“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

GasliniGasliniGasliniGaslini

Istituto Istituto Istituto Istituto

Giannina Giannina Giannina Giannina

GasliniGasliniGasliniGaslini

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 ageCharlottesville, VA

Heymann PW. JACI 2004: 114: 239-47.

Rhino

Rhino

Rhino

Rhino

Rhino

RSV

RSVRSV

RSV infection: Replication & Spreading

G Protein(adhesion)

F Protein(fusion)

A. RSV penetration B. RSV replication C. 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

RNATRANSLATION

SYNCYTIA

FORMATION

RSV is a highly cytotoxic virus

Respiratory

2

RespiratorySyncytial

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 leakOedema

Vasodilation

New vessels

Bronchoconstriction

Neurogenicreflexes

• 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

Rhinovirus3

• Rhinovirus (RV) infection is characterized by stimulation of the secretion ofmediators 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

Ast

hm

a a

t 6

ye

ars

(%

)100 -

80 -

60 -

OR=9.8OR=10.0

Viral infections in the first 3 years of life

RhinovirusRSVA

sth

ma

at

6 y

ea

rs (

%)

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 asthmaAllergic asthma

Sampath D. J. Clin. Invest 1999; 103: 1353-1361 .

Rhinovirus infection induces expression of

its own receptor ICAM-1

ICAM-1HRV 16

Papi A. J Biol Chem 1999; 274: 9707-20.

Thus …

ICAM-1

ICAM-1HRV 16

In conclusion

Infants &

Young children

Extensive damage

to the airways

Long lasting bronchial

hyperreactivity

Short epidemic

seasonRSV

Prematurity

Low Chronological

Age

Neurogenic

Inflammation

More Inducer

than Trigger

Infants, Children

& Adolescents

Every

seasonAtopy 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 CellsICAM-1

Mastromarino 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 cellsEffect 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 othermolecules thus facilitating stability, mucoadesivityand 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

MIPH2O2

Dectin-1Complement 3

Neutrophils Macrophages

Dendritic cells

TNF-αααα

IFN-γγγγ

ββββ-Glucan

Dectin-1Receptor

Complement 3Receptor

T- Lymphocytes

B- Lymphocytes NK- Cells

Airway Epithelial

Cells

Virus busters …

Resveratrol

ββββ - glucan