supplementary appendix 20062018 final

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SupplementaryAppendix

TableofContents

Inclusioncriteria

Exclusioncriteria

Studydrugadministration

Drugstorageandsupply

Co-primaryendpoints–definitions

Dosefrequencycommittee(DFC)charter

Supplementarytables

TableS1:Doseandfrequencyallocationaccordingtocohort

TableS2:Doseandfrequencyallocationaccordingtopopulation

TableS3:Adverseeventsbyseverityandtypeinthesafetypopulation(N=37)

TableS4:Safetyparameters(safetypopulation)

TableS5:Clinicalfluorescence-activatedcellsorting(FACS)

TableS6:AICforallmodels(analysispopulation)

TableS7:Estimatedtargetdose/frequency

TableS8:Bestestimatedallowabledoes/frequencyaccordingtoMahalanobisdistanceandjointprobability–Lineardose,quadraticfrequency,interaction(analysispopulation)

TableS9:Bestestimatedallowabledose/frequencyaccordingtoMahalanobisdistanceandjointprobability-Lineardose,quadraticfrequency,interaction(evaluablepopulation)

Supplementalfigures

FigureS1:Eosinophiliaforindividualparticipants(safetypopulation)

FigureS2:TPOantibodiesandTSHreceptorantibodies

FigureS3:Clinicalimagesandimmunohistochemicalstainingofpsoriasisinoneparticipantreceivingtreatmentwithaldesleukin0.32x106IU/m2everythreedays

FigureS4:PercentagechangeofCD19%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS5:PercentagechangeofCD19totalBcellsinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS6:PercentagechangeofCD3%inindividualparticipantsbydoseandfrequency(safetypopulation)

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FigureS7:PercentagechangeofCD3totalTcellsinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS8:PercentagechangeofCD4%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS9:PercentagechangeofCD4total%inindividualpatientsbydoseandfrequency(safetypopulation)

FigureS10:PercentagechangeofCD56%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS11:PercentagechangeofCD56totalNKcellsinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS12:PercentagechangeofCD8%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS13:PercentagechangeofCD8totalinindividualpatientsbydoseandfrequency(safetypopulation)

FigureS14:Percentagechangeoflymphocytescountinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS15:PercentagechangeofTreg%CD4Tcellsinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS16:PercentagechangeofTregabsoluteinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS17:PercentagechangeofTregCD25MeanFIinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS18:PercentagechangeofTregCD25MEFinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS19:PercentagechangeofNon-TregNaïve%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS20:PercentagechangeofNon-TregNaïveabsoluteinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS21:PercentagechangeofNon-TregTemra%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS22:PercentagechangeofNonTregTemraabsoluteinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS23:PercentagechangeofNonTregtotalmemory%inindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS24:PercentagechangeofNonTregtotalmemoryabsoluteinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS25:PercentagechangeofNonTregnaïve/memoryratioinindividualparticipantsbydoseandfrequency(safetypopulation)

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FigureS26:IL-2levelsinindividualparticipantsbydoseandfrequency(safetypopulation)

FigureS27:HbA1c(%)inindividualparticipantsbydoseandfrequency(analysispopulation)

StatisticalAnalysisPlan(SAP)

4

Inclusioncriteria

• Type1diabetes

• 18-70yearsofage

• Durationofdiabeteslessthan60monthsfromdiagnosis

• Writteninformedconsenttoparticipate

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Exclusioncriteria

• Hypersensitivitytoaldesleukinoranyoftheexcipients

• Historyofseverecardiacdisease

• Historyofmalignancywithinthepast5years(withtheexceptionoflocalizedcarcinomaoftheskinthathadbeenresectedforcureorcervicalcarcinomainsitu

• Historyorconcurrentuseofimmunosuppressiveagentsorsteroids.

• Historyofunstablediabeteswithrecurrenthypoglycaemia

• ActiveautoimmuneHyperorhypothyroidism

• Historyoflivevaccinationtwoweekspriortofirsttreatment

• Activeclinicalinfection

• Majorpre-existingorgandysfunctionorpreviousorganallograft

• Femaleswhoarepregnant,lactatingorintendtogetpregnantduringthestudy

• Maleswhointendtofatherapregnancyduringthestudy

• Donationofmorethan500mlofbloodwithin2monthspriortoaldesleukinadministration

• Participationinaprevioustherapeuticclinicaltrialwithin2monthspriortoaldesleukinadministration

• AbnormalECG

• Abnormalfullbloodcount,chronicrenalfailure(Stage3,4,5)and/orevidenceofseverelyimpairedliverfunction(ALT/AST>3xULNatscreening;alkalinephosphataseandbilirubin2xULNatscreening(isolatedbilirubin>2xULNisacceptableifbilirubinisfractionatedanddirectbilirubin<35%)).

• PositiveHBsAgorHepCserologyorHIVtest

• Anymedicalhistoryorclinicallyrelevantabnormalitythatisdeemedbytheprincipalinvestigator/delegateand/ormedicalmonitortomakethepatientineligibleforinclusionbecauseofasafetyconcern.

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Studydrugadministration

Thetrialdrugwillbethestandardcommercialstockofaldesleukin(MarketingAuthorisationHolder:Novartis Pharmaceuticals UK Limited). Aldesleukin is produced by recombinant DNA technologyusinganEscherichiacolistrain,whichcontainsageneticallyengineeredmodificationofthehumaninterleukin2(IL-2)gene.

Thedrugisprovidedaspowderforsolutionforinjectionorinfusion.Forallinjections,aldesleukinisfirst reconstituted under aseptic conditions as per Proleukin® SmPC. Solutions are then furtherdiluted to the required concentration in glucose 5% and the appropriate volume drawn up intosterileU-100insulinsyringes.

The first dose of aldesleukin will be self-administered by participants at the trial site followinginstructionbyanappropriately traineddelegatedmemberof the trial team.Participants can thenadminister further doses either at the trial site or in the community. Alternatively, dosesmay beadministered by an appropriately qualifiedmember of the trial team. The date and time of eachdoseadministeredatthetrialsitewillberecordedinthesourcedocuments.Inthecommunity,thedateandtimeofeachdoseadministeredwillberecorded intheparticipant’sstudydiarythatwillbecome part of the source documents. The study diary, injection sites and count of unusedmedicationwillbecarriedoutateachstudyvisitwhileontreatmenttoconfirmcompliance.

Drugstorageandsupply

Commercialsupplyofaldesleukin,obtainedviausualhospitalroute,willbeusedforthisstudy.VialswillbestoredasperSmPC.at2ºCto8ºC(inarefrigerator)intheoriginalpackageinordertoprotectfrom light. The vials must not be frozen. Vials will be stored securely with access restricted toappropriateindividuals.

Previous (unlicensed)datasupportsstabilityandsterilityof reconstituteddiluted IL-2preparations(reconstitutedwithWaterforInjectionaspertheProleukin®SmPCandfurtherdilutedwithglucose5%) for up to 14 days at 2-8ºCwhen syringes are preparedby qualified health-care professionalsunderasepticconditions.Amaximumshelflifeof7dayswillbeassignedtotheprepareddosesforthisstudy.

The pharmacy at the participating site will dispense aldesleukin for this trial upon receipt of asuitablysignedtrialspecificprescription.Foreachindividualparticipant,multipleindividualdosesofa single concentration of the aldesleukin may be prepared under aseptic conditions foradministration in individual insulin syringes foreach7daysof the treatmentperiod.Alldosesnotusedwithinthe7-dayperiodwillbereturnedtothepharmacyatparticipatingsiteandarecordoftheirdisposalwillbemade.

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Co-primaryendpoints-DefinitionsofTregs,TeffsandCD25expressionwithintheCD3+CD4+fluorescence-activatedcellsorting(FACS)gate

Tregs:Tregs%CD25highCD127lowwithintheCD3+CD4+gate

CD25expressiononTregs:Meanfluorescenceintensity(MFI)ofCD25allophycocyanin(APC)withintheTreg(CD3+CD4+CD25highCD127low)gate

Teffs:Teffpopulations(non-Tregs)accountforalltheotherCD3+CD4+cellsthatarenotdefinedasTregswithintheCD3+CD4+gate:

• Effectormemory%CD45RA−CD62L−

• Centralmemory%CD45RA−CD62L+

• NaïveTeffs%CD45RA+CD62L+

• EffectormemoryRA+(TEMRA)CD45RA+CD62L−

• Centralmemory%+Effectormemory%=TotalMemoryeffectors%

• ChangeintheratioNaïveeffectors%:TotalMemoryeffectors%definestheTeffprimaryendpoint.

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9

10

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Supplementarytables

TableS1DoseandfrequencyallocationaccordingtocohortLEAR

NING

COHORT1

0.09x106IU/m2

0.47x106IU/m2

0.20x106IU/m2

0.32x106IU/m2

14daily 2 2

10daily 2

5daily 2

2daily 2 2

COHORT2

10daily 1

5daily 2

4daily 1 1

3daily 1 2

CONFIRM

ING COHORT3

3daily 5 4

COHORT4

3daily 8

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TableS2Doseandfrequencyallocationaccordingtopopulation.

SAFETYPOPULATION

0.09x106IU/m2

0.47x106IU/m2

0.20x106IU/m2

0.32x106IU/m2

14daily 2 2 0 0

10daily 1 2 0 0

5daily 2 2 0 0

4daily 1 1 0 0

3daily 1 2 5 12

2daily 2 2 0 0

ANALYSISPOPULATION

14daily 1 2 0 0

10daily 1 2 0 0

5daily 2 2 0 0

4daily 1 0 0 0

3daily 1 2 4 8

2daily 1 1 0 0

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TableS3.Adverseeventsbyseverityandtypeinthesafetypopulation(N=37)

TypeofAdverseevent

Category Events Participants

TotalAEs MostcommonAEs(>10%)- Hypoglycaemia- Injectionsiteerythema- Injectionsitenodule

94(13.5%)230(33.1%)234(33.7%)

20(54.1%)35(94.6%)33(89.2%)

Adverseevents- Unexpected- Expectedadverseevents- Related- Unrelated

127(18.3%)567(81.7%)479(69.1)215(31.0%)

34(91.9%)37(100%)

Unexpected Unexpectedadverseeventsbyseverity(N/%)- Mild- Moderate

108(85%)19(15%)

32(86.5%)11(29.7%)

Mostcommon(>10%)unexpectedAEs- Headache- Nasopahryngitis

22(17.3%)18(14.2%)

12(32.4%)16(43.2%)

Unexpectedadverseevents(N/%)byrelatedness- Unrelated- Related

120(94.5%)7(5.5%)

Expected Expectedadverseeventsbyseverity(N/%)- Mild- Moderate

562(99.1%)5(0.9%)

37(100%)3(8.1%)

MostcommonexpectedAEs(>10%AEs)- Hypoglycaemia- Injectionsiteerythema- Injectionsitenodule

94(16.6%)230(40.6%)234(41.3%)

20(54.1%)35(94.6%)33(89.2%)

ExpectedAEs- Related- Unrelated

472(83.2%)95(16.8%)

UnrelatedAEs Unrelatedadverseeventsbyseverity(N/%)- Mild- Moderate

196(91.2%)19(8.8%)

35(94.59%)11(29.73%)

MostcommonunrelatedAEs(>10%)- Headache- Hypoglycaemia

22(10.2%)94(43.7%)

12(32.4%)20(54.1%)

Unrelatedadverseevents(N/%)- Expected- Unexpected

95(44.2%)120(55.8%)

21(56.7%)34(91.9%)

RelatedAEs Drug-relatedAEsaccordingtocohortsCohort1

- Injectionsiteerythema- Injectionsitenodule- Nauseaandvomitingsymptoms

Cohort2- Acne- Eosinophilia- Injectionsiteerythema- Injectionsitenodule

55(49.1%)56(50.0%)1(0.9%)2(2.0%)1(1.0%)50(49.0%)

11(91.7%)10(83.3%)1(8.3%)2(25.0%)1(12.5%)7(87.5%)

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- Malaise- Migraineheadaches

Cohort3- Injectionsiteerythema- Injectionsitenodule- Oropharyngealpain

Cohort4- Astheniccondition- Dryeye- Injectionsiteerythema- Injectionsitenodule- Nasopharyngitis

Nauseaandvomitingsymptoms

47(46.1%)1(1.0%)1(1.0%)68(55.3%)54(43.9%)1(0.8%)3(2.1%)1(0.7%)57(40.1%)77(54.2%)3(2.1%)1(0.7%)

6(75.0%)1(12.5%)1(12.5%)9(100%)9(100%)1(11.1%)3(37.5%)1(12.5%)8(100%)8(100%)3(37.5%)1(12.5%)

Drugrelatedinjectionsitereactionspernumberofdoses

- Injectionsiteerythemapernumberofdose

- Injectionsitenodulepernumberofdose

230/323(71.2%)234/323(72.4%)

DrugrelatedinjectionsitereactionspernumberofdosespercohortCohort1

- Injectionsiteerythemapernumberofdose

- InjectionsitenodulepernumberofdoseCohort2

- Injectionsiteerythemapernumberofdose

- InjectionsitenodulepernumberofdoseCohort3

- Injectionsiteerythemapernumberofdose

- InjectionsitenodulepernumberofdoseCohort4

- Injectionsiteerythemapernumberofdose

- Injectionsitenodulepernumberofdose

55/78(70.5%)56/78(71.8%)50/80(62.5%)47/80(58.8%)68/85(80.0%)54/85(63.5%)57/80(71.3%)77/80(96.3%)

11(91.7%)10(83.3%)7(87.5%)6(75.0%)9(100%)9(100%)8(100%)8(100%)

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TableS4.Safetyparameters(safetypopulation)

Parameter Baseline Followup Normalrange

hsCRP(mg/L)

2.55(±5.72) 1.90(±2.47) ≤3

Sodium(mmol/L)

138.75(±2.32) 138.6(±2.4) 133-146

Potassium(mmol/L)

4.36(±0.25) 4.35(±0.28) 3.5-5.3

Albumincorrectedcalcium(mmol/L)

2.32(±0.09) 2.36(0.13±) 2.08-2.65

Urea(mmol/L)

5.55(±1.6) 5.69(±1.34) 2.5-7.8

Creatinine(umol/L)

72.39(±14.92) 71.3(±11.3) F:44-97M:62-115

Alaninetransaminase(U/L)

21.97(±15.42) 24.9(±22.7) 7-40

Aspartateaminotransferase(U/L)

20.31(±7.23) 22.71(±12.76) ≤34

GGT(U/l)

17.11(±12.04) 20.9(±23.8) F:0-37M:0-72

Alkalinephosphatase(U/L)

74.11(±20.25) 80.8(±23.9) 30-130

TSH(mU/L)

1.55(±0.92) 1.85(±1.06) 0.35-5.50

FreeT4(pmol/L)

15.42(±2.4) 15.06(±1.74) 10.0-19.8

Redbloodcells(109/l) 4.79(±0.42) 4.74(±0.40) F:3.90-5.20M:4.00-5.65

Haemoglobin 144.9(±12.2) 142.0(±13.3) F:120-156M:135-172

Haematocrit(L/L) 0.42(±0.03) 0.41(±0.03) F:0.355-0.455M:0.395-0.505

Meancellhaemoglobin(pg) 30.21(±1.53) 29.93(±1.8) F:27.0-33.5M:27.0-33.5

Redcelldistributionwidth(%) 13.2(±0.54) 13.05(±0.71) 11.0-16.0

Plateletcount(109/l) 224.4(±44.4) 234.5(±51.6) 150-370

Whitebloodcells(109/l) 5.71(±1.76) 5.57(±1.65) 3.90-10.20

Neutrophils(109/l) 3.49(±1.61) 3.36(±1.36) 1.50-7.70

Lymphocytes(109/l) 1.53(±0.37) 1.56(±0.34) 1.50-7.70

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Monocytes(109/l) 0.41(±0.14) 0.42(±0.16) 0.10-0.90

Eosinophils(109/l) 0.16(±0.16) 0.15(±0.10) 0.10-0.90

Basophils(109/l) 0.05(±0.04) 0.04(±0.02) 0.00-0.20

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TableS5.ClinicalFACSanalysis(safetypopulation)

Parameter Baseline Followup

CD19% 12.2(±3.9) 12.3(±4.3)

CD19TotalBcells(*109/L) 0.2168(±0.17) 0.19(±0.08)

CD3% 74.8(±6.1) 75.1(±5.6)

CD3TotalTcells(*109/L) 1.16(±0.30) 1.19(±0.27)

CD4% 47.1(±7.5) 48.1(±8.1)

CD4Total(*109/L) 0.73(±0.23) 0.76(±0.21)

CD56% 12.4(±5.8) 12.1(±5.6)

CD56(*109/L) 0.19(±0.09) 0.18(±0.08)

CD8% 25.4(±6.6) 24.9(±6.7)

CD8Total(*109/L) 0.39(±0.13) 0.39(±0.14)

Lymphocytescount(*109/L) 1.53(±0.37) 1.58(±0.33)

Treg%CD4Tcells 7.13(±1.66) 7.28(±1.87)

TregAbsoulte(*109/L) 0.05(±0.02) 0.05(±0.02)

TregCD25MeanFI 2401.8(±794.7) 2292.4(±748.1)

TregCD25MEF 2863.8(±754.9) 2558.7(±520.5)

Non-Treggatedpanel Naïve% 42.4(±15.1) 39.6(±15.3)

Naïveabsolute(*109/L) 0.28(±0.14) 0.28(±0.15)

Temra% 1.01(±1.95) 0.7(±1.7)

Temraabsolute(*109/L) 0.006(±0.013) 0.004(±0.01)

Totalmemory% 56.6(±15.9) 59.9(±15.9)

Totalmemoryabsolute(*109/L) 0.38(±0.16) 0.41(±0.14)

Naïve/memoryratio 0.94(±0.84) 0.84(±0.79)

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TableS6.AICforallmodels(analysispopulation)

Model TregAIC CD25AIC TeffAIC

Lineardose,linearferquency 245.8 235.93 243.89

Quadraticdose,linearfrequency

243.55 236.14 245.85

Lineardose,quadraticfrequency

245.72 219.52 240.79

Quadraticdose,quadraticfrequency

245.27 215.0 238.63

Logdose,linearfrequency 244.26 235.04 243.95

Lineardose,logfrequency 244.27 228.82 244.73

Logdose,logfrequency 242.88 228.29 244.71

Lineardose,quadraticfrequency,interaction

246.66 213.44 236.82

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TableS7.Estimatedtargetdose/frequency

Model Bestdose

DoseSE

DoseCI

BestFrequency

FrequencySE

FrequencyCI

Lineardose,linearfrequency

-0.082 1.927 -3.858,3.694

-2.9 31.9 65.4,59.6

Quadraticdose,linearfrequency

0.133 0.154 -0.17,0.435

-0.7 10.1 -20.4,19.0

Lineardose,quadraticfrequency

0.240 0.114 0.017,0.463

3.0 0.5 1.7,3.5

Quadraticdose,quadraticfrequency

0.186 0.048 0.093,0.279

2.6 0.5 1.7,3.5

Logdose,linearfrequency

0.067 0.420 -0,757,0.891

-2.5 27.4 -56.3,51.2

Lineardose,logfrequency

0.170 0.482 -0.775,1.115

2.4 2.7 -2.8,7.7

Logdose,logfrequency

0.168 0.252 -0.327,0.662

2.5 2.2 -1.8,6.9

Lineardose,quadraticfrequency,interaction

0.239 0.126 -0,008,0.485

2.9 0.8 1.4,4.4

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TableS8.Bestestimatedallowabledoes/frequencyaccordingtoMahalanobisdistanceandjointprobability–Lineardose,quadraticfrequency,interaction(analysispopulation)

Dose Frequency MahalanobisDistance

JointProbability

0.26

3 0.044 0.742

0.25 3 0.062 0.7370.27 3 0.111 0.7160.24 3 0.134 0.7020.28 3 0.186 0.661

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TableS9.Bestestimatedallowabledose/frequencyaccordingtoMahalanobisdistanceandjointprobability-Lineardose,quadraticfrequency,interaction(evaluablepopulation)

Dose Frequency MahalanobisDistance

Joint

Probability

0.21 3 0.124 0.696

0.22 3 0.12 0.694

0.2 3 0.16 0.663

0.23 3 0.151 0.658

0.19 3 0.213 0.603

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SupplementaryFigures

FigureS1.Eosinophiliaforindividualparticipants(safetypopulation)

Oneparticipantonaldesleukin0.47x106IU/m2every4daysdevelopedtransienteosinophilia(red);pre-existingeosinophiliaresolvedintwoparticipantsreceiving0.32x106IU/m2every3days(purple),and0.09x106IU/m2every2days(blue),respectively.

1 2 3 4 5 6 7 8 9 10 11 120.0

0.5

1.0

1.5

Visit number

Eosi

noph

ils (1

09 /L)

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FigureS2.TPOantibodiesandTSHreceptorantibodies.Datashownforindividualpatientsinsafetypopulation.

NoparticipantdevelopeddenovoTPOantibodies(A)orTSHreceptorantibodies(B)duringthetrial.Tickedlineindicatesupperlimitofnormalrange.

1 1210

100

1000

Visit number

Log1

0 TP

O a

ntib

odie

s (IU

/ml)

1 120.0

0.5

1.0

1.5

Visit number

TSH

rece

ptor

ant

ibod

ies

(IU/L

)

A B

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FigureS3.Clinicalimagesandimmunohistochemicalstainingofpsoriasisinoneparticipantreceivingtreatmentwithaldesleukin0.32x106IU/m2everythreedays.

(A-F)Clinicalimages:Therewasnochangeinplaquesize,andplaquenumber(threelesionsonrightelbow, abdomen, and left ear; data not shown) during treatment. Image A to E show plaque onelbowat subsequentdosing visits 3 (A), 4 (B), 8 (C), 9 (D), and10 (E).At followup, anadditionalplaquewasobservedontheabdomen,whilethesizeofpre-existingplaquesremainedstable(F).

(G-L)Immunohistochemicalstaining:skinpunchbiopsiesofthepsoriaticplaqueonrightelbowweretakenatvisit10andatfollowupvisit12.HistopathologysectionswerestainedforCD3before(G)andafter (J) treatment,CD4before(H)andafter (K)treatment,andFOXP3before(I)andafter (L)treatment.Imagesareshown10xenlarged.CD3,CD4andFOXP3positivecellswerecountedinfiverandomfieldsofviewat100xmagnification.FOXP3toCD3ratio(0.17before,0.17after)andFOXP3toCD4ratio(0.13before,0.15after)remainedunchanged.

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FigureS4:PercentagechangeofCD19%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS5:PercentagechangeofCD19totalBcellsinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS6:PercentagechangeofCD3%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS7:PercentagechangeofCD3totalTcellsinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS8:PercentagechangeofCD4%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS9:PercentagechangeofCD4totalinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS10:PercentagechangeofCD56%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS11:PercentagechangeofCD56totalNKcellsinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS12:PercentagechangeofCD8%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS13:PercentagechangeofCD8totalinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS14:Percentagechangeoflymphocytescountinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS15:PercentagechangeofTreg%CD4Tcellsinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS16:PercentagechangeofTregabsoluteinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS17:PercentagechangeofTregCD25MeanFIinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS18:PercentagechangeofTregCD25MEFinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS19:PercentagechangeofNon-TregNaïve%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

41

FigureS20:PercentagechangeofNon-TregNaïveabsoluteinindividualparticipants(eachline)bydoseandfrequency(safetypopulation

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FigureS21:PercentagechangeofNon-TregTemra%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS22:PercentagechangeofNon-TregTemraabsoluteinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

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FigureS23:PercentagechangeofNon-Tregtotalmemory%inindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

45

FigureS24:PercentagechangeofNon-Tregtotalmemoryabsoluteinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

46

FigureS25:PercentagechangeofNon-Tregnaïve/memoryratioinindividualparticipants(eachline)bydoseandfrequency(safetypopulation)

47

FigureS26:IL-2levelsinindividualparticipantsbydoseandfrequency(safetypopulation)

0.001

0.01

0.1

1

10

0 5 10 15

0.09 x106 IU/m2, 2−day

0.001

0.01

0.1

1

10

0 10 20 30

0.09 x106 IU/m2, 3−day

0.001

0.01

0.1

1

10

0 10 20 30 40

0.09 x106 IU/m2, 4−day

0.001

0.01

0.1

1

10

0 10 20 30 40

0.09 x106 IU/m2, 5−day

0.001

0.01

0.1

1

10

0 20 40 60 80 100

0.09 x106 IU/m2, 10−day

0.001

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0.1

1

10

0 20 40 60

0.09 x106 IU/m2, 14−day

0.001

0.01

0.1

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0 10 20 30

0.20 x106 IU/m2, 3−day

0.001

0.01

0.1

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0.32 x106 IU/m2, 3−day

0.001

0.01

0.1

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0.47 x106 IU/m2, 2−day

0.001

0.01

0.1

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10

0 10 20 30

0.47 x106 IU/m2, 3−day

0.001

0.01

0.1

1

10

0 10 20 30 40

0.47 x106 IU/m2, 4−day

0.001

0.01

0.1

1

10

0 10 20 30 40 50

0.47 x106 IU/m2, 5−day

0.001

0.01

0.1

1

10

0 10 20 30 40

0.47 x106 IU/m2, 10−day

0.001

0.01

0.1

1

10

0 20 40 60

0.47 x106 IU/m2, 14−day

IL−2

(IU

/ml)

Days

48

FigureS27:HbA1c(%)inindividualparticipants(eachline)bydoseandfrequency(analysispopulation)

5

7

9

11

13

−10 0 10 20 30 40 50

0.09 x106 IU/m2, 2−day

5

7

9

11

13

−10 0 10 20 30 40 50 60

0.09 x106 IU/m2, 3−day

5

7

9

11

13

−20 −10 0 10 20 30 40 50 60 70

0.09 x106 IU/m2, 4−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70

0.09 x106 IU/m2, 5−day

5

7

9

11

13

−20 0 20 40 60 80 100 120

0.09 x106 IU/m2, 10−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70 80 90

0.09 x106 IU/m2, 14−day

5

7

9

11

13

−20 −10 0 10 20 30 40 50 60

0.20 x106 IU/m2, 3−day

5

7

9

11

13

−30 −20 −10 0 10 20 30 40 50 60

0.32 x106 IU/m2, 3−day

5

7

9

11

13

−10 0 10 20 30 40 50

0.47 x106 IU/m2, 2−day

5

7

9

11

13

−10 0 10 20 30 40 50 60

0.47 x106 IU/m2, 3−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70

0.47 x106 IU/m2, 4−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70 80

0.47 x106 IU/m2, 5−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70

0.47 x106 IU/m2, 10−day

5

7

9

11

13

−10 0 10 20 30 40 50 60 70 80 90

0.47 x106 IU/m2, 14−day

HbA

1c (%

)

Days