Transcript IL-4

Immunomodulation by Probiotics
in Animal Model of Food Allergy
Soo-Young Lee, MD
Department of Pediatrics
Ajou University School of Medicine
Suwon, Korea
Contents
1. Basic immuno-pathogenesis and the goals of
immunomodulation in allergic disease
2. Innate immune responses of human cells to
bacteria and its experimental use
3. Probiotics(Lactic acid bacteria) and its
immunomodulatory effect in peanut allergic
mice
Basic Immuno-pathogenesis of Allergy
APC
IL-12, IL-18
IFN-a
Event in Target Tissues
IL-4
IL-4
IFN-g(-)
IL-12(-)
Plasma
cell
B
IFN-g(-)
Th1
Th2
IL-4
IL-10, 4(-)
IFN-g
IL-2
TNF-a
IgE secretion
FcR induction
MHC II increase
Th3
IL-5
IL-4, IL-5
IL-13, IL-9
(-)
(-)
TGF-b
IL-10
IL-4
(-)
IL-10
from Tr-1
IL-3
Th2
IL-4
Mast
cell
IL-3
IL-5
Eosino
-phil
Differentiation
Proliferation
FcR increase
Mediators release
Allergic inflammation
Differentiation
Survival, FcR increase
Mediator production
Cellular infiltration
Tissue damage
Positive & Negative Regulation of Cytokines
in Development of Effector Functions of T-helper Cells
APC
IL-12, IL-18
IFN-a
IL-4
IL-4
IFN-g(-)
IL-12(-)
IFN-g(-)
Th1
IFN-g
IL-2
TNF-a
(-)
Th2
Th3
IL-10(-)
IL-4(-)
IL-4, IL-5
IL-13, IL-9
IL-10
from Tr-1
(-)
TGF-b
IL-10
IL-4
(-)
Goal of Immunomodulation in Allergy
1. Can we re-direct the already existing deviation of Th2screwed immune environment, especially during perinatal or
early years in life ?
 We do actually want !
2. Are we trying to prevent allergic diseases in the real field ?
 Yes, we are trying to.
3. Can we reverse or treat the allergic sensitization or
expressed clinical allergies with certain type of
immunotherapy of immune modulation ?
 Partly yes, and somehow we might can.
• Bacteria and bacterial components :
one of candidate for immunomodulation
Innate Immune Responses of Human Neonatal Cell
to Bacteria from Normal GI Flora
Karlsson H etl al., Infect Immunol, 2002
Adult PBMC and cord blood mononuclear cell
Culture with bacteria : B. adolescnetis, E. faecalis, B. vulgatus, L. plantarum, S. mitis, C.
minutissimum, C. perfringnes, E. coli, P. aeruginosa
Cytokine production pattern:
IL-12, TNF-a : similar in CD and adult PBMC
G+ bacteria : produce more higher level of IL-12 in both CB and adult cells
IL-6 : significantly higher in CB cells than adult cells
IL-10 : similar in CB and adult cell
G+ bacteria : produce higher level of IL-12 and IL-10,
G- bacteria : produce less higher level of IL-12, but same level of higher IL-10
DC maturation pattern:
L. plantarum : bound or signaled through CD14, TLR-2, and TLR-4,
E. Coli : signaled minly through CD14 and TLR4
Microbial products in allergy prevention and therapy
- Matricardi PM et al. Allergy 2003, Position paper-
Bacterial extract
Route: early SQ injection  recently PO
Experiment : oral bacterial extract  spontaneous or PHA-induced TNF-a, IL12, IFN-g production of NK cells
In human volunteers, oral bacterial extracts induced an increased of IFN-a, IgA,
IL-2 concentrations in the BAL fluid, and increased serum total IgG, and
decreased total IgE
Proposed rationale : “modulators” of the host local immune response
Effect of bacterial immunotherapy : 75 – 87% of childhood asthmatics showed
therapeutic effect, but there was no DBPC study.
Side effect : minor
Conclusions and perspectives : effect is minimal, need dose adjustment and
interval
Microbial products in allergy prevention and therapy
- Matricardi PM et al. Allergy 2003, Position paper-
Mycobacteria(M. vaccae)
BCG
Immunostimulatory sequence of bacterial DNA(ISS-ODN)
Lipopolysaccharide(LPS) derivatives of G(-) bacteria
In this position paper, there are large amount details about many experimental
data, proposed rationale, and clinical studies relating above microbiota and their
products. However, those issues will discuss later in another sessions.
Microbial products in allergy prevention and therapy
- Matricardi PM et al. Allergy 2003, Position paper-
Probiotics
Term: living or inactivated organisms that are claimed to exertbeneficial effects
on health when ingested
Experiment: volunteer study, oral ingestion  triggers IFN-g production by
blood leukocyte / lactobacilli stimulate ample of IL-12 producton in human
macrophages in vitro * New experimental data will discuss later.
Proposed rationale (epidemiologic aspect)
- Early colonization of enterococcus, lactobacilli, eubacteria  low allergy, and
higher turn over of E. coli strains (Bennet R et al. 1991;Seep E et al. 1997; Adlerberth I et al.
1999)
- Swedish and Estonian study ; coli form bacteria were higher in the atopic
children while bifidobacteria were more prevalent in controls by 2 years (Bjorksten
B et al. 1999)
Finland : developing atopic sensitization at 12 mo of age –
related with more clostridia and fewer bifidobacteria at 3 week of life (Kalliomaki
- Prospective study in
M et al.2001)
Bacteria-Based immunomodulation
1. Heat-Killed Listeria or E. coli
- Studies using animal model of food allergy •
Engineered recombinant peanut protein and heat-killed L. monocytogenes
coadministration protects against peanut induced anaphylaxis in a murine
model.
Xiu-Min Li et. al, JACI 2003
•
Persistent protective effect of heat-killed E. coli producing “engineered”,
recombinant peanut proteins in a murine model of peanut allergy.
Xiu-Min Li et. al. JACI 2003
•
Effects of lysed E. faecalis FK-23 on allergen-induced serum antibody
responses and active cutaneous anaphylaxis in mice.
Shimada T et al.,Clin Exp Allergy, 2004
•
Allergen immunotherapy with heat-killed L. monocytogenes alleviates
peanut and food-induced anaphylaxis in dogs.
Frick IL et al., Allergy, 2005
Bacteria-Based immunomodulation(2)
2) Lactic acid bacteria
-Introductions•
Lactic acid bacteria(LAB) : Lactobacilli, Bifidobacteria
•
Normal intestinal flora
•
Maintaining the physiologic inflammation  prevent inflammatory
diseases(including allergy) in the gut
•
Possibly increased IgA, IL-10, IL-12, IFN-g
•
Suppression of TNF-a, and inhibition of T-cell activation
•
Improves severity of atopic dermatitis in infants with milk allergy
•
Prevents development of atopy in at-risk infants
•
Oral delivery, generally safe and well-tolerated
Role of intestinal flora in the development of allergy
Kalliomaki M, Isolauri E.
Curr Opin Allergy Clin Immunol. 2003 Feb;3(1):15-20.
Recent findings :
1) Alterations in intestinal microbiota have been detected both in
infants suffering from allergic disease and in those later
developing the disorder.
2) Delay in the compositional development of gut microflora was a
general finding in allergic children.
(cont.) Role of intestinal flora in the development of allergy.
Subsequent studies :
3) Perinatal administration of lactobacilli halved the later
development of atopic eczema during the first 2 years of life.
4) Specific strains of the healthy gut microbiota have been shown
to induce the production of IL-10 and TGF-b
5) Probiotics also strengthen gut defense barrier mechanisms and
reduce antigen load in the gut.
6) In the intestinal epithelial and antigen-presenting cells , Pattern
recognition receptors have been demonstrated to mediate a
continuing dialogue between host and gut-microbes
7) Clinical effects has been shown in allergic rhinitis and asthma,
and cow’s milk allergy
Dendritic cell and Probiotics
Innate immunity
Adaptive immunity
Lactic acid bacteria (+)
Figure from Kapsenberg. Nature Review immunol 2003
Dendritic Cells and Probiotics(1)
“ By responding differently to different type of microbes, DC
can signal the nature of the pathogen(or antigen)”
IL-12
E.col
DC
LPS
P. gingivalis
Candida albicans
Murine CD8aa+DC
Th2
No IL-12
Unicellular yeat
Hypae
Th1
DC
IL-12
Th1
Murine DC line
Th2
IL-4
Theiler’s virus
IL-12
Wild-type
DC
Variant
Th1
Murine spleen DC
IL-10
Th2
Dendritic Cells and Probiotics(2)
Lactobacilli differently modulate expression of cytokines and
maturation surface markers in murine dendritic cells
Christensen HR et al., J Immunol, 2002
6 lactobacilli strains : L. reuteri, L. plantarum, L. fermentum, L. casei,
L. alactus, L jonsonii
DC: bone marrow derived murine DC (C57BL/6), harvest enriched DC (8 day)
Cytokine and surface marker induction : DC culture with irradiated lactobacilli
Result :
1) IL-12, TNF-a productions : substantial differences among lactobacilli strains
2) IL-6, IL-10 : less pronounced differences
3) MHC class II, B7.2(CD86) : up-regulated by all kinds of lactobacilli
4) Amount of IL-12 production correlated with DC maturation markers
5) Different species of Lactobacillus exert very different patterns of DC
activation, and one species may have inhibiting activities of other species
(L. casei inhibit the IL-12 production by L. reuteri, and vice versa)
 potential for Th1/Th2/Th3-driving capacities of the gut DC to be modulated
according to composition of cut microflora
Dendritic Cells and Probiotics(3)
Lactobacilli activate human dendritic cells that screw T cells toward
T helper 1 polarization
Mohamadzadeh M et al., PNAS, 2005
Human myeloid DC : from PBMC, and enriched and harvested
Lactobacilli : L. gasseri, L. johnsonii, L. reuteri / killed by 15 min UV exposure
Result : Lactobacilli exposed DCs
1) Up-regulated HLA-DR, CD83, CD40, CD80, CD86
2) Induced bioactive IL-12 and IL-18, but not IL-10
* IL-12 production did not inhibited by LPS from E. coli
whereas, LPS induced IL-10 production was greatly inhibited
* Amount and pattern of IL-12 or IL-10 productions are strain dependent
3) Induced of T cell proliferation and activation (CD4+CD8+ Th1 cell)
4) Up-regulated TLR-2 by lactobaculli
Dendritic Cells and Probiotics(4)
Debating points on the modulatory effect of lactobacilli on DCs
1. IL-10 induction
2. IL-12 induction
3. IgG1, IgG2a regulation vs. IgE regulation
4. Half-time of effect, amount of dose, species
5. Non-specific vs. antigen specific
For example, regarding the cytokine production ;
Drakes et al., Infect Immunol, 2004 : human DC produce IL-10, not IL-12p70
Mohamadzadeh et al., PNAS, 2005 : human DC, produce IL-12p70, but not IL-12
Conclusions
1. Recentrly, immunomodulatory effect of live or killed bacteria(especially
probiotics) have been extensively evaluated and reviewed in experimental
studies.
2. However, the concept to prevent or modify already existing atopic disease
via administration of live bacteria or bacterial components remains to be
tested in more randomized and controlled clinical studies before any
conclusion can be made on this important matter.
3. We would expect that these types of immunomodulation could be a good
candidate for prevention or treatment of allergic disease, especially in
infants and children, in near future.
Acknowledgement
Ajou University School of Medicine
Se-Jo Oh, Msc, Gye-Ree Jeon, MD, nahm-Hyang Koo, MD
Yonsei University School of Medicine
Prof. Kyu-Earn Kim, Prof. Myung-Hyun Sohn,
Seoul National Universisy
Prof. GE Ji
Mount Sinai Medical Center,
Xiu-Min Li, MD, Sampson HA, MD
Gi-Kang Whang, Msc
Thank you for your attention !
2005.6.
Soo-Young Lee, MD.PhD.
Department of Pediatrics Ajou University School of Medicine
Suwon, Republic of Korea