A nonclassical non-Valpha14Jalpha18 CD1d-restricted (type II) NKT cell is sufficient for down-regulation of tumor immunosurveillance - PubMed (original) (raw)

Comparative Study

A nonclassical non-Valpha14Jalpha18 CD1d-restricted (type II) NKT cell is sufficient for down-regulation of tumor immunosurveillance

Masaki Terabe et al. J Exp Med. 2005.

Abstract

The importance of immunoregulatory T cells has become increasingly apparent. Both CD4+CD25+ T cells and CD1d-restricted NKT cells have been reported to down-regulate tumor immunity in mouse tumor models. However, the relative roles of both T cell populations have rarely been clearly distinguished in the same tumor models. In addition, CD1d-restricted NKT cells have been reported to play a critical role not only in the down-regulation of tumor immunity but also in the promotion of the immunity. However, the explanation for these apparently opposite roles in different tumor models remains unclear. We show that in four mouse tumor models in which CD1d-restricted NKT cells play a role in suppression of tumor immunity, depletion of CD4+CD25+ T cells did not induce enhancement of immunosurveillance. Surprisingly, among the two subpopulations of CD1d-restricted NKT cells, Valpha14Jalpha18+ (type I) and Valpha14Jalpha18- (type II) NKT cells, type I NKT cells were not necessary for the immune suppression. These unexpected results may now resolve the paradox in the role of CD1d-restricted NKT cells in the regulation of tumor immunity, in that type II NKT cells may be sufficient for negative regulation, whereas protection has been found to be mediated by alpha-galactosylceramide-responsive type I NKT cells.

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Figures

Figure 1.

Figure 1.

T reg cells are not necessary for down-regulation of tumor immunosurveillance. (A) 106 15-12RM tumor cells were injected subcutaneously into BALB/c mice on day 0. 1 mg anti-CD25 (PC61; closed squares) or control rat IgG (open diamonds) was injected i.v. on days −4 and −2. 500 μg anti-CD4 (GK1.5; closed circles) was injected i.p. on days 0, 1, 2, 6, and 10. The results are representative of two experiments (n = 5). (B) T reg cell depletion is effective against subcutaneous CT26 but not CT26-L5 variant tumors. Mice challenged subcutaneously with 104 CT26 or CT26-L5 cells were given 1 mg PC61 i.p. on days −4 and −1 of tumor challenge. n = 5 for all groups except those receiving control Ig, in which n = 4. Mice were culled when tumors exceeded 1 cm in diameter, and data are represented as percent survival. (C) 5 × 105 CT26 tumor cells were injected i.v. into BALB/c mice on day 0. 1 mg anti-CD25 was injected i.v. on days −5 and −3. On day 14 or 15, mice were killed and pulmonary tumor metastases were enumerated. CD1d KO mice had a significantly smaller number of metastases (P < 0.002 by one-way ANOVA). The results are pooled from two independent experiments, and additional results from these experiments are shown in Fig. 4 C. The horizontal lines indicate the means.

Figure 2.

Figure 2.

Antigen presentation by CD1d is necessary for down-regulation of tumor immunosurveillance. 106 15-12RM tumor cells were injected into BALB/c mice subcutaneously on day 0. 200 μl anti-CD1d ascites (closed circles) was injected i.v. every other day for 1 wk from day 0. 500 μg anti-CD4 (open triangles) was injected i.p. on days 0, 1, 2, 6, and 10. P < 0.05 by log-rank test between control and anti-CD1d–treated or anti-CD4–treated groups. The results are representative of two experiments (n = 5).

Figure 3.

Figure 3.

Tumor cell lines do not express CD1d in vitro or in situ. (A) 15-12-CD1d (CD1d-transfected positive control), 15-12RM, CT26, 4T1, and CT26-L5 tumor cell lines and spleen cells of CD1d KO mice (negative control) were stained with FITC-labeled anti-CD1d mAb and analyzed by flow cytometry. Shaded graphs indicate cells stained with isotype-matched control antibody; red and bold lines indicate cells stained with anti-CD1d antibody. (B) Frozen sections of subcutaneous CT26-L5 tumors (b, d, and f) or spleens (a, c, and e) were stained with control IgG (a and b), anti-CD1d (c and d), or anti-CD11b (e and f). The sections were counter stained with hemotoxylin (×100).

Figure 4.

Figure 4.

Vα14Jα18**+** (type I) NKT cells are not necessary for down-regulation of tumor immunosurveillance. (A) The wild-type (open diamonds), CD1d KO (closed triangles), and Jα18 KO (closed circles) BALB/c mice were injected subcutaneously with 106 15-12RM tumor cells on day 0, and tumor size was monitored over time. P < 0.05 by log-rank test between CD1d KO and wild-type or Jα18 KO mice. The results are representative of two experiments (n = 5). (B) The wild-type, CD1d KO, and Jα18 KO BALB/c mice were inoculated subcutaneously with 104 CT26-L5 cells and subsequently monitored for survival (n = 5). Data are representative of two experiments. (C) The wild-type, CD1d KO, and Jα18 KO BALB/c mice were injected i.v. with 5 × 105 CT26 cells. After 14 d, pulmonary metastases were enumerated. In one group, mice were treated with PC61 as in Fig. 1 C. CD1d KO mice had significantly fewer metastases (P < 0.002 by one-way ANOVA). The results are pooled from two independent experiments. The horizontal lines indicate the means. The data in Fig. 4 C and in Fig. 1 C were all obtained within the same larger experiments.

Figure 5.

Figure 5.

Vα14Jα18**+** (type I) NKT cells are not necessary for down-regulation of tumor immunosurveillance against lung metastases of 4T1 breast tumors, and primary tumors are not affected by CD1d-restricted NKT cells. (A) The wild-type (open diamonds), Jα18 KO (closed circles), and CD1d KO (closed triangles) BALB/c mice were inoculated in the mammary gland on day 0 with 7,000 4T1 tumor cells, and mice were followed for primary tumor growth. Data are from seven Jα18 KO, seven wild-type, and three CD1d1 KO mice. (B) The wild-type (open diamonds), CD1d KO (closed triangles), and Jα18 KO (closed circles) BALB/c mice were injected with 7,000 4T1 cells in the mammary fat pad. Primary 4T1 tumors were surgically removed 14 d after tumor injection, and mice were subsequently monitored for survival. In all three experiments, CD1d KO mice survived significantly longer than wild-type and Jα18 KO mice (P < 0.02 by log-rank test). In experiments 1 and 3, Jα18 KO mice survived significantly longer than wild-type mice (P < 0.02).

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