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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 the  HA110-120–specific TcH by  various numbers of dendritic and  B cells. Various numbers of brachial and axillary lymph node,  dendritic cells, or B cells were  purified by cell sorting (a) from  BALB/c mice immunized in the  biceps and scapular muscles with  pC and cultured (b) with TcH (2  × 105) for 24 h in the presence  of HA110-120 peptide (25 μg/ ml). Dendritic or B cells isolated  from mice immunized with  pVH–TB or pC were cultured for  24 h with TcH (2 × 105) in the  absence of exogenous antigen (c).  The percentage of TcH-activated cells was determined by  FACS® analysis.
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Figure 2: Activation of the HA110-120–specific TcH by various numbers of dendritic and B cells. Various numbers of brachial and axillary lymph node, dendritic cells, or B cells were purified by cell sorting (a) from BALB/c mice immunized in the biceps and scapular muscles with pC and cultured (b) with TcH (2 × 105) for 24 h in the presence of HA110-120 peptide (25 μg/ ml). Dendritic or B cells isolated from mice immunized with pVH–TB or pC were cultured for 24 h with TcH (2 × 105) in the absence of exogenous antigen (c). The percentage of TcH-activated cells was determined by FACS® analysis.

Mentions: To assess the role of professional APCs in priming specific T cells after the intramuscular immunization with naked DNA, we studied the ability of sorted B cells (CD11c−B220+) and dendritic cells (CD11c+B220−) from mice immunized with pVH–TB to activate the HA110-120–specific TcH. Mice were immunized in the biceps and scapular muscles with 30 μg of pVH–TB or pC on days 0 and 2, and were killed 2 d after the last immunization. The high purity of the sorted APCs is shown in Fig. 2 a.


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 the  HA110-120–specific TcH by  various numbers of dendritic and  B cells. Various numbers of brachial and axillary lymph node,  dendritic cells, or B cells were  purified by cell sorting (a) from  BALB/c mice immunized in the  biceps and scapular muscles with  pC and cultured (b) with TcH (2  × 105) for 24 h in the presence  of HA110-120 peptide (25 μg/ ml). Dendritic or B cells isolated  from mice immunized with  pVH–TB or pC were cultured for  24 h with TcH (2 × 105) in the  absence of exogenous antigen (c).  The percentage of TcH-activated cells was determined by  FACS® analysis.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199124&req=5

Figure 2: Activation of the HA110-120–specific TcH by various numbers of dendritic and B cells. Various numbers of brachial and axillary lymph node, dendritic cells, or B cells were purified by cell sorting (a) from BALB/c mice immunized in the biceps and scapular muscles with pC and cultured (b) with TcH (2 × 105) for 24 h in the presence of HA110-120 peptide (25 μg/ ml). Dendritic or B cells isolated from mice immunized with pVH–TB or pC were cultured for 24 h with TcH (2 × 105) in the absence of exogenous antigen (c). The percentage of TcH-activated cells was determined by FACS® analysis.
Mentions: To assess the role of professional APCs in priming specific T cells after the intramuscular immunization with naked DNA, we studied the ability of sorted B cells (CD11c−B220+) and dendritic cells (CD11c+B220−) from mice immunized with pVH–TB to activate the HA110-120–specific TcH. Mice were immunized in the biceps and scapular muscles with 30 μg of pVH–TB or pC on days 0 and 2, and were killed 2 d after the last immunization. The high purity of the sorted APCs is shown in Fig. 2 a.

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