Longitudinal single-cell cytokine responses reveal recurrent autoimmune myelin reactivity in relapsing-remitting multiple sclerosis patients

Abstract

The relationship between multiple sclerosis (MS) disease activity and myelin protein-induced cytokine responses over time is not elucidated. We addressed this relationship by examining longitudinal cytokine responses to myelin proteins every three months for one year, in the context of gadolinium (gad)-enhancing brain lesions and of clinical relapses. The ELISPOT assay was used to determine the ex vivo cytokine production in response to nine amino acid long peptides spanning the entire proteolipid protein (PLP) and myelin basic protein (MBP) molecules in relapsing—remitting (RR) MS patients and matched healthy controls. We identified three longitudinal levels of myelin-induced cytokine secretion by adding up the positive responses for all PLP or MBP peptides obtained for five timepoints, at three- month intervals: low reactivity (<200 cumulative cytokine-secreting cells), isolated peptide reactivity (201-450 cumulative cytokine- secreting cells) and recurrent protein-wide bursts of cytokine reactivity (> 451 cumulative cytokine-secreting cells). The majority of MS patients showed recurrent bursts to PLP and MBP. In contrast, controls showed a more even distribution between all levels of cytokine reactivity. The majority of patients with gad-enhancing lesions showed PLP/IFNg and MBP/IFNg recurrent burst responses. This is the first longitudinal study on MS patients in which nine amino acid long myelin peptides are used to reveal the broad range of PLP- and MBP- peptide cytokine reactivity across the whole molecule of these two major myelin proteins. This study also reveals the extremely dynamic nature of the immune reactivity to numerous regions of myelin, which can fluctuate dramatically over time. Such fluctuation could hamper the efficacy of antigen-based therapies for MS.

Keywords

ELISPOT IFNγ IL-10 longitudinal cytokine reactivity multiple sclerosis myelin basic protein proteolipid protein

Get full access to this article

View all access options for this article.

References

Raine CS , Scheinberg LC. On the immunopathology of plaque development and repair in multiple sclerosis . J Neuroimmunol 1988; 20: 189-201 .

Martin R , McFarland HF , McFarlin DE. Immunological aspects of demyelinating diseases . Annu Rev Immunol 1992; 10: 153-187 .

Pette M , Fujita K , Wilkinson D , Altmann DM , Trowsdale J , Giegerich G et al. Myelin autoreactivity in multiple sclerosis: recognition of myelin basic protein in the context of HLA-DR2 products by T lymphocytes of multiple-sclerosis patients and healthy donors . Proc Natl Acad Sci USA 1990; 87: 7968-7972 .

Valli A , Sette A , Kappos L , Oseroff C , Sidney J , Miescher G et al. Binding of myelin basic protein peptides to human histocompatibility leukocyte antigen class II molecules and their recognition by T cells from multiple sclerosis patients . J Clin Invest 1993; 91: 616-628 .

Allegretta M , Nicklas JA , Sriram S , Albertini RJ. T cells responsive to myelin basic protein in patients with multiple sclerosis . Science 1990; 247: 718-721 .

Johnson D , Hafler DA , Fallis RJ , Lees MB , Brady RO , Quarles RH et al. Cell-mediated immunity to myelin-associated glycoprotein, proteolipid protein, and myelin basic protein in multiple sclerosis . J Neuroimmunol 1986; 13: 99-108 .

Kerlero DR , Hoffman M , Mendel I , Yust I , Kaye J , Bakimer R et al. Predominance of the autoimmune response to myelin oligodendrocyte glycoprotein (MOG) in multiple sclerosis: reactivity to the extracellular domain of MOG is directed against three main regions . Eur J Immunol 1997; 27: 3059-3069 .

Lehmann PV , Forsthuber T , Miller A , Sercarz EE. Spreading of T-cell autoimmunity to cryptic determinants of an autoantigen . Nature 1992; 358: 155-157 .

Tuohy VK , Yu M , Weinstock-Guttman B , Kinkel RP. Diversity and plasticity of self recognition during the development of multiple sclerosis . J Clin Invest 1997; 99: 1682-1690 .

10.

Vanderlugt CJ , Miller SD. Epitope spreading . Curr Opin Immunol 1996; 8: 831-836 .

11.

Perry LL , Barzaga-Gilbert E , Trotter JL. T cell sensitization to proteolipid protein in myelin basic protein-induced relapsing experimental allergic encephalomyelitis . J Neuroimmunol 1991; 33: 7-15 .

12.

Miller SD , Tan LJ , Pope L , McRae BL , Karpus WJ. Antigen-specific tolerance as a therapy for experimental autoimmune encephalomyelitis . Int Rev Immunol 1992; 9: 203-222 .

13.

Lehmann PV , Sercarz EE , Forsthuber T , Dayan CM , Gammon G. Determinant spreading and the dynamics of the autoimmune T-cell repertoire . Immunol Today 1993; 14: 203-208 .

14.

Lisak RP , Zweiman B , Waters D , Korpowski H , Pleasure DE. Cell-mediated immunity to measles, myelin basic protein, and central nervous system extract in multiple sclerosis. A long-itudinal study employing direct buffy coat migration inhibition assays . Neurology 1978; 28: 798-803 .

15.

Calabresi PA , Fields NS , Farnon EC , Frank JA , Bash CN , Kawanashi T et al. ELI-spot of Th-1 cytokine secreting PBMC’s in multiple sclerosis: correlation with MRI lesions . J Neuroimmunol 1998; 85: 212-219 .

16.

Pender MP , Csurhes PA , Greer JM , Mowat PD , Henderson RD , Cameron KD et al. Surges of increased T cell reactivity to an encephalitogenic region of myelin proteolipid protein occur more often in patients with multiple sclerosis than in healthy subjects . J Immunol 2000; 165: 5322-5331 .

17.

Hellings N , Gelin G , Medaer R , Bruckers L , Palmers Y , Raus J et al. Longitudinal study of antimyelin T-cell reactivity in relapsing-remitting multiple sclerosis: association with clinical and MRI activity . J Neuroimmunol 2002; 126: 143-160 .

18.

Arbour N , Holz A , Sipe JC , Naniche D , Romine JS , Zyroff J et al. A new approach for evaluating antigen-specific T cell responses to myelin antigens during the course of multiple sclerosis . J Neuroimmunol 2003; 137: 197-209 .

19.

Rudick RA , Fisher E , Lee JC , Simon J , Jacobs L. Use of the brain parenchymal fraction to measure whole brain atrophy in relapsing-remitting MS. Multiple Sclerosis Collaborative Research Group . Neurology 1999; 53: 1698-1704 .

20.

Fisher E , Cothren RM , Tkach J , Masaryk TJ , Cornhill JF. Knowledge-based 3D segmentation of MR images for quantitative MS lesion tracking . SPIE Medical Imaging 1997; 3034: 19-25 .

21.

Pelfrey CM , Rudick RA , Cotleur AC , Lee JC , Tary-Lehmann M , Lehmann PV. Quantification of self-recognition in multiple sclerosis by single-cell analysis of cytokine production . J Immunol 2000; 165: 1641-1651 .

22.

Moldovan IR , Rudick RA , Cotleur AC , Born SE , Lee JC , Karafa MT , Pelfrey CM. Interferon gamma responses to myelin peptides in multiple sclerosis correlate with a new clinical measure of disease progression . J Neuroimmunol 2003; 141: 132-140 .

23.

Pelfrey CM , Trotter JL , Tranquill LR , McFarland HF. Identification of a novel T cell epitope of human proteolipid protein (residues 40-60) recognized by proliferative and cytolytic CD4+ T cells from multiple sclerosis patients . J Neuroimmunol 1993; 46: 33-42 .

24.

Pelfrey CM , Trotter JL , Tranquill LR , McFarland HF. Identification of a second T cell epitope of human proteolipid protein (residues 89-106) recognized by proliferative and cytolytic CD4+ T cells from multiple sclerosis patients . J Neuroimmunol 1994; 53: 153-161 .

25.

Tuohy VK. Peptide determinants of myelin proteolipid protein (PLP) in autoimmune demyelinating disease: a review . Neurochem Res 1994; 19: 935-944 .

26.

Greer JM , Csurhes PA , Cameron KD , McCombe PA , Good MF , Pender MP. Increased immunoreactivity to two overlapping peptides of myelin proteolipid protein in multiple sclerosis . Brain 1997; 120: 1447-1460 .

27.

Burns J , Rosenzweig A , Zweiman B , Lisak RP. Isolation of myelin basic protein-reactive T-cell lines from normal human blood . Cell Immunol 1983; 81: 435-440 .

28.

Goebels N , Hofstetter H , Schmidt S , Brunner C , Wekerle H , Hohlfeld R. Repertoire dynamics of autoreactive T cells in multiple sclerosis patients and healthy subjects: epitope spreading versus clonal persistence . Brain 2000; 123: 508-518 .

29.

Hellings N , Baree M , Verhoeven C , D’hooghe MB , Medaer R , Bernard CC et al. T-cell reactivity to multiple myelin antigens in multiple sclerosis patients and healthy controls . J Neurosci Res 2001; 63: 290-302 .

30.

Correale J , de los Milagros Bassani Molinas . Time course of T-cell responses to MOG and MBP in patients with clinically isolated syndromes . J Neuroimmunol 2003; 136: 162-171 .

31.

Vergelli M , Mazzanti B , Traggiai E , Biagioli T , Ballerini C , Parigi A et al. Short-term evolution of autoreactive T cell repertoire in multiple sclerosis . J Neurosci Res 2001; 66: 517-524 .

32.

Adorini L , Muller S , Cardinaux F , Lehmann PV , Falcioni F , Nagy ZA. In vivo competition between self peptides and foreign antigens in T-cell activation . Nature 1988; 334: 623-625 .

33.

Gammon G , Geysen HM , Apple RJ , Pickett E , Palmer M , Ametani A et al. T cell determinant structure: cores and determinant envelopes in three mouse major histocompatibility complex haplotypes . J Exp Med 1991; 173: 609-617 .

34.

Targoni OS , Lehmann PV. Endogenous myelin basic protein inactivates the high avidity T cell repertoire . J Exp Med 1998; 187: 2055-2063 .

35.

Tuohy VK , Yu M , Yin L , Kawczak JA , Johnson JM , Mathisen PM et al. The epitope spreading cascade during progression of experimental autoimmune encephalomyelitis and multiple sclerosis . Immunol Rev 1998; 164: 93-100 .

36.

Tuohy VK , Yu M , Yin L , Kawczak JA , Kinkel PR. Regression and spreading of self-recognition during the development of autoimmune demyelinating disease . J Autoimmun 1999; 13: 11-20 .