| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
* Departments of Pediatric Dentistry and Microbiology, Immunobiology Vaccine Center, University of Alabama at Birmingham, Birmingham, AL 35294;
Research Center for Biologicals, Kitasato Institute, Saitama, Japan;
Department of Gastroenterology, Research Institute, International Medical Center of Japan, Tokyo, Japan;
Cine-Science Laboratory Co. Ltd., Tokyo, Japan; and
¶ Division of Mucosal Immunology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan
Nasal application of native cholera toxin (nCT) as a mucosal adjuvant has potential toxicity for the CNS through binding to GM1 gangliosides in the olfactory nerves. Although mutants of cholera toxin (mCTs) have been developed that show mucosal adjuvant activity without toxicity, it still remains unclear whether these mCTs will induce CNS damage. To help overcome these concerns, in this study we created new double mutant CTs (dmCTs) that have two amino acid substitutions in the ADP-ribosyltransferase active center (E112K) and COOH-terminal KDEL (E112K/KDEV or E112K/KDGL). Confocal microscopic analysis showed that intracellular localization of dmCTs differed from that of mCTs and nCTs in intestinal epithelial T84 cells. Furthermore, both dmCTs exhibited very low toxicity in the Y1 cell assay and mouse ileal loop tests. When mucosal adjuvanticity was examined, both dmCTs induced enhanced OVA-specific immune responses in both mucosal and systemic lymphoid tissues. Interestingly, although both dmCT E112K/KDEV and dmCT E112K/KDGL showed high Th2-type and significant Th1-type cytokine responses by OVA-specific CD4+ T cells, dmCT E112K/KDEV exhibited significantly lower Th1-type cytokine responses than did nCT and dmCT E112K/KDGL. These results show that newly developed dmCTs retain strong biological adjuvant activity without CNS toxicity.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This research was supported by National Institutes of Health Grants DC 04976, DE 12242, AI 18958, and AI 43197 and Grants-in-Aid from the Ministry of Health and Labor, Japan, the Ministry of Education, Science and Sports, Japan, and CREST of Japan Science and Technology Corporation.
2 Address correspondence and reprint requests to Dr. Kohtaro Fujihashi, Department of Pediatric Dentistry, Immunobiology Vaccine Center, University of Alabama at Birmingham, 761 Bevill Biomedical Research Building, 845 19th Street South, Birmingham, AL 35294-2170. E-mail address: kohtarof{at}uab.edu
3 Abbreviations used in this paper: S-IgA, secretory IgA; AFC, Ab forming cell; CHO, Chinese hamster ovary; CLN, cervical lymph node; CT, cholera toxin; nCT, native CT; CT-A, A subunit of nCT; CT-B, B subunit of nCT; dmCT, double mutant CT; mCT, mutant CT; DD, dimer of an Ig binding element; ER, endoplasmic reticulum; LT, heat-labile enterotoxin; nLT, native LT; NALT, nasopharyngeal-associated lymphoreticular tissue; NP, nasal passage; OB, olfactory bulb; ON/E, olfactory nerves and epithelium; SMG, submandibular gland.
This article has been cited by other articles:
|
|
 |
|
  S. Sekine, K. Kataoka, Y. Fukuyama, Y. Adachi, J. Davydova, M. Yamamoto, R. Kobayashi, K. Fujihashi, H. Suzuki, D. T. Curiel, et al. A Novel Adenovirus Expressing Flt3 Ligand Enhances Mucosal Immunity by Inducing Mature Nasopharyngeal-Associated Lymphoreticular Tissue Dendritic Cell Migration J. Immunol., June 15, 2008; 180(12): 8126 - 8134. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
