Probiotics & Vaccines

Prof. N. K. Ganguly

Former DG, ICMR

Visiting Professor of Eminence,

Translational Health Science and Technology Institute

India

March, 2015

We have come a long way to understand Probiotics

Élie Metchnikoff first suggested the possibility of colonizing the gut with beneficial flora in the early 20th century: Circa1907.

The World Health Organization's 2001 definition of probiotics is "live micro-organisms which, when administered in adequate amounts, confer a health benefit on the host"

The term ‘probiotics’ was originally used by Lilly andStillwell (1965) for a substance that stimulates the growth ofother microorganisms.

• Lactic acid bacteria (LAB) are microbial communities normally present in the intestine of most animals.

• They are helpful in the treatment and prevention of disease as well as improving the digestion and absorption of nutrients.

Role of Probiotics Preventative & Therapeutic aspects

Probiotics Benefits

• The effectiveness of vaccines is based on immunological memory

• Heightened immune response directed against a previously encountered microorganism determined by genetics and nutrition

The best way to measure the function of the immune system is to ask it to respond to an infection. It is, however, not acceptable to experimentally give real infections to people to test the immune system.

A well-accepted surrogate of the response to infection, is the immune response to vaccines against the common infectious diseases.

Vaccine efficacy is measured by the ability to induce serum antibody responses and the level of specific serum antibody elicited by a vaccine usually correlates with protection.

Studying vaccine-induced antibody responses in individuals receiving a probiotic is a gold standard way to determine if probiotics can modulate immune responses.

• Through mucosal immunity

• By acting as adjuvants for mucosal vaccines

• By influence of resident intestinal micro flora on the development and functions of the intestinal-associated lymphoid tissue

• By modulation of the immune response of the immunosuppressed host,

• By modulation of cytokine expression by lactobacilli and its possible therapeutic use

• As bioactive peptides from fermented foods

• Through immuno-stimulation by lactic acid bacteria, and

• Using probiotic bacteria as live oral vaccines, Lactobacillus as the versatile delivery vehicle

Probiotics and Vaccines: A synergistic act

Key roles of adjuvants in modern vaccines

Several pattern recognition receptors (PRRs) that activate an innate immune response can be targeted by adjuvants, and details of their downstream signalingpathways are shown. TLRs, located at the cell surface (TLR1, TLR2, TLR4, TLR5 etc.)

Nature Medicine19, 1597–1608

(2013)

Putative mechanisms for enhancement ofsystemic immune response.

• By activating dendritic cells in Peyer’s patches, probiotic bacteria may cause cytokine release into the circulation

• Alternatively, fragments of probiotics may enter the bloodstream and have a direct but weak adjuvant effect at a distant lymph node

Source:Macdonald and Bell , Proceedings of the Nutrition Society (2010)

Interaction between probiotic bacteria and the epithelial & immune cells

2010 Federation of European Microbiological SocietiesPublished by Blackwell Publishing Ltd.

These live bacteria to elicit an immune response or

carry a vaccine component used in development of the vaccine

Effects of probiotic bacteria & yeast on intestinal

epithelial barrier function

Ohland C L , and MacNaughton W K Am J Physiol

Gastrointest Liver Physiol 2010;298:G807-G819

Cross-talk between the probiotic bacteria & the intestinal mucosa..

2010 Federation of European Microbiological SocietiesPublished by Blackwell Publishing Ltd.

The advantage • Live bacterial vaccines mimic

natural infection, have intrinsic adjuvant properties and can be given orally.

• Components of pathogenic and non-pathogenic food-related microorganisms are currently being evaluated as candidates for oral vaccines

Adjuvant Properties of Probiotic Bacteria:Mucosally Administered vaccines

Discov Med. 2011 Dec;12(67):525-33

Adjuvant Properties of Probiotic Bacteria: Parentally administered vaccine

Discov Med. 2011 Dec;12(67):525-33

Probiotic Lactobacillus strains in vaccine development : some examples.

Cultivable levels of Lactobacillus gasseri throughout the human mucosa

FEMS Microbiol Rev 37 (2013) 915–935

Strategies using Lactic Acid Bacteria against various diseases

FEMS Microbiol Rev 37 (2013) 915–935

• Young adult volunteers were given an oral polio vaccine. • Probiotic or placebo treatment started a week before vaccination and• continued for another month and two different probiotics were used

• Lactobacillus rhamnosus GG • or L. acidophilusCRL431.

• Serum antibody responses were measured up to 7 weeks later and it was clear that the probiotics induced significantly higher levels of neutralising antibodies and

• higher levels of anti-polio serum IgG and IgA 2

2 De Vrese M, Rautenberg P, Laue C et al. (2005) Probiotic bacteria stimulate virus-specific neutralizing antibodies following a booster polio vaccination. Eur J Nutr 44, 406–413.

Effect on probiotics on oral vaccines- Polio

In 1995, it was first shown that giving infants a single dose of Lactobacillus casei GG just before the oral administration of a rotavirus vaccine followed by 5 d of probiotic or placebo treatment, resulted in higher serum IgM, IgG and IgA anti-viral titres on day1

P values ,however, did not achieve statistical significance.

1. Isolauri E, Joensuu J, Suomalainen H et al. (1995) Improved immunogenicity of oral D

x RRV reassortant rotavirus vaccine by Lactobacillus casei GG. Vaccine 13,310–312.

Effect on probiotics on oral vaccines: Rotavirus

Lactobacillus casei has been used in Finland for recovery from acute rotavirus diarrhea in children (Gomboova et al., 1986)

Children with rotavirus diarrhoea recovered 1.5 days after treatment with Lactobacillus reuteri vs. 2.5 days in matched control infants (Korik et al., 1968). It also reinforces the local immune defenses through a specific IgA response to rotavirus

Rotavirus Vaccination & Probiotics: reduction in infantile diarrhea

Source:

Walter J et al. PNAS 2011;108:4645-4652

A complex study where a combination of four different bacteria

• L. rhamnosus GG, • L. rhamnosus LC705, • Bifidobacterium breve Bbi99 • Propionibacterium freudenreicherii ssp. shermanii

and a prebiotic were given to mothers pre-term and their children for 6 months after birth

Children received routine diphtheria, tetanus and pertussis (DTP) vaccines at 3, 4 and 5 months and the Hemophilus influenza type b (Hib) vaccine at 4 months and serum antibodies were measured.

No effect was seen with DTP, but significantly higher protective Hib titres were seen in children receiving the complex mixture.

Probiotics can affect parenteral vaccines too

Kukkonen K, Nieminen T, Poussa T et al. (2006) Effect of probiotics on vaccine antibody responses in infancy – a

randomized placebo-controlled double-blind trial. Pediatr Allergy Immunol 17, 416–421.

Effective Safe Vaccine Against HIV : A distant dream

Multiple strategies to produce an immunogenic HIV vaccine have been developed (Ke-Qin et al., 2003): HIV subunit peptide vaccines, DNA as a vaccine, recombinant virus-vector vaccines (using viral vectors such as vaccinia virus, adenovirus, rabies virus, flavivirus, Friend murine leukemiavirus, Venezuelan equine encephalitis virus and adeno associated virus), and bacteria as a vector, such as Bacille Calmette–Guerin.

Each of these strategies showed some promise in animal models

FEMS Immunol Med Microbiol 58 (2010) 299–306

HIV vaccine development using probiotic bacteria

Induced humoral and cell-mediated immune response is sufficient to provide immunity against a HIV Env-expressing vaccinia virus challenge in mice (Aldovini & Young, 1991)

Oral administration of recombinant L. lactis encoding the V2–V4 loop of the HIV env gene can induce HIV-specific mucosal and systemic immunity in Mice and hence recombinant L. lactis is now an appropriate candidate for HIV vaccine development

HIV

L .Lactis

FEMS Immunol Med Microbiol 58 (2010) 299–306

Trichomoniasis

Trichomoniasis vaccination & LAB

Lactobacillus

T. vaginalis

FEMS Immunol Med Microbiol 58 (2010) 299–306

Probiotic Provide Protection Against Pneumococcal Disease

• Serotype replacement is considered the most significant problem in the post-PCV7 era. A prerequisite for pneumococcal disease is adherence of the bacterium to host nasopharyngeal epithelium leading to colonization (carriage).

• Elimination of vaccine- serotype from carriage has provided new challenge in prevention of pneumoccoal disease, particularly for resource-poor settings.

• Research on probiotics preventing pneumococcal disease in early life will make the vaccination schedules more effective

PloS Pathogens June 2012 | Volume 8 | Issue 6 | e1002652

• Commensal and/or probiotic bacteria can prevent pathogens (pneumococci) from attaching to and colonizing the respiratory epithelium by associating with specific cell surface receptors

• By enhancing mucus secretion and the production of secretory IgA• Probiotic bacteria interact with immune cells and trigger variety of effector cells and

maintain barrier integrity

PloS Pathogens June 2012 | Volume 8 | Issue 6 | e1002652

Proposed mechanism of action of probiotic bacteria in protection against pneumococcal infection

Gut probiotics present innovative tools for malaria

prevention & lead the way to novel types of vaccination

strategies

Probiotic Based Malaria Vaccines

(1) Produce toxic substances against malaria midgut stages (2) or stimulate mosquito immune responses to secrete antimicrobial

molecules into the midgut lumen, which target both bacteria and plasmodia.(3) In the human, gut bacteria can result in the production of protective

antibodies, particularly of class IgM (I), which target malaria sporozoites once deposited in the human dermis (II) by the blood-feeding mosquito

Trends in Parasitology, January 2015, Vol. 31, No. 1

Probiotics based malaria vaccines

• In the mosquito, gut microbiota can form a physical barrier which prevents ookinetes from penetrating the gut epithelium

• Protective antigens ( MSP1) can be delivered to the mucosal surfaces using live bacteria with plasmids that are responsible for the expression of specific antigens.

• The immunogenicity of soluble protein is low when administered orally, but this can be enhanced using genetically engineered bacteria, so that low-level expression of MSP119 in Lactococcus lactis was still able to confer strong protection against the malarial parasite (Mercenier et al., 2000)

Clinical Case Studies Probiotic And Child Vaccination

Probiotics in MMRV, mumps, measles, Rubella & Varicellavaccination

(Assaf Harofeh Medical Center in Israel, January 2008 to August 2009)

Youngster et al, Arch Dis Child 2011;96:345–349.

Number of study participants failing to reach protectiveIgG antibody titres 3 months after vaccination for mumps,

measles, rubella and varicella

Arch Dis Child 2011;96:345–349.

Conclusion: Oral probiotics do not interfere with the immune response to mumps, measles, rubella and varicella vaccine, and may improve sero-conversion rates.

Study participants reporting vaccine relatedadverse events, 10 days following vaccination for mumps,

measles, rubella and varicella

Arch Dis Child 2011;96:345–349.

Vaccine response & probiotic administration in elderly are very few to

draw conclusionsStudy 1Lactobacillis paracasei (NCC 2461) 1x109 cfu and 6g prebiotic fructo-oligosaccharide was used as a part of the daily nutritional formulae with Parental trivalent influenza vaccine and pneumococcal vaccine containing 23 serotypes

No effect was seen on antibody response to vaccines but a significantly lower incidenceof infection after 12 months , in particular respiratory illnesses , in treatment vs control

groups(P=0.034) ( Bunout et al.2004)

Study 2Lactobacillus paracasei ssp. ( Acimel®) 1010 cfu /100 g bottle twice daily w s used with parental inactivated trivalent influenza vaccine

There was a trend for higher virus-specific antibody titres in probiotic vs. control groupSignificantly high greater sero-conversion rate for B strain in main study at 3,67 9 weeks post vaccination in probiotic vs.placebo group ( Boge et al. 2009)

Probiotic: Safety Status

• Several studies in infants on modulation of vaccine response with concomitant probiotic administration have shown probiotics are safe & effective in enhancing immune response

Vaccine response with probiotic administration in adults show increase vaccine specific serum IgA

More studies are needed to study the seroprotection/seroconversion

Br J Clin Pharmacol / 75:3 / 663–670 /

WAY FORWARD

• Creating cataloguing probiotic repository

• Promoting clinical studies probiotic –vaccine co administration

• Promotion of research into probiotics & prebiotics and long term immune surveillance of vaccinated population to assess the true potential of benefits of probiotics

Time will come when for vaccinating your child you will need to make him drink a smoothie