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CD1-reactive natural killer T cells are required for development of systemic tolerance through an immune-privileged site.

Sonoda KH, Exley M, Snapper S, Balk SP, Stein-Streilein J - J. Exp. Med. (1999)

Bottom Line: Therefore, this model for immune-privileged site-mediated tolerance provided us with an excellent format for studying the role of NKT cells in the development of tolerance.Significantly, CD1-reactive NKT cells were not required for intravenously induced systemic tolerance, thereby establishing that different mechanisms mediate development of tolerance to antigens inoculated by these routes.A critical role for NKT cells in the development of systemic tolerance associated with an immune-privileged site suggests a mechanism involving NKT cells in self-tolerance and their defects in autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.

ABSTRACT
Systemic tolerance can be elicited by introducing antigen into an immune-privileged site, such as the eye, or directly into the blood. Both routes of immunization result in a selective deficiency of systemic delayed type hypersensitivity. Although the experimental animal model of anterior chamber-associated immune deviation (ACAID) occurs in most mouse strains, ACAID cannot be induced in several mutant mouse strains that are coincidentally deficient in natural killer T (NKT) cells. Therefore, this model for immune-privileged site-mediated tolerance provided us with an excellent format for studying the role of NKT cells in the development of tolerance. The following data show that CD1-reactive NKT cells are required for the development of systemic tolerance induced via the eye as follows: (a) CD1 knockout mice were unable to develop ACAID unless they were reconstituted with NKT cells together with CD1(+) antigen-presenting cells; (b) specific antibody depletion of NKT cells in vivo abrogated the development of ACAID; and (c) anti-CD1 monoclonal antibody treatment of wild-type mice prevented ACAID development. Significantly, CD1-reactive NKT cells were not required for intravenously induced systemic tolerance, thereby establishing that different mechanisms mediate development of tolerance to antigens inoculated by these routes. A critical role for NKT cells in the development of systemic tolerance associated with an immune-privileged site suggests a mechanism involving NKT cells in self-tolerance and their defects in autoimmunity.

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Generation of ACAID correlates with increased number of NKT cells in the spleen. ACAID was induced in mice as described previously. In brief, B6 mice were killed 7 d after ac inoculation of OVA. Column-enriched splenic T cells were harvested from ac-inoculated and uninoculated mice by application to IMMULAN™ columns, and stained for analysis by flow cytometry to determine the ratio of NK and NKT cells present within the lymphocyte gate for individual animals. Fluorescence for the TCR β chain (CyChrome 5) and the NK1.1 marker (PE) are shown on the ordinate and abscissa, respectively. The percentage of cells within the NK cell quadrant (rectangle) and the NKT cell quadrant (square) are indicated for the representative experiment shown. Absolute number (mean ± SEM) of NK and NKT cells was calculated from the precounted viable cell numbers in individual animals (n = 5), and is shown below the abscissa. Significant differences (P ≤ 0.05) are indicated by an asterisk.
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Figure 1: Generation of ACAID correlates with increased number of NKT cells in the spleen. ACAID was induced in mice as described previously. In brief, B6 mice were killed 7 d after ac inoculation of OVA. Column-enriched splenic T cells were harvested from ac-inoculated and uninoculated mice by application to IMMULAN™ columns, and stained for analysis by flow cytometry to determine the ratio of NK and NKT cells present within the lymphocyte gate for individual animals. Fluorescence for the TCR β chain (CyChrome 5) and the NK1.1 marker (PE) are shown on the ordinate and abscissa, respectively. The percentage of cells within the NK cell quadrant (rectangle) and the NKT cell quadrant (square) are indicated for the representative experiment shown. Absolute number (mean ± SEM) of NK and NKT cells was calculated from the precounted viable cell numbers in individual animals (n = 5), and is shown below the abscissa. Significant differences (P ≤ 0.05) are indicated by an asterisk.

Mentions: B6 mice were inoculated (ac) with OVA, and 7 d later the spleens were extirpated, cells dissociated, and the numbers of NK and NKT cells were analyzed by flow cytometry after staining for the TCR β chain and the NK1.1 molecule. Analysis was performed on five individual mice per group. The flow cytometry data showed that the ratio of NKT cells to total gated lymphocytes from spleens (depleted of B cells and macrophages) was increased in all ACAID mice compared with naive mice (Fig. 1), subcutaneously or intravenously inoculated mice (data not shown) at 7 d after ac inoculation. In contrast to NKT cells, the number of NK cells did not change in the spleen during ACAID induction. Both the percent and the absolute number of NKT cells in the spleen began to increase as early as day 3 (data not shown), and peaked at day 7 (Fig. 1). These data show an association of splenic accumulation of NKT cells, not NK cells, with the induction of ACAID.


CD1-reactive natural killer T cells are required for development of systemic tolerance through an immune-privileged site.

Sonoda KH, Exley M, Snapper S, Balk SP, Stein-Streilein J - J. Exp. Med. (1999)

Generation of ACAID correlates with increased number of NKT cells in the spleen. ACAID was induced in mice as described previously. In brief, B6 mice were killed 7 d after ac inoculation of OVA. Column-enriched splenic T cells were harvested from ac-inoculated and uninoculated mice by application to IMMULAN™ columns, and stained for analysis by flow cytometry to determine the ratio of NK and NKT cells present within the lymphocyte gate for individual animals. Fluorescence for the TCR β chain (CyChrome 5) and the NK1.1 marker (PE) are shown on the ordinate and abscissa, respectively. The percentage of cells within the NK cell quadrant (rectangle) and the NKT cell quadrant (square) are indicated for the representative experiment shown. Absolute number (mean ± SEM) of NK and NKT cells was calculated from the precounted viable cell numbers in individual animals (n = 5), and is shown below the abscissa. Significant differences (P ≤ 0.05) are indicated by an asterisk.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195676&req=5

Figure 1: Generation of ACAID correlates with increased number of NKT cells in the spleen. ACAID was induced in mice as described previously. In brief, B6 mice were killed 7 d after ac inoculation of OVA. Column-enriched splenic T cells were harvested from ac-inoculated and uninoculated mice by application to IMMULAN™ columns, and stained for analysis by flow cytometry to determine the ratio of NK and NKT cells present within the lymphocyte gate for individual animals. Fluorescence for the TCR β chain (CyChrome 5) and the NK1.1 marker (PE) are shown on the ordinate and abscissa, respectively. The percentage of cells within the NK cell quadrant (rectangle) and the NKT cell quadrant (square) are indicated for the representative experiment shown. Absolute number (mean ± SEM) of NK and NKT cells was calculated from the precounted viable cell numbers in individual animals (n = 5), and is shown below the abscissa. Significant differences (P ≤ 0.05) are indicated by an asterisk.
Mentions: B6 mice were inoculated (ac) with OVA, and 7 d later the spleens were extirpated, cells dissociated, and the numbers of NK and NKT cells were analyzed by flow cytometry after staining for the TCR β chain and the NK1.1 molecule. Analysis was performed on five individual mice per group. The flow cytometry data showed that the ratio of NKT cells to total gated lymphocytes from spleens (depleted of B cells and macrophages) was increased in all ACAID mice compared with naive mice (Fig. 1), subcutaneously or intravenously inoculated mice (data not shown) at 7 d after ac inoculation. In contrast to NKT cells, the number of NK cells did not change in the spleen during ACAID induction. Both the percent and the absolute number of NKT cells in the spleen began to increase as early as day 3 (data not shown), and peaked at day 7 (Fig. 1). These data show an association of splenic accumulation of NKT cells, not NK cells, with the induction of ACAID.

Bottom Line: Therefore, this model for immune-privileged site-mediated tolerance provided us with an excellent format for studying the role of NKT cells in the development of tolerance.Significantly, CD1-reactive NKT cells were not required for intravenously induced systemic tolerance, thereby establishing that different mechanisms mediate development of tolerance to antigens inoculated by these routes.A critical role for NKT cells in the development of systemic tolerance associated with an immune-privileged site suggests a mechanism involving NKT cells in self-tolerance and their defects in autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.

ABSTRACT
Systemic tolerance can be elicited by introducing antigen into an immune-privileged site, such as the eye, or directly into the blood. Both routes of immunization result in a selective deficiency of systemic delayed type hypersensitivity. Although the experimental animal model of anterior chamber-associated immune deviation (ACAID) occurs in most mouse strains, ACAID cannot be induced in several mutant mouse strains that are coincidentally deficient in natural killer T (NKT) cells. Therefore, this model for immune-privileged site-mediated tolerance provided us with an excellent format for studying the role of NKT cells in the development of tolerance. The following data show that CD1-reactive NKT cells are required for the development of systemic tolerance induced via the eye as follows: (a) CD1 knockout mice were unable to develop ACAID unless they were reconstituted with NKT cells together with CD1(+) antigen-presenting cells; (b) specific antibody depletion of NKT cells in vivo abrogated the development of ACAID; and (c) anti-CD1 monoclonal antibody treatment of wild-type mice prevented ACAID development. Significantly, CD1-reactive NKT cells were not required for intravenously induced systemic tolerance, thereby establishing that different mechanisms mediate development of tolerance to antigens inoculated by these routes. A critical role for NKT cells in the development of systemic tolerance associated with an immune-privileged site suggests a mechanism involving NKT cells in self-tolerance and their defects in autoimmunity.

Show MeSH
Related in: MedlinePlus