Collection of nasal lavage fluid from experimentally infected healthy human volunteers for the generation of a GMP batch of HRV16

Asthma and chronic obstructive pulmonary disease (COPD) are chronic
inflammatory disorders of the airways. Patients with asthma or COPD frequently
suffer from exacerbations, which are manifested by episodes of acute worsening
of symptoms, such as shortness of breath, cough, wheezing and chest tightness
in conjunction with airways obstruction. The frequent exacerbations contribute
to faster deterioration of lung function compared to that in healthy
individuals and thus attenuate the health status and quality of life of these
patients. The mechanisms that underlie these exacerbations are poorly
understood, which hampers the development of adequate prophylaxis and/or
treatment of these exacerbations. Exacerbations are triggered in particular by
viral respiratory infections. Of all human respiratory viruses, rhinovirus
species are most frequently causing exacerbations in asthma and COPD patients.

Relevance:
These preparations are crucial to establish a new batch of HRV16, prepared
according to GMP standards. This batch will enable researchers worldwide to
study exacerbations of asthma and COPD, and extent that to other respiratory
diseases.

Experimental infections of humans with rhinoviruses, in particular rhinovirus
16 (HRV16) have proven to be a highly relevant and safe approach to study
exacerbations in patients with asthma and COPD. The worldwide availability of
stocks of HRV16 for experimental infections is decreasing and thus there is a
need for a new batch. Various groups have joined forces in a European Union
project, U-BIOPRED, amongst others to prepare a state of the art HRV16 stock.
The major aim of this study is to collect samples of healthy individuals after
infection HRV16 that can serve as a stock to generate a GMP batch of HRV16,
which will be made available to multiple research groups currently using this
experimental exacerbation model.

Passage of rhinovirus in cell lines or animal models bears the risk of reducing
the potency of rhinovirus to infect humans. Therefore, in vivo passage of
rhinovirus in humans is required to obtain viable virus that is also infectious
for humans. Six, extensively screened healthy humans will be exposed to 10
TCID50 HRV16 (low dose). Daily, i.e. up to day 8, throat swabs and nasal lavage
samples will be obtained, each of which will be processed and stored
separately. The swabs serve to screen for other respiratory viruses and
bacteria. In nasal lavage samples we will determine HRV16 viability and load
(TCID50), number of viral particles (quantitative PCR) as well as execute an
extensive screening for other micro-organisms. In three nasal samples that
contain high and viable HRV16 loads, and that are free of any contaminating
micro-organisms, we will confirm HRV16 identity by sequencing the viral genome.
By using deep sequencing we will in parallel re-assess any other RNA- and
DNA-containing contaminants. When confirmed, these samples are considered fit
to be used in the preparation of the GMP batch of HRV16 by Charles River, a
project that will be conducted by the U-BIOPRED consortium.

Exposed individuals will develop a mild cold. The nasal washings and the throat
swabs are a minor burden to the participants. This experimetnal infection
protocol has been used in several hundres of individuals without any unexpected
effects and thus is considered a safe procedure..