Immunostimulatory DNA sequences necessary for effective intradermal gene immunization

Science

Volumn 273, Issue 5273, 1996, Pages 352-354

  Sato, Yukio ^a   Roman, Mark ^a   Tighe, Helen ^a   Lee, Delphine ^a   Corr, Maripat ^a   Nguyen, Minh Duc ^a   Silverman, Gregg J ^a   Lotz, Martin ^a   Carson, Dennis A ^a   Raz, Eyal ^a  

^a UNIVERSITY OF CALIFORNIA SAN DIEGO   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; DNA SEQUENCE; GENE EXPRESSION; GENETIC CODE; IMMUNE RESPONSE; IMMUNIZATION; IMMUNOGENICITY; IMMUNOSTIMULATION; PLASMID; PRIORITY JOURNAL; RECEPTOR DOWN REGULATION;

EID: 8944253295     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5273.352     Document Type: Article

References (31)

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19. The sequences of the annealed, blunt-end, double-stranded oligonucleotides used for transfection are as follows: ISS oligonucleotide, 5′-TCATTGGAAAACGTTCTTCGGGGCG-3′, from the ampR gene in the pUC19 sequence (nucleotides 2288 to 2312); and ISS-deficient oligonucleotide, 5′-TCATTGGAAAAGGTTCTTGGGGGGG-3′. Bold nucleotides indicate the ISS.
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28. R limulus amebocyte lysate (LAL) assay (Association Cape Cod, Woods Hole, MA) and were less than 5 ng per milligram of DNA. Female BALB/c mice were intradermally injected at the base of the tail three times, 10 days apart, with the various pDNAs dissolved in normal saline. Mice were immunized with β-Gal protein in alum as previously described (3). All animal studies were approved by the University of California, San Diego, animal welfare committee.
29. The enzyme-linked immunosorbent assay (ELISA) for antibody to β-Gal was described previously (3). To quantitate the relative amount of antibody, we compared titration curves for individual sera with a standard curve on each plate using DeltaSOFT II v. 3.66 software (BioMetallics, Princeton, NJ). Additional experiments showed that the effects of the ISS were not attributable to polyclonal activation (Y. Sato et al., data not shown).
30. + splenocytes were assessed as previously described (3) except that commercial ELISAs were used (Endogen, Cambridge, MA).
31. We thank J. A. Dudler, Y. Geng, and J. Quach for providing the pACB vector, IL-12 p40 primer, and fresh human monocytes. We also thank P. Charos, A. Ronaghy, and S. Malek for their technical assistance and N. Noon for editorial assistance. Supported in part by grants AI37305, AR41897, and AI36214 from NIH. Y.S. was supported in part by the Japan Rheumatism Foundation. M.R. was supported by an NIH training grant (AI07384). H.T. was supported in part by an award from the CaPCURE Foundation.