A First-in-Human Safety and Dose-Finding Study of New Type-16 Human Rhinovirus (RG-HRV16) Inoculum in Healthy Volunteers

Rhinoviruses are the most frequently cause of the common cold. HRV16 (Family Picornaviridae
Genus Rhinovirus type 16) has been used extensively to induce colds in studies of
experimentally inoculated volunteers that are designed to study the pathogenesis of colds and
effects of antiviral medications.

Experimental inoculation with human rhinovirus type 16 (HRV16) administered intranasally via
aerosolization has been used at the University of Wisconsin for over 30 years, and has proven
to be a safe tool to reproducibly induce symptomatic colds. HRV has been linked with
exacerbations of asthma and COPD, and this model has been used to evaluate inflammatory
mechanisms and to test the efficacy of treatments for the common cold. Recent refinements in
the technology available to produce and safety test reagents that are intended to be
administered to human volunteers as part of research protocols has prompted us to produce a
new lot of HRV16 in accordance with standards of current Good Manufacturing Procedures
(cGMP). For this inoculum, we have used a cDNA clone (reverse genetics) to generate source
virus, thus this new virus inoculum will be referred to as RG-HRV16.

This approach has two main advantages over using viruses isolated from nasal secretions.
First, several "new" respiratory viruses (e.g. metapneumovirus, bocavirus, SARS, rhinovirus
group C) have been discovered in the past 10 years, and there is little doubt that additional
viruses will be discovered. Therefore, it is impossible to ensure that nasal secretions that
are chosen for isolation of "seed virus" contain only the pathogen of interest. This problem
is minimized through the use of virus derived from a cDNA clone that was produced in E. coli.
Second, RNA viruses, such as HRV, mutate as the virus grows because their RNA polymerases
have no error-correcting function. The cDNA clone, reproduced by the much more accurate E.
coli DNA polymerase, provides a stable source of virus sequence for production of future
inocula.

This study represents a first-in-human, phase 1 study to assess the safety of RG-HRV16 in
humans and identify the dose needed to produce moderate-to-severe colds in 75% of
HRV16-seronegative human volunteers.

Inclusion Criteria:

1. Capable and willing to grant written informed consent (in English) and cooperate with
study procedures and requirements including willingness to avoid cold symptom
medications during the study.

2. Age between 18 and 50 years (children and older adults will be excluded from the study
due to the possibility of greater morbidity associated with upper respiratory
infections in these age groups).

3. Absence of HRV16 neutralizing antibody.

4. Safety laboratory assessments within normal University of WI Hospital and Clinics
(UWHC) ranges or grade 1 (mild) laboratory abnormalities which are deemed not
clinically significant in the judgment of the PI. Labs to include CBC with
differential and platelets, BUN, creatinine, AST, ALT and IgA and IgG serum
immunoglobulins. Any lymphopenia outside of the normal range will be an absolute
exclusion.

5. Subjects must be willing to refrain from taking nasal decongestants, antihistamines or
cough/cold preparations (this includes dietary supplements and homeopathic
preparations used specifically for cold/flu e.g., vitamin C, zinc, echinacea) within
the 7 days prior to Visit 1 and throughout the study until after the completion of
Visit 5.

Exclusion Criteria:

1. A chronic medical condition, which may increase risk or interfere with the conduct of
the study, including, but not limited to heart disease, Type I of II diabetes
mellitus, asthma, COPD, other chronic lung disease, perennial rhinitis and chronic
rhinosinusitis.

2. Subjects with household contacts who are pregnant or planning a pregnancy during the
main subject's participation, who have chronic respiratory disease, who are children
under the age of 2 years, or who are adults over 50 years of age.

3. Subjects who provide healthcare services or work with elderly or children (e.g.
daycare provider, senior citizen care giver).

4. Subjects with seasonal allergies will be excluded only if allergy symptoms are present
at baseline, or are anticipated to occur during the study period.

5. Smoking within the past 6 months.

6. Subjects who have received immunosuppressive treatment within the last 12 months.

7. Upper respiratory infection (URI or sinusitis) in the past 4 weeks.

8. Subjects with a history of a significant adverse reaction or intolerance to a previous
live, attenuated vaccine.

9. Pregnant or breastfeeding women or a woman who has a planned pregnancy during the
course of the study.

10. Unwillingness of study participants to use adequate birth control methods during the
course of the study. Adequate birth control methods include oral contraceptives,
injections such as Depo-Provera, an intrauterine device or double-barrier
contraception (combination of condom and contraceptive sponge or cervical cap and
spermicide or another combination approved in writing by the Principal Investigator).