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Regulation of experimental autoimmune encephalomyelitis by natural killer (NK) cells.

Zhang B, Yamamura T, Kondo T, Fujiwara M, Tabira T - J. Exp. Med. (1997)

Bottom Line: The disease enhancement was associated with augmentation of T cell proliferation and production of Th1 cytokines in response to MOG35-55.We further showed that NK cells inhibit T cell proliferation triggered by antigen or cytokine stimulation.Taken together, we conclude that NK cells are an important regulator for EAE in both induction and effector phases.

View Article: PubMed Central - PubMed

Affiliation: Department of Demyelinating Disease and Aging, National Institute of Neuroscience, Tokyo, Japan.

ABSTRACT
In this report, we establish a regulatory role of natural killer (NK) cells in experimental autoimmune encephalomyelitis (EAE), a prototype T helper cell type 1 (Th1)-mediated disease. Active sensitization of C57BL/6 (B6) mice with the myelin oligodendrocyte glycoprotein (MOG)35-55 peptide induces a mild form of monophasic EAE. When mice were deprived of NK cells by antibody treatment before immunization, they developed a more serious form of EAE associated with relapse. Aggravation of EAE by NK cell deletion was also seen in beta 2-microglobulin-/- (beta 2m-/-) mice, indicating that NK cells can play a regulatory role in a manner independent of CD8+ T cells or NK1.1+ T cells (NK-T cells). The disease enhancement was associated with augmentation of T cell proliferation and production of Th1 cytokines in response to MOG35-55. EAE passively induced by the MOG35-55-specific T cell line was also enhanced by NK cell deletion in B6, beta 2m-/-, and recombination activation gene 2 (RAG-2)-/- mice, indicating that the regulation by NK cells can be independent of T, B, or NK-T cells. We further showed that NK cells inhibit T cell proliferation triggered by antigen or cytokine stimulation. Taken together, we conclude that NK cells are an important regulator for EAE in both induction and effector phases.

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Effect of insolubilized anti-NK1.1 mAb, IFN-γ, and anti–IFN-γ on ZB-1 line proliferation. (A) Anti-NK1.1 and control M-11 mAb dissolved in PBS were added into relevant wells at various concentrations (shown in micrograms per milliliter), incubated overnight, and then washed with  PBS intensively. ZB-1 line cells (4 × 104/well) were stimulated with MOG35-55 (25 μg/ml) in the presence of irradiated spleen APCs (8 × 105/well) in  the wells coated with M-11 (ins. M11) or with anti-NK1.1 mAb (ins.NK1.1). (B) ZB-1 line cells were cultured with spleen cell APCs in the absence of  the MOG peptide in the antibody-coated wells in parallel with experiment A. (C) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in  the presence of different concentrations of recombinant mouse IFN-γ (PharMingen). (D) ZB-1 line cells were stimulated with MOG35-55 using spleen  APCs in the presence of different concentrations of anti–mouse IFN-γ mAb (PharMingen). All the data represent mean cpm ± SD of triplicate cultures.
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Figure 10: Effect of insolubilized anti-NK1.1 mAb, IFN-γ, and anti–IFN-γ on ZB-1 line proliferation. (A) Anti-NK1.1 and control M-11 mAb dissolved in PBS were added into relevant wells at various concentrations (shown in micrograms per milliliter), incubated overnight, and then washed with PBS intensively. ZB-1 line cells (4 × 104/well) were stimulated with MOG35-55 (25 μg/ml) in the presence of irradiated spleen APCs (8 × 105/well) in the wells coated with M-11 (ins. M11) or with anti-NK1.1 mAb (ins.NK1.1). (B) ZB-1 line cells were cultured with spleen cell APCs in the absence of the MOG peptide in the antibody-coated wells in parallel with experiment A. (C) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in the presence of different concentrations of recombinant mouse IFN-γ (PharMingen). (D) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in the presence of different concentrations of anti–mouse IFN-γ mAb (PharMingen). All the data represent mean cpm ± SD of triplicate cultures.

Mentions: We next examined the effect of insolubilized anti-NK1.1 mAb which can trigger activation of NK cells via cross-linking NKR-P1 molecule (37). In a reciprocal manner with the results in Fig. 9, ZB-1 cell proliferation in response to MOG was significantly inhibited when spleen NK cells were stimulated by anti-NK1.1 mAb (Fig. 10 A). In contrast, insolubilized control mAb had no effect, further supporting the inhibitory role of NK cells against T cell proliferation. The background proliferation of ZB-1 cells was not inhibited by insolubilized anti-NK1.1 (Fig. 10 B). We also explored the possible involvement of IFN-γ as a downregulatory factor, since this Th1 cytokine is the major product of NK cells. However, with the addition of exogenous IFN-γ, the proliferation of ZB-1 line cells was slightly but significantly enhanced (Fig. 10 C), whereas anti– IFN-γ mAb inhibited the cell proliferation (Fig. 10 D). In addition, depressed proliferative response induced by insolubilized anti-NK1.1 mAb was further inhibited by anti– IFN-γ mAb (data not shown). These results indicate that NK cell–mediated T cell suppression is not mediated by IFN-γ.


Regulation of experimental autoimmune encephalomyelitis by natural killer (NK) cells.

Zhang B, Yamamura T, Kondo T, Fujiwara M, Tabira T - J. Exp. Med. (1997)

Effect of insolubilized anti-NK1.1 mAb, IFN-γ, and anti–IFN-γ on ZB-1 line proliferation. (A) Anti-NK1.1 and control M-11 mAb dissolved in PBS were added into relevant wells at various concentrations (shown in micrograms per milliliter), incubated overnight, and then washed with  PBS intensively. ZB-1 line cells (4 × 104/well) were stimulated with MOG35-55 (25 μg/ml) in the presence of irradiated spleen APCs (8 × 105/well) in  the wells coated with M-11 (ins. M11) or with anti-NK1.1 mAb (ins.NK1.1). (B) ZB-1 line cells were cultured with spleen cell APCs in the absence of  the MOG peptide in the antibody-coated wells in parallel with experiment A. (C) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in  the presence of different concentrations of recombinant mouse IFN-γ (PharMingen). (D) ZB-1 line cells were stimulated with MOG35-55 using spleen  APCs in the presence of different concentrations of anti–mouse IFN-γ mAb (PharMingen). All the data represent mean cpm ± SD of triplicate cultures.
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Related In: Results  -  Collection

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Figure 10: Effect of insolubilized anti-NK1.1 mAb, IFN-γ, and anti–IFN-γ on ZB-1 line proliferation. (A) Anti-NK1.1 and control M-11 mAb dissolved in PBS were added into relevant wells at various concentrations (shown in micrograms per milliliter), incubated overnight, and then washed with PBS intensively. ZB-1 line cells (4 × 104/well) were stimulated with MOG35-55 (25 μg/ml) in the presence of irradiated spleen APCs (8 × 105/well) in the wells coated with M-11 (ins. M11) or with anti-NK1.1 mAb (ins.NK1.1). (B) ZB-1 line cells were cultured with spleen cell APCs in the absence of the MOG peptide in the antibody-coated wells in parallel with experiment A. (C) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in the presence of different concentrations of recombinant mouse IFN-γ (PharMingen). (D) ZB-1 line cells were stimulated with MOG35-55 using spleen APCs in the presence of different concentrations of anti–mouse IFN-γ mAb (PharMingen). All the data represent mean cpm ± SD of triplicate cultures.
Mentions: We next examined the effect of insolubilized anti-NK1.1 mAb which can trigger activation of NK cells via cross-linking NKR-P1 molecule (37). In a reciprocal manner with the results in Fig. 9, ZB-1 cell proliferation in response to MOG was significantly inhibited when spleen NK cells were stimulated by anti-NK1.1 mAb (Fig. 10 A). In contrast, insolubilized control mAb had no effect, further supporting the inhibitory role of NK cells against T cell proliferation. The background proliferation of ZB-1 cells was not inhibited by insolubilized anti-NK1.1 (Fig. 10 B). We also explored the possible involvement of IFN-γ as a downregulatory factor, since this Th1 cytokine is the major product of NK cells. However, with the addition of exogenous IFN-γ, the proliferation of ZB-1 line cells was slightly but significantly enhanced (Fig. 10 C), whereas anti– IFN-γ mAb inhibited the cell proliferation (Fig. 10 D). In addition, depressed proliferative response induced by insolubilized anti-NK1.1 mAb was further inhibited by anti– IFN-γ mAb (data not shown). These results indicate that NK cell–mediated T cell suppression is not mediated by IFN-γ.

Bottom Line: The disease enhancement was associated with augmentation of T cell proliferation and production of Th1 cytokines in response to MOG35-55.We further showed that NK cells inhibit T cell proliferation triggered by antigen or cytokine stimulation.Taken together, we conclude that NK cells are an important regulator for EAE in both induction and effector phases.

View Article: PubMed Central - PubMed

Affiliation: Department of Demyelinating Disease and Aging, National Institute of Neuroscience, Tokyo, Japan.

ABSTRACT
In this report, we establish a regulatory role of natural killer (NK) cells in experimental autoimmune encephalomyelitis (EAE), a prototype T helper cell type 1 (Th1)-mediated disease. Active sensitization of C57BL/6 (B6) mice with the myelin oligodendrocyte glycoprotein (MOG)35-55 peptide induces a mild form of monophasic EAE. When mice were deprived of NK cells by antibody treatment before immunization, they developed a more serious form of EAE associated with relapse. Aggravation of EAE by NK cell deletion was also seen in beta 2-microglobulin-/- (beta 2m-/-) mice, indicating that NK cells can play a regulatory role in a manner independent of CD8+ T cells or NK1.1+ T cells (NK-T cells). The disease enhancement was associated with augmentation of T cell proliferation and production of Th1 cytokines in response to MOG35-55. EAE passively induced by the MOG35-55-specific T cell line was also enhanced by NK cell deletion in B6, beta 2m-/-, and recombination activation gene 2 (RAG-2)-/- mice, indicating that the regulation by NK cells can be independent of T, B, or NK-T cells. We further showed that NK cells inhibit T cell proliferation triggered by antigen or cytokine stimulation. Taken together, we conclude that NK cells are an important regulator for EAE in both induction and effector phases.

Show MeSH
Related in: MedlinePlus