Definition of „good bacteria“ Origin, Effects and … · 2018. 12. 10. · Several...

Definition of „good bacteria“

Origin, EffectsEffects and EffectiveEffective ComponentsComponents

T bi A O l hlTobias A. OelschlaegerInstitut für Molekulare InfektionsbiologieUniversität Würzburg

ZENTRUM FÜR INFEKTIONSFORSCHUNG INSTITUT FÜR MOLEKULARE INFEKTIONSBIOLOGIE

A) Definition of Good bacteria“A) Definition of „Good bacteria“

Good bacteria Group 1

Not to be sick is good. Therefore, bacteria which do colonize the host but do notNot to be sick is good. Therefore, bacteria which do colonize the host but do not cause disease because they lack virulence genes are good bacteria. These goodbacteria are harmless commensals.

Good bacteria Group2 or the „even better bacteria“

Properties as group 1 but in addition they excert a (health) benefit for the human host due to certain components/properties not present in group 1 bacteria.These good bacteria are termed probiotics.


) f d bB) Origin of good bacteria

Fermented milk (products): e.g. kefir, koumiss

H i iHuman origin


C) EffectsEffects on the host and EffectiveEffective Components Components of Probiotics

C.1 Nutritional Physiology

C.1.1 AbilityAbility toto breackbreack down a down a widewiderangerange ofof nutritional nutritional constituentsconstituents thatthatcannotcannot bebe metabolizedmetabolized byby thethe hosthost:cannotcannot bebe metabolizedmetabolized byby thethe hosthost:

Lactose, Raffinose, Stachyose , Verbascose , Fructose polymers e.g. inulin

Inulin improves calcium and magnesium absorption (in rats) and increases theproportion of SCFA in cecal contents (Velazques et al. 1997)

C.1.2 KatabolicKatabolic inactivationinactivation ofof somesome antinutritional antinutritional factorsfactors:

Tannins (proanthocanidines, poly‐hydoxy‐phenols)Ph (H h h i id f i i )Phytates (Hexa‐phosphoric acid of myo‐inosit)These factors chelate minerals including iron, zinc, and calciumTannins (tea, wine, fruits) can be broken down by Lactobacillus plantarum, Lb. pentosus and Lb paraplantarum (tannin acylhydrolase)


pentosus and Lb. paraplantarum (tannin acylhydrolase)Phytates are broken down by several Lactobacilli


C.1 Nutritional PhysiologyC.1.3 Modification of the host gut physiology byg p y gy y

increasingincreasing productionproduction ofof growthgrowth factorsfactors

Spermidin and spermin are recognized as growth factors in eukyrotes and theyhave been proven to be involved in intestinal maturation in the rat.Lb. hilgardi and Lb. buchneri are able to produce putrescinputrescin, the precursor ofsuch growth factors (Alberto et al. 2007).

C.1.4 Synthesis Synthesis ofof VitaminsVitamins

Intake of the probiotic strain Lb. johnsonii Lb. johnsonii NCC533 is one of the rare Lb.NCC533 increasedincreased thethe precentageprecentage ofof plasmaplasmafolatefolate by 55.9 + 19.6 % in children(Mohammad et al. 2006).

species able to increaseincrease plasmaplasma cobalamincobalamin by52 + 15.6 % in children (Mohammad et al.2006).



C.2 Prevention of Disease

C.2.1 Anti Anti CarcinogenesisCarcinogenesis EffectsEffects

• Increase of the detoxifying glutathioneglutathione SS transferasetransferase (GTS)(GTS) in Caco‐2 cells by Lb.fermentum I5007 (Yang et al 2007)fermentum I5007 (Yang et al. 2007).

•• DecreasingDecreasing thethe amountamount ofof growthgrowth factorsfactors. VSL#3 and Lb. brevis CD2 decrease ornithindecarboxylase (OCD) activity in the host. This enzyme is involved in polyamine growthy ( ) y y p y gfactor synthesis (e.g. putrescin)(Linsalata et al. 2004, 2005).

• MetabolicalMetabolical inactivationinactivation ofof mutagenicmutagenic substancessubstances ((Lb., bifidobacteria, probiotic E. colistrain Nissle 1917 EcN)strain Nissle 1917 = EcN)

•• ReductionReduction ofof ROSROS may reduce risk of mutation (Mood and Hassan 1982). Isoflavones – aclass of flavonoids‐ require deglycosylation to exert their antioxidant properties (Wangand Ho 2009). Increase in isoflavone acglycones in soymilk fermented with Lb.acidophilus L10, B. lactis B94, and Lb. casei L26 (Donkor and Shah 2008).

•LAB can absorbabsorb carcinogenscarcinogens on their cellcell wallswalls prevent DNA damage in the colon and•LAB can absorbabsorb carcinogenscarcinogens on their cellcell wallswalls, prevent DNA damage in the colon andliver of rats after ingestion of certain LAB (Zsivkonvits et al. 2003).




C.2.2 Benefical Effects of Probiotics againstagainst CardiovascularCardiovascular DiseaseDiseaseagainstagainst CardiovascularCardiovascular DiseaseDisease

Angiotensin converting enzyme inhibitor (ACEi)Angiotensin can increase blood pressure. InIn vitrovitro ACEiACEi activityactivity could beAngiotensin can increase blood pressure. InIn vitrovitro ACEiACEi activityactivity could bedemonstrated for yoghurt extracts after inoculation with Lb. delbrueckiisubsp. bulgaricus, Streptococcus thermophilus, Lb. paracasei subsp. paracasei(Papadimitriou et al. 2007).( p )

PreventionPrevention ofof hypercholesterolemiahypercholesterolemia by promoting the utilization of livercholesterol due to the bilebile saltsalt hydrolasehydrolase activityactivity of certain probiotics (Klaverand van der Meer 1993).




C.2.3 BeneficalBenefical EffectsEffects of Probiotics in InflammatoryInflammatory BowelBowel DiseasesDiseases

ProlongedProlonged remissionremission time time of colitis ulcerosa patients by e.g. probiotic E. coli Nissle1917 (EcN) (reviewed by Do et al. 2010).




C 2 4 ReductionReduction inin PainPain PerceptionPerceptionC.2.4 ReductionReduction in in PainPain PerceptionPerception

Lb. reuteri (109 CFU/day) ingestion by rats actedacted onon anan ionion channelchannel inin entericentericsensorysensory nervesnerves resulting in modification of gut motility and pain perceptionsensorysensory nervesnerves resulting in modification of gut motility and pain perception(Kunze et al. 2009).

HT29 cells treated with Lb. acidophilus NCFM resulted in an increaseincrease inin thetheptranscriptiontranscription ofof thethe genegene encodingencoding thethe mumu‐‐opioidopioid receptorreceptor MORMOR11 involved inanalgesis functions and of the cannabinoidcannabinoid receptorsreceptors CBCB22 involved in paintransmission (Rousseaux et al. 2007).




C.2.5 Benefical Effects of Probiotics on on AllergyAllergy(Modulation(Modulation ofof thethe Immune System)Immune System)(Modulation (Modulation ofof thethe Immune System)Immune System)

Lb. rhamnosus (1010 CFU/day) GG reducedreduced thethe frequencyfrequency ofofeczemaeczema in 2in 2‐‐yearyear‐‐oldold childrenchildren (Kalliomaki et al 2001)eczemaeczema in 2in 2 yearyear old old childrenchildren (Kalliomaki et al. 2001).




C.2.6 Modulation Modulation ofof thethe Immune System Immune System by Probiotics

Strengthening of the gut barrier

ChangeChange ofof proteinprotein kinasekinase CC signallingsignalling resulting in amplificationamplification ofof expressionexpression andandChangeChange ofof proteinprotein kinasekinase CC signallingsignalling resulting in amplificationamplification ofof expressionexpression andandredistributionredistribution ofof ZOZO‐‐22 in T84 cells by EcN which protects against destruction of aT84 monolayers by EPEC (Zyrek et al. 2007).

EcN caused in the murine model of dextran sulfate induced colitis not onlyincreasedincreased expressionexpression ofof ZOZO‐‐11 but also protectedprotected thethe animalsanimals by reducing loss ofbody weight and colon shortening as a consequence of increased intestinaly g g qbarrier function (Ukena et al. 2007).

Lb. rhamnosus GG secretes 2 proteins (pp4040,, pp7575) which modifymodify thethe distributiondistribution ofofoccludinoccludin,, EE‐‐cadherincadherin andand ββ‐‐catenincatenin viavia mitogenmitogen activatedactivated proteinprotein (MAP)(MAP) andandproteinprotein kinasekinase CC (PKC)(PKC) (Shet et al. 2008).





Strenghthening of the gut barrier

Th d i il l i h i MUC2MUC2 dd MUC3MUC3 i ii i iiThe predominant ileocolonic human mucins, MUC2 MUC2 andand MUC3, MUC3, transcriptiontranscription isisincreasedincreased in HT29 cells and inhibits adhesion of EPEC by Lb. plantarum 299v (Mack et al. 1999). Other Lactobacillus strains induceinduce MUC3 MUC3 genegene expressionexpressionthrough the action of a 3360 AA3360 AA proteinprotein encoded by sdr (de Vries et al 2006)through the action of a 3360 AA 3360 AA proteinprotein encoded by sdr (de Vries et al. 2006).





flfl ffffAntiAnti‐‐inflammatoryinflammatory effectseffectsThe DD‐‐alaninalanin content of Lb. plantarum NCIMB8826 lipoteichoic acid (LTA) is responsible for ILIL‐‐10 10 inductioninduction:

a dlt mutant with much less D‐Ala in its LTA inducesinduces higherhigher ILIL‐‐1010 andand decreasesdecreasesILIL‐‐1212,, TNFTNF‐‐αα andand IFNIFNγγ productionproduction inin peripheralperipheral bloodblood mononuclearmononuclear cellscellscompared to the WT These effects are TLR2 dependent Furthermore thecompared to the WT. These effects are TLR2 dependent . Furthermore, themutant was also moremore protectiveprotective inin aa murinemurine colitiscolitis modelmodel (Grangette et al.2005).

EcN modulatesmodulates TT‐‐cellcell cyclingcycling andand expansionexpansion via TLR2 signalling (Sturm et al. 2005).

A secretedsecreted factorfactor of EcN supressessupresses thethe TNFTNFαα‐‐inducedinduced ILIL‐‐88 transactivationtransactivation inA secretedsecreted factorfactor of EcN supressessupresses thethe TNFTNFαα inducedinduced ILIL 88 transactivationtransactivation inHCT15 cells however not via NFkB activation, nuclear translocation and DNAbinding or even activation of other transciptional factors (Kamada et al. 2008).




C.2.6 Modulation Modulation ofof thethe Immune System Immune System by Probiotics y

InductionInduction ofof defensinsdefensins

EcN inducesinduces via its flagellaflagella ((FliCFliC)) humanhuman ββ‐‐defensindefensin 22 (hBD(hBD22)) in Caco‐2 cells(Schlee et al. 2007).Not flagella mediated hBDhBD22 synthesissynthesis andand secretionsecretion was reported for Lb.acidophilus PZ1138, Lb. fermentum PZ1162, Pediococcus pentosaceusATCC25745 and the mixture VSL#3 in Caco‐2 cells (Schlee et al. 2008).

F h l ii h l hh l h hh ll hBDhBD22 i d ii d i bFurthermore, also inin healthyhealthy humanhuman volunteersvolunteers hBDhBD22 inductioninduction by non‐flagellated E. coli strains (Symbioflor 2) and its fecal excretion could bedemonstrated (Möndel et al. 2008).





Some probiotics are able to alteralter cytokinecytokine productionproduction in vitro by modulatingcellular signalling.

They eitherblockblock degradationdegradation ofof thethe inhibitorinhibitor IIκκBB byby inhibitinginhibiting thethe ubiquitinationubiquitination ofof thisthisinhibitorinhibitor (in vitro S. boulardii, Sougioultzis et al. 06: SAIFSAIF isis << 11kDa,kDa, heatheatstablestable andand waterwater solublesoluble)orinterfereinterfere withwith proteasomeproteasome functionfunction (in vitro VSL#3, Petrof et al. 2004)orinfluencinginfluencing RelARelA localisationlocalisation via the peroxisome proliferator activatedreceptor γ‐(PPAR‐γ)dependent signal cascade (in vitro Bacteroidesthetaiotaomicron Kelly et al 2004)thetaiotaomicron, Kelly et al. 2004).


D) EffectsEffects on on otherother MOsMOs and EffectiveEffective Components Components of Probiotics

D.1 D.1 FungalFungal InhibitionInhibition

Lb. plantarum MiLAB393 inhibitsinhibits growthgrowth ofof variousvarious mouldsmoulds in vitro by producing33 h ll tih ll ti idid ll (L(L PhPh LL P )P ) dd ll (L(L PhPh tt 44 OHOH LL P )P ) (St33‐‐phenyllacticphenyllactic acidacid cyclocyclo (L(L‐‐PhePhe‐‐LL‐‐Pro)Pro) andand cyclocyclo (L(L‐‐PhePhe‐‐transtrans‐‐44‐‐OHOH LL‐‐Pro)Pro) (Stromet al. 2002).

Lb sanfrancisco CB1 inhibits moulds by aceticacetic acidacid production (Corsetti et alLb. sanfrancisco CB1 inhibits moulds by aceticacetic acidacid production (Corsetti et al.1998).

Lb pentosus TV35b produces a pentocinpentocin that inhibitsinhibits CC albicansalbicans in vitro (Okkers etLb. pentosus TV35b produces a pentocinpentocin that inhibitsinhibits CC.. albicansalbicans in vitro (Okkers etal. 1999).

Several Lactobacillus strains were reported to inhibitinhibit fungifungi in vitro by phenyllacticphenyllacticSeveral Lactobacillus strains were reported to inhibitinhibit fungifungi in vitro by phenyllacticphenyllacticacidacid andand pp‐‐hydroxyphenyllactichydroxyphenyllactic acidacid production (Lavermicocca et al. 2000).



herpesherpes simplexsimplex virusvirus type 1type 1herpesherpes simplexsimplex virusvirus type 1type 1

D.2 AntiAnti‐‐viral viral EffectsEffects

BacteriocinBacteriocin bacSTbacST284284BZBZ produced by Lb. paracasei ST284BZ has antianti‐‐herpesherpessimplexsimplex virusvirus typetype 11 activityactivity Anti‐viral activity was determined by infectingsimplexsimplex virusvirus typetype 11 activityactivity. Anti viral activity was determined by infectingmonkey kidney vero cells in the presence and absence of bacteriocin andsubsequent determination of viral titers obtained (Todorov et al. 2008).



LowLow MolecularMolecular WeightWeight BacteriocinsBacteriocins

D.3 Killing of (pathogenic) bacteria byby BacteriocinsBacteriocins

LowLow MolecularMolecular WeightWeight BacteriocinsBacteriocinsare antimicrobial peptides producedby e.g. many lactobacilli :

Lantibiotics (e.g. nisin, subtilin)Heat stable non‐lantibioticsCyclic antimicrobial peptidesCyclic antimicrobial peptides

BacteriocinsBacteriocins withwith MW > 20 MW > 20 kDakDaProduced e.g. by variousBifidobacterium strains(Cheikhyoussef et al. 2008)



D.4 Bacteriostatic / bactericidal Effects

Many Probiotics produce

OragnicOragnic acidsacids e.g. e.g. lacticlactic acidacid

HH22OO2222 22

DeconjugatedDeconjugated bilebile saltssalts (these show a stronger antimicrobial acitivity comparedto the bile salts synthezied by the host)



D 5 Competitive Exclusion

By competitioncompetition forfor nutrientsnutrients

D.5 Competitive Exclusion

By competitioncompetition forfor nutrientsnutrients

InhibitionInhibition ofof colonizationcolonization by Helicobacter pylori by e.g. Lb. brevis is due to Lb.brevis deamninasedeamninase activity. H. pylori requires arginine, the amount of whichbrevis deamninasedeamninase activity. H. pylori requires arginine, the amount of whichis decreased by the deaminase (Rousseau et al. 2005).



D.5 Competitive Exclusion

By competitioncompetition forfor bindingbinding sitessites

FimbrialFimbrial and nonfimbrialnonfimbrial adhesinsadhesins and other surface structures of Gram‐negative probiotics

AdhesinsAdhesins of Gram‐positive probiotics: for lactobacilli there are 5 classes ofbinding proteins known:

AnchorlessAnchorless housekeepinghousekeeping proteinsproteins (GAPDH, EF(GAPDH, EF‐‐Tu, Tu, GroELGroEL))SurfaceSurface layerlayer proteinsproteins ((SlpASlpA, , CbsACbsA))LPXTGLPXTG‐‐motifmotif proteinsproteins ((MubMub, , MsaMsa))TranspoterTranspoter proteinsproteins ((CnBpCnBp, , MapAMapA))Other Other proteinsproteins ((FbpAFbpA))

b li i d li l id ( i l G 1)Receptors can be gangliotri‐ and gangliotetra‐osylceramides (asialo‐GM1) e.g. for Lb. johnsonii La1 (Neeserr et al. 2000)



Saccharomyces boulardiiD.6 Antitoxin Activity

ii b i i di i d b bi i dToxinToxin can be inactivatedinactivated by probiotics due to

Degradation Degradation ofof toxintoxinS h b l dii t tt hi h d t t i A fSaccharomyces boulardii secretes a proteaseprotease which can destroy toxin A ofClostridium difficile (Castagliuolo et al. 1996).



D 6 A i i A i iD.6 Antitoxin Activity

BindingBinding ofof thethe toxintoxin to the probioticprobiotic cellcell wallwallBinding Binding ofof thethe toxintoxin to the probioticprobiotic cellcell wallwall

Certain Lactobacillus strains are able to bind mycotoxins (e.g. deoxynivalenolby Lb. rhamnosus GG; aflaltoxin by Lb. rhamnosus LC‐705), modulateby b. rhamnosus GG; aflaltoxin by b. rhamnosus C 705), modulateintestinal absorption, increase fecal excretion and therefore lowerlower toxicitytoxicity ininthethe ratrat modelmodel (Gratz et al. 2006; Turner et al. 2008).




Inhibition of toxin expression

Bifidobacterium breve Yakult inhibitsinhibits shigashiga toxintoxin expressionexpression in E. coli (STEC)O157:H7 and protectedprotected allall micemice challanged with STEC whereas 90 % of miceO157:H7 and protectedprotected allall micemice challanged with STEC whereas 90 % of micein the control group died. This effect might be due to high concentration ofaceticacetic acidacid produced by strain Yakult (Asahara et al. 2004). Further 15different probiotic lactobacilli strains inhibitedinhibited shigatoxinshigatoxin 22 expressionexpression viadifferent probiotic lactobacilli strains inhibitedinhibited shigatoxinshigatoxin 22 expressionexpression viaproduction of organicorganic acidsacids at subbactericidal concentrations for EHECO157:H7 (Carey et al. 2008).




InductionInduction ofof a a toxintoxin‐‐specificspecific immune immune responseresponse

S. boulardii induces a specific anti‐C. difficile toxin A IgA immune responsein mice (Qamar et al. 2001).

InterferenceInterference withwith toxintoxin inducedinduced inflammatoryinflammatory signalsignal cascadecascade

S. boulardii interferes with the C. difficile toxin A –induced signal cascadeactivating Erk1/2 and JNK/SAPK pathways (Chen et al. 2006).


Summary 1: Benefical Effects on the Host

Nutritional Nutritional PhysiologyPhysiology

Absorption Absorption ofof MineralsMineralse.g. Cae.g. Ca2+2+, Mg, Mg2+2+ MetabolisationMetabolisation

ofof resistantresistant CarbohydratesCarbohydratesInduktion Induktion ofofGrowth Growth FactorsFactorssese

Deto

Deto

Degradation Degradation ofofantianti‐‐Nutritional Nutritional FactorsFactors

yy

EnzymesEnzymes

Anti

Anti CaCaarar

Disea

Disea

olases

olases

EE

oxificationoxification

Good = Probiotic Bacteria

rcinogenrcinogen

diovascul

diovascul

Enzymes

Enzymes

ACEi

ACEi

leleSalt

Salt Hydr o

HydroEnzym

esEnzym

es

GTS

GTS

IIAA

DefensinDefensin??

hh

[D[D‐‐Ala] Ala] ofof LTALTA

P40, p75 P40, p75 ofof L.rL.r..

FlagellaFlagella3360 AA3360 AAL. L. sppspp..

nesisnesis

Anti

Anti‐‐C

ard

Card BiBi

ncreasedncreasedApoptosis

Apoptosis

?

InductionInductionProinflammatoryProinflammatoryCatokinesCatokines

AntiinflammatoryAntiinflammatoryCytokinesCytokines StrengtheningStrengthening

Gut Gut BarrierBarrier

MucusMucusProductionProduction

TightTightJunctionsJunctions

AA

Modulation Modulation ofof thethe Immune System Immune System andand anti IBD anti IBD ActivityActivity


Summary 2: Direct Effects on Other Microorganisms

AntiAnti‐‐BacterialBacterial EffectsEffectsCompetitionCompetitionCompetitionCompetitionforfor NutrientsNutrientse.g. Arginin

EnzymesEnzymes

CompetitionCompetitionforfor Binding SitesBinding Sites

Killing Killing ofof ((PathogenicPathogenic) ) BacteriaBacteria

BacteriocinsBacteriocins HH22OO22

)) EnzymesEnzymese.g. e.g. DeaminaseDeaminase

AdhesinsAdhesins AceticAcetic AcidAcid, , LacticLactic AcidAcid

DeconjugatedDeconjugatedBileBile SaltsSalts

ion

ion

ee‐‐LL‐‐Pro)

Pro)

ss‐‐44‐‐OH

OH‐‐LL‐‐ProPro

Ant

Ant

HSV

HSVBacBacbacbac

Good = Probiotic Bacteria

alalInhibit

Inhibit

etic

eticAcid

Acid

cyclo

cyclo(L(L‐‐Ph ePhe

(L(L‐‐PhePhe‐‐trans

trans

titi‐‐Viral

Viral

V Type1

V Type1

cteriocincteriocincST284BZcST284BZ

ProteasesProteases CellCell WallWallBindingBinding

OrganicOrganic AcidsAcids

Inhibition of

Funga

Funga

Ace

Ace

Phenyllactic

Phenyllactic

llactic

llactic cyclo

cyclo

AntiAnti‐‐Toxin Toxin EffectsEffects

Toxin‐Expression33‐‐PP

33‐‐Ph

enyl

Phenyl


Juliusspital Institut für Molekulare Infektionsbiologie


FIG. 2. Antifungal compounds isolated from supernatant of L. plantarum MiLAB 393 grown in MRS broth

Strom, K. et al. 2002. Appl. Environ. Microbiol. 68(9):4322-4327

Definition of „good bacteria“ Origin, Effects and … · 2018. 12. 10. · Several...

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