NK-cells develop specific memory to haptens. (a) Hapten-specific CHS responses in naïve Rag2−/−Il2rg−/− recipients of hepatic CD45+NK1.1+Thy1+ NK-cells (1×105) from naïve (vehicle-exposed), DNFB or OXA sensitized Rag1−/− donors. Recipients were challenged 24h or four months post transfer. Hapten-specific ear swelling was determined after 24hrs by subtracting background swelling in naïve mice from that in NK recipients. n=10–15 recipients/group. *p<10−2; **p<10−3 (b) Survival and expansion of adoptively transferred NK-cells is not altered by prior sensitization. Two weeks after the 4 month challenge, recipient mice shown in a were analyzed by FACS and the number of liver-resident CD45+ NK1.1+ cells was determined. Results were similar for DNFB or OXA challenged mice, so data were pooled. No NK1.1+ cells were detected in mock recipient Rag2−/−Il2rg−/− mice (not shown). (c) Liver-restricted memory NK-cells arise in the presence of T and B-cells. Sorted CD45+NK1.1+CD3−Thy1+ NK-cells (105) from actin-GFP transgenic donors were transferred to naïve C57BL/6 mice; recipients were challenged six weeks later and analyzed as in a. n=10–15 recipients/group. *p<10−3; **p<10−4 (d) Recruitment of memory NK-cells to sites of challenge is Ag-specific. Hepatic CD45+NK1.1+Thy1+ NK-cells from naïve CD45.1+ WT donors (C57BL/6) and from CD45.2+ DNFB or OXA sensitized WT or actin-GFP transgenic donors were mixed (105 each) and adoptively transferred into naïve Rag2−/−Il2rg−/− recipients. One month post adoptive transfer, recipient ears were challenged with either DNFB or OXA. Livers and ears were harvested at 24, 48 and 72 h, and analyzed for the presence of NK-cells whose origin was distinguished by congenic/fluorescent markers using FACS. No NK-cells were found in acetone challenged control ears (not shown). The mean of all mice analyzed at 24, 48 and 72 h is shown (n=6–7 recipients/group). *p<10−11.

Mentions: Previous experiments inRag2−/−mice showed that NK-cells retain memory to DNFB up to four weeks after skin sensitization17. However, haptens form covalent adducts with self proteins, which may persist in the body for weeks27, so it was unclear if NK memory was maintained by residual Ag. To address this, we adoptively transferred sensitized hepatic Thy1+ NK-cells into Ag-free recipients. Rag1−/− mice were either left naïve (i.e. treated with solvent) or sensitized with DNFB or oxazolone (OXA) and used as donors for adoptive transfers of FACS-sorted NK-cells into Rag2−/−Il2rg−/− recipients, which were chosen because this strain cannot produce lymphocytes, including NK-cells19, 28, allowing for easy recovery and analysis of donor NK-cells. One day or four months later, recipients were challenged by painting one ear with hapten, the other with solvent, and ear swelling was measured after 24h (Fig. 1a). Whereas recipients of sensitized splenic NK-cells (not shown) or naïve hepatic NK-cells were unresponsive, the challenged ears of DNFB- or OXA-primed hepatic NK-cell recipients became markedly inflamed at both time points. A response was only elicited when animals were challenged with the sensitizing hapten, but not the other hapten. The difference in recall capacity between transferred NK-cell subsets cannot be explained by differential survival, as all groups harbored similar NK-cell numbers four months after transfer (Fig. 1b). Indeed, consistent with earlier reports29, the transferred NK-cells expanded ~5-7-fold in the lymphopenic Rag2−/−Il2rg−/−recipients irrespective of their organ of origin or immunization history.


Critical role for the chemokine receptor CXCR6 in NK cell-mediated antigen-specific memory of haptens and viruses.

Paust S, Gill HS, Wang BZ, Flynn MP, Moseman EA, Senman B, Szczepanik M, Telenti A, Askenase PW, Compans RW, von Andrian UH - Nat. Immunol. (2010)

Liver NK-cells develop specific memory to haptens. (a) Hapten-specific CHS responses in naïve Rag2−/−Il2rg−/− recipients of hepatic CD45+NK1.1+Thy1+ NK-cells (1×105) from naïve (vehicle-exposed), DNFB or OXA sensitized Rag1−/− donors. Recipients were challenged 24h or four months post transfer. Hapten-specific ear swelling was determined after 24hrs by subtracting background swelling in naïve mice from that in NK recipients. n=10–15 recipients/group. *p<10−2; **p<10−3 (b) Survival and expansion of adoptively transferred NK-cells is not altered by prior sensitization. Two weeks after the 4 month challenge, recipient mice shown in a were analyzed by FACS and the number of liver-resident CD45+ NK1.1+ cells was determined. Results were similar for DNFB or OXA challenged mice, so data were pooled. No NK1.1+ cells were detected in mock recipient Rag2−/−Il2rg−/− mice (not shown). (c) Liver-restricted memory NK-cells arise in the presence of T and B-cells. Sorted CD45+NK1.1+CD3−Thy1+ NK-cells (105) from actin-GFP transgenic donors were transferred to naïve C57BL/6 mice; recipients were challenged six weeks later and analyzed as in a. n=10–15 recipients/group. *p<10−3; **p<10−4 (d) Recruitment of memory NK-cells to sites of challenge is Ag-specific. Hepatic CD45+NK1.1+Thy1+ NK-cells from naïve CD45.1+ WT donors (C57BL/6) and from CD45.2+ DNFB or OXA sensitized WT or actin-GFP transgenic donors were mixed (105 each) and adoptively transferred into naïve Rag2−/−Il2rg−/− recipients. One month post adoptive transfer, recipient ears were challenged with either DNFB or OXA. Livers and ears were harvested at 24, 48 and 72 h, and analyzed for the presence of NK-cells whose origin was distinguished by congenic/fluorescent markers using FACS. No NK-cells were found in acetone challenged control ears (not shown). The mean of all mice analyzed at 24, 48 and 72 h is shown (n=6–7 recipients/group). *p<10−11.
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Figure 1: Liver NK-cells develop specific memory to haptens. (a) Hapten-specific CHS responses in naïve Rag2−/−Il2rg−/− recipients of hepatic CD45+NK1.1+Thy1+ NK-cells (1×105) from naïve (vehicle-exposed), DNFB or OXA sensitized Rag1−/− donors. Recipients were challenged 24h or four months post transfer. Hapten-specific ear swelling was determined after 24hrs by subtracting background swelling in naïve mice from that in NK recipients. n=10–15 recipients/group. *p<10−2; **p<10−3 (b) Survival and expansion of adoptively transferred NK-cells is not altered by prior sensitization. Two weeks after the 4 month challenge, recipient mice shown in a were analyzed by FACS and the number of liver-resident CD45+ NK1.1+ cells was determined. Results were similar for DNFB or OXA challenged mice, so data were pooled. No NK1.1+ cells were detected in mock recipient Rag2−/−Il2rg−/− mice (not shown). (c) Liver-restricted memory NK-cells arise in the presence of T and B-cells. Sorted CD45+NK1.1+CD3−Thy1+ NK-cells (105) from actin-GFP transgenic donors were transferred to naïve C57BL/6 mice; recipients were challenged six weeks later and analyzed as in a. n=10–15 recipients/group. *p<10−3; **p<10−4 (d) Recruitment of memory NK-cells to sites of challenge is Ag-specific. Hepatic CD45+NK1.1+Thy1+ NK-cells from naïve CD45.1+ WT donors (C57BL/6) and from CD45.2+ DNFB or OXA sensitized WT or actin-GFP transgenic donors were mixed (105 each) and adoptively transferred into naïve Rag2−/−Il2rg−/− recipients. One month post adoptive transfer, recipient ears were challenged with either DNFB or OXA. Livers and ears were harvested at 24, 48 and 72 h, and analyzed for the presence of NK-cells whose origin was distinguished by congenic/fluorescent markers using FACS. No NK-cells were found in acetone challenged control ears (not shown). The mean of all mice analyzed at 24, 48 and 72 h is shown (n=6–7 recipients/group). *p<10−11.
Mentions: Previous experiments inRag2−/−mice showed that NK-cells retain memory to DNFB up to four weeks after skin sensitization17. However, haptens form covalent adducts with self proteins, which may persist in the body for weeks27, so it was unclear if NK memory was maintained by residual Ag. To address this, we adoptively transferred sensitized hepatic Thy1+ NK-cells into Ag-free recipients. Rag1−/− mice were either left naïve (i.e. treated with solvent) or sensitized with DNFB or oxazolone (OXA) and used as donors for adoptive transfers of FACS-sorted NK-cells into Rag2−/−Il2rg−/− recipients, which were chosen because this strain cannot produce lymphocytes, including NK-cells19, 28, allowing for easy recovery and analysis of donor NK-cells. One day or four months later, recipients were challenged by painting one ear with hapten, the other with solvent, and ear swelling was measured after 24h (Fig. 1a). Whereas recipients of sensitized splenic NK-cells (not shown) or naïve hepatic NK-cells were unresponsive, the challenged ears of DNFB- or OXA-primed hepatic NK-cell recipients became markedly inflamed at both time points. A response was only elicited when animals were challenged with the sensitizing hapten, but not the other hapten. The difference in recall capacity between transferred NK-cell subsets cannot be explained by differential survival, as all groups harbored similar NK-cell numbers four months after transfer (Fig. 1b). Indeed, consistent with earlier reports29, the transferred NK-cells expanded ~5-7-fold in the lymphopenic Rag2−/−Il2rg−/−recipients irrespective of their organ of origin or immunization history.

Bottom Line: We report here that hepatic NK cells, but not splenic or naive NK cells, also developed specific memory of vaccines containing antigens from influenza, vesicular stomatitis virus (VSV) or human immunodeficiency virus type 1 (HIV-1).NK cell memory of haptens and viruses depended on CXCR6, a chemokine receptor on hepatic NK cells that was required for the persistence of memory NK cells but not for antigen recognition.Thus, hepatic NK cells can develop adaptive immunity to structurally diverse antigens, an activity that requires NK cell-expressed CXCR6.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
Hepatic natural killer (NK) cells mediate antigen-specific contact hypersensitivity (CHS) in mice deficient in T cells and B cells. We report here that hepatic NK cells, but not splenic or naive NK cells, also developed specific memory of vaccines containing antigens from influenza, vesicular stomatitis virus (VSV) or human immunodeficiency virus type 1 (HIV-1). Adoptive transfer of virus-sensitized NK cells into naive recipient mice enhanced the survival of the mice after lethal challenge with the sensitizing virus but not after lethal challenge with a different virus. NK cell memory of haptens and viruses depended on CXCR6, a chemokine receptor on hepatic NK cells that was required for the persistence of memory NK cells but not for antigen recognition. Thus, hepatic NK cells can develop adaptive immunity to structurally diverse antigens, an activity that requires NK cell-expressed CXCR6.

Show MeSH
Related in: MedlinePlus