Rev. Bras. Hematol. Hemoter. vol.39 número4

w w w . r b h h . o r g

Revista

Brasileira

de

Hematologia

e

Hemoterapia

Brazilian

Journal

of

Hematology

and

Hemotherapy

Original

article

Lack

of

association

between

Kidd

blood

group

system

and

chronic

kidney

disease

Tiago

Verri

Capriolli,

Jeane

Eliete

Laguila

Visentainer,

Ana

Maria

Sell

UniversidadeEstadualdeMaringá(UEM),Maringá,PR,Brazil

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Articlehistory:

Received27March2017 Accepted30May2017 Availableonline28June2017

Keywords:

Kiddbloodgroupsystem Chronickidneyfailure Serotyping

Bloodureanitrogen Ureatransporter

a

b

s

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t

Background:TheKiddbloodgroupsystemhasthreeantigens,Jka_,Jkb_{andJk3,foundonred}

bloodcellsandonendothelialcellsoftheinnerliningofbloodvesselsintherenalmedulla. Theseare knownasureatransporter B(UT-B).Researchershavefoundthatindividuals carryingtheJk(a−b−)orJk-null(UT-Bnull)phenotypeshavealowerurine-concentrating capabilityandriskofsevererenalimpairment.Thisstudyevaluatedthedistributionofthe KiddphenotypesinpatientswithchronickidneydiseaseandapossibleassociationofKidd antigenswiththedevelopmentofrenaldisease.

Methods:Jka_andJkb_{antigenswerephenotypedusingthegelcolumnagglutinationtest}

(ID-cardsBio-RAD)in197patientswithchronickidneydiseaseand444blooddonors,asthe controlgroup.Thephenotypeandantigenfrequenciesbetweenpatientsandcontrolswere evaluatedusingtheChi-squaremethodwithYatescorrectionandlogisticregressionafter adjustmentsforgenderandage.

Results:NodifferenceswereobservedbetweentheKiddphenotypesfrequencydistribution betweenpatientswithchronickidneydiseaseandblooddonors[Jk(a−b+)=22.3%and27.2%; Jk(a+b−)=30.5%and24.3%;Jk(a+b+)=47.25%and48.4%,respectively].

Conclusion: ThedistributionofKiddphenotypesfoundinthestudiedpopulationisexpected forCaucasians;Jka_andJkb_{antigensandphenotypeswerenotfoundtoberelatedto}

sus-ceptibilityforchronickidneydisease.

©2017Associac¸ ˜aoBrasileiradeHematologia,HemoterapiaeTerapiaCelular.Published byElsevierEditoraLtda.ThisisanopenaccessarticleundertheCCBY-NC-NDlicense (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

Theredbloodcell(RBC)membranecontainsmanyanchored surfaceproteinsandproteinsthatcrossthelipidbilayer car-ryingdifferentbloodgroupantigens.Currently,36systems1_of

RBCgroupshavebeendescribedaccordingtotheInternational

∗ _{Correspondingauthorat:UniversidadeEstadualdeMaringá(UEM),Av.Colombo,5790,blocoT20,87020-900Maringá,PR,Brazil.}

E-mails:anamsell@gmail.com,amsell@uem.br(A.M.Sell).

SocietyofBloodTransfusion(ISBT)(http://www.isbtweb.org). Amongthem,theKiddbloodgroupsystem(JK;ISBT009)has beenrecognizedasclinicallyimportantsinceitsidentification in1951.2

AntigensoftheKiddbloodgroupsystemareexpressedon type3glycoproteins, alsoknownas theurea transporterB

http://dx.doi.org/10.1016/j.bjhh.2017.05.007

(UT-B).Thisproteincontains389aminoacidsandpassesten timesthroughthelipidbilayerwithboththeNterminusandC terminusbeingintracellular.Threeantigenshavebeenfound (Jka_,Jkb_{andJk3)ontheneighboringfourthextracellular} loop-ingandthreephenotypes,Jk(a+b−),Jk(a−b+),andJk(a+b+), arecommonamongdifferentpopulations.TheJk(a−b−) phe-notypeis rarein mostpopulations. Itwas first foundin a FilipinaofSpanish andChineseancestrywiththe antibod-iesusuallybeingdetectedaftertransfusionsorpregnancies. Afterimmunization,anti-Jk3canbefoundinpatientswith theJk(a−b−)recessivephenotype,causingacuteanddelayed hemolytictransfusionreactions.3,4

TheJkglycoproteiniscodedbytheSolutecarrierfamily14, memberA1(SLC14A1)gene,amemberoftheurea-transporter genefamily,locatedonchromosome18q12-q21andorganized into11exons.Thetwomajorcodominantallelesofthegene, JK*AandJK*B,havesimilar frequenciesinCaucasian popu-lations(0.51and0.49,respectively)anddefinetheJka_andJkb antigens,respectively.TheJka_/Jkb_{polymorphismisdefinedby} 838A>Ginexon9(Asp280Asnsubstitution),theothertwo sin-glenucleotidepolymorphisms(SNPs)causenochangestothe aminoacidsequence.5

The Kidd antigens or UT-B are expressed on RBCs and theendotheliumofthevasarectaandepithelialsurfacesof therenalinnermedulla,aswellasinnon-renaltissuesand endothelialcells.6–8 _{TheKiddproteinisamajortransporter}

of urea across the RBC membrane and this rapid process helps maintainthe osmoticstability of the cell. Null phe-notypeindividuals lackthis transporter.WithintheKidney, theureatransporterenablestherenalmedullatomaintaina highconcentrationofbothureaandurine,aswellasconserve water.IndividualswiththeJk(a−b−)phenotypehavelower urine-concentratingability.9–11 _{InUT-B-nullmice,long-term}

UTBdeficiencywasassociatedtosevererenaldysfunctionand structuraldamage.12_{UT-Bisoformsarealsoimportantin}

sev-eralcellularfunctions,includingureanitrogensalvageinthe colon,nitricoxidepathwaymodulationinthehippocampus, andnormalizationofthecardiacconductionsystem.13

Chronic kidney disease (CKD) is a major public health problem,definedasabnormalitiesofkidneystructureand/or function,presentforatleastthreemonthswithimplications on health.14 _{The adverse outcomes of the disease include}

lossofkidneyfunction,cardiovasculardiseaseandpremature death.Besidesenvironmentalfactors,geneticabnormalities arealsoinvolved15_{includingvariationsintheMYH9(encoding}

non-muscle myosin IIAheavy chain),16 _APOL1

(apolipopro-teinL1),17–19_{NPHS1(nephrin),}20_{andSHROOM3(shroomfamily}

member3)21_genes.

TherelationshipbetweentheKiddantigensand chronic kidneydiseaseremainsunknown.Therefore,theaimofthis studywastoinvestigatethedistributionofKiddphenotypes inpatientswith thechronic kidney disease anda possible associationofKiddantigenswiththedevelopmentofrenal disease.

Methods

Theethicaland methodological aspectsofthis study were approved by the Research Ethics Committee on Human

Beings from the Maringa State University (COPEP-UEM # 1.141.385/2014,CAAE43117115.0.0000.0104,accordingtothe ResolutionoftheBrazilianCouncilonHealth-CNS466/12.

Subjects

Thisretrospectivecase–controlstudyenrolled197unrelated patients with chronic kidney disease (CKD group) and 444 unrelated blood donors as a control group, living in the same geographical area as the patients. The individuals were attended and immunophenotypedbetween 2013 and 2015attheRegional BloodBank ofPatoBranco,southwest regionofParana(locatedinthesouthernregionofBrazilat 26◦₁₃′₄₆′′_–09′′_Sand52◦₄₀′₁₆′′_–09′′_W).

Serologictests

RedbloodcellphenotypingforKiddbloodgroupsystemswas performedonagelcard (ID-PerfilII–k-Kpa_-Kpb_-Jka_-Jkb_-ctl) usingmonoclonalantibodiesaccordingtothemanufacturer’s instructions (Diamed ID-Cards, DiaMed® AG, Switzerland). RBCsweresuspendedinBromelinsolution(BioRadID-Diluent 1)atafinalconcentrationof1:21or5%.

Statisticalanalysis

Theantigenandphenotypefrequencieswereestimatedand the data was tested for their fit to the Hardy–Weinberg equilibrium22 _{by calculating the expected frequencies of}

the genotypes and comparing them with the observed values. The Student’s t-test was used to compare differ-encesbetweengroups.Statisticalcomparisonsbetweenthese groups were performedand the estimatedrisk of develop-ing CKD in individuals who have genetic polymorphisms was calculated by determining the Odds Ratio (OD) with a 95% of confidence interval (CI) adjusted for gender and age. Association tests were carried out to identify the codominant, dominant, recessive, overdominant and log-additive geneticinheritance models. p-Values≤0.05 by the Chi-square test with Yates correction and logistic regres-sion were considered statisticallysignificant. All statistical analyses were performed using the software OpenEpi pro-gramVersion2.3.1(http://www.openepi.com)andSNPStats23

(http://bioinfo.iconcologia.net/index.php).

Results

The Kidd phenotype frequency distribution in the stud-ied populations was in Hardy–Weinberg equilibrium (p -value=0.48:CKDandp-value=0.51:controls).

Table1–CharacteristicsoftheCKDpatientandcontrolindividuals.

Patients(n=197) Controls(n=444)

Total(>45yearsold) Total <45yearsold >45yearsold

n=197 n=444 n=349 n=94

n(%) n(%) n(%) n(%)

Gendera

Male 110(55.8) 219(49.3) 163(46.7) 56(59.6)

Female 87(44.2) 225(50.7) 186(53.3) 38(40.4)

Age(years)

Mean 62.8± 13.9 30.6 ± 11.1b _{31.1± 7.2}b _{52.5± 5.4}b

Ethnicgroup

Caucasian 104(100) 437(98.4) 342(98.0) 94(100)

Black 0 6(1.3) 6(1.7) 0

Mulatto 0 1(0.3) 1(0.28) 0

a _{p-Value>0.05(between-groupcomparisonby}2_test).

b _{p-Value<0.0001(betweenpatientsandcontrols.Student’st-test).}

Table2–DistributionofJkantigenandphenotypefrequenciesinCKDpatientsandcontrols(totaland>45yearsold).

CKDPatients

n=197

n(%)

Control Total(n=444)

n(%)

Control >45yearsold

(n=94)

n(%)

Phenotypes

Jk(a−b+) 44(22.3) 121(27.2)a _29(30.8)e

Jk(a+b−) 60(30.5) 108(24.3)b _23(24.5)f

Jk(a+b+) 93(47.2) 215(48.4)c _42(44.7)g

Jkantigens

Jka _{213(54.0) 431(49.0)}d _90(48.0)h

Jkb _{181(46.0) 457(51.0) 98(52.0)}

a_{p-Value=0.32;}b_{p-Value=0.55;}c_p-Value=1;d_{p-Value=0.16(CKDpatientsvs.controls−totalofsamples).}

e_{p-Value=0.55;}f_{p-Value=0.18;}g_p-Value=1;h_{p-Value=0.16(CKDpatientsvs.controls>45yearsold).}

(n=94)foranalyses,consideringtheminimumagereported bypatients.Inthisgroup,themeanagewas52.54±5.36years (p-value<0.0001),and58.9%were malesand 41.1%females withallofthembeingself-declaredCaucasians.

Thedistributionsofantigenandphenotypefrequenciesfor theKiddbloodgroupsystemsinCKDpatientsandcontrols (totalgroupandgroup>45 yearsold)areshowninTable2. ThephenotypefrequenciesfortheKiddbloodgroupsystem inCKDpatientswere22.3%forJk(a−b+),30.5%forJk(a+b−) and47.2%forJk(a+b+).Forcontrols(totaland>45yearsold, respectively)thephenotypefrequencieswere27.2%and30.8% forJk(a−b+), 24.3%and 24.5%forJk(a+b−),and 48.4%and 44.7%forJk(a+b+).Thenullphenotypewasnotobservedin anyofthegroups.

DifferencesindistributionsoftheJka_andJkb_antigenand phenotypefrequencieswerenotobservedwhencaseand con-trolgroupswerecomparedaccordingtoinheritancemodelsor afterlogisticregressionstratifiedbyage.

Discussion

Inthisassociation study,patientswithCKDon hemodialy-siswereanalyzedinordertoinvestigatethedistributionof JKphenotypesinpatientswithCKDandapossibleinfluence

ofJka _andJkb_{antigensandphenotypesinthedevelopment} ofrenaldisease.However,differencesbetweenJka_orJkb anti-genandphenotypefrequencieswerenotobservedbetween patientsandcontrols.

controlgroupwithanageofunder45arepossiblecandidates fordevelopingrenaldysfunctioninthefuture.Ontheother hand,incase–controlstudies,ideallythereshouldbeatleast onecontrolpercase,24 _{andthecontrolgroupofindividuals}

olderthan45yearshadfewerpatientsthantheCKDGroup. However,theJka _orJkb _{antigenand phenotypefrequencies} inboth control groupsof this study were similar tothose frequenciesfoundinanotherBrazilianpopulation,25_thereby

validatingtheresultsinthisstudy.

Theurea transporter in the kidney enables its medulla to maintain a high concentration of both urea and urine, as well as conserve water. Individuals with the Jk(a−b−) phenotypehad lowerurine-concentratingability.Inanimal models,long-termUTBdeficiencywasassociatedwithsevere renaldysfunctionandstructuraldamage.Ontheotherhand, geneticvariationsoftheSLC14A1genewereassociatedwith bladder cancer and,some genotypes were associated with highermorbidity,26–29 _{andrejectionafterrenaltransplant.}30

However,wedidnotfindanynullallelesinCKDpatientsor controlsthatcouldbeidentifiedasariskfactorforthedisease. Inthis study,morethan 50% ofthe CKDpatients were homozygous carrying either Jk(a+b−) or Jk(a−b+) pheno-types,inagreementwiththeresultsreportedintheliterature forCaucasians.25_{Furthermore,nodifferenceswereobserved}

betweenpatientsandcontrolsregardingthedistributionofJka orJkb_{antigenandphenotypefrequencies.Thus,Kidd} pheno-typeswerenotassociatedtoCKD.

Conclusion

ThedistributionofKiddphenotypesfoundinthestudied pop-ulationwasasexpectedforCaucasiansandtherefore,Jka_and Jkb_{antigensandphenotypeswerenotfoundtobeassociated} tothedevelopmentofCKD.

Conflicts

of

interest

Theauthorsdeclarenoconflictsofinterest.

Acknowledgements

Theauthorsappreciatetheparticipationandcooperationof allvolunteers(patientsandcontrols),aswellasoftheRegional BloodBankofPatoBrancoandHEMEPAR-Centrode Hematolo-giaeHemoterapiadoParaná,andImmunogeneticsLaboratory (LIG-UEM).

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