India presentation final b pot

1. The faith of the microbiota at older age…
Bruno Pot
Vrije Universiteit Brussels
Brussels, Belgium

2. Age, we all know the positive curves!

3. Age, a ‘growing’ issue for the near future?
CPI DP 07 (heritage.org)
NOW

4. While the total number is decreasing,
34% of Japan's population will be 65
years or older by 2055.
This is higher than European
countries United States, China, India,
Korea or Singapore.
Janet Lord, Reumatology conference 2014, Liverpool, UK
E.g. Japan has an issue to consider

5. Autoimmune (arthritis) disease with age
Duggal et al 2013 Aging Cell
65 85

6. Janet Lord, Reumatology conference 2014, Liverpool, UK
A longer life but not necessarily more healthy…
HEALTHY life expectancy
THEORETICAL life expectancy
How to avoid the
gap increases?

7. Prevalence* of Self-Reported Obesity Among U.S. Adults by State
and Territory, BRFSS, 2013
The gut microbiota and obesity: from correlation to causality
Liping Zhao, NATURE REVIEWS MICROBIOLOGY, 2013
In the United States, it is estimated that the total health care costs attributable to obesity or overweight patients will
double every decade to reach US$860.7–956.9 billion by 2030, accounting for 16–18% of the total US health care
costs.
Not really easy!

8. Ageing deeply affects the structure of
the human gut microbiota, as well as
its homeostasis with the host’s
immune system.
Because of its crucial role in the host
physiology and health status, age-
related differences in the gut
microbiota composition may be
related to the progression of diseases
and frailty in the elderly population.
Biagi E, Nylund L, Candela M, Ostan R, Bucci L, et al. (2010)
Through Ageing, and Beyond: Gut Microbiota and Inflammatory Status in Seniors and Centenarians.
PLoS ONE 5(5): e10667. doi:10.1371/journal.pone.0010667
Age is inherently linked to decreased fitness
Immunosenescence

9. Microbiota: a lifetime dynamics
Bacteroidetes
Firmicutes
Acinobacteria
Proteobacteria
Variable/other
AD Kostic et al.; Genes Dev. 2013
MJ Claesson et al.; Nature 2012
The human intestinal microbiota is
• is individual-specific at the level of the OTUs
• stable over time in healthy adults
• required for development and homeostasis of
the immune system
+ 65 years:
• increase in Bacteroidetes
• extremely variable between individuals
• different from the core microbiota and
diversity levels of younger adults

10. • changing diet
Alterations in oral / intestinal microbiota composition,
which are associated with:
 Affecting the intestinal microbiota
 Cause or consequence often not so clear
 Linked to chronic conditions
• obesity
• inflammatory diseases (IBD, arthritis, …)
• irritable bowel syndrome
 levels of frailty
• deterioration in dentition
• salivary function
• digestion
• intestinal transit time
• nutritional status
• compositional changes
• metabolic changes
• immune changes
However, links between diet, microbiota
composition and health in large human
cohorts are unclear…
Mai, V et al.; Nutr. J. 2009
Muegge BD et al. Science 2011
De Filippo C et al.; PNAS 2010

11. Eldermet study
Measuring intestinal microbiota composition and diet together with
immune and metabolic parameters in elderly from different community
situations

12. • 178 elderly (mean age = 78 yrs; ranging from 64 to 102), from different residence locations in the
community
• community dwellers, N=83;
• attending an out-patient day hospital, N=20
• in shortterm (<6 weeks) rehabilitation hospital care, N=15
• long-term residential care (long-stay), N=60
• 13 young adults with a mean age of 36 yrs.
• Parameters measured
• faecal microbiota
• dietary intake information
• faecal water metabolites
• inflammation
• psychological parameters
Target population

13. Microbiota

14. Diet
Polarization continues

15. Cluster analysis reveals
4 Dietary Groups
DG1: low fat/high fibre
DG2: moderate fat/high fibre
DG3: moderate fat/low fibre
DG4: high fat/low fibre
98% of the community and
day hospital subjects
83% of the long-stay
subjects.

16. Metabolic parameters in faecal water
Elipses represent the 95%
confidence interval obtained for
random permutation tests
1H NMR spectra
Community subjects (green) versus
long-stay subjects (red)
Community subjects (green) versus
rehabilitation subjects (orange)
Major separating metabolites
glutarate and butyrate
community subjects
glucose, glycine and lipids
long-stay
community subjects
acetate, propionate and valerate,
were also more abundant in
community dwellers.
Metagenomic genes predicted for
butyrate, acetate and propionate
production were significant higher
in rehabilitation or community
subjects compared to long-stay
subjects.

17. Metabolic parameters in faecal water: the genera involved
Candidate genera associated include Ruminococcus and Butyricicoccus for
butyrate production.
Requires validation in larger cohorts though.
Diet MetabolismMicrobiotaimpacts impacts
But what about the immune parameters?

18. The immune system: a lifetime dynamics?
Systematic differences at older age:
immunosenescence!

19. What is immunosenescence?
Immunosenescence is the functional deterioration of the immune system
during natural aging. It is evidenced by
• increased susceptibility to bacterial infections
• decreased NK cell activity
• persistent NF-kB-mediated inflammation (also sign of inflamm-aging)
• chronic activation of the innate and adaptive immune system
• increased pro-inflammatory cytokine levels (although also often
encountered in young adults)
• changes to neutrophil responses

20. Sapey E et al 2014 Blood
Increased serum Neutrophil Elastase activity in Healthy Elderly
Changed neutrophil responses

21. Sapey E et al 2014 Blood
Neutrophil migration (chemokinesis)
YOUNG
ELDERLY
Chemokinesis and chemotaxis towards IL-8

22. Sapey E et al 2014 Blood
Neutrophil migration
The problem starts around the age of 60!

23. Burns et al, Physiol Rev (2003)
1.3µm
Area of tissue damage
Increased tissue damage during migration  increased inflammation?
Elderly neutrophils cause 40% more tissue damage during migration.

24. In the Eldermet study 8 groups rather than 4
diet groups were discriminated for global
health analysis, including immune status,
Eldermet study
IL-6 and IL-8 levels were lower in community , whereas CRP
levels were higher in the lower path in longstay- only analysis

25. Prof. Janet Lord, Reumatology conference 2014, Liverpool, UK

26. Diet MetabolismMicrobiotaimpacts impacts

27. Diet MetabolismMicrobiotaimpacts impacts
Immunity (senescens; inflammaging)
Physical activity
impacts

28. Diet MetabolismMicrobiotaimpacts impacts
Immunity (senescens; infammaging)
Physical activity
impacts
DNA damage
Obesity
Oxidative stresses
Years of exposure to
inflammatory proteins

29. Diet MetabolismMicrobiotaimpacts impacts
Immunity (senescens; infammaging)
Physical activity
impacts
DNA damage
Obesity
Oxidative stresses
Years of exposure to
inflammatory proteins
Exacerbation of
chronic diseases
Exacerbation of
aging
CVD
Cancer
Frailty
Cognitive dysfunction
Musculoskeletal decline …

30. Diet MetabolismMicrobiotaimpacts impacts
Immunity (senescens; infammaging)
Physical activity
impacts
DNA damage
Obesity
Oxidative stresses
Years of exposure to
inflammatory proteins
Exacerbation of
chronic diseases
Exacerbation of
aging
CVD
Cancer
Frailty
Cognitive dysfunction
Musculoskeletal decline …

31. So an anti-inflammatory diet is what we need?

32. EAT MORE EAT LESS
Fruits
Red/black/purple fruits, all berries inc strawberries,
raspberries, blackberries, blueberries, elderberries,
blackcurrants, citrus, plums, cherries, grapes
There are no fruits we should eat
less of
Veg
Broccoli, chard, spinach, cabbage, collards, kale,
onions, carrots, sweet potatoes, garlic, peppers,
mushrooms, courgettes (zucchini), celery, asparagus
Potatoes or potato products, corn
or corn products, unless you are
very active physically
Legumes/Legume
products
Lentils, beans, peas
Tofu (from soybeans), dhal (from lentils), hummus
(from chickpeas)
Herbs and Spices
Turmeric, garlic, ginger, cayenne, chilli, curry powder,
basil, thyme, black pepper, cinnamon, oregano,
rosemary, nutmeg
Salt
Fats and oils Olive or rapeseed (canola) oil
Other vegetable and palm oils inc
sunflower. Hard margarines.
Fish
Salmon (if wild), herring, tuna, mackerel, sardines,
pilchards, trout, oysters
Deep-fried fish, fish fingers
Meat
Game, grass-fed beef, mutton & lamb, free range
chicken
Intensively farmed beef, pork or
poultry. Sausages, burgers, bacon,
cured meats such as hot dogs,
salami
Anti and pro-inflammatory foods

33. EAT MORE EAT LESS
Dairy products
Real cheeses especially green & blue, plain yoghurt,
particularly “live”
Sweetened yoghurt, ice cream
Breads
Wholemeal & rye in moderation, although physically
active people can eat more
White (refined) flour products
Cereals Bran cereals, no added sugar muesli, porridge oats Cornflakes, all sugared cereals
Biscuits and snacks Crisps, chips, pretzels, biscuits, cookies and pies
Pasta and grains Wholemeal pasta, brown rice, quinoa White rice, white pasta, gnocchi
Nuts and seeds
Eat in moderation – they are full of omega 6 fatty
acids
Salted and roasted nuts
Sweeteners
Prefer intense natural sweeteners such as stevia if
needed.
Sugar, honey, syrup, molasses
Desserts and sweets Dark chocolate
Most sweets and desserts, ice
cream, baked pastries
Drinks
Fruit and vegetable juices, tea, coffee, milk, moderate
red wine
Sugar-sweetened soft drinks, colas,
spirits
Anti and pro-inflammatory foods
http://www.nutrishield.com/.../inflamm-ageing-ebook/

34. Could probiotics do the job?

35. Cytokine profiles after LAB stimulation of PBMC
IL-10
TNFa
IL-12
IFNg
0
500
1000
1500
2000
2500
3000
3500
4000
4500pg/ml
0
5000
10000
15000
20000
25000
30000
35000
40000
45000
50000
pg/ml
0
200
400
600
800
1000
1200
1400
pg/ml
0
50000
100000
150000
200000
250000
pg/ml

36. 0
20
40
60
80
100
0
5
10
15
20
25
ratioIl10/IL12
Ratio IL-10 / IL-12 (PBMC)
Anti-inflammatory strains?
Strain specificity !

37. 0
20
40
60
80
100
IL-10 / IL-12 ratio (PBMC)
0
5
10
15
20
25
ratioIl10/IL12
Protection%
***
**
***
**
*** *
**
**
***
*
-50
-40
-30
-20
-10
0
10
20
30
40
50
60
70
80
90
100
Protection in the TNBS colitis model

38. Heat-killed Lactobacillus gasseri can enhance immunity in the
elderly in a double-blind, placebo-controlled clinical study
DOI: http://dx.doi.org/10.3920/BM2014.0108
Abstract
This double-blind, placebo-controlled clinical trial was conducted to test whether Lactobacillus gasseri TMC0356 (TMC0356)
can modify the immune response in the elderly. Heat-killed TMC0356 or placebo was orally administered to 28 healthy
subjects aged 50-70 years old for 4 weeks at a dosage of 1.0×109 cfu/day.
Peripheral blood mononuclear cells (PBMCs) were collected from the subjects before and after the study completion, together
with general health and blood examination records. Isolated PBMCs were examined for the number of T cells, CD8+CD28+ cells,
native T cells, B cells, natural killer (NK) cells and the ratios of CD4/CD8 T cells and native/memory T cells. NK cell activation
and concanavalin A-induced lymphocyte transformation of the isolated PBMCs were also examined.
The number of CD8+ T cells significantly increased in the subjects after TMC0356 oral administration (P<0.05). Furthermore,
the population of CD8+CD28+ T cells and the amount of lymphocyte transformation both significantly decreased in PBMCs from
the placebo group (P<0.05). However, such changes were not observed in the subjects exposed to TMC0356. These results
suggest that TMC0356 can increase the number of CD8+ T cells and reduce CD28 expression loss in CD8+ T cells of the elderly.
The effect of TMC0356 on immune responses in the elderly may enhance their natural defense mechanisms against
pathogenic infections.
K. Miyazawa, M. Kawase, A. Kubota , K. Yoda, G. Harata , M. Hosoda, F. He
2015

39. The effects of non-viable Lactobacillus paracasei on immune function in the elderly: a randomised, double-blind,
placebo-controlled study
Abstract
Forty-two participants in two nursing homes who were ≥65 years of age were randomised to receive a jelly containing 10
billion heat-killed Lactobacillus paracasei MCC1849 cells (LP group) or a placebo jelly without lactobacilli (placebo group)
for 6 weeks.
Three weeks after beginning jelly intake, all subjects received an influenza vaccination (A/H1N1, A/H2N3 and B). Blood
samples were collected before and after the treatment period. There were no significant differences in immune parameters,
including in antibody responses against the vaccination, between the groups. In the subgroup of the oldest old, defined
as ≥85 years of age (n = 27), the antibody responses to the A/H1N1 and B antigens, which were impaired in the placebo
group, were improved in the LP group. No significant effects of non-viable L. paracasei MCC1849 were observed in the
elderly. A possible beneficial effect in the oldest old should be explored in further large-scale studies.
Keywords: Antibody titre, immunosenescence, Lactobacillus paracasei, oldest old, vaccination
Michio Maruyama, Ryoji Abe, Tomohiro Shimono, Noriyuki Iwabuchi, Fumiaki Abe & Jin-Zhong Xiao
http://dx.doi.org/10.3109/09637486.2015.1126564

40. Conclusion regarding elderly people (eldermet study)
• Collectively, the data support a relationship between diet, microbiota and health
status.
• The healthiest people live in a community setting, eat better and have a
microbiota distinct from long-term residential care people.
• Measures of increased inflammation and increased frailty support a diet–
microbiota link regarding accelerated ageing.
• The intestinal microbiota of older people is associated with inflammation.
• Many studies argue in favor of an approach of modulating the microbiota with
dietary interventions, designed to promote healthier ageing.
• Dietary supplements with defined food ingredients that promote particular
components of the microbiota may prove useful for maintaining health in older
people; this could be pro- and /or prebiotics

41. Diet MetabolismMicrobiotaimpacts impacts
Immunity (senescens; infammaging)
Physical activity
impacts
DNA damage
Obesity
Oxidative stresses
Years of exposure to
inflammatory proteins
Exacerbation of
chronic diseases
Exacerbation of
aging
CVD
Cancer
Frailty
Cognitive dysfunction
Musculoskeletal decline … The importance of the holistic approach

42. Probiotics are our
invisible friends!
Thank you for yor
attention!