“Rinite, Virus e Asma”
Giovanni A. Rossi U.O. di Pneumologia e Allergologia Pediatrica I.R.C.C.S. G. Gaslini, Genova giovannirossi@ospedale-gaslini.ge.it
Fondazione Fondazione Fondazione Fondazione Gerolamo Gerolamo Gerolamo Gerolamo
Gaslini Gaslini Gaslini Gaslini Istituto
Istituto Istituto Istituto Giannina Giannina Giannina Giannina
Gaslini Gaslini Gaslini Gaslini
Which respiratory virues can induce rhinitis, wheezing and asthma?
Airway in Asthma
1
Mucus Swelling Tight Muscles
Influenza Rhinovirus RSV
Other viruses
RSV and Rhinovirus are key agents in the exacerbation of childhood wheeze
Viral pathogens in nasal secretions among children hospitalized for wheezing in relation to age
Charlottesville, VA
Heymann PW. JACI 2004: 114: 239-47.
Rhino
Rhino
Rhino
Rhino
Rhino RSV
RSV
RSV
RSV infection: Replication & Spreading
G Protein
(adhesion)
F Protein
(fusion)
A.
RSV penetrationB.
RSV replicationC.
Membrane fusion& Syncytia formation
G Protein
(adhesion)
RSV penetration in epithelial cells due to G-GLYCOPROTEIN
interaction with its receptors FUSION PROTEIN
VIRAL PROTEIN SYNTHESIS
& ASSEMBLY
TRANSCRIPTION RNA
TRANSLATION
SYNCYTIA FORMATION
RSV is a highly cytotoxic virus
Respiratory
2
Respiratory Syncytial
Virus
Response to RSV-induced airway epithelial cell injury
Silvestri M et al. Paediatr Respir Rev 2004; 5: S81-7
Inflammation & neutrophil recruitment and activation
Release of arachidonic acid metabolites
Upregulation of neurogenic inflammation
Immune response to viral antigens (Th1 or Th2)
Disruption of epithelial cells
Increased sensitivity to neurotransmitters [Substance P and Neurokinins A and B]
Plasma leak Oedema Vasodilation
New vessels
Bronchoconstriction
Neurogenic reflexes
• The first viral episode may cause later bronchial obstructive
episodes by:
a) damaging the growing lung & inducing structural changes b) altering the host immune responses
c) producing airway neuro-muscular tone instability
RSV infection and pre-school wheeze
a … “soft” virus” inducing airway inflammation
Rhinovirus 3
• Rhinovirus (RV) infection is characterized by stimulation of the secretion of mediators of inflammation by airway structural/inflammatory cells
• Epithelial cell cytotoxicity does not appear to play a major role in the
pathogenesis of Rhinovirus (RV) infection
The risk of asthma at age 6 yrs (odds ratio) in children who wheezed during because of RSV and RV infections
Asthma at 6 years (%)
100 - 80 - 60 -
OR=9.8
OR=10.0
Viral infections in the first 3 years of life
Rhinovirus RSV
Asthma at 6 years (%)
60 - 40 - 20 -
0 - RSV RV RV&
Neither only only RSV Wheezing illness
OR=1.0
OR=2.6
Jackson DJ. AJRCCM 2008; 178: 667–672.
Seasonality and etiology of wheezing episodes in 285 children in the 1st year of life
RV
Madison, WI
Gern JE. JACI 2006;117:72-8.
RV
RV
RV
RV
HRV-A, HRV-B and HRV-C are very widespread and continuously co-circulating on all continents throughout the world
RSV
RSV
The prevalence of RSV & HRV bronchiolitis
• The prevalence of HRV-associated wheezing (20–40% in emergency department) increases steadily with age, the
breaking point in the dominance between HRV and RSV being around age of 12 mo in young hospitalized wheezing children
Turku, Finland
• The susceptibility to HRV bronchiolitis seems to be linked to predisposition, because its prevalence has
Jartti T. PAI 2011; 22: 350-355.
• Infants with HRV bronchiolitis present more often atopic dermatitis (54%) and blood eosinophilia (54%) than did infants with RSV bronchiolitis
linked to predisposition, because its prevalence has
been 50–80% during the first year of life in infants
with recurrent wheezing from atopic families
Is there a link between Rhinovirus
infection and allergic sensitization ?
4
The major receptor for HRV-A and HRV-B is ICAM-1, an adhesion molecule that is overexpressed by
airway epithelial cells in allergic asthma
ICAM-1
ICAM-1
Sacco O. Ann Allergy Asthma Immunol 1999; 83: 49-54
HRV
ICAM-1 expression in airway epithelial tissue of control indivituals and allergic asthma patients
Control Control
San Francisco, CA
Allergic asthma
Allergic asthma
Sampath D. J. Clin. Invest 1999; 103: 1353-1361 .
Rhinovirus infection induces expression of its own receptor ICAM-1
ICAM-1
HRV 16
Papi A. J Biol Chem 1999; 274: 9707-20.
Thus …
ICAM-1
ICAM-1
HRV 16
In conclusion
Infants &
Young children
Extensive damage to the airways
Long lasting bronchial hyperreactivity Short epidemic
season RSV
Prematurity Low Chronological
Age
Neurogenic Inflammation
More Inducer than Trigger
Infants, Children
& Adolescents
Every season
Atopy as facilitator
factor
Pro-inflammatory cell activation &
limited damage Recurrent
infection HRV
Long lasting bronchial hyperreactivity More Trigger
than Inducer
We really need …
We really need effective virus-busters …
Is Resveratrol an effective virus-busters …
5
Resveratrol
Resveratrol
• Resveratrol is a phytoalexin present in the peel of grapes, produced naturally by several plants when under attack by pathogens such as bacteria or fungi
• The activities of this molecule include: a) inhibition of
the replication of a variety of viruses; b) antioxidant and anti-infiammatory effects; c) inhibition of LDL
• When taken orally, Resveratrol appears to be well-absorbed by humans, but its bioavailability is relatively low because it is rapidly metabolized and the side effects of a long-term treatment
with this molecules are not known
anti-infiammatory effects; c) inhibition of LDL
peroxidation and platelet aggregation
Resveratrol-Mediated Reduction of Viral Replication and Airway Inflammation in RSV-Infected
Cyclophosphamide-Treated Mice
A. Pulmonary virus titers C. BAL cell type proportions
B. BAL cell numbers
Zang N. J Virol 2011; 85: 13061-13068
The Effects of Resveratrol, Dexamethasone or
Ribavirin on the Cytopathologic Effect of RSV on 9HTEO Cells
Xiao-hong Xie. Inflammation, 2012; 35: 1392-1401.
Resveratrol-Mediated Reduction of Viral Replication in Human Nasal Epithelial Cells
Infected by Human Rhinovirus (HRV)
HRV titers
Mastromarino P. 2013 (in press)
Apical cells Total cells
Resveratrol inhibits HRV-induced ICAM-1 Expression by Human Nasal
Epithelial Cells
ICAM-1Mastromarino P. 2013 (in press)
Inhibition of Influenza A Virus Replication by Resveratrol
Palamara AT. J Infect Dis 2005 May 15;191(10):1719-29.
Resveratrol (20 mg/mL) was added to cell cultures immediately after Influenza A Virus challenge (hour 0) and removed at different time points (1, 3, 6, 9, or 24 h) after infection.
The viral yields is reported for 24 and 48 h
Resveratrol treatment on late (24 h)
Influenza A Virus proteins expression and ribonucleoprotein trafficking
Infected
Nuclei Viral protein Merge
Palamara AT. J Infect Dis 2005 May 15;191(10):1719-29.
Infected + Resveratrol
Nuclei Viral protein Merge
Resveratrol as an antioxidant
Resveratrol acts as an antioxidant by:
• Inhibiting production of reactive oxygen species by inflammatory cells
Effect of Resveratrol on superoxide and hydrogen peroxide produced by murine macrophages stimulated with LPS (1 mg/mL, ) or PMA (1 mM, ) for 1 hr.
• Scavenging free radicals, related to the availability of hydroxyl groups (-OH) and the
availability of hydroxyl groups (-OH) and the presence of conjugated double bonds
• Stimulating the biosynthesis of endogenous
antioxidants by mechanisms such as stimulation
of nuclear erythroid-related factor activity
What about adding β - glucan to Resveratrol
6
Resveratrol +
ββββ - glucan
β -Glucans
• β-Glucans are chains of polysaccharides, that can vary with respect to molecular mass, solubility, viscosity, and three-dimensional
configuration, mode of action, and overall biological activity
• They occur most commonly as cellulose in plants, in cereal grains, the cell wall of certain fungi and bacteria
• 1,3- β -Glucan gels have been used in drug delivery to create helical micelles encapsulating other
molecules thus facilitating stability , mucoadesivity and solubility
• β-Glucans can act also as “biological response modifiers”
because of their ability to activate the immune system
thus reducing the risk of serious infections
Cells expressing receptors for β -Glucans
H2O2 MIP
Dectin-1 Complement 3
Neutrophils Macrophages
Dendritic cells
TNF-αααα IFN-γγγγ
ββββ-Glucan Dectin-1
Receptor
Complement 3 Receptor
T- Lymphocytes
B- Lymphocytes NK- Cells
Airway Epithelial Cells
