Microbiologie Appliquée

Vaccins

Prof Gillet Nicolas

Université de Namur
Louis Pasteur
1. Un peu d’histoire
Immunisation = procédé qui déclenche une réaction immunitaire permettant à l'organisme
de se défendre contre un élément étranger (substance ou micro-organisme)

Variolation en Chine dès le 11ème siècle

ancêtre des vaccins inactivés

2
1. Un peu d’histoire

Vaccination, 1796, Edward Jenner

Cowpox/Horsepox pour vacciner contre Smallpox
premier vaccin vivant

3
1. Un peu d’histoire

Variole éradiquée par vaccination avec le Vaccinia virus (dernier cas humain reporté en 1977)

4
Production de vaccins contre le smallpox (Lab à Rockville, Maryland, Strategic National Stockpile)
5
Horsepox recréé par biologie synthétique en 2017 (David Evans, University of Alberta, Canada,
6
assemblage de séquences génétiques reçues par la poste)
1. Un peu d’histoire
Vaccin contre le choléra des poules (Pasteurella multocida), 1879, Pasteur
Vaccin vivant atténué, atténuation délibérée et (+ ou -) contrôlée du pathogène

7
 with Dr. Vulpian and Dr. Grancher (pro-
1. Un peu d’histoire fessor at the Faculty of Medicine, University of
Paris), Pasteur inoculated the boy with mate-
rial from a rabid rabbit spinal cord that had
Vaccin contre la rage (Rabies virus), 1885, Pasteur been dehydrated for 15 days (since June 21), as
indicated in the handwritten table (shown in
Vaccin inactivé, le virus est « mort » the image and transcribed into English below).

Ouvre la voie aux vaccins à base de microbes From July 7 to July 16, Pasteur continued
to inoculate Meister with rabid rabbits’ spinal
tués ou de fragments de microbes cord material that had been air dried for in-
creasingly shorter periods (15, 14, 12, 11, 9, 8,
7, 6, 5, 4, 3, 2, 1 day). The entire immuniza-
tion procedure is reported in Pasteur’s hand-
Image of Pasteur’s handwritten table of the rabies vaccination procedure. English trans- written table.
In parallel, Pasteur had inoculated rabbits
lation version of the table is below. See Fig. S1 for complete image of page. Image courtesy
with the same material used for Joseph
of Bibliothèque nationale de France.
Meister and had found that the spinal
material used on July 6, 7, 8, 9, and 10 was
Half a syringe not virulent, but the others were increasingly
virulent and, therefore with the inoculations
Marrow exposed Marrow
of July 15 and 16, he had exposed Meister to
to dry air on: dehydration for:
fully virulent virus. Pasteur concluded that
6 July 8 pm 21 June 15 d* the boy had not only survived the rabies from
7 July 9 am 23 June 14 d* the bites of a rabid dog, but also direct
7 July 6 pm 25 June 12 d* inoculation of a more virulent virus.
8 July 9 am 27 June 11 d*
8 July 6 pm 29 June 9 d*
9 July 11 am 1 July 8 d* 1 Pasteur L (1885) Méthode pour prévenir la rage après morsure.
10 July 11 am 3 July 7 d* Comptes Rendus des Séances de l’Académie des Sciences. Séance
11 July 11 am 5 July 6 d** du lundi 26 octobre 1885.
12 July 11 am 7 July 5 d**
13 July 11 am 9 July 4 d**
14 July 11 am 11 July 3 d** Author contributions: R.R. wrote the paper.
15 July 11 am 13 July 2 d** The author declares no conflict of interest.
16 July 11 am 15 July 1 d** 1
Email: rino.rappuoli@novartis.com.
8
*Nonvirulent in rabbits. This article contains supporting information online at www.pnas.org/
**Virulent in rabbits. lookup/suppl/doi:10.1073/pnas.1414226111/-/DCSupplemental.
1. Un peu d’histoire
Clostridium tetani et Corynebacterium
diphtheriae produisent des toxines
responsables du tétanos et de la diphtérie

Ramon, Descombey et Zoller, 1922-1927

Vaccins anti-tétanique et anti-diphtérique sont
des toxoïdes (= toxine inactivée/dénaturée)
(premiers vaccins sous-unitaires)

Glenny, 1926, ajout d’hydroxyde d’aluminium

améliore l’efficacité du vaccin, le 1er adjuvant
(ad juvare = pour aider)
impératif pour vaccins inactivés ou sous-
unitaires
9
1. Un peu d’histoire
1890, Von Behring et Kitasato

Immunité d’un animal peut être

transféré à un autre en injectant le
sérum du premier au second

Pionniers de l’immunisation passive

par des antitoxines (= anticorps)

10
 the baby to benefit from
2. Immunisation the broad immune experience of the mother.
passive
This passive protection falls to low levels at about 6 months of age as the Protection from Infection or Protection
Transfert
baby’s own d’anticorps
immune response(confère une
takes over. protection/traitement
Total antibody concentrations temporaire)
Disease?
are low from about 6 months to 1 year after birth, a property that may
Antitoxine
• increase diphtérique/tétanique
susceptibility (= anticorps
to disease. Premature infants contre
are particularly at les toxines diphtérique
It is important ou tétanique)
to consider that there are di
Ac monoclonaux
• risk contre
for infections because protéine
the level deIgG
of maternal Spike du and
is lower SARS-CoV-2
their (Regeneron,
outcomes Eli Lilly)
following vaccine administration.
immune system remains underdeveloped. The time course of production the antibody and memory T cells established
Administrations
• of various isoforms of d’Ig aux(IgG,
antibody patients
IgM, andimmunosupprimés
IgA) synthesized by the
De lais indicated.
• baby mère auAdapted
fœtusfrom et au nouveau-né
C. A. Janeway, Jr., et al., Immunobiolo- are maintained for long periods, and their mob
gy: the Immune System in Health and Disease (Current Biology Limited, sufficient to stop a subsequent infection befor
Garland Publishing Inc., New York, NY, 2001), with permission. spread beyond the site of entry. Disease is pre
the virus cannot reproduce or spread. In oth
Fraction of adult values

Months Years reproduction and spread may not be blocke

100
lgM Such infections can only be cleared by the coo
of vaccine-induced immune effectors and inf
Passively
transferred immune responses (e.g., interferon productio
lgG
maternal
lgA
disease may not be prevented, but its onset c
lgG or its severity lessened. In a third, less optima
0 virus will not be eliminated because the host’s
0 3 6 9/0 3 6 9 1 2 3 4 5 vaccine or to subsequent infection (or both)
Conception Birth Adult Consequently, disease is not prevented and va
Serum immunoglobulin levels confer only a modest delay in the appearance o
11
3. Immunisation active
Inoculation avec pathogène rendu non ou très faiblement pathogénique ou avec des antigènes
pour éliciter une réponse immune protectrice

12
3. Immunisation active
3.1. vaccins vivants atténués

13
 3. Immunisation active
3.1. vaccins vivants atténués
ModeChapter
70 d’emploi
8 (méthode “artisanale”) pour atténuer un virus et le transformer en vaccin

Pathogenic virus is isolated The cultured virus is used The virus acquires many The virus no longer grows well
from a patient and grown to infect monkey cells mutations that allow it to in human cells and may be a
in human cultured cells grow well in monkey cells candidate for a vaccine

Figure 8.11 Viruses specific for humans may become attenuated by passage in nonhuman
cell lines. The four panels show the process of producing an attenuated human virus by repeated transfers
in cultured cells. The first panel depicts isolation of the virus from human cells (yellow). The second panel 14
shows passage of the new virus in monkey cells (lavender). During the first few passages in nonhuman
3. Immunisation active
3.1. vaccins vivants atténués
Poliovirus: ssRNA(+), groupe IV, famille: picornaviridae, genre: enterovirus, non-enveloppé
Transmis par matières fécales et sécrections respiratoires
Asymptomatique, ou poliomyélite quand atteinte de la moelle épinière -> paralysie des membres
inférieurs, du système respiratoire

15
3. Immunisation active
3.1. vaccins vivants atténués Attenuation of poliovirus neurovirul
Oral Polio Vaccines (OPV) - Sabin’s vaccines

Virus atténués qui se repliquent dans le tractus

intestinal (comme le virus sauvage) et y
induisent une réponse immune protectrice de
longue durée (à vie), administration orale

Présence du vaccin dans matières fécales,

transmission du vaccin possible pendant
plusieurs semaines après inoculation

16
3. Immunisation active
3.1. vaccins vivants atténués
Oral Polio Vaccines (OPV) - Sabin’s vaccines

Immunisation de masse dès 1961 aux USA avec

OPV trivalent (contre virus type 1, 2 et 3)

Dans 1 cas sur 3 millions: paralysie associée au

vaccin (9 cas par an de 1961 à 1989 aux USA)
-> Réversion 17
 3. Immunisation active
3.1. vaccins vivants atténués

inants of Sabin vaccine strain attenuation

Virus Mutation

5’-UTR nt 480
VP1 aa 1106
P1/Sabin VP1 aa 1134
Moins sujet à VP3 aa 3225
réversion VP4 aa 4065
5’-UTR nt 481
P2/Sabin
VP1 aa 1143

5’-UTR nt 472
P3/Sabin
VP3 aa 3091

bia University

Virology Lectures 2017 • Prof. Vincent Racaniello • Columbia University 18 Princ

3. Immunisation active
3.1. vaccins vivants atténués

Réversion du Sabin type 3

La seule reversion dans l’IRES (U -> Reversion of re-transforme
C position 472) P3/Sabin le vaccin en virus virulent

Time of isolation Histological

Virus Base at 472
after vaccination lesion score

Sabin vaccine U 0.36

DM1 U 24 h ND
DM2 U 31 h 1.58
DM3 U/C 35 h ND
DM4 C 47 h 2.48
DM38 C 18 da ND
P3/119 C 3-4 weeks 3.34
19
Virology Lectures 2017 • Prof. Vincent Racaniello • Columbia University Evans et al., Nature 314:548 (1985)
3. Immunisation active
3.1. vaccins vivants atténués

Réversion du Sabin type 2

Vaccin encore utilisé dans pays du Sud

mOPV2 (monovalent oral PV2) peut facilement circuler dans population non immunisée et
évoluer pour regagner de la virulence
Le vaccin reverté (circulating Vaccine-Derived PolioVirus type 2 = cVDPV2) peut être transmis par
matière fécale et sécrétions respiratoires
-> + de 1000 cas de cVDPV2 dans le monde en 2020 (majoritairement en Afrique)
-> cVDPV2 retrouvés dans eaux usées de Londres et NY en 2022

Remplacement par un nouveau vaccin oral atténué contre le virus de type 2, contenant
notamment une mutation dans la RdRp la rendant plus fidèle: nOPV2 (novel Oral PolioVaccine
type 2)
nOPV2 « réverte » plus rarement que la Sabin mOPV2 mais est toujours responsable de rares cas
de polio 20
3. Immunisation active
3.1. vaccins vivants atténués
Isolate pathogenic virus

Vaccins vivants atténués par ingénierie génétique

Clone genome
• Clonage du génome viral dans un plasmide (ou
plusieurs si virus segmenté)
• Plusieurs mutations Virulence gene
• Délétions plus ou moins longues
• Virus recombinants et/ou réassortants
Isolate virulence gene

Mutate Delete
virulence virulence
gene gene

The resulting virus is viable and immunogenic

but not virulent. It may be used as a vaccine.
21
Figure 8.13 Construction of attenuated viruses by using
3. Immunisation active
3.1. vaccins vivants atténués

Vaccin bilié de Calmette-Guérin (BCG)

• Contre la tuberculose (induit par Mycobacterium tuberculosis)
• Souche atténuée de Mycobactérium bovis (239 sous-cultures d’une souche virulente de M.
bovis sur 13 ans !, après chaque sous-culture test sur animaux de laboratoire)
• 1ère utilisation chez l’Homme en 1921

22
3. Immunisation active
3.1. vaccins vivants atténués

Avantages
• Réponse immune efficace et durable -> immunité humorale (Ac) et cellulaire (CTL)

Désavantages
• Mutations vers une forme virulente (rare)
• Inadaptés aux personnes immunosupprimées (importance de l’immunité collective)
• (parfois necessité de stockage à basse T, stabilité)

23
3. Immunisation active
3.2. vaccins inactivés - tués
Inactivation par agent chimique (ou gamma-irradiation)

(IPV)

24
3. Immunisation active
3.2. vaccins inactivés - tués
Inactivated Polio Vaccine (IPV) - Salk’s vaccine

Virus inactivé chimiquement (formaldéhyde)

Ne se replique pas mais induit une réponse immune dans le sang (et pas dans la muqueuse
intestinale)
Pas de cas de paralysie associée au vaccin

Utilisé dès 1955 aux USA, remplacé par l’OPV de 1961 à 89, puis réintroduit en 89
Seul vaccin obligatoire de nos jours en Belgique
Europe indemne de polio depuis 2003 (mais risque de resurgence à cause d’une couverture
vaccinale déficiente et de cas de vaccine-derived polio)

25
3. Immunisation active Inactivated virus vaccine
3.2. vaccins inactivés - tués

Avantages

Immune response
Initial Second Third
• Sûr dose dose dose
• Généralement stable

Désavantages Time

• Moins efficace que vaccins atténués (ne se Replication competent virus vaccine
réplique pas)
• Doivent être injectés
• Nécessite plusieurs rappels

Immune response
Initial
dose
Amplification of
injected dose

Time 26
3. Immunisation active
3.3. vaccins sous unitaire (subunit vaccine)

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3. Immunisation active
3.3. vaccins sous unitaire (subunit vaccine)
On casse le virus ou la bactérie et on immunise On clone un gène du pathogène et on l’exprime
avec ses composants purifiés en cellules (bactéries, levures, cellules
d’insectes) et on purifie la protéine

Subunit
Subunit vaccines
vaccines

L’antigène est généralement une protéine de capside du virus, d’enveloppe, une toxine
• Break virus
bactérienne…. into components, immunize with purified components
(toxoïde)
Break virus into components, immunize with purified compone
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3. Immunisation active
3.3. vaccins sous unitaire – protéine virale

Vaccin contre réactivation de VZV (Varicella Zoster Virus) causant le zona

(Shingrix – GSK)
Protéine de surface de VZV (glycoprotéine E) + Adjuvant

Vaccin maladie de Lyme chez le chien

Outer Surface Protein A (OspA) de Borrelia burgdorferi + Adjuvant
(hydroxyde d’aluminium)

Candidat vaccin COVID (GSK Sanofi Pasteur)

Proteine de surface du SARS-CoV-2 (protéine spike produite en cellule
eucaryote) + Adjuvant -> immunogénicité trop faible dans les premiers
essais de phase I/II
29
3. Immunisation active
3.4. Virus Like Particules (VLPs)

30
3. Immunisation active
3.4. Virus Like Particules (VLPs)
Human PapillomaViruses (HPVs)
• dsDNA, groupe I, non-enveloppé, 8kb
• Plusieurs dizaines de génotypes différents
• Certains infectent la peau et sont
transmis par contacts cutanés, peuvent
induire des verrues
• Certains sont sexuellement
transmissibles et induisent des tumeurs
du col de l’uterus, cou, gorge
(notamment HPV16 et HPV18, leur
prévalence est très elevée)

31
 3. Immunisation active
3.4. Virus Like Particules
papillomavirus vaccines (VLPs)

Cancer
Cervarix vaccines
(GSK): HPV 16 et 18, produit en
cellules d’insecte

k): types9 (Merck):

Gardasil 6, 11, 16,HPV
18 16
produced in31,
et 18, 6, 11, S.33,
45, 52 et 58 produit en levure
erck): types 6, 11, 16, 18, 31, 33, 45, 52,
Administration avant les premiers rapports
sexuels
oSmithKline): types 16, 18 produced in

en before becoming sexually active

32
bia University
3. Immunisation active
3.4. Virus Like Particules (VLPs)

RTS,S/AS01, GSK Mosquirix

• 1er vaccin anti-malaria
• Fusion Circumsporozoïd protein – HBsAg
• Efficacité de ~30% (faible)
• Réduit le nombre d’épisodes paludiques

• Doit être adjuvanté

• 3-deacyl monophosphoryl lipid A (dérivé
du LPS)
• une saponine
-> induction locale et transitoire d’une
réponse immune innée

33
3. Immunisation active
3.3/3.4 vaccins sous unitaire - VLPs

Novavax COVID vaccine =

« Vaccin à nanoparticules
recombinantes »
(à mi-chemin entre vaccin sous-unitaire
VLPs)

Protéines de Spike produites en cellule

d’insecte puis inserées dans une
nanoparticule lipidique de 40nm de
diamètre et adjuvantées par la
saponine (une douzaine de protéines
de spike par nanoparticule)

34
3. Immunisation active
3.3/3.4 vaccins sous unitaire et VLPs

Avantages
• Sûr
• (Généralement stable)

Désavantages
• Moins efficace (ne se réplique pas)
• Doivent être injectés
• Relativement couteux (recombinants)
• Nécessite plusieurs rappels et adjuvants

35
3. Immunisation active
3.5. Adjuvants
Substance qui stimule la réponse immune innée de manière transitoire et locale, permettant
ainsi l’organisation d’une réponse immune spécifique contre les Ag accompagnant cet adjuvant

1923, Ramon, ajout du pus à la toxoïde diphtérique

1926, Glenny, sels d’aluminium
1927, adjuvant de Freund = émulsion contenant des fragments de bactéries

36
3. Immunisation active
3.5. Adjuvants
Substance qui stimule la réponse immune innée de manière transitoire et locale, permettant
ainsi l’organisation d’une réponse immune spécifique contre les Ag accompagnant cet adjuvant

Sels d’aluminium
-> inflammation locale

Squalène (émulsion huile dans eau)

-> recrutement de cellules innées, facilite l’absorption des Ag par les APC

3-deacyl monophosphoryl lipid A (dérivé du LPS)

-> TLR4

Virosome (bicouche phospholipidique dans laquelle sont insérés les Ag)

-> facilite l’absorption des Ag par les APC
37
3. Immunisation active
3.6. Virus chimérique

Virus chimérique Hippogriff

est un virus créé par ingénierie génétique

(recombinant) dont le génome est composé
d’informations provenant de plusieurs virus
3. Immunisation active
3.6. Virus chimérique
Dengue
• Groupe IV, ssRNA(+), Famille: Flaviviridae, Genre: Flavivirus, DENV1, 2, 3, 4, 5
• Fièvre bénigne, 100 millions de cas/an
• Fièvre hémorragique, 500.000/an, dont 25.000 morts
• Pour être immunisé contre la dengue, il faut avoir contracté les 5 sérotypes différents. Chaque
fois, les symptômes sont plus importants
3. Immunisation active
3.6. Virus chimérique
Dengvaxia, vaccin vivant atténué recombinant (chimérique), réplicatif
• Le 1er vaccin contre la dengue, tétravalent
• Backbone = Yellow Fever 17D, virus vivant souche atténuée, vaccin contre la fièvre jaune, très
bon safety records, Famille: Flaviviridae, Genre: Flavivirus
• Transgènes E et prM des virus de la dengue 1, 2, 3 et 4
3. Immunisation active
3.7. Vecteur

Cheval de Troie

41
3. Immunisation active
3.7.1. Vecteur viral

Vecteur viral (cheval de Troie)

est un virus réplicatif (qui peut répliquer son génome et produire de nouvelles particules virales
infectieuses) ou non-réplicatif

créé par ingénierie génétique (recombinant) dont le génome est composé d’information
provenant de plusieurs virus (organismes). C’est aussi une chimère.

Le vecteur a pour but d’infecter des cellules et d’obliger ces cellules à transcrire et traduire le
transgène (l’information génétique qu’il y a dans le ventre du cheval de Troie).

42
3. Immunisation active 1
2 Clathrin-

3.7.1. Vecteur viral coated

vesicle
Cytoplasm
3
Early
endosome
Nucleus
5 Protein VII
4
TP
Immediate-
early 6
7
5’ c
5’ c
8
9
E1A 10a 10b

5’ c
11 VA RNA-1
5’ c 5’ c 14
5’ c
12
Early
Pol 13
DBP
Pre-TP Pre-TP 14

15
IVa2; L4 33
and 22 kDa
16 IVa2; L4 33 and 22 kDa
E1B,
17 L1 52/55 kDa
5’ c E4 5’ c
5’ c 5’ c
Late 18 20
VA RNA-1,
L4 100 kDa
19
L3 protease
21
Structural proteins
L4 100 kDa

ADP

22

43
ted grown in cell culture
3. Immunisation active
3.7.1. Vecteur viral
AstraZeneca-Oxford ChAdOx1 nCoV-19 (AZD1222), vecteur adénoviral simien non-réplicatif

• Vaccin contre le SARS-CoV-2

• Backbone = adenovirus de chimpanzé (ChAdY25-E) modifié génétiquement

• par la délétion de l’oncogène E1 (empêche la réplication du virus)
• par
g VLPs la délétion
carry dubut
no genome gène E3h(empêche l’évasion de lavector
Replication-incompetent réponse immune)
display the spike protein on vaccines cannot propagate in the
their surface cells of the
• Transgènes = gène de la protéine Spike duvaccinated
SARS-CoV-2individual but
sous promoteur CMV
express the spike protein within them

Spike
gene

44
 3. Immunisation active
3.7.1. Vecteur viral
f Recombinant
RBD-based
vaccines
protein
vector vaccines carry k ProtéineldeRNA
ke protein on their surface
emically inactivated
g VLPs carry no genome but
display the spike protein on
their surface Cheval =
Adenovirus (de express
Ulysse =
DNA vaccines consist of plasmid
DNA encoding the spike gene under
a mammalian promoter
h Replication-incompetent vector
vaccines cannot propagate in the
cells of the vaccinated individual but
Chimpanzé)thegénétiquement
spike proteinmodifié
within them
Spike
gene
Spikevaccines consist of RNA encoding
the spike protein and are typically
packaged in LNPs
45
3. Immunisation active
3.7.1. Vecteur viral
Autres vaccins SARS-CoV-2 basés sur la même
approche

• Johnson & Johnson / Janssen COVID

vaccine: 1 seule dose avec un adénovirus
recombinant de sérotype D26

• Sputnik V: prime (1ère dose) avec un

adénovirus recombinant de sérotype D26 et
boost (2nd dose) avec un adénovirus
recombinant sérotype C5

46
3. Immunisation active
3.7.1. Vecteur viral

VOL. 77, 2003 Ad35-BASED VECTORS BYPASS PREEXISTING IMMUNITY 8265

47
DOI: 10.1128/jvi.77.15.8263-8271.2003
3. Immunisation active
3.7.1. Vecteur viral
Les adénovirus ne sont pas les seuls à pouvoir servir de vecteur
Les poxvirus sont aussi notamment utilisés (/ex vaccin contre Ebola, voir cours vecteur)

Virus Advantages Disadvantages

Retrovirus Long-term gene expression
Long-term gene expression, Infects non-
Lentivirus
dividing and dividing cells
High immunogenicity Safety: used as a
Vaccinia virus (Poxvirus)
smallpox vaccine, High titer production
High immunogenicity Safety: used in
Adenovirus
many clinic trails, High titer production
Long-term gene expression, Non-
Adeno-associated virus
pathogenic virus
Induces a unique CTL response, Protects
Cytomegalovirus
against SIV infection in an animal model
Sendai virus High immunogenicity
Generation of replication-competent
virus, Potential for tumorigenesis, Infects
dividing cells only
Generation of replication-competent
virus, Potential for tumorigenesis
Pre-existing immunity
Pre-existing immunity
Low titer production
Pre-existing immunity, Risk of
pathogenesis in specific individuals
48
Pre-existing immunity
3. Immunisation active
3.7.1. Vecteur viral

Avantages
• Réponse humorale (Ac neutralisants) mais aussi réponse cellulaire (CTL)
• C’est une plateforme de vaccination, il suffit de changer le transgène
• Production aisée (titre élevé de virus)
• Vaccin assez stable (4°C)

Inconvénients
• Immunité contre le vecteur

49
3. Immunisation active
3.7.2. DNA vaccine

Très peu utilisé, seulement quelques applications vétérinaires

50
3. Immunisation active
3.7.2. DNA vaccine

West Nile virus, genre: Flavivirus, ssRNA(+)

chez l’homme et chevaux provoque la fièvre du Nil occidental (fièvre, méningite)
Transmis par piqûre d’insecte
Réservoir aviaire

West Nile-Innovator DNA (Fort Dodge)

Injection intramusculaire d’ADN plasmidique codant pour prM/E du WNV
Arrêt de la commercialisation en 2009

Avantage: stabilité de l’ADN permet une conservation facile du vaccin

Mais jusqu’ici succès très mitigé car
• faible immunogénicité (dégradation du plasmide par effecteurs innés?)
• risque d’insertion de l’ADN dans le génome de l’hôte pour applications chez l’Homme

Chez l’Homme, ces vaccins sont déjà dépassés par les vaccins à ARNm 51
 Unwanted Avoid Phosphatase Generate 5'OH RNA
3. Immunisation
product active
PRR
3.7.3. mRNA HPLC STAR
vaccine – La nouvelle
recognition purification Remove dsRNA

ARNm entourée d’une lipid nanoparticle (LNP)

Pure Cap-1
mRNA
(D) mRNA - lipid nanoparticle complex

PEG -phospholipids

Cholesterol
Pure Cap-1
mRNA
Phospholipids

Ionizable lipids
52
3. Immunisation active and the latest mRNA optimization strategies.

3.7.3. mRNA vaccine – La nouvelle STAR

mRNA Capping
The 50 cap of mRNAs is characteristic for eukaryotic mRNA and consists of the addition of N7-m
ylated guanosine to the first nucleotide of these molecules [27]. Besides its functions in pre-mR

ARNm contenant des nucléosides modifiés splicing, polyadenylation, nuclear export, and protection against exonucleases, other key asp

• Coiffe, 5’UTR, ORF, 3’UTR, queue poly A 5'Cap 5'UTR ORF 3'UTR Poly-A
• Dont certains U sont des Pseudo-uridine
pour éviter une dégradation du transcrit
par immunité innée Biological relevance

Ψ ou pseudoU est le nucléotide modifié le

Cap 5' UTR ORF 3'UTR Poly A
-Eukaryotic modification -Recognition by -Sequence encoding -Important for translation -Important for mRNA
-Important for translation translation machinery. the gene of interest initiation and mRNA stability.

plus abondant dans l’ARN, modification initiation, mRNA stability,

nuclear export.
-When suboptimal,
-Recognized and
scanned by ribosomes.
-Important for mRNA
(GOI), in mRNA vaccine stability.
encodes the antigen.
-Recognition by Poly-A
binding proteins (PABP)
and recruitment of

post traductionnelle recognized as PAMPs

by the innate immunity.
-RNA Closed-Loop.
translation and stability.
-RNA Closed-Loop.
translation factors.
-Important for translation
initiation
(RNA Closed-Loop).

-Natural Cap-1 to avoid
PRR recognition and
enhance translation.
-Enzymatic capping for
higher capping efficiency.
Region optimization
-Inclusion of Kozak -Codon optimization -Optimal sequences -Poly-A sequences of 120
sequence. increases translation. derived from highly units.
-No strong secondary -Low optimal codons may stable mRNA ( -Globin). -Adding a poly-U
structures. be important for -2x copies in tandem sequence, providing a
-No other start codon. adequate folding. -RNA Closed-Loop dsRNA in the poly-A,
-Polysome profiling to increases adjuvant effect.
count the ribosome
loading in sequences
in-silico.

Whole molecule optimization

-Avoiding binding sites of miRNAs present in the target cells.
-Uridine depletion to avoid recognition by innate immunity. 2+
53 dsRNA levels.
-Production at high temperature (50ºC) using a thermostable polymerase and/or low Mg concentration, to decrease

Uridine Ψ ou pseudoU -mRNA purification using HPLC to decrease dsRNA amount.

-Avoidance of highly stable and long secondary structures that could activate PRRs.
 Unwanted Avoid Phosphatase Generate 5'OH RNA
3. Immunisation
product active
PRR
3.7.3. mRNA vaccine
recognition HPLC purification Remove dsRNA

Fonction de la LNP
LNP est endocytosée, protège l’ARNm dans Pure Cap-1
mRNA
l’endosome, évite la reconnaissance par TLRs et
(D) mRNA
permet - lipid
la libération nanoparticle
de l’ARNm complex
dans cytoplasme

Composition de la LNP
PEG -phospholipids
• Double couche de phospholipides
• Cholestérol qui stabilise la double couche
• Lipides cationiques complexent les molécules
Cholesterol
d’ARNm chargées négativement et permettent
Pure Cap-1
l’assemblage autonome en particules
mRNA de la
Phospholipids
taille d’un virus (~100 nm)
• Polyéthylène glycol (PEG) lié à un lipide
Ionizable
augmente lipids
la durée de vie de vaccin dans sa
fiole 54
3. Immunisation active
3.7.3. mRNA vaccine

Pfizer-BioNTech BNT162b2
ARNm encodant la protéine S stabilisée
dans sa conformation de préfusion par
deux substitutions (S-2P)

Moderna mRNA-1273

CureVac, échec du premier candidat

probablement car ARN non modifié (pas de
pseudo uridine)

55
3. Immunisation active
3.7.3. mRNA vaccine
Futures évolutions des vaccins à ARNm
Plusieurs ARNm différents peuvent être utilisés, codant pour plusieurs protéines virales
Par exemple pour la création d’un vaccin « universel » contre Influenza
Figure 1. mR
Utilized for th
Influenza Vir
(A) Schematic
vaccine techn
idine-modified
vesicle for effi
(B) Diagrams i
this study. A
mRNA coding

cine, conser
man influen
sequences f
selected for
1918, to cov
subtype, th
while the h
ability (Fig
ports.30,31 T
conservatio
56 tion with a
https://doi.org/10.1016/j.ymthe.2020.04.018. cross-reacti
3. Immunisation active production in the cytoplasm
and the consequent formation
3.7. Fonctionnement
of foreign des vaccins
antigens as proteinsvecteur viraux / vaccin ADN / vaccin ARNm
Exogenous
or as peptide strings. The cell MHC II antigens
and humoral immunity by DNA vaccines
provides endogenous MHC I
Vaccine formulation post-translational
and adjuvants modifications to antigens that
Optimized gene Muscle site
of interest reproduce native protein
of inoculation
conformations and the cell 5
customizes the antigens in a similar Afferent
Next-generation lymphatic
delivery methods manner to the pathways induced by live
vessel
infection with recombinant vectors. 6
These host-synthesized antigens then Draining lymph node
can become the subject of immune Direct and cross presentation
1 surveillance2 in the context of both major of antigens in the draining
Direct transfection Direct transfection
histocompatibility complex class I (MHC I) lymph node
of myocytes of APC
and MHC II proteins of the vaccinated
CD8 T
individual. APCs have a dominant role in TCR
MHC I the induction of immunityMHC of DNA
I
Myocyte APC
vaccines by presenting vaccine-derived
endogenous peptides onEndogenous MHC I Antigen
CD4 8
4 3
molecules. This can follow either direct
antigens CD4 T
Apoptotic or
Efferent
Shed exogenous
antigens
transfection by the plasmid vaccine (2) or
necrotic bodies BCR lymphatic
cross-presentation of cell-associated TCR vessel
exogenous antigens; for example, owing to B cell
APC engulfment of apoptotic transfected cells MHC II
Exogenous 7
MHC II (3). In addition, antigens
APCs mediate the display of
MHC I
peptides on MHC II molecules after secreted
Activated
protein antigens that have been shed from lymphocytes
57
transfected cells are captured and processed within the
3. Immunisation active
3.7.3. mRNA vaccine

Avantages
• Réponse humorale (Ac neutralisants) mais aussi bonne réponse cellulaire (CTL)
• Production de l’ARNm par transcription in vitro (sans cellule, sans œufs embryonnés), faible
cout de production et débit élevé (/ex vaccins anti-COVID Pfizer)
• C’est une plateforme de vaccination, il suffit de changer l’ORF

Inconvénients
• Stockage du vaccin à basse T (-80°C, -20°C) car ARN est fragile
• L’antigène doit être une protéine
• Prix relativement élevé

Inconnues
• Le tropisme cellulaire des LNPs est large et peu défini (est-ce un inconvénient?)
• Nécessité d’avoir un transgène qui code pour une protéine qui sera secrétée et/ou
cytotoxique (?) 58
3. Immunisation active
3.8. Replicon vaccines – SAM (self amplifying mRNA)
Groupe IV: ssRNA(+), famille: Togaviridae, genre: Alphavirus, linéaire, non-segmenté, 12kb,
enveloppé
e.g. Semliki forest virus, transmission par piqûre insecte, généralement asymptomatique sauf si
individu immunodéprimé (encéphalite)

59
3. Immunisation active
3.8. Replicon vaccines – SAM (self amplifying mRNA)
Groupe IV: ssRNA(+)
genre: Alphavirus
Production d’un ARN
subgénomique codant pour les
protéines de structure

60
3. Immunisation active
3.8. Replicon vaccines – SAM
Groupe IV: ssRNA(+)
genre: Alphavirus
Production d’un ARN
subgénomique codant pour les
protéines de structure (étape 5)

61
3. Immunisation active
3.8. Replicon vaccines - SAM
Les gènes de structures sont remplacés par un gène codant pour une ou des protéines du virus
contre lequel on veut vacciner

62
 Cytotoxic
3. Immunisation active Cytotoxic
T cell b T cell
3.8. Replicon vaccines - SAM
response response
Replicon
Le vaccin est administré sous forme
d’ARN POI une LNP
(ou d’ADN) dans POI
In vivo In vivo

L’ARN génomiquePOI est répliqué/amplifié POI

dans la cellule POI POI
• permettantPOI la production d’une grande
EUS
quantité de protéine d’intérêt (POI) qui
RNA
élicitera une GOI
réponse adaptative
P26S
• induisant unePlease
réponse
mRNA innée par TLR3
cite this article in press as: Maruggi et al., mRNA as a Transformative Technology for Vaccine Development
senseur de dsRNA P26S
Therapy (2019), https://doi.org/10.1016/j.ymthe.2019.01.020
GOI
RNA
On CYTOPLASM
parle de SAM Self-Amplifying mRNA replication
dsRNA
www.moleculartherapy.org danger signal
P26S P26S
Review nsPs GOI nsPs GOI
T cell–
mediated RNA63
DNA immunity
3. Immunisation active
3.8. Replicon vaccines - SAM
Le vaccin est
administré sous
forme de VRP (viral
replicon particles)

L’ARNg du replicon
viral est empaqueté
par des protéines de
structures fournies en
trans (helper E1 et E2)

64
3. Immunisation active
3.8. Replicon vaccines - SAM

Highly pathogenic swine influenza virus strain H3N2

VRP basées sur l’alphavirus Venezuelan Equine Encephalitis avec comme GOI l’Hemagglutinine
(SirraVax technology, Harrisvaccines)

VEE backbone où les gènes de

structures ont été remplacés par
l’HA d’influenza

Replicon vaccines: technologie prometteuse et facilement déclinable contre de nombreux

virus/pathogènes, il « suffit » de changer le GOI (-> plateforme de vaccination) 65
3. Immunisation active
3.9. Planning de vaccination en Belgique
Seule la vaccination contre la poliomyélite est légalement obligatoire en Belgique depuis 1967
D’autres vaccinations sont fortement recommandées
CALENDRIER DE VACCINATION
2019-2020 Nourrissons
8 sem. 12 sem. 16 sem.
(2 mois) (3 mois) (4 mois)
12
mois
15
mois
1
Enfants et adolescents
5-6
ans
11-12
ans
13-14
ans
Adultes
15-16 Femmes Tous les
ans enceintes 10 ans
65
ans
Poliomyélite Ce calendrier est
Diphtérie susceptible d’être

Hexavalent
modifié au fil des ans.
Tétanos Votre médecin pourra
Coqueluche éventuellement
l’adapter à votre
Haemophilus influenza de type b enfant, n’hésitez pas
Hépatite B à discuter vaccination
avec lui.
Rougeole

RRO
Rubéole
Oreillons
Méningocoque C
1
Pneumocoque ( )
2
Rotavirus (vaccin oral) ( )
3
Papillomavirus (HPV) Pour plus d’infos :
Consultez votre médecin
Grippe (Influenza) traitant ou le site
www.vaccination-info.be
Recommandé à tous et gratuit Femmes enceintes : Vaccination à partir de 24 semaines et idéalement
avant 32 semaines de grossesse.
Recommandé à tous
Hexavalent : Vaccin qui confère une protection contre 6 maladies
Vaccin combiné (une seule injection)
1 Pour les bébés nés avant 37 semaines de grossesse ou 2 En 2 ou 3 doses en fonction du vaccin administré.
ayant un poids de naissance inférieur à 2.5 kg, une dose 3 Vaccination en 2 doses à 6 mois d’intervalle
supplémentaire de vaccin contre le pneumocoque est (minimum 5 mois d’écart).
recommandée à 3 mois et les vaccins prévus à 15 mois
seront administrés à 13 mois (hexavalent et méningo 66
-coque C).
3. Immunisation active
3.10. Etapes de developpement d’un vaccin

Traditional development
Evidence de
réponse immune
qq dizaines
individus
Clinical trials
(5–7 years total)
Design and Process development IND BLA Regulatory Large-scale
exploratory preclinical, submitted submitted review by FDA, production
preclinical studies toxicology studies Phase I Phase II Phase III EMA etc. and distribution
(years) (2–4 years) (1–2 years) (2 years) (2–3 years) (1–2 years)
15 years
or longer

SARS-CoV-2 vaccine development Toxicité à Efficacité et

court termeClinical trials
innocuité
qq dizaines (months) qq dizaines de
individus milliers d’individus
Phase III

Phase I
Design and Process development IND BLA Regulatory 67
Après la mise sur
exploratory le marché
preclinical, et la vaccination
submitted Phase II
en masse, vientreview
submitted la phase
by FDA, IV de pharmacovigilance
 (5–7 years total)
3.Design
Immunisation
and active
Process development IND BLA Regulatory
submitted submitted
exploratory
3.10.
preclinical studies
Etapes de developpement
preclinical,
toxicology studies
d’un
Phase vaccin
I Phase II Phase III
review by FDA,
EMA etc. a
(years) (2–4 years) (1–2 years) (2 years) (2–3 years) (1–2 years)

SARS-CoV-2 vaccine development

Clinical trials
(months)

Phase III

Phase I
Design and Process development IND BLA Regulatory
exploratory preclinical, submitted Phase II submitted review by FDA,
preclinical studies toxicology studies EMA etc.
(months) (months) (1–2 months)
10 months to
Pre-existing from Partially pre-existing Overlapping Production Review on
1.5 years total
SARS-CoV and parallel clinical (at risk) a rolling
and MERS-CoV development phases basis?

Fig. 1 | Traditional and accelerated vaccine-development pipelines. vaccine is licensed. After that point, large-scale prod
Traditional vaccine development can take 15 years or more, starting with a development for SARS-CoV-2 is following an accelera
lengthy discovery phase in which vaccines are designed and exploratory knowledge gained from the initial development of va
preclinical experiments are conducted. This is usually followed by a phase in MERS-CoV, the discovery phase was omitted. Existin
68
Aprèsmore
which la mise
formal sur le marché
preclinical et laand
experiments vaccination en masse,
toxicology studies are vientand
la phase
phase I/IIIV dewere
trials pharmacovigilance
started. Phase III trials were
3. Immunisation active a ChAdOx1 nCoV-19 prime
ChAdOx1 nCoV-19 prime–boost
b
3.10. Etapes de developpement d’un vaccin ChAdOx1 GFP
*
** ** * **
Essais préclinique vaccin COVID AstraZeneca 15

(AZD1222)

Clinical score
10

Moins de virus dans Même charge virale

5
poumons dans le nasopharynx
(BAL=Broncho Alveolar 0
• Challenge 1 mois après la vaccination 1 2 3 4 5 6 7
Lavage) second
Time dose
(days after of ChAdOx1
inoculation)
• Pas de pneumonie chez
a singes vaccinés
ChAdOx1 nCoV-19 prime
ChAdOx1 nCoV-19 prime–boost
b
ChAdOx1 GFP c ously published study, i
* **
** ** * ** * 8 with malaria vaccine 8 ca

sgRNA load in nose swabs

** *

gRNA load in nose swabs

• N’induit pas d’immunité15stérilisante 8 8

sgRNA load in BAL fluid

gRNA load in BAL fluid
decrease in spike- and v

(log10[copies per ml])

(log10[copies per ml])

(log10[copies per ml])

(log10[copies per ml])

*
(charge virale dans le nez10 identique entre 6 between 14 days befor 6
Clinical score

6 6
prime–boost group. Lon
animaux vaccinés et ceux ayant reçu le 4 4 4 investigate whether4 this d
5
placebo) Two recently publishe
2 2 2 2
0
3 5 3 5 0
similar results: the first
1 2 3 4 5 6 7 1 3 5 7 0
Time (days after inoculation) Time (days after inoculation) Time (dayshigh dose of
after inoculation)
whole ina
Tim
sus macaques from SAR
c d Fig.
** 3 | Clinical signs andvaccination
viral load in rhe
two-dose re
8 8 a ChAdOx1 nCoV-19
10 **** prime
SARS-CoV-2 after vaccination b with ChA
nose swabs
nose swabs

spike protein, which sig

ung tissues
ChAdOx1 nCoV-19 ****prime–boost

s per ml])
s per ml])

s per ml])

ChAdOx18 GFP NHPs.**** Boxes show the 25th to 75th percen

fluid and nasal swabs10. T
6 6 *
6 whiskers range from the 5th to 95th perce **
** ** locations
* ** using different
3. Immunisation active
3.10. Etapes de developpement d’un vaccin
Primary efficacy analysis:
Essai clinique de Phase III (AZD1222) SD/SD or LD/SD vaccination
• Efficacité du vaccin = 70% (sur une période
0·06 Treatment group
de 2 mois après la seconde injection)
ChAdOx1 nCoV-19

Cumulative proportion of participants

• 30 COVID19 / 5807 dans le groupe 0·05 MenACWY
vacciné
• 101 COVID19 / 5829 dans le groupe 0·04
contrôle
• Pas d’effets secondaires graves à court terme 0.03

Exclusion period
0·02
• Ne semble pas réduire la proportion de
porteurs asymptomatiques 0·01
• 29 asymptomatiques / 3288 dans le
groupe vacciné 0
• 40 asymptomatiques / 3350 dans le 0 20 40 60 80 100
groupe contrôle Days since second dose
Number at risk
(number censored) https://doi.org/10.1016/S0140-6736(20)32661-1
4. Immunité collective/de troupeau - Herd immunity
Phénomène par lequel la propagation d'une maladie contagieuse peut être enrayée si un certain
pourcentage des individus est immunisé

71
4. Immunité collective/de troupeau - Herd immunity

72
5. Nombre de reproduction et couverture vaccinale

Nombre de reproduction de base R0 (R naught / R nought)

• nombre moyen de personnes infectées par une personne contagieuse
• dans des conditions où la population est entièrement susceptible (pas d’immunité préalable)
et sans mesure de mitigation (gestes barrières, distanciation physique)

• R0 Influenza ≈ 1 à 2
• R0 SARS-CoV-2 Wuhan ≈ 2 à 2,5, R0 Delta ≈ 7, R0 Omicron ≈ 10
• R0 Measles (rougeole) ≈ 12 à 18

73
5. Nombre de reproduction et couverture vaccinale

Nombre de reproduction à un instant t (dans une population précise) Rt est fonction de

• R0
• diminue avec la proportion de personnes immunisées
• diminue avec les gestes barrières et (surtout) avec la distanciation physique

74
5. Nombre de reproduction et couverture vaccinale

En l’absence de gestes barrières et de distanciation physique

Rt = R0 x S où S est la proportion de la population qui est susceptible

Au début d’une épidémie Rt = R0 x 1 car tous les individus sont susceptibles

75
5. Nombre de reproduction et couverture vaccinale

Pour endiguer l’épidémie on doit avoir

Rt < 1
R0 x S < 1
S < 1/R0
1-S > 1-1/R0
NS > 1-1/R0 où NS est la proportion de personnes Non Susceptibles
V > (1-1/R0)/E où V est la proportion de personnes vaccinées et E l’efficacité du vaccin

Attention, ce calcul est une simplification de la réalité, il ne tient pas compte

• de la durée de l’immunité (efficacité du vaccin diminue avec le temps car le système
immunitaire oublie progressivement)
• de l’évolution génétique du virus (le virus peut évader la réponse induite par le vaccin)
• et assume une définition particulière de l’efficacité: E est la proportion de personnes
vaccinées qui ne développent pas la maladie ET qui ne sont pas contagieux (un vaccin peut
protéger contre les signes cliniques mais permettre la réplication du virus et donc sa
transmission) 76
5. Nombre de reproduction et couverture vaccinale

77
5. Nombre de reproduction et couverture vaccinale
Pour obtenir interrompre la transmission du SARS-CoV-2 par immunité collective
V > (1-1/R0)/E

Si R0 = 2,5 et E=100% alors V=60%

Si R0 = 2,5 et E=90% alors V=66%
Si R0 = 2,5 et E=50% alors V=120% (impossible)

Si R0 = 7 et E=100% alors V=86%

Si R0 = 7 et E=90% alors V=95%
Si R0 = 7 et E=50% alors V=171% (impossible)

Si R0 = 10 et E=100% alors V=90%

Si R0 = 10 et E=90% alors V=100% (impossible)
Si R0 = 10 et E=50% alors V=180% (impossible)

Attention V est une moyenne, il peut toujours y avoir des sous-populations où V est faible/nul
(antivax ou personnes qui ne peuvent pas recevoir le vaccin) 78
6. Abattage de masse
Pour arreter une épidémie chez animaux d’élevage
• Vaccination
• Abattage de masse (si vaccination pas disponible/efficace/rentable economiquement, ou si
cela pose des problèmes à l’export)

79
6. Abattage de masse
Epidemie de fièvre aphteuse (foot and mouth disease) au Royaume Uni en 2001
• Vaccin disponible mais non-utilisé
• Abattage de tous les animaux (infectés ou pas) dans une zone entourant le foyer
• Décontamination des fermes
• 6 millions d’animaux abattus pour 2000 cas confirmés (9 mois, 8 milliard de livres)

80
7. Les anti-vax

81
7. Les anti-vax

Doctrine: la vaccination (tous les vaccins) est

inutile voire nuisible (responsable de la
sclérose en plaques, l'autisme…)

Motivations:
• “Ne pas être un mouton”, en savoir plus que
les moutons, devenir un “lanceur d’alerte”,
un “leader d’opinion”
• Financières (parfois)

82
7. Les anti-vax

83
7. Les anti-vax

84
7. Les anti-vaccins – technique n°1: le dénigrement

85
86
7. Les anti-vaccins – technique n°2: la pseudo-science

87
7. Les anti-vaccins – technique n°2: la pseudo-science

88
7. Les anti-vaccins

89
7. Les anti-vaccins

90
7. Les anti-vaccins

91
7. Les anti-vaccins – technique n°3: des messages simples - simplistes

92
7. Les anti-vaccins

93
7. Les anti-vaccins – technique n°4: « la science »
“Les vaccins causent l’autisme”, “le vaccin ROR (Rougeole-Oreillons-Rubeole, ou MMR for
Measles-Mumps-Rubella) cause l’autisme”

MMR est un vaccin composé de 3 virus vivants atténués (atténuation par passages successifs en
cellules humaines et animales)

Andrew Wakefield, médecin gastro-entérologue, étudie un possible lien entre le vaccin MMR et
maladies intestinales en faisant comme hypothèse qu'une infection persistante par le virus
vaccinal causerait une perturbation de la muqueuse intestinale, ce qui conduirait à une maladie
intestinale et à une maladie neuropsychiatrique (en particulier l'autisme).
EARLY REPORT

Wakefield et al. Lancet 1998 Early report

12 enfants présentant des troubles autistiques Ileal-lymphoid-nodular hyperplasia, non-specific colitis, and
Développement des troubles autistiques chez 8 pervasive developmental disorder in children

enfants /12 après la vaccination ROR

A J Wakefield, S H Murch, A Anthony, J Linnell, D M Casson, M Malik, M Berelowitz, A P Dhillon, M A Thomson,
P Harvey, A Valentine, S E Davies, J A Walker-Smith

Summary
Background We investigated a consecutive series of
children with chronic enterocolitis and regressive
Introduction
We saw several children who, after a94 period of apparent
normality, lost acquired skills, including communication.
They all had gastrointestinal symptoms, including
7. Les anti-vaccins – technique n°4: « la science »

Le papier est un “case report”, très peu de cas

sans groupe controle

95

7. Les anti-vaccins – technique n°4: « la science »
Le papier est un “case report”, très peu de cas
sans groupe controle
S’en suivront de nombreuses publications
étudiant le lien entre ROR et autisme basées
sur des données épidémiologiques larges –
sans résultat
Measles, mumps, and rubella vaccination and bowel
problems or developmental regression in children with
autism: population study
Brent Taylor, Elizabeth Miller, Raghu Lingam, Nick Andrews, Andrea Simmons, Julia Stowe
Abstract
Objectives To investigate whether measles, mumps,
and rubella (MMR) vaccination is associated with
bowel problems and developmental regression in
children with autism, looking for evidence of a “new
variant” form of autism.
Design Population study with case note review linked
to independently recorded vaccine data.
Setting Five health districts in north east London.
Participants 278 children with core autism and 195
with atypical autism, mainly identified from
computerised disability registers and born between
1979 and 1998.
Main outcome measures Recorded bowel problems
lasting at least three months, age of reported
regression of the child’s development where it was a
feature, and relation of these to MMR vaccination.
Results The proportion of children with
developmental regression (25% overall) or bowel
symptoms (17%) did not change significantly (P value
for trend 0.50 and 0.47, respectively) during the 20
years from 1979, a period which included the
introduction of MMR vaccination in October 1988.
No significant difference was found in rates of bowel
problems or regression in children who received the
MMR vaccine before their parents became concerned
about their development (where MMR might have
caused or triggered the autism with regression or
bowel problem), compared with those who received it
only after such concern and those who had not
received the MMR vaccine. A possible association
between non-specific bowel problems and regression
in children with autism was seen but this was
unrelated to MMR vaccination.
Conclusions These findings provide no support for
an MMR associated “new variant” form of autism with
developmental regression and bowel problems, and
further evidence against involvement of MMR vaccine
in the initiation of autism.
Introduction
The suggestion that the measles, mumps, and rubella
(MMR) vaccine causes or triggers autism centres on the
putative existence of “new variant” autism where devel-
opmental regression is reported to follow shortly after
MMR vaccination, typically accompanied by bowel
symptoms.1 Epidemiological studies designed to inves-
tigate such a causal association have found no
correspondence between temporal trends in MMR
uptake and the incidence of autism, nor any evidence
of clustering of onset of behavioural disturbance,
including regression, shortly after vaccination.2–6 How-
ever, it is now postulated that the onset of MMR
induced “regressive autism” with “autistic enterocolitis”
may occur after a prolonged induction interval and, in
BMJ VOLUME 324 16 FEBRUARY 2002 bmj.com
Papers
BMJ: first published as 10.1136/bmj.324.7334.393 on 16 February 2002. Downloaded from http://www.bmj.com/ on 13 September 2019 by guest. Protected by copyright.
Centre for
Community Child
Definitions Health, Royal Free
and University
Childhood autism College Medical
Children with symptoms before the age of 3 years that School, University
meet the necessary criteria* under each section of the College London
Royal Free Campus,
diagnostic triad for autism: communication difficulties,
London NW3 2PF
problems with social interaction, and behaviour
problems such as stereotyped repetitions
Brent Taylor
professor of
Atypical autism community child
health
Cases with many of the features of childhood autism
Raghu Lingam
but not quite meeting the required criteria for that research fellow
diagnosis, or with atypical features such as onset of
Andrea Simmons
symptoms after age 3 years (also known as pervasive research fellow
developmental disorder not otherwise specified) Julia Stowe
Developmental regression research associate
A documented deterioration in any aspect of Immunisation
development or reported loss of skills, however Division, Public
transient Health Laboratory
Service,
*ICD-10 (international classification of diseases, 10th
revision) and DSMIV (Diagnostic and Statistical Manual of Communicable
Mental Disorders, 4th ed) Disease
Surveillance Centre,
London NW9 5EQ
Elizabeth Miller
addition, requires the presence of cofactors such as an head
intercurrent infection, receipt of antibiotics, a history of Nick Andrews
statistician
atopy, a strong family history of autoimmune disease,
or MMR or rubella immunisation of the mother Correspondence to:
B Taylor
shortly before, during, or after pregnancy.7 This modi- b.taylor@rfc.
fied hypothesis could thereby explain the negative ucl.ac.uk
findings of the epidemiological studies. Whatever the
postulated induction interval or trends in incidence of BMJ 2002;324:393–6
autism, the present hypothesis requires that the
proportion of autistic children with regression and
bowel symptoms be higher in children given MMR
vaccine before parents became concerned about their
child’s development and that the pattern of bowel
problems and regression in autism should have
changed after MMR was introduced.
We have published epidemiological evidence that
showed no causal association between MMR vaccina-
tion and autism.2 3 We repeated that survey in late 2000
and early 2001 after an interval of two and a half years
in five of the original eight study districts to examine
further the relation between MMR vaccine and the
onset of autism, particularly autism associated with
regression, and to collect information on bowel
problems in children with autism.
Methods
We identified children with childhood and atypical
autism born between 1979 and 1998 from computer-
ised health registers of children with disabilities in the
community and from special school and child psychia-
try records, using the same methods and classifications
as in our earlier study.2 3 We abstracted information
from the clinical notes then linked the information to
independent computerised vaccination records (the 96
393
7. Les anti-vaccins – technique n°4: « la science »
Journal of Child Psychology and Psychiatry 46:6 (2005), pp 572–579 doi: 10.1111/j.1469-7610.2005.01425.x

No effect of MMR withdrawal on the incidence

of autism: a total population study

Le papier est un “case report”, très peu de cas 1

Hideo Honda,1 Yasuo Shimizu,1 and Michael Rutter2
Yokohama Rehabilitation Center, Yokohama, Japan; 2Institute of Psychiatry, London, UK

sans groupe controle Background: A causal relationship between the measles, mumps, and rubella (MMR) vaccine and
occurrence of autism spectrum disorders (ASD) has been claimed, based on an increase in ASD in the
USA and the UK after introduction of the MMR vaccine. However, the possibility that this increase is
coincidental has not been eliminated. The unique circumstances of a Japanese MMR vaccination pro-
gram provide an opportunity for comparison of ASD incidence before and after termination of the
program. Methods: This study examined cumulative incidence of ASD up to age seven for children
born from 1988 to 1996 in Kohoku Ward (population approximately 300,000), Yokohama, Japan. ASD

S’en suivront de nombreuses publications

cases included all cases of pervasive developmental disorders according to ICD-10 guide-
lines. Results: The MMR vaccination rate in the city of Yokohama declined significantly in the birth
cohorts of years 1988 through 1992, and not a single vaccination was administered in 1993 or there-

étudiant le lien entre ROR et autisme basées

after. In contrast, cumulative incidence of ASD up to age seven increased significantly in the birth
cohorts of years 1988 through 1996 and most notably rose dramatically beginning with the birth cohort
of 1993. Conclusions: The significance of this finding is that MMR vaccination is most unlikely to be a
main cause of ASD, that it cannot explain the rise over time in the incidence of ASD, and that withdrawal

sur des données épidémiologiques larges –

of MMR in countries where it is still being used cannot be expected to lead to a reduction in the incidence
of ASD. Keywords: MMR vaccine, autism, incidence, etiology, regression, total population
study. Abbreviations: ISCO-68: International Standard Classification of Occupations, 1968; DIS-
COVERY: Detection and Invention System in the COmmunity for VERy Young children with develop-

sans résultat
mental disorders; YACHT: Young Autism and other developmental disorders Checkup Tool.

Publication of a study claiming a causal relationship
between the measles, mumps, and rubella (MMR)
vaccine and autism spectrum disorders (ASD)
(Wakefield, 1999; Wakefield et al., 1998) sparked a
heated debate, primarily in the USA and UK, which
relates not only to the etiology of ASD, but also im-
pacts immunization policy. Among data used to ar-
gue the case are ASD frequency studies. Recent US
and British research examining the relationship of
trends in MMR vaccination rates and ASD frequency
have noted a dramatic increase in ASD frequency
most strikingly in the late 1980s (Croen, Grether,
Hoogstrate, & Selvin, 2002; Dales, Hammer, &
Smith, 2001; Department of Developmental Ser-
vices, 1999; Hillman, Kanafani, Takahashi, & Miles,
2000; Kaye, der Mar Melero-Montes, & Jick, 2001;
Lingam, Simmons, Andrews, Miller, & Stowe, 2003;
Powell et al., 2000; Taylor et al., 1999), although
evidence of a rise appeared before then (Gillberg,
Steffenburg, & Schaumann, 1991; Taylor et al.,
1999; Webb, Lobo, Hervas, Scourfield, & Fraser,
1997). However, because there was no stepwise in-
crease in the frequency of ASD after MMR was
introduced, and because the frequency of ASD con-
tinued to rise while the rate of MMR vaccination re-
mained stably high, there have been major doubts
about the claimed causal relationship between the
MMR vaccine and ASD occurrence.
Nevertheless, these epidemiological studies, al-
though not providing any support for the causal
hypothesis, are inconclusive because they do not
! Association for Child Psychology and Psychiatry, 2005.
Published by Blackwell Publishing, 9600 Garsington Road, Oxford OX4 2DQ, UK and 350 Main Street, Malden, MA 02148, USA
test what happens when the postulated risk factor,
namely MMR, ceases to be operative. When exam-
ining causal hypotheses regarding effects of a risk
factor on time trends in the frequency of a disorder, it
is highly desirable to study the effects of withdrawal
of the risk factor on frequency of disorder as well as
the effects on frequency following introduction of the
risk factor (Rutter & Smith, 1995). With respect to
MMR, the Japanese situation provides just such a
test.
In Japan, the Immunization Law had provided
measles vaccination of children aged 12 months to
72 months since 1978 and rubella vaccination of
junior high school female students since 1977; then
an MMR vaccination program was launched in April
1989. In contrast to the practice in many countries
where an MMR booster is administered a few years
following the initial vaccination, in Japan only one
vaccination was administered to children between
12 and 72 months of age, with the majority vaccin-
ated between 12 and 18 months of age. However,
due to a high frequency of reports of aseptic menin-
gitis, a suspected side effect of the mumps vaccine
(Urabe strain), the program was terminated in April
1993. Subsequently, only monovalent vaccines were
administered. Following a reform of the Immuniza-
tion Law in 1994, measles and rubella vaccinations
were each specified for children between the ages of
12 and 90 months (the measles vaccine was recom-
mended between 12 and 24 months of age and the
rubella vaccine between 12 and 36 months). The
97
7. Les anti-vaccins – technique n°4: « la science »
Le papier est un “case report”, très peu de cas
sans groupe controle
S’en suivront de nombreuses publications
étudiant le lien entre ROR et autisme basées
sur des données épidémiologiques larges –
sans résultat
Cohorte d’un
demi-million
d’enfants
The New England
Journal of Medicine
C o py r ig ht © 2 0 0 2 by t he Ma s s ac h u s e t t s Me d ic a l S o c ie t y
VOLUME 347
A POPULATION-BASED STUDY OF MEASLES, MUMPS, AND RUBELLA
VACCINATION AND AUTISM
KREESTEN MELDGAARD MADSEN, M.D., ANDERS HVIID, M.SC., MOGENS VESTERGAARD, M.D., DIANA SCHENDEL, PH.D.,
JAN WOHLFAHRT, M.SC., POUL THORSEN, M.D., JØRN OLSEN, M.D., AND MADS MELBYE, M.D.
ABSTRACT
Background It has been suggested that vaccina-
tion against measles, mumps, and rubella (MMR) is
a cause of autism.
Methods We conducted a retrospective cohort study
of all children born in Denmark from January 1991
through December 1998. The cohort was selected on
the basis of data from the Danish Civil Registration
System, which assigns a unique identification num-
ber to every live-born infant and new resident in Den-
mark. MMR-vaccination status was obtained from the
Danish National Board of Health. Information on the
children’s autism status was obtained from the Danish
Psychiatric Central Register, which contains informa-
tion on all diagnoses received by patients in psychiat-
ric hospitals and outpatient clinics in Denmark. We
obtained information on potential confounders from
the Danish Medical Birth Registry, the National Hospi-
tal Registry, and Statistics Denmark.
Results Of the 537,303 children in the cohort (rep-
resenting 2,129,864 person-years), 440,655 (82.0 per-
cent) had received the MMR vaccine. We identified
316 children with a diagnosis of autistic disorder and
422 with a diagnosis of other autistic-spectrum disor-
ders. After adjustment for potential confounders, the
relative risk of autistic disorder in the group of vac-
cinated children, as compared with the unvaccinated
group, was 0.92 (95 percent confidence interval, 0.68
to 1.24), and the relative risk of another autistic-spec-
trum disorder was 0.83 (95 percent confidence inter-
val, 0.65 to 1.07). There was no association between
the age at the time of vaccination, the time since vac-
cination, or the date of vaccination and the develop-
ment of autistic disorder.
Conclusions This study provides strong evidence
against the hypothesis that MMR vaccination causes
autism. (N Engl J Med 2002;347:1477-82.)
Copyright © 2002 Massachusetts Medical Society.
N Engl J Med, Vol. 347, No. 19 · November 7, 2002 · www.nejm.org · 1477
N O V E M B E R 7, 2002 NUMBER 19
I
T has been suggested that the measles, mumps,
and rubella (MMR) vaccine causes autism.1-4 The
widespread use of the MMR vaccine has report-
edly coincided with an increase in the incidence
of autism in California,5 and there are case reports of
children in whom signs of both developmental regres-
sion and gastrointestinal symptoms developed shortly
after MMR vaccination.1 Measles virus has been found
in the terminal ileum in children with developmental
disorders and gastrointestinal symptoms but not in de-
velopmentally normal children with gastrointestinal
symptoms.6 The measles virus used in the MMR vac-
cine is a live attenuated virus that normally causes no
symptoms or only very mild ones. However, wild-type
measles can infect the central nervous system and even
cause postinfectious encephalomyelitis, probably as a
result of an immune-mediated response to myelin pro-
teins.7-9
Studies designed to evaluate the suggested link be-
tween MMR vaccination and autism do not support
an association, but the evidence is weak and based on
case-series, cross-sectional, and ecologic studies. No
studies have had sufficient statistical power to detect
an association, and none had a population-based co-
hort design.10-16 The World Health Organization and
other organizations have requested further investiga-
tion of the hypothetical association between the MMR
vaccine and autism.2,17-20 We evaluated the hypothesis
in a cohort study that included all children born in
Denmark in 1991 through 1998.
From the Danish Epidemiology Science Center, Department of Epidemi-
ology and Social Medicine, Århus, Denmark (K.M.M., M.V., P.T., J.O.); the
Danish Epidemiology Science Center, Department of Epidemiology Research,
Statens Serum Institute, Copenhagen, Denmark (A.H., J.W., M.M.); and
the National Center on Birth Defects and Developmental Disabilities, Centers
for Disease Control and Prevention, Atlanta (D.S.). Address reprint requests
to Dr. Madsen at the Danish Epidemiology Science Center, Department of
Epidemiology and Social Medicine, Vennelyst Blvd. 6, DK-8000, Aarhus C,
Denmark, or at kmm@dadlnet.dk.
98
7. Les anti-vaccins – technique n°4: LA FRAUDE
Le papier est un “case report”, très peu de cas
sans groupe controle
S’en suivront de nombreuses publications
étudiant le lien entre ROR et autisme basées
sur des données épidémiologiques larges –
sans résultat
Le papier de Andrew Wakefield sera finalement
rétracté en 2000 pour fraude, les données
rapportées avaient en partie été fabriquées…
Wakefield fera alors ‘carrière’ comme leader
anti-vax aux US (livres, conférences…)
Early report
Ileal-lymphoid-nodular hyperplasia, non-specific colitis, and
pervasive developmental disorder in children
A J Wakefield, S H Murch, A Anthony, J Linnell, D M Casson, M Malik, M Berelowitz, A P Dhillon, M A Thomson,
P Harvey, A Valentine, S E Davies, J A Walker-Smith
Summary
Background We investigated a consecutive series of
children with chronic enterocolitis and regressive
developmental disorder.
Methods 12 children (mean age 6 years [range 3–10], 11
boys) were referred to a paediatric gastroenterology unit
with a history of normal development followed by loss of
acquired skills, including language, together with diarrhoea
TE
and abdominal pain. Children underwent
gastroenterological, neurological, and developmental
assessment and review of developmental records.
Ileocolonoscopy and biopsy sampling, magnetic-resonance
imaging (MRI), electroencephalography (EEG), and lumbar
puncture were done under sedation. Barium follow-through
radiography was done where possible. Biochemical,
haematological, and immunological profiles were
AC
examined.
Findings Onset of behavioural symptoms was associated,
by the parents, with measles, mumps, and rubella
vaccination in eight of the 12 children, with measles
infection in one child, and otitis media in another. All 12
children had intestinal abnormalities, ranging from
lymphoid nodular hyperplasia to aphthoid ulceration.
Histology showed patchy chronic inflammation in the colon
R
in 11 children and reactive ileal lymphoid hyperplasia in
seven, but no granulomas. Behavioural disorders included
autism (nine), disintegrative psychosis (one), and possible
postviral or vaccinal encephalitis (two). There were no
ET
focal neurological abnormalities and MRI and EEG tests
were normal. Abnormal laboratory results were significantly
raised urinary methylmalonic acid compared with age-
matched controls (p=0·003), low haemoglobin in four
children, and a low serum IgA in four children.
Interpretation We identified associated gastrointestinal
disease and developmental regression in a group of
previously normal children, which was generally associated
R
in time with possible environmental triggers.
Lancet 1998; 351: 637–41
See Commentary page
Inflammatory Bowel Disease Study Group, University Departments
of Medicine and Histopathology (A J Wakefield FRCS, A Anthony MB,
J Linnell PhD, A P Dhillon MRCPath, S E Davies MRCPath) and the
University Departments of Paediatric Gastroenterology
(S H Murch MB, D M Casson MRCP, M Malik MRCP,
M A Thomson FRCP, J A Walker-Smith FRCP,), Child and Adolescent
Psychiatry (M Berelowitz FRCPsych), Neurology (P Harvey FRCP), and
Radiology (A Valentine FRCR), Royal Free Hospital and School of
Medicine, London NW3 2QG, UK
Correspondence to: Dr A J Wakefield
THE LANCET • Vol 351 • February 28, 1998
EARLY REPORT
Introduction
We saw several children who, after a period of apparent
normality, lost acquired skills, including communication.
They all had gastrointestinal symptoms, including
abdominal pain, diarrhoea, and bloating and, in some
D
cases, food intolerance. We describe the clinical findings,
and gastrointestinal features of these children.
Patients and methods
12 children, consecutively referred to the department of
paediatric gastroenterology with a history of a pervasive
developmental disorder with loss of acquired skills and intestinal
symptoms (diarrhoea, abdominal pain, bloating and food
intolerance), were investigated. All children were admitted to the
ward for 1 week, accompanied by their parents.
Clinical investigations
We took histories, including details of immunisations and
exposure to infectious diseases, and assessed the children. In 11
cases the history was obtained by the senior clinician (JW-S).
Neurological and psychiatric assessments were done by
consultant staff (PH, MB) with HMS-4 criteria.1 Developmental
histories included a review of prospective developmental records
from parents, health visitors, and general practitioners. Four
children did not undergo psychiatric assessment in hospital; all
had been assessed professionally elsewhere, so these assessments
were used as the basis for their behavioural diagnosis.
After bowel preparation, ileocolonoscopy was performed by
SHM or MAT under sedation with midazolam and pethidine.
Paired frozen and formalin-fixed mucosal biopsy samples were
taken from the terminal ileum; ascending, transverse,
descending, and sigmoid colons, and from the rectum. The
procedure was recorded by video or still images, and were
compared with images of the previous seven consecutive
paediatric colonoscopies (four normal colonoscopies and three
on children with ulcerative colitis), in which the physician
reported normal appearances in the terminal ileum. Barium
follow-through radiography was possible in some cases.
Also under sedation, cerebral magnetic-resonance imaging
(MRI), electroencephalography (EEG) including visual, brain
stem auditory, and sensory evoked potentials (where compliance
made these possible), and lumbar puncture were done.
Laboratory investigations
Thyroid function, serum long-chain fatty acids, and
cerebrospinal-fluid lactate were measured to exclude known
causes of childhood neurodegenerative disease. Urinary
methylmalonic acid was measured in random urine samples from
eight of the 12 children and 14 age-matched and sex-matched
normal controls, by a modification of a technique described
previously.2 Chromatograms were scanned digitally on
computer, to analyse the methylmalonic-acid zones from cases
and controls. Urinary methylmalonic-acid concentrations in
patients and controls were compared by a two-sample t test.
Urinary creatinine was estimated by routine spectrophotometric
assay.
Children were screened for antiendomyseal antibodies and
boys were screened for fragile-X if this had not been done
99
637
7. Les anti-vaccins – technique n°5: la théorie du complot

THE PIRBRIGHT INSTITUTE

The present invention provides a live, attenuated coronavirus
comprising a variant replicase gene encoding polyproteins comprising a
mutation in one or more of non-structural protein(s) (nsp)-10, nsp-14,
nsp-15 or nsp-16. The coronavirus may be used as a vaccine for treating
and/or preventing a disease, such as infectious bronchitis, in a subject.

FIELD OF THE INVENTION

The present invention relates to an attenuated coronavirus comprising
a variant replicase gene, which causes the virus to have reduced
pathogenicity. The present invention also relates to the use of such a
coronavirus in a vaccine to prevent and/or treat a disease.

BACKGROUND TO THE INVENTION

Avian infectious bronchitis virus (IBV), the aetiological agent of
infectious bronchitis (IB), is a highly infectious and contagious pathogen
of domestic fowl that replicates primarily in the respiratory tract but
also in epithelial cells of the gut, kidney and oviduct. IBV is a member of
the Order Nidovirales, Family Coronaviridae, Subfamily Corona virinae
and Genus Gammacoronavirus; genetically very similar coronaviruses
cause disease in turkeys, guinea fowl and pheasants…

100
8. Vaccin et ratio bénéfice/risque

Un vaccin idéal est

• Sûr
• Efficace, conférant une immunité de longue durée
• Facilement administrable (oral, une seule dose)
• Thermostable
• Pas cher
• Multivalent
• Traçable pour médecine vétérinaire (différentiation entre individus vaccinés vs infectés -
DIVA)

Un vaccin est un objet pharmaceutique complexe

Balance bénéfice (protection de l’individu et de la société) / cout (risque pour l’individu)

101
DIVA = Differentiating Infected from Vaccinated Animals

102
 8. Vaccin et ratio bénéfice/risque
8.1. Antibody-dependent enhancement = ADE

Infection facilitée par les Ac (PAS AVEC TOUS LES VIRUS! C’est RARE)
Mécanisme bien connu pour le virus de la Dengue (flavivirus) et le Feline Infectious Peritonitis
URE MICROBIOLOGY
Virus (coronavirus) PERSPECTIV

Macrophage-tropic viruses: dengue virus, FIPV

FcγRIIa

Non-neutralizing
antibody Monocyte/macrophage Enhanced viral replication 103
8. Vaccin et ratio bénéfice/risque
8.1. Antibody-dependent enhancement = ADE

Early Death after Feline Infectious Peritonitis Virus Challenge due to Recombinant Vaccinia
Virus Immunization

« Cas d’école de l’ADE »

Vaccin recombinant
• vecteur viral = virus de la vaccine
• Gene d’intérêt = protéine de Spike du coronavirus félin

Injection à des chatons qui vont produire des Ac neutralisants avec un faible titre
Les chatons vaccinés vont mourir plus vite que les contrôles

104
8. Vaccin et ratio bénéfice/risque
8.1. Antibody-dependent enhancement = ADE

Antibody-Dependent Enhancement 1318 Occurs Upon Re-Infection with the Identical

T. TAKANO ET AL. Serotype Virus
in Feline Infectious Peritonitis Virus Infection

Le transfert d’Ac anti-FIPV (Feline Infectious

Peritonitis Virus) ne protège pas contre le FIPV
mais au contraire en favorise sa réplication et
donc sa virulence

Fig. 3. Survival rates of cats inoculated with type I FIPV after passive
105
immuni-
zation with serum and purified ascitic IgG from type I FIPV-infected cats.
8. Vaccin et ratio bénéfice/risque
8.1. Antibody-dependent enhancement = ADE

Dengvaxia, vaccin contre la Dengue

2014, Dengvaxia passe la phase III

“adverse events” children—hospitalized at a
higher rate, 20/2099 (0.99%) than controls,
2/1005 (0.2%)

2016, apprové au Mexique, Indonésie, Brésil,

Péru, Thaïlande…

2017
OMS recommande de restreindre l’administration de Dengvaxia aux seuls individus ayant déjà
été infectés par la Dengue

Dengvaxia reste un vaccin très utile, démontre l’importance de la pharmacovigilance (phase IV)
8. Vaccin et ratio bénéfice/risque
8.2. Enhanced Respiratory Disease = ERD

Accentuation de maladie respiratoire

1967, vaccin inactivé (RSV fixé au formaldéhyde) contre le virus respiratoire syncytial (RSV)
administré à nourrissons
Au lieu de protéger, le vaccin a induit une forme accrue de maladie
Hospitalisations fréquentes et deux enfants immunisés sont décédés des suites d'une infection
par le RSV

107
 FcγRIIa
8. Vaccin et ratio bénéfice/risque
8.2. Enhanced Respiratory Disease = ERD

Accentuation de maladie respiratoire

Le vaccin induisait la production d’Ac de faible affinité (absence de maturation d’affinité des Ac).
Après une infection avec le RSV, le vaccin conduisait à une réponse Th2 entrainant le
Non-neutralizing
recrutement
antibody
d'éosinophiles, dépôt de complexes immuns et
Monocyte/macrophage
activation du complément dans
Enhanced viral replication

b les poumons
Respiratory viruses: RSV, measles

Immune complex
formation

Immune cell Complement Pro-inflammatory

recruitment cascade activation cytokine secretion

Causes inflammation
and airway obstruction 108
8. Vaccin et ratio bénéfice/risque
8.2. Enhanced Respiratory Disease = ERD

2023, nouveaux vaccins RSV mis sur le marché (vaccins sous-unitaire: prefusion F protein +
adjuvant, GSK et Pfizer) pour femmes enceintes et personnes âgées

109
 the shift of the mild (mMDV) to virulent (vMDV) classical approaches.
8. Vaccin
ains et ratio
is thought bénéfice/risque
to have occurred in the late 1950s. MD is a good example where genetic resistance to the
is shift in 8.3. Pression
virulence, whichdeoccurred
sélection
evenexercée par le vaccin
before the et évolution
development of tumours duhaspathogène
been well documented and
tiation of the vaccination programmes, is believed inbred chicken lines showing susceptibility or resistance
be driven by the move to the intensive poultry pro-
Virus
ction de la maladie
practices de Marek
at that time. Subsequent shifts in the
• Gallid
ulence havealphaherpesvirus
been attributed to 2the
(GaHV-2)
introduction of
Rispens
ccessive generations of MD vaccines (Witter, 1997a).
• Virus oncogénique chez le poulet,
e first shift in the virulence after the introduction of

Relative virulence
Lymphome T en qq semaines et mort
D vaccines became apparent in he early 1980s after Bivalent
vv+MDV

Très of
e •reports contagieux,
higher thanvirions produits
expected frequencyparof MD vvMDV
follicules plumeux,withtransmission par 1983).
HVT
ses in flocks vaccinated HVT (Witter,
mpared to the previous isolates, the MDV strains
inhalation vMDV
ch as ALA-1, Md-5 and RB-1B obtained during this
riod were more virulent for HVT-vaccinated chickens
d Augmentation
these isolates de la virulence
were designated very virulent mMDV
Mild ->
vMDV) Virulent (Witter,
pathotypes -> Very1983).
Virulent -> Very the
Subsequently,
pearance of the very virulent plus (vv+MDV) patho-
Virulent + 1940 1960 1980 2000 2020 2040
pes in the late 1980s heralded a further shift in the
Fig. 6. Step-wise evolution of virulence of MDV. Relationship
ulence, and such strains continued to cause major between the virulence increase and the introduction of different
Pourquoi?
ses until the more effective CVI988 vaccines were vaccines is shown. HVT, herpes virus of turkeys; bivalent, HVT and
roduced. serotype 2 (SB-1) vaccines; Rispens, CVI988 strain.
110
8. Vaccin et ratio bénéfice/risque REVIEWS
8.3. Pression de sélection exercée par le vaccin et évolution du pathogène
Virulent MDV Very virulent MDV Very virulent plus MDV

Imperfect Imperfect Imperfect

Vaccins qui n’induisent pas d’immunité vaccination vaccination vaccination

stérilisante

Le vaccin protège du développement de

lymphomes mais le virus peut infecter
l’hôte, s’y répliquer et se propager à
d’autres hôtes

L’augmentation de la virulence a été induite

par la vaccination pour permettre au virus
de continuer à se répandre
Moderate mortality of High mortality of Very high mortality of
Les vaccins qui empêchent l’infection sont unvaccinated chickens unvaccinated chickens unvaccinated chickens

moins susceptibles de sélectionner pour une

augmentation de la virulence que les vaccins
qui se contentent de réduire le taux de
réplication
Low High 111
virulence virulence
8. Vaccin et ratio bénéfice/risque
8.4. VITT = Vaccine-induced Immune Thrombotic Thrombocytopenia
Day of Vaccination Post B-Cell Maturation

Thrombose et thrombocytopénie sévère après

administration des vaccins COVID AZ (et dans une
moindre mesure Johnson et Johnson)
Vaccination
B-cell matures and
secretes PF4/ChAdOx1 antibodies

Fc RIIa
Effet secondaire très rare mais sévère (fréquemment
fatal, 4 morts en Belgique suite à la campagne de Vector enters the blood
Antibody/PF4 immuno-aggregates
vaccination) activate platelets

Association de PF4 (Platelet Factor 4) sur la capside du

vecteur Electrostatic association of

RI contre le PF4 PF4 and adenovirus vector

Activated platelets release
further PF4

Chez individus ayant préalablement des Ac anti-PF4

DOI: 10.1126/sciadv.abl8213
-> Pistes pour augmenter la sécurité des vecteurs
The PF4/AdV complex is
adénoviraux: modification de la structure de la capside phagocytosed, trafficked to
lymph nodes, and stimulates Increasing numbers of activated

pour réduire sa charge négative, changement de PF4 memory B-cells platelets stimulate clot formation

Fig.S10: Cartoon representation of a proposed mechanism by which ChAdOx1 association with PF4 might

sérotype result in thrombotic thrombocytopenic syndrome. Following intramuscular vaccination with ChAdOx1 nCoV-19
small quantities of viral vector may enter the blood where they could interact with PF4 and form a complex through the
mechanisms described in this study. This complex can then be taken up by monocytes and transported to the lymph
nodes where it may stimulate the proliferation of pre-existing PF4 specific B-cells. After maturation of these B-cells, >5
days later, 112
4 IgG will be secreted which can form aggregates with PF4 circulating in the blood. These aggregates
can stimulate activation of platelets by binding to , stimulating further PF4 release. This could trigger a positive
feedback loop culminating in clot formation and NETs, as in HITT.
1. Vaccins (https://youtu.be/uqqCqagOs28)
Immunisation passive par transfert d’anticorps, protection temporaire
Immunisation active par administration d’antigènes
• Vaccins vivants
• Vaccins vivants atténués
• Vaccins inactivés, sous unitaires, VLPs et leurs Adjuvants
• Vaccins par virus chimérique
• Vaccins par vecteur viraux
• Vaccins ARNm dans Lipid NanoParticle (LNP)
• Vaccins ADN (déjà dépassés par ARNm)
• Vaccins à réplicons – SAM (en développement, dit « ARNm de second génération »)

Immunité collective, Nombre de reproduction de base (R0), Couverture vaccinale ((1-1/R0)/E )

Un bon vaccin est sûr, efficace, conférant une immunité durable, multivalent, facilement
administrable, peu cher, thermostable (traçable dit ‘DIVA’ pour application VT)
Balance bénéfice (pour l’individu et la société) / cout (pour l’individu)
Antibody-dependent enhancement, Enhanced Respiratory Disease, Sélection de souches 113
plus
virulentes/contagieuses, VITT
1. Vaccins (https://youtu.be/uqqCqagOs28)

114
1. Vaccins (https://youtu.be/uqqCqagOs28)

115
1. Vaccins (https://youtu.be/uqqCqagOs28)

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