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Infection and Immunity, June 2007, p. 2826-2832, Vol. 75, No. 6
0019-9567/07/$08.00+0     doi:10.1128/IAI.00127-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Transcutaneous Immunization with Clostridium difficile Toxoid A Induces Systemic and Mucosal Immune Responses and Toxin A-Neutralizing Antibodies in Mice{triangledown}

Chandrabali Ghose,1,2 Anuj Kalsy,1 Alaullah Sheikh,1 Julianne Rollenhagen,1,2 Manohar John,1,2 John Young,1 Sean M. Rollins,1,2 Firdausi Qadri,3 Stephen B. Calderwood,1,2,4 Ciaran P. Kelly,5 and Edward T. Ryan1,2,6*

Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts,1 Department of Medicine, Harvard Medical School, Boston, Massachusetts,2 ICDDR,B Centre for Health and Populations Studies, Dhaka, Bangladesh,3 Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts,4 Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts,5 Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts6

Received 25 January 2007/ Returned for modification 2 March 2007/ Accepted 13 March 2007

Clostridium difficile is the leading cause of nosocomial infectious diarrhea. C. difficile produces two toxins (A and B), and systemic and mucosal anti-toxin A antibodies prevent or limit C. difficile-associated diarrhea. To evaluate whether transcutaneous immunization with formalin-treated C. difficile toxin A (CDA) induces systemic and mucosal anti-CDA immune responses, we transcutaneously immunized three cohorts of mice with CDA with or without immunoadjuvantative cholera toxin (CT) on days 0, 14, 28, and 42. Mice transcutaneously immunized with CDA and CT developed prominent anti-CDA and anti-CT immunoglobulin G (IgG) and IgA responses in serum and anti-CDA and anti-CT IgA responses in stool. Sera from immunized mice were able to neutralize C. difficile toxin A activity in an in vitro cell culture assay. CDA itself demonstrated adjuvant activity and enhanced both serum and stool anti-CT IgA responses. Our results suggest that transcutaneous immunization with CDA toxoid may be a feasible immunization strategy against C. difficile, an important cause of morbidity and mortality against which current preventative strategies are failing.

* Corresponding author. Mailing address: Division of Infectious Diseases, Massachusetts General Hospital, Jackson 504, 55 Fruit Street, Boston, MA 02114. Phone: (617) 726-3812. Fax: (617) 726-7416. E-mail: etryan{at}partners.org

{triangledown} Published ahead of print on 19 March 2007.

Editor: W. A. Petri, Jr.

Infection and Immunity, June 2007, p. 2826-2832, Vol. 75, No. 6
0019-9567/07/$08.00+0     doi:10.1128/IAI.00127-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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