Current Update in Immunology

Gut Immune System and Gut Microbiota in

the Post-Antibiotic Era Medicine

Parameth Thiennimitr, M.D., Ph.D.

Department of Microbiology

Faculty of Medicine, Chiang Mai University

2016

Learning Objectives
- Update the new trend and knowledge in
immunology that relevant to human health &
diseases
- Example of knowledge and information from
peer-reviewed scientific journals

Traditional Medical Microbiology & Immunology

Study Pathogens
Study How to fight
against pathogens

The New Concepts of

Medical Microbiology & Immunology

Study How our body

response to it
Study Good microbes (microbiota)

Were our microbes

human

(Picture from www.nature.com)

microbiota

Shaping of Our Gut Microbiome

Tamburini, et al., Nat. Med., 2016

Gut immune system in fetus

Presence of secondary lymphoid structures:
- Peyers patch and Mesenteric lymph node (MLN) in gut lamina propria
- Small numbers of immune cells

Tamburini, et al., Nat Med., 2016

Development of infant gut immune system by gut microbiota

Microbialassociated
molecular patterns

DC

1. M-cells sampling
luminal commensal
bacteria into gut lymphoid
organs

2. DCs present Ag into

immature T and B cells

3. Mature B cells
secrete dimeric IgA

4. Mature T cells
(TH2 and Treg) secrete
anti-inflammatory cytokines
(IL-10, TGF-)
Tamburini, et al., Nat Med., 2016

Gut dysbiosis leads to Gut Inflammation

1. Some factors (mostly
antibiotic) cause gut
dysbiosis an imbalance
of gut microbiota
population-

commensal bacteria
pathogenic bacteria
2. PAMPs activate TH1 and
TH17 to secrete proinflammatory cytokines:
IL-1, IL-17, TNF-, IFN-
3. PAMPs activate B-cell
to secrete IgG

Tamburini, et al., Nat Med., 2016

Gut inflammation

Four major phyla of human gut microbiota

Balanced gut microbiota

Obligate
anaerobes

4 dominant bacterial phyla

Firmicutes
Bacteroidetes
Actinobacteria
Proteobacteria

1. Firmicutes

- Gram+ bacteria
- e.g. Clostridium spp, Lactobacillus spp..

2. Bacteroidetes

- Gram- bacteria
- Very well-adapted to GI environment
- Ferment indigestible carbohydrates producing short
chain fatty acid (SCFA)
- e.g. Bacteroides spp.

3. Actinobacteria

- Gram+ bacteria
- Probiotic, Mucosal-protective effects
- e.g. Bifidobacterium spp.

4. Proteobacteria

- Gram- bacteria
- Expanded in gut dysbiosis, e.g. Escherichia coli.

Inflammation of lower GI tract associated with

Imbalanced gut microbiota (Dysbiosis)
Balanced gut microbiota

Imbalanced gut microbiota

(Dysbiosis)

Obligate
anaerobes

4 dominant bacterial phyla

Inflammatory
host innate
immune
response

Firmicutes

Proteobacteria

Antibiotic treatment
Infection with enteric pathogens

Bacteroidetes

HIV enteropathy

Actinobacteria

Chemically induced colitis

Proteobacteria
Adapted from Winter SE. and Bumler AJ, Cellular Microbiology, 2014

Diet-induced obesity

Gut Inflammation Promotes

Growth of Proteobacteria

E. coli uses iNOS-induced nitrate respiration

to overgrow obligate anaerobic gut microbiota in the inflamed gut

Inducible nitric oxide synthase

Winter, et al., Science, 2013

Mucus layer

Gut lumen

iNOS

Oxidases

E. coli
RNS & ROS

NO3Nitrate

2[H]

Nitrate
respiration

NO2Nitrite

Inflamed
gut
Gut microbiota
Lamina propria

(Obligate anaerobe)

Gut Inflammation Leads to

Systemic Inflammation and Obesity

Kim KA. et al., PLoS one, 2012

Gut lumen

G- LPS containingbacteria

G+
bacteria
Bifidobacteria

High Fat Diet

LPS

TLR-4

High Fat Diet

Dysbiosis

Intestinal Detoxified
alkaline
LPS
phosphatase
(IAP)

Enterobacteriaceae

Increased gut
permeability

Pro-inflammatory
cytokines:
IL-6, TNF-a, IL-1b

LPS
TLR-4

tight junction
proteins

TLR-4

macrophage

neutrophil
Blood vessel

adipose tissue

High Fat Diet

Metabolic
endotoxemia
(systemic LPS)

Pro-inflammatory
cytokines
(IL-6, TNF-a, IL-1b)

Gut mucosa

High fat diet causes gut dysbiosis by enhancing growth of

Enterobacteriaceae
Kim KA., et al., PLoS one, 2012

Low-fat diet
12 wks

High-fat diet

WT C57BL6
mice
Enterobacteriaceaeselective agar
(DHL agar)

Bifidobacteriaselective agar
(BL agar)

High fat diet induces inflammation in both systemic and local (gut) sites
Kim KA. et al., PLoS one, 2012

Low-fat diet

High-fat
diet
WT C57BL/6
mice

12 wks

Systemic (plasma) inflammation

Local (gut) inflammation

The first scientific evidence that link gut microbiota and obesity together.

Bckhed F., & Ding H. et al., PNAS, 2004

Gut microbiota increases body and epididymal fat weights in mouse

C57BL/6

Cecal content

2 weeks of
conventionalization
2 weeks on

standard chow
CONV-R mouse
Gut microbiota +

GF (Germ-free) mouse
Gut microbiota -

Epididymal fat pads

Gut microbiota -

Gut microbiota +

CONV-D mice
GF + Gut microbiota

(57% carbohydrate, 5% fat)

Cani P., et al., Diabetes, 2007

High fat diet increases plasma LPS and body weight in mouse
C57BL/6

Normal
diet

Plasma LPS level

LPS

Subcutaneous
infusion for 4 wks

High-fat
diet

Metabolic
endotoxemia
High-fat
fed mice
Normal
diet-fed
mice
Normal
endotoxemia

Gut microbiota modulation by dietary manipulations in obesity

Kovatcheva-Datchary et al., Best Pract. Res. Clin. Gastroenterol. 2013

Antibiotic Treatment Induces

Gut Microbiota Changes that
Correlate With Diseases

Antibiotic exposure during an early life increases risk of several diseases

Becattini et al., Trends Mol Med, (June 2016)

Broad use and misuse of

antibiotics in the past decades

More antibiotic
resistant bacteria
(superbug)

More allergic ,
metabolic
syndromes
and more..

Nature (May 2016)

Science (April 2016)

Nature (June 2016)

Broad Prophylactic or Therapeutic Intervention

On Gut Microbiota-Depleted Subjects
Fecal microbiota
transplantation
(FMT)

Colonization resistance
Gut microbiota
mucus
Gut epithelium
Goblet cell

Becattini et al., Trends Mol Med, (June 2016)

Selective Pathogen Targeting

Intervention Strategies

pathogen

microbiota

ABX = Antibiotics
Ab = antibody
CRISPR/Cas9 is a bacterial adaptive
immunity system that has been used in
modern biomedical research.

microbiota

Becattini et al., Trends Mol Med, (June 2016)

References
1. Becattini S, Taur Y, Pamer EG. Antibiotic-Induced Changes in the Intestinal
Microbiota and Disease. Trends Mol Med. 2016;22(6):458-78.
2. Backhed F, Ding H, Wang T, Hooper LV, Koh GY, Nagy A, et al. The gut
microbiota as an environmental factor that regulates fat storage. Proc Natl
Acad Sci U S A. 2004;101(44):15718-23.
3. Kim KA, Gu W, Lee IA, Joh EH, Kim DH. High fat diet-induced gut
microbiota exacerbates inflammation and obesity in mice via the TLR4
signaling pathway. PLoS One. 2012;7(10):e47713.
4. Kovatcheva-Datchary P, Arora T. Nutrition, the gut microbiome and the
metabolic syndrome. Best Pract Res Clin Gastroenterol. 2013;27(1):59-72.
5. Tamburini S, Shen N, Wu HC, Clemente JC. The microbiome in early life:
implications for health outcomes. Nat Med. 2016;22(7):713-22.
6. Winter SE, Winter MG, Xavier MN, Thiennimitr P, Poon V, Keestra AM, et al.
Host-derived nitrate boosts growth of E. coli in the inflamed gut. Science.
2013;339(6120):708-11.