The faith of the microbiota at older age…

Bruno Pot

Vrije Universiteit Brussels

Brussels, Belgium

Age, we all know the positive curves!

Age, a ‘growing’ issue for the near future?

CPI DP 07 (heritage.org)

NO

W

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

Autoimmune (arthritis) disease with age

Duggal et al 2013 Aging Cell

65 85

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?

Prevalence* of Self-Reported Obesity Among U.S. Adults by State and Territory, BRFSS, 2013

The gut microbiota and obesity: from correlation to causalityLiping 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!

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

Microbiota: a lifetime dynamics

BacteroidetesFirmicutesAcinobacteria Proteobacteria Variable/other

AD Kostic et al.; Genes Dev. 2013MJ Claesson et al.; Nature 2012

relatively low diversity, with

great inter-individual variability

3 Yrs: more diverse

‘‘adult-like’’ structure

increase in Actinobacteria and Proteobacteria

and increased diversity, but the gut microbiota

returns to its original structure some time after

delivery

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

• 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. 2009Muegge BD et al. Science 2011De Filippo C et al.; PNAS 2010

Eldermet studyMeasuring intestinal microbiota composition and diet together with immune and metabolic parameters in elderly from different community situations

• 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

Microbiota

Diet

Polarization continues

Cluster analysis reveals 4 Dietary Groups

DG1: low fat/high fibreDG2: moderate fat/high fibre DG3: moderate fat/low fibreDG4: high fat/low fibre

98% of the community andday hospital subjects

83% of the long-stay subjects.

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 lipidslong-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.

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?

The immune system: a lifetime dynamics?

Systematic differences at older age: immunosenescence!

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

Sapey E et al 2014 Blood

Increased serum Neutrophil Elastase activity in Healthy Elderly

Changed neutrophil responses

Sapey E et al 2014 Blood

Neutrophil migration (chemokinesis)

YOUNG

ELDERLY

Chemokinesis and chemotaxis towards IL-8

Sapey E et al 2014 Blood

Neutrophil migration

The problem starts around the age of 60!

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.

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

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

Diet MetabolismMicrobiotaimpacts impacts

Diet MetabolismMicrobiotaimpacts impacts

Immunity (senescens; inflammaging)

Physical activityim

pact

s

Diet MetabolismMicrobiotaimpacts impacts

Immunity (senescens; infammaging)

Physical activityim

pact

s

DNA damage

Obesity

Oxidative stresses

Years of exposure to inflammatory proteins

Diet MetabolismMicrobiotaimpacts impacts

Immunity (senescens; infammaging)

Physical activityim

pact

s

DNA damage

Obesity

Oxidative stresses

Years of exposure to inflammatory proteins

Exacerbation of chronic diseases

Exacerbation of aging

CVDCancerFrailtyCognitive dysfunctionMusculoskeletal decline …

Diet MetabolismMicrobiotaimpacts impacts

Immunity (senescens; infammaging)

Physical activityim

pact

s

DNA damage

Obesity

Oxidative stresses

Years of exposure to inflammatory proteins

Exacerbation of chronic diseases

Exacerbation of aging

CVDCancerFrailtyCognitive dysfunctionMusculoskeletal decline …

So an anti-inflammatory diet is what we need?

EAT MORE EAT LESS

FruitsRed/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

VegBroccoli, 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, peasTofu (from soybeans), dhal (from lentils), hummus (from chickpeas)

Herbs and SpicesTurmeric, 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

EAT MORE EAT LESSDairy 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 cerealsBiscuits and snacks Crisps, chips, pretzels, biscuits, cookies and piesPasta and grains Wholemeal pasta, brown rice, quinoa White rice, white pasta, gnocchiNuts 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/

Could probiotics do the job?

Cytokine profiles after LAB stimulation of PBMC

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Anti-inflammatory strains?

Strain specificity !

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Protection in the TNBS colitis model

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

AbstractThis 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

The effects of non-viable Lactobacillus paracasei on immune function in the elderly: a randomised, double-blind, placebo-controlled study

AbstractForty-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

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

Diet MetabolismMicrobiotaimpacts impacts

Immunity (senescens; infammaging)

Physical activity

impa

cts

DNA damage

Obesity

Oxidative stresses

Years of exposure to inflammatory proteins

Exacerbation of chronic diseases

Exacerbation of aging

CVDCancerFrailtyCognitive dysfunctionMusculoskeletal decline … The importance of the holistic approach

Probiotics are our invisible friends!

Thank you for yor attention!