Mucosal Immunization of Cynomolgus Macaques with the VSVDG/Z EBOVGP Vaccine Stimulates Strong Ebola GP-Specific Immune Responses Xiangguo Qiu1., Lisa Fernando1., Judie B. Alimonti1,3, P. Leno Melito1, Friedericke Feldmann1¤, Daryl Dick1, Ute Stro¨ her1,3, Heinz Feldmann1,3¤, Steven M. Jones1,2,3* 1 Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada, 2 Department of Immunology, University of Manitoba, Winnipeg, Manitoba, Canada, 3 Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada Abstract Background: Zaire ebolavirus (ZEBOV) produces a lethal viral hemorrhagic fever in humans and non-human primates. Methodology/Principal Findings: We demonstrate that the VSVDG/ZEBOVGP vaccine given 28 days pre-challenge either intranasally (IN), orally (OR), or intramuscularly (IM) protects non-human primates against a lethal systemic challenge of ZEBOV, and induces cellular and humoral immune responses. We demonstrated that ZEBOVGP-specific T-cell and humoral responses induced in the IN and OR groups, following an immunization and challenge, produced the most IFN-c and IL-2 secreting cells, and long term memory responses. Conclusions/Significance: We have shown conclusively that mucosal immunization can protect from systemic ZEBOV challenge and that mucosal delivery, particularly IN immunization, seems to be more potent than IM injection in the immune parameters we have tested. Mucosal immunization would be a huge benefit in any emergency mass vaccination campaign during a natural outbreak, or following intentional release, or for mucosal immunization of great apes in the wild. Citation: Qiu X, Fernando L, Alimonti JB, Melito PL, Feldmann F, et al. (2009) Mucosal Immunization of Cynomolgus Macaques with the VSVDG/ZEBOVGP Vaccine Stimulates Strong Ebola GP-Specific Immune Responses. PLoS ONE 4(5): e5547. doi:10.1371/journal.pone.0005547 Editor: Derya Unutmaz, New York University School of Medicine, United States of America Received February 3, 2009; Accepted April 27, 2009; Published May 14, 2009 This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. Funding: The project received funding from the Public Health Agency of Canada (PHAC), and the Chemical, Biological, Radiological, and Nuclear Research and Technology Initiative (CRTI). Grant # CRTI-05-0078RTD URL:http://www.css.drdc-rddc.gc.ca/crti/index-eng.asp. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: steven_jones@phac-aspc.gc.ca ¤ Current address: Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, Montana, United States of America . These authors contributed equally to this work. Introduction Ebola virus (EBOV) and Marburg virus (MARV) are members of the Filoviridae family causing a lethal viral haemorrhagic fever (HF). Zaire ebolavirus (ZEBOV), is the most virulent species in the Ebola genus with human case fatality rates of over 80%, [1–5]. Sporadic outbreaks of ZEBOV have occurred in Central Africa since 1976 causing more than 1,800 human cases, as well as epidemics among chimpanzees and great apes [6–9]. Populations of wild apes in Central Africa have been decimated and there is a significant risk that these populations could become extinct because of ongoing outbreaks [7]. Importantly in the bordering region between Gabon and the Republic of Congo, human outbreaks have been shown to correspond to outbreaks in great apes, and so preventing disease in these animals could potentially prevent many human cases. Experimental infections of non-human primates (NHPs) with ZEBOV by intramuscular injection or mucosal exposure results in a disease with symptomology similar to humans [10–12]. Transmission of ZEBOV is generally through exposure to infected bodily fluids via either breaks and abrasions of the skin, or through mucosal surfaces. It is currently unknown which plays a more prominent role in the spread of disease. However, infection can occur via aerosol administration [13]. In addition, there is evidence that ZEBOV can be transmitted between experimentally inoculated monkeys and na¨ıve controls probably by virus-laden droplets secreted or excreted leading to mucosal infection [14]. For this reason there is concern of EBOV being incorporated into a biological weapon. In addition, research was conducted into the use of weaponized MARV with the most likely means of dispersal through aerosol generation [15]. The severity and high fatality of the disease, in addition to the possibility of EBOV or MARV to be used as a bioterrorist weapon has necessitated the research and development of vaccines for both EBOV and MARV. Currently there is no preventative vaccine or post-exposure treatment option approved for human or animal use. Early attempts at creating a vaccine included c-irradiated MARV [16] or ZEBOV [10]; or formalin-inactivated ZEBOV [17]. These vaccines were able to provide protection in guinea pigs, but in the NHP model the vaccines failed to protect. Recent attempts focusing PLoS ONE | www.plosone.org 1 May 2009 | Volume 4 | Issue 5 | e5547