Letters in Peptide Science

, Volume 6, Issue 1, pp 51–59 | Cite as

Synthesis of lipopeptides of the immunodominant epitope hMBP(83–99) containing amide or C-C bond linked hydrophobic chains for the study of T cell response

  • Silvia Mazzucco
  • Elena Nardi
  • Mario Chelli
  • Mauro Ginanneschi
  • Gianfranco Rapi
  • Anna M. Papini
  • Marco Vergelli
  • Benedetta Mazzanti
  • Luca Massacesi
  • Luigi Amaducci
Article

Summary

We previously demonstrated that the lipopeptide of the myelin basic protein (MBP) immunodominant epitope in Lewis rat Palm-GpMBP(74-85) (Gp: guinea pig), which induced experimental autoimmune encephalomyelitisin vivo strongly increased the T cell proliferative responsein vitro. We extended this study to the human immunodominant epitope hMBP(83-99), synthesizing different lipophilic peptides bearing a hydrophobic chain linked through an amide or a C-C bond. To this aim, we developed a synthetic pathway for (±)-N-Fmoc-Ahd-OH (Ahd: 2-aminohexadecanoic acid) which was used to synthesize diastereomeric peptides which were successfully separated by reverse-phase high-performance liquid chromatography. MBP-specific T cell lines recognizing the immunodominant epitope hMBP(83-99) have been generated from patients affected by multiple sclerosis. Their proliferative response to the native peptide and to some lipoderivatives has been investigated. In contrast to the animal model, none of the investigated lipopeptides exhibited ‘superagonist’ activity.

Key words

2-aminohexadecanoic acid hMBP lipidic α-amino acid lipopeptides T cell response 

Abbreviations

Ahd

2-aminohexadecanoic acid

Boc

tert-butyloxycarbonyl

CNS

central nervous system

CPM

count per minute

DCC

dicyclohexylcarbodiimide

DCM

dichloromethane

DMAP

dimethylaminopyridine

DMF

N,N-dimethylformamide

EAE

experimental autoimmune encephalomyelitis

ESI-MS

electrospray ionization mass spectrometry

FAB-MS

fast-atom-bombardment mass spectrometry

FCC

flash column chromatography

Fmoc

fluorenylmethoxycarbonyl

Fmoc-OSu

N-(9-fluorenylmethoxycarbonyloxy)succinimide

GpMBP

guinea pig MBP

HATU

O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

hMBP

human MBP

HPLC

high-performance liquid chromatography

MBP

myelin basic protein

MHC

major histocompatibility complex

NMM

N-methylmorpholine

Palm

palmitoyl

PBL

peripheral blood lymphocytes

Pfp

pentafluorophenyl ester

RP

reverse phase

SPPS

solid-phase peptide synthesis

TCL

T cell line

TCR

T cell receptor

TFA

trifluoroacetic acid

TLC

thin layer chromatography

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References

  1. 1.
    BenMohamed, L., Gras-Masse, H., Tartar, A., Daubersies, P., Brahimi, K., Bossus, M., Thomas, A. and Druilhe, P., Eur. J. Immunol. 27 (1997) 1242.PubMedGoogle Scholar
  2. 2.
    Martin, R., McFarland, H.F. and McFarlin, D.E., Annu. Rev. Immunol., 10, (1992) 153.PubMedCrossRefGoogle Scholar
  3. 3.
    Papini, A.M., Mazzucco, S., Pinzani, D., Biondi, M., Chelli, M., Ginanneschi, M., Rapi, G., Mazzanti, B., Vergelli, M., Massacesi, L., and Amaducci, L., In Tam, J.P. and Kaumaya, P.T.P. (Eds), Peptides: Chemistry, Structure and Biology (Proceedings of the 15th American Peptide Symposium), Kluwer Academic Publishers, Dordrecht, 1998, pp. 700–702.Google Scholar
  4. 4. a.
    Martin, R., Howell, M.D., Jaraquemada, D., Flerlage, M., Richert, J., Brostoff, S., Long, E.O., McFarlin, D.E., and McFarland, H.F. J. Exp. Med. 173 (1991) 19.PubMedCrossRefGoogle Scholar
  5. 4. b.
    Ota, K., Matsui, M., Milford, E.L., Mackin, G.A., Weiner, H.L. and Hafler, D.A., Nature, 346 (1990) 183.PubMedCrossRefGoogle Scholar
  6. 4. c.
    Pette, M., Fujita, K., Wilkinson, D., Altmann, D.M., Trowsdale, J., Giegerich, G., Hinkkanen, A., Epplen, J.T., Kappos, L. and Wekerle, H., Proc. Natl. Acad. Sci. USA, 87 (1990) 7968.PubMedCrossRefGoogle Scholar
  7. 5.
    Branton, W.D., Fields, C.G., Van Drisse, V.L. and Fields, G.B., Tetrahedron Lett. 34 (1993) 4885.CrossRefGoogle Scholar
  8. 6.
    Khare, R.K., Becker, J.M. and Naider, F.R., J. Med. Chem., 31 (1988) 650.PubMedCrossRefGoogle Scholar
  9. 7.
    Deprez, B., Sauzet, J.P., Boutillon, C., Martinon, F., Tartar, A., Sergheraert, C., Guillet, J.G., Gomard, E. and Gras-Masse, H., Vaccine, 14 (1996) 375.PubMedCrossRefGoogle Scholar
  10. 8.
    Papini, A.M., Mazzucco, S., Nardi, E., Chelli, M., Ginanneschi, M. and Rapi, G., In Tam, J.P. and Kaumaya P.T.P. (Eds.), Peptides: Chemistry, Structure and Biology (Proceedings of the 15th American Peptide Symposium), Kluwer Academic Publishers, Dordrecht, 1998, pp. 345–346.Google Scholar
  11. 9.
    Perrin, D.D. and Armarego, W.L.F., In Purification of Laboratory Chemicals, 3rd ed., Pergamon, Oxford, 1988, p. 145.Google Scholar
  12. 10.
    Gibbons, W.A., Hughes, R.A., Charalambous, M., Christodoulou, M., Szeto, A., Aulabaugh, A.E., Mascagni, P. and Toth, I., Liebigs Ann. Chem., (1990) 1175.Google Scholar
  13. 11.
    Vergelli, M., Hemmer, B., Utz, U., Vogt, A., Kalbus, M., Tranquill L., Conlon, P., Ling, N., Steinman, L., McFarland, H.F. and Martin, R., Eur. J. Immunol., 26 (1996) 2624.PubMedGoogle Scholar
  14. 12.
    Carpino, L.A., and Han, G.Y., J. Am. Chem. Soc., 92 (1970) 5748.CrossRefGoogle Scholar
  15. 13.
    Atherton, E., Bury, C., Sheppard, R.C. and Williams, B.J., Tetrahedron Lett., 32 (1979) 3041.CrossRefGoogle Scholar
  16. 14.
    Mori, K. and Funaki, Y., Tetrahedron, 41 (1985) 2369.CrossRefGoogle Scholar
  17. 15.
    Koppitz, M., Huenges, M., Gratias, R. and Kessler, H., Helv. Chim. Acta, 80 (1997) 1280.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Silvia Mazzucco
    • 1
  • Elena Nardi
    • 1
  • Mario Chelli
    • 1
  • Mauro Ginanneschi
    • 1
  • Gianfranco Rapi
    • 1
  • Anna M. Papini
    • 1
  • Marco Vergelli
    • 2
  • Benedetta Mazzanti
    • 2
  • Luca Massacesi
    • 2
  • Luigi Amaducci
    • 2
  1. 1.Dipartimento di Chimica Organica ‘Ugo Schiff’ and Centro di Studio sulla Chimica e la Struttura dei Composti Eterociclici e loro ApplicazioniUniversità di FirenzeFlorenceItaly
  2. 2.Dipartimento di Scienze Neurologiche e PsichiatricheUniversità di FirenzeFlorenceItaly

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