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Impairment of antigen-presenting cell function in mice lacking expression of OX40 ligand.

Murata K, Ishii N, Takano H, Miura S, Ndhlovu LC, Nose M, Noda T, Sugamura K - J. Exp. Med. (2000)

Bottom Line: OX40L expression was demonstrated on splenic B cells after CD40 and anti-immunoglobulin (Ig)M stimulation, while only CD40 ligation was capable of inducing OX40L on dendritic cells.The impaired T cell priming was also accompanied by a concomitant reduction of both T helper type 1 (Th1) and Th2 cytokines.Collectively, these results provide convincing evidence that OX40L, expressed on APCs, plays a critical role in antigen-specific T cell responses in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Tohoku University School of Medicine, Sendai 980-8575, Japan.

ABSTRACT
OX40 expressed on activated T cells is known to be an important costimulatory molecule on T cell activation in vitro. However, the in vivo functional significance of the interaction between OX40 and its ligand, OX40L, is still unclear. To investigate the role of OX40L during in vivo immune responses, we generated OX40L-deficient mice and a blocking anti-OX40L monoclonal antibody, MGP34. OX40L expression was demonstrated on splenic B cells after CD40 and anti-immunoglobulin (Ig)M stimulation, while only CD40 ligation was capable of inducing OX40L on dendritic cells. OX40L-deficient and MGP34-treated mice engendered apparent suppression of the recall reaction of T cells primed with both protein antigens and alloantigens and a significant reduction in keyhole limpet hemocyanin-specific IgG production. The impaired T cell priming was also accompanied by a concomitant reduction of both T helper type 1 (Th1) and Th2 cytokines. Furthermore, antigen-presenting cells (APCs) derived from the mutant mice revealed an impaired intrinsic APC function, demonstrating the importance of OX40L in both the priming and effector phases of T cell activation. Collectively, these results provide convincing evidence that OX40L, expressed on APCs, plays a critical role in antigen-specific T cell responses in vivo.

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Impaired T cell priming and cytokine production in OX40L-deficient mice and MGP34-treated mice. (A) OX40L-deficient mice have impaired recall proliferative responses to protein antigens. OX40L-deficient (□) or wild-type (▪) mice (four per group) were immunized with KLH, HEL, or OVA in the hind footpads. 9 d after immunization, draining lymph nodes were extracted and subjected to an in vitro challenge of the various protein antigens. After culturing for 3 d, their [3H]thymidine uptake was measured. (B) Impairment of recall proliferative response of the CD4+ T cells of OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes of the OX40L-deficient (□) or wild-type mice (▪) were assayed for their recall proliferation in response to KLH in the presence of APCs from wild-type mice. (C) Defective APC function in OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes from wild-type mice primed with KLH were assayed for their recall proliferation in response to KLH in the presence of OX40L-deficient (□) or wild-type (▪) irradiated spleen cells used as APCs. (D) MGP34-treated mice have similarly impaired lymph node recall proliferation to KLH. C57BL/6 mice (four per group) were immunized in the hind footpads with KLH in CFA. On days 0, 3, and the 6, mice received 500 μg anti-OX40L (○) or rat IgG (•) intraperitoneally. The recall proliferative responses of the draining lymph nodes (left) were tested in the same manner as described above. Purified CD4+ T cells from the draining lymph nodes of the MGP34-treated (○) or control IgG–treated (•) mice were assayed for their recall proliferation in response to KLH in the presence of APCs from C57BL/6 mice (right). (E and F) Absence of OX40L or MGP34 treatment inhibits recall cytokine production by lymph nodes in response to KLH. Production of Th1 (E) and Th2 (F) cytokines by the draining lymph nodes of the mice in the four groups (□, ▪, ○, •) immunized with KLH and further subjected to an in vitro challenge with KLH were measured.
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Figure 4: Impaired T cell priming and cytokine production in OX40L-deficient mice and MGP34-treated mice. (A) OX40L-deficient mice have impaired recall proliferative responses to protein antigens. OX40L-deficient (□) or wild-type (▪) mice (four per group) were immunized with KLH, HEL, or OVA in the hind footpads. 9 d after immunization, draining lymph nodes were extracted and subjected to an in vitro challenge of the various protein antigens. After culturing for 3 d, their [3H]thymidine uptake was measured. (B) Impairment of recall proliferative response of the CD4+ T cells of OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes of the OX40L-deficient (□) or wild-type mice (▪) were assayed for their recall proliferation in response to KLH in the presence of APCs from wild-type mice. (C) Defective APC function in OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes from wild-type mice primed with KLH were assayed for their recall proliferation in response to KLH in the presence of OX40L-deficient (□) or wild-type (▪) irradiated spleen cells used as APCs. (D) MGP34-treated mice have similarly impaired lymph node recall proliferation to KLH. C57BL/6 mice (four per group) were immunized in the hind footpads with KLH in CFA. On days 0, 3, and the 6, mice received 500 μg anti-OX40L (○) or rat IgG (•) intraperitoneally. The recall proliferative responses of the draining lymph nodes (left) were tested in the same manner as described above. Purified CD4+ T cells from the draining lymph nodes of the MGP34-treated (○) or control IgG–treated (•) mice were assayed for their recall proliferation in response to KLH in the presence of APCs from C57BL/6 mice (right). (E and F) Absence of OX40L or MGP34 treatment inhibits recall cytokine production by lymph nodes in response to KLH. Production of Th1 (E) and Th2 (F) cytokines by the draining lymph nodes of the mice in the four groups (□, ▪, ○, •) immunized with KLH and further subjected to an in vitro challenge with KLH were measured.

Mentions: To examine the in vivo effect of OX40L on antigen-specific T cell activation, we immunized OX40L-deficient or wild-type mice with KLH and tested their in vitro T cell recall proliferative response. A significantly reduced response was observed in OX40L-deficient mice, and the antigen dose–response curve was shifted by several orders of magnitude (Fig. 4 A). Similarly reduced responses were seen with two other protein antigens, hen egg lysozyme (HEL) and OVA, in OX40L-deficient mice (Fig. 4 A). These results suggest that OX40L plays an important role in antigen-specific T cell priming. The reduced response in OX40L-deficient mice may be due to a defect at the level of the APCs, since the intrinsic potential of OX40L-deficient T cells in response to antigens is preserved (data not shown) and since OX40L expression was restricted to activated APCs but not T cells.


Impairment of antigen-presenting cell function in mice lacking expression of OX40 ligand.

Murata K, Ishii N, Takano H, Miura S, Ndhlovu LC, Nose M, Noda T, Sugamura K - J. Exp. Med. (2000)

Impaired T cell priming and cytokine production in OX40L-deficient mice and MGP34-treated mice. (A) OX40L-deficient mice have impaired recall proliferative responses to protein antigens. OX40L-deficient (□) or wild-type (▪) mice (four per group) were immunized with KLH, HEL, or OVA in the hind footpads. 9 d after immunization, draining lymph nodes were extracted and subjected to an in vitro challenge of the various protein antigens. After culturing for 3 d, their [3H]thymidine uptake was measured. (B) Impairment of recall proliferative response of the CD4+ T cells of OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes of the OX40L-deficient (□) or wild-type mice (▪) were assayed for their recall proliferation in response to KLH in the presence of APCs from wild-type mice. (C) Defective APC function in OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes from wild-type mice primed with KLH were assayed for their recall proliferation in response to KLH in the presence of OX40L-deficient (□) or wild-type (▪) irradiated spleen cells used as APCs. (D) MGP34-treated mice have similarly impaired lymph node recall proliferation to KLH. C57BL/6 mice (four per group) were immunized in the hind footpads with KLH in CFA. On days 0, 3, and the 6, mice received 500 μg anti-OX40L (○) or rat IgG (•) intraperitoneally. The recall proliferative responses of the draining lymph nodes (left) were tested in the same manner as described above. Purified CD4+ T cells from the draining lymph nodes of the MGP34-treated (○) or control IgG–treated (•) mice were assayed for their recall proliferation in response to KLH in the presence of APCs from C57BL/6 mice (right). (E and F) Absence of OX40L or MGP34 treatment inhibits recall cytokine production by lymph nodes in response to KLH. Production of Th1 (E) and Th2 (F) cytokines by the draining lymph nodes of the mice in the four groups (□, ▪, ○, •) immunized with KLH and further subjected to an in vitro challenge with KLH were measured.
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Related In: Results  -  Collection

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Figure 4: Impaired T cell priming and cytokine production in OX40L-deficient mice and MGP34-treated mice. (A) OX40L-deficient mice have impaired recall proliferative responses to protein antigens. OX40L-deficient (□) or wild-type (▪) mice (four per group) were immunized with KLH, HEL, or OVA in the hind footpads. 9 d after immunization, draining lymph nodes were extracted and subjected to an in vitro challenge of the various protein antigens. After culturing for 3 d, their [3H]thymidine uptake was measured. (B) Impairment of recall proliferative response of the CD4+ T cells of OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes of the OX40L-deficient (□) or wild-type mice (▪) were assayed for their recall proliferation in response to KLH in the presence of APCs from wild-type mice. (C) Defective APC function in OX40L-deficient mice. Purified CD4+ T cells from the draining lymph nodes from wild-type mice primed with KLH were assayed for their recall proliferation in response to KLH in the presence of OX40L-deficient (□) or wild-type (▪) irradiated spleen cells used as APCs. (D) MGP34-treated mice have similarly impaired lymph node recall proliferation to KLH. C57BL/6 mice (four per group) were immunized in the hind footpads with KLH in CFA. On days 0, 3, and the 6, mice received 500 μg anti-OX40L (○) or rat IgG (•) intraperitoneally. The recall proliferative responses of the draining lymph nodes (left) were tested in the same manner as described above. Purified CD4+ T cells from the draining lymph nodes of the MGP34-treated (○) or control IgG–treated (•) mice were assayed for their recall proliferation in response to KLH in the presence of APCs from C57BL/6 mice (right). (E and F) Absence of OX40L or MGP34 treatment inhibits recall cytokine production by lymph nodes in response to KLH. Production of Th1 (E) and Th2 (F) cytokines by the draining lymph nodes of the mice in the four groups (□, ▪, ○, •) immunized with KLH and further subjected to an in vitro challenge with KLH were measured.
Mentions: To examine the in vivo effect of OX40L on antigen-specific T cell activation, we immunized OX40L-deficient or wild-type mice with KLH and tested their in vitro T cell recall proliferative response. A significantly reduced response was observed in OX40L-deficient mice, and the antigen dose–response curve was shifted by several orders of magnitude (Fig. 4 A). Similarly reduced responses were seen with two other protein antigens, hen egg lysozyme (HEL) and OVA, in OX40L-deficient mice (Fig. 4 A). These results suggest that OX40L plays an important role in antigen-specific T cell priming. The reduced response in OX40L-deficient mice may be due to a defect at the level of the APCs, since the intrinsic potential of OX40L-deficient T cells in response to antigens is preserved (data not shown) and since OX40L expression was restricted to activated APCs but not T cells.

Bottom Line: OX40L expression was demonstrated on splenic B cells after CD40 and anti-immunoglobulin (Ig)M stimulation, while only CD40 ligation was capable of inducing OX40L on dendritic cells.The impaired T cell priming was also accompanied by a concomitant reduction of both T helper type 1 (Th1) and Th2 cytokines.Collectively, these results provide convincing evidence that OX40L, expressed on APCs, plays a critical role in antigen-specific T cell responses in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Tohoku University School of Medicine, Sendai 980-8575, Japan.

ABSTRACT
OX40 expressed on activated T cells is known to be an important costimulatory molecule on T cell activation in vitro. However, the in vivo functional significance of the interaction between OX40 and its ligand, OX40L, is still unclear. To investigate the role of OX40L during in vivo immune responses, we generated OX40L-deficient mice and a blocking anti-OX40L monoclonal antibody, MGP34. OX40L expression was demonstrated on splenic B cells after CD40 and anti-immunoglobulin (Ig)M stimulation, while only CD40 ligation was capable of inducing OX40L on dendritic cells. OX40L-deficient and MGP34-treated mice engendered apparent suppression of the recall reaction of T cells primed with both protein antigens and alloantigens and a significant reduction in keyhole limpet hemocyanin-specific IgG production. The impaired T cell priming was also accompanied by a concomitant reduction of both T helper type 1 (Th1) and Th2 cytokines. Furthermore, antigen-presenting cells (APCs) derived from the mutant mice revealed an impaired intrinsic APC function, demonstrating the importance of OX40L in both the priming and effector phases of T cell activation. Collectively, these results provide convincing evidence that OX40L, expressed on APCs, plays a critical role in antigen-specific T cell responses in vivo.

Show MeSH
Related in: MedlinePlus