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Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility complex class II-restricted viral epitope.

Casares S, Inaba K, Brumeanu TD, Steinman RM, Bona CA - J. Exp. Med. (1997)

Bottom Line: We used DNA that encodes an A/PR/8/34 influenza peptide for CD4 T cells and that elicits protective antiviral immunity.When DNA was injected into muscles or skin, and antigen-presenting cells were isolated from either the draining lymph nodes or the skin, dendritic, but not B, cells presented antigen to T cells and carried plasmid DNA.We suggest that the uptake of DNA and/or the protein expressed by dendritic cells triggers immune responses to DNA vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York 10029, USA.

ABSTRACT
Intramuscular and intracutaneous immunization with naked DNA can vaccinate animals to the encoded proteins, but the underlying mechanisms of antigen presentation are unclear. We used DNA that encodes an A/PR/8/34 influenza peptide for CD4 T cells and that elicits protective antiviral immunity. DNA-transfected, cultured muscle cells released the influenza polypeptide, which then could be presented on the major histocompatibility complex class II molecules of dendritic cells. When DNA was injected into muscles or skin, and antigen-presenting cells were isolated from either the draining lymph nodes or the skin, dendritic, but not B, cells presented antigen to T cells and carried plasmid DNA. We suggest that the uptake of DNA and/or the protein expressed by dendritic cells triggers immune responses to DNA vaccines.

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Activation of a HA110-120–specific TcH by enriched Langerhans cells transfected in vivo with pVH– TB plasmid. Langerhans cells (2 × 105)  were obtained from mice immunized  intracutaneously with pVH–TB plasmid  and cultured for 24 h with 14.3.1 TcH  (2 × 105). FACS® analysis shows that  Langerhans cells from animals immunized with pVH–TB activated 26% of  the HA110-120–specific TcH (a), but  Langerhans cells from animals immunized with pC did not activate any of  the TcH (b). (c), PCR analysis of pVH– TB and pC plasmids using T7 and Sp6 primers in skin cells from mice immunized intracutaneously. Lane 1, molecular markers; lane 2, purified pVH–TB  plasmid; lane 3, purified pC plasmid; lane 4, skin cells from mice immunized with pC; lane 5, skin cells from mice immunized with pVH–TB; lane 6,  MHC class II–positive Langerhans cells from mice immunized with pVH–TB; lane 7, MHC class II–negative skin cells from mice immunized with pVH– TB; lane 8, negative control.
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Figure 4: Activation of a HA110-120–specific TcH by enriched Langerhans cells transfected in vivo with pVH– TB plasmid. Langerhans cells (2 × 105) were obtained from mice immunized intracutaneously with pVH–TB plasmid and cultured for 24 h with 14.3.1 TcH (2 × 105). FACS® analysis shows that Langerhans cells from animals immunized with pVH–TB activated 26% of the HA110-120–specific TcH (a), but Langerhans cells from animals immunized with pC did not activate any of the TcH (b). (c), PCR analysis of pVH– TB and pC plasmids using T7 and Sp6 primers in skin cells from mice immunized intracutaneously. Lane 1, molecular markers; lane 2, purified pVH–TB plasmid; lane 3, purified pC plasmid; lane 4, skin cells from mice immunized with pC; lane 5, skin cells from mice immunized with pVH–TB; lane 6, MHC class II–positive Langerhans cells from mice immunized with pVH–TB; lane 7, MHC class II–negative skin cells from mice immunized with pVH– TB; lane 8, negative control.

Mentions: Both intracutaneous and intradermal routes of administration are currently used for the immunization with naked DNA (16). The major APCs in skin are the epidermal dendritic, or Langerhans, cells (17). To investigate the cellular mechanism of subcutaneous DNA immunization, Langerhans cells were isolated from the ears of B6 × D2 F1 (H-2b × H-2d) mice. 6 h after the injection of 30 μg pVH– TB or pC, the ear skin was explanted and cultured for 4 d, during which time cells emigrated from the skin. The emigrated cells were tested for the ability to activate HA110-120–specific TcH. A substantial activation was obtained when Langerhans cells from mice immunized with pVH–TB were cultured with TcH in the absence of exogenous antigen. This activation was measured by the production of IL-2 (Fig. 4 a), IFN-γ, and IL-4 (Table 1, group 7). Langerhans cells from mice immunized with pC failed to activate the TcH in absence of exogenous antigen (Fig. 4 b; Table 1, group 5).


Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility complex class II-restricted viral epitope.

Casares S, Inaba K, Brumeanu TD, Steinman RM, Bona CA - J. Exp. Med. (1997)

Activation of a HA110-120–specific TcH by enriched Langerhans cells transfected in vivo with pVH– TB plasmid. Langerhans cells (2 × 105)  were obtained from mice immunized  intracutaneously with pVH–TB plasmid  and cultured for 24 h with 14.3.1 TcH  (2 × 105). FACS® analysis shows that  Langerhans cells from animals immunized with pVH–TB activated 26% of  the HA110-120–specific TcH (a), but  Langerhans cells from animals immunized with pC did not activate any of  the TcH (b). (c), PCR analysis of pVH– TB and pC plasmids using T7 and Sp6 primers in skin cells from mice immunized intracutaneously. Lane 1, molecular markers; lane 2, purified pVH–TB  plasmid; lane 3, purified pC plasmid; lane 4, skin cells from mice immunized with pC; lane 5, skin cells from mice immunized with pVH–TB; lane 6,  MHC class II–positive Langerhans cells from mice immunized with pVH–TB; lane 7, MHC class II–negative skin cells from mice immunized with pVH– TB; lane 8, negative control.
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Figure 4: Activation of a HA110-120–specific TcH by enriched Langerhans cells transfected in vivo with pVH– TB plasmid. Langerhans cells (2 × 105) were obtained from mice immunized intracutaneously with pVH–TB plasmid and cultured for 24 h with 14.3.1 TcH (2 × 105). FACS® analysis shows that Langerhans cells from animals immunized with pVH–TB activated 26% of the HA110-120–specific TcH (a), but Langerhans cells from animals immunized with pC did not activate any of the TcH (b). (c), PCR analysis of pVH– TB and pC plasmids using T7 and Sp6 primers in skin cells from mice immunized intracutaneously. Lane 1, molecular markers; lane 2, purified pVH–TB plasmid; lane 3, purified pC plasmid; lane 4, skin cells from mice immunized with pC; lane 5, skin cells from mice immunized with pVH–TB; lane 6, MHC class II–positive Langerhans cells from mice immunized with pVH–TB; lane 7, MHC class II–negative skin cells from mice immunized with pVH– TB; lane 8, negative control.
Mentions: Both intracutaneous and intradermal routes of administration are currently used for the immunization with naked DNA (16). The major APCs in skin are the epidermal dendritic, or Langerhans, cells (17). To investigate the cellular mechanism of subcutaneous DNA immunization, Langerhans cells were isolated from the ears of B6 × D2 F1 (H-2b × H-2d) mice. 6 h after the injection of 30 μg pVH– TB or pC, the ear skin was explanted and cultured for 4 d, during which time cells emigrated from the skin. The emigrated cells were tested for the ability to activate HA110-120–specific TcH. A substantial activation was obtained when Langerhans cells from mice immunized with pVH–TB were cultured with TcH in the absence of exogenous antigen. This activation was measured by the production of IL-2 (Fig. 4 a), IFN-γ, and IL-4 (Table 1, group 7). Langerhans cells from mice immunized with pC failed to activate the TcH in absence of exogenous antigen (Fig. 4 b; Table 1, group 5).

Bottom Line: We used DNA that encodes an A/PR/8/34 influenza peptide for CD4 T cells and that elicits protective antiviral immunity.When DNA was injected into muscles or skin, and antigen-presenting cells were isolated from either the draining lymph nodes or the skin, dendritic, but not B, cells presented antigen to T cells and carried plasmid DNA.We suggest that the uptake of DNA and/or the protein expressed by dendritic cells triggers immune responses to DNA vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York 10029, USA.

ABSTRACT
Intramuscular and intracutaneous immunization with naked DNA can vaccinate animals to the encoded proteins, but the underlying mechanisms of antigen presentation are unclear. We used DNA that encodes an A/PR/8/34 influenza peptide for CD4 T cells and that elicits protective antiviral immunity. DNA-transfected, cultured muscle cells released the influenza polypeptide, which then could be presented on the major histocompatibility complex class II molecules of dendritic cells. When DNA was injected into muscles or skin, and antigen-presenting cells were isolated from either the draining lymph nodes or the skin, dendritic, but not B, cells presented antigen to T cells and carried plasmid DNA. We suggest that the uptake of DNA and/or the protein expressed by dendritic cells triggers immune responses to DNA vaccines.

Show MeSH
Related in: MedlinePlus