Introduction

1
Peptide labelling with fluor-19 and Peptide labelling with fluor-19 and fluor-18 : fluor-18 : OVA 323-339 and OVA 257-264 for the OVA 323-339 and OVA 257-264 for the study of dendritic cells migration by study of dendritic cells migration by PET Scan PET Scan Laïque Salma [1] , Lazarova Elena [2] , Monclus Michel [1] , Jacquemotte Françoise [3] , Communi David [4] , Flamand Véronique [2] , Goldman Serge [1] [1] PET/Biomedical Cyclotron Unit, ULB, Hopital Erasme, Route de Lennik 808, 1070 Anderlecht, BELGIUM [2] Institute for Medical Immunology, ULB, Rue A. Bolland, B 6041 Gosselies, BELGIUM [3] Département des Substances Naturelles, Institut Meurice, 50 Avenue Emile Gryzon, 1070 Anderlecht, BELGIUM [4] I.R.I.B.H.M. , Bldg. C, ULB, Route de Lennik 808, 1070 Anderlecht, BELGIUM OVA 323-339 : NH 2 – Ile – Ser – Gln – Ala – Val – His – Ala – Ala – His – Ala – Glu – Ile – Asn – Glu – Ala – Gly – Arg – OH Introduction OVA 257-264 : NH 2 – Ser – Ile – Ile – Asn – Phe – Glu – Lys – Leu – OH Dendritic cells (DC) migration is important to evaluate in immunotherapy protocoles. Ovalbumin 323-339 and Ovalbumin 257-264 are known to be recognised by MHC II complex of DC and to induce T lymphocytes proliferation (1). We propose to radiolabel these fragment with [ 18 F]fluoroethyl moiety to image by PET pulsed DC injected into mice. We here present the biological results of the non radioactive tracer, N-[ 19 F]fluoro-ethyl-OVA 323-339 (FEthOVA 323-339 ) and our preliminary tests of radiolabelling on OVA 257-264 . 0.0 0.2 0.4 0.6 0.8 0.0 2.0e+4 4.0e+4 6.0e+4 8.0e+4 1.0e+5 1.2e+5 1.4e+5 DC-W ild Type D C -LPS activated W T-O VA-F 19 loaded W T-O VA loaded LPS act.-O VA-F 19 loaded LPS act.-O VA loaded Biology Non radioactive peptides Peptide synthesis : on solid phase Principle of SPPS In vitro comparaison between OVA 323-339 and FEtOVA 323-339 , measured by 3 H-thymidin incorporation after 48h MLR FEtOTs synthesis (2) Coupling reaction Conclusions: FEthOVA 323-339 holds antigenic properties of OVA 323-339 . The F-18-labeled form of this peptide should therefore be evaluated for PET imaging of pulsed-DC migration. Radioactive peptides 1,2-ditosyloxyethane protected OVA 257-264 NH-CH 2 -CH 2 - 18 F (+ 18 FEtOTs) OVA 257-264 NH-CH 2 -CH 2 - 18 F 18 FEtOTs protected OVA 257-264 + basic conditions TFA/TIS/ thioanisole 18 FEtOTs (+ 18 F - + 1,2-ditosyloxyethane) 18 F - /K 2.2.2. / K 2 CO 3 purification 18 FEtOT s QMA Protected peptide : OVA 257-264 : NH 2 – Ser – Ile – Ile – Asn (Ttr) – Phe – Glu (OtBu) – Lys (Boc) – Leu – OH X : protecting groups Validation by analyitical HPLC LC-MS OVA 257-264 NH-CH 2 -CH 2 - 18 F scavengers suppression preparative HPLC purificat ion protected OVA 257-264 NH-CH 2 -CH 2 - 18 F (1) Wilson AA, Dasilva JN, Houle S. Synthesis of two radiofluorinated cocaine analogues using distilled 2-[ 18 F]fluoroethylbromide. Appl. Radiat. Isot. 46(8):765-770,1995 (2) Roberston JM, Jensen PE, Evavold BD. DO11.10 and OT.II T cells recognise a C-terminal Ovalbumin 323-339 epitope. J. Immunol. 164:4706- 4712, 2000 References : [email protected] : 0032-2-555.47.11 Contacts

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Peptide labelling with fluor-19 and fluor-18 : OVA 323-339 and OVA 257-264 for the study of dendritic cells migration by PET Scan. Laïque Salma [1] , Lazarova Elena [2] , Monclus Michel [1] , Jacquemotte Françoise [3] , Communi David [4] , Flamand Véronique [2] , Goldman Serge [1] - PowerPoint PPT Presentation

Transcript of Introduction

Page 1: Introduction

Peptide labelling with fluor-19 and fluor-18 : Peptide labelling with fluor-19 and fluor-18 : OVA 323-339 and OVA 257-264 for theOVA 323-339 and OVA 257-264 for the

study of dendritic cells migration by PET Scanstudy of dendritic cells migration by PET Scan

Laïque Salma [1], Lazarova Elena [2], Monclus Michel [1], Jacquemotte Françoise [3], Communi David [4], Flamand Véronique [2], Goldman Serge [1]

[1] PET/Biomedical Cyclotron Unit, ULB, Hopital Erasme, Route de Lennik 808, 1070 Anderlecht, BELGIUM[2] Institute for Medical Immunology, ULB, Rue A. Bolland, B 6041 Gosselies, BELGIUM[3] Département des Substances Naturelles, Institut Meurice, 50 Avenue Emile Gryzon, 1070 Anderlecht, BELGIUM[4] I.R.I.B.H.M. , Bldg. C, ULB, Route de Lennik 808, 1070 Anderlecht, BELGIUM

OVA 323-339 : NH2– Ile – Ser – Gln – Ala – Val – His – Ala – Ala – His – Ala – Glu – Ile – Asn – Glu – Ala – Gly – Arg – OH

Introduction

OVA 257-264 : NH2 – Ser – Ile – Ile – Asn – Phe – Glu – Lys – Leu – OH

Dendritic cells (DC) migration is important to evaluate in immunotherapy protocoles. Ovalbumin323-339 and Ovalbumin257-264 are known to be recognised by MHC II complex of DC and to induce T lymphocytes proliferation (1).We propose to radiolabel these fragment with [18F]fluoroethyl moiety to image by PET pulsed DC injected into mice. We here present the biological results of the non radioactive tracer, N-[19F]fluoro-ethyl-OVA323-339 (FEthOVA323-339) and our preliminary tests of radiolabelling on OVA257-264.

0.0 0.2 0.4 0.6 0.8

0.0

2.0e+4

4.0e+4

6.0e+4

8.0e+4

1.0e+5

1.2e+5

1.4e+5

DC-Wild TypeDC-LPS activatedWT-OVA-F19 loadedWT-OVA loadedLPS act.-OVA-F19 loadedLPS act.-OVA loaded

Biology

Non radioactive peptides

Peptide synthesis : on solid phase

Principle of SPPS In vitro comparaison between OVA323-339

and FEtOVA323-339, measured by 3H-thymidin incorporation after 48h MLR

FEtOTs synthesis (2)

Coupling reaction

Conclusions: FEthOVA323-339 holds antigenic properties of OVA323-339. The F-18-labeled form of this peptide should therefore be evaluated for PET imaging of pulsed-DC migration.

Radioactive peptides

1,2-ditosyloxyethane

protected OVA257-264NH-CH2-CH2-18F (+ 18FEtOTs)

OVA257-264NH-CH2-CH2-18F

18FEtOTs protected OVA257-264+

basic

conditions

TFA/TIS/thioanisole

18FEtOTs (+18F- + 1,2-ditosyloxyethane)18F-/K2.2.2./K2CO3

purification

18FEtOTs

QMA

Protected peptide :

OVA 257-264 : NH2 – Ser – Ile – Ile – Asn (Ttr) – Phe – Glu (OtBu) – Lys (Boc) – Leu – OH X : protecting groups

Validation byanalyitical HPLCLC-MS

OVA257-264NH-CH2-CH2-18F

scavengers suppression

preparative HPLC

purification

protected OVA257-264NH-CH2-CH2-18F

(1) Wilson AA, Dasilva JN, Houle S. Synthesis of two radiofluorinated cocaine analogues using distilled 2-[18F]fluoroethylbromide. Appl. Radiat. Isot. 46(8):765-770,1995(2) Roberston JM, Jensen PE, Evavold BD. DO11.10 and OT.II T cells recognise a C-terminal Ovalbumin 323-339 epitope. J. Immunol. 164:4706-4712, 2000

References : [email protected] : 0032-2-555.47.11

Contacts

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