Multiple Sclerosis and the Immune Response By: Terra McMillan

1. Multiple Sclerosis and the Immune ResponseBy: Terra McMillan Hermans, Guy, Ulrike Denzer, Ansgar Lohse, Jef Raus, and Piet Stinissen. 1999. Cellular and Humoral Immune Responses Against Autoreactive T cells in Multiple Sclerosis Patients After T cell Vaccination. Journal of Autoimmunity 13, 233-246.

2. The Purpose • Study the safety, immune responses and clinical effects of T cell vaccination in 49 multiple sclerosis patients. • Gain insight on the mechanisms of autoreactive T cells

3. What is Multiple Sclerosis? • Autoimmune disease • T cell mediated • ~250,000 people have MS • Women 2:1 ratio with men

4. Basic Physiology of the Nervous System

5. What triggers Multiple Sclerosis?

7. Common Vocabulary • Antigen-A substance that cause the production of specific antibodies and combine with those antibodies. • Antibodies-Protein produced by the body to specifically react with a foreign substance • EAE- experimental autoimmune encephalomyelitis • Cytokines-Protein released in response to an antigen which influences other cells.

8. Vocabulary Cont’d • PBMC-peripheral blood mononuclear cells • MBP-myelin basic protein • TCR-T cell receptor • CD markers-clusters of differentiation • MHC-major histocompatibility complex

9. Materials and Methods • MBP reactive T cells were isolated from the peripheral blood and cloned • T cell clones was analyzed for TCR, rearrangements, and CDR3 sequence • Inactivation by irradiation • 49 MS patients were injected subcutaneously with 10x106 cells for T cell vaccination • 3 immunizations were performed at 2-month intervals

10. Materials and Methods Cont’d. • Mononuclear cells isolated from blood samples and irradiated with T cell vaccine • Incubated for 4 days • Purified, label with radioactivity, and stored at –20oC for cytokine analysis • Results recorded in Stimulation Indexes (SI) • Net production of cytokine

11. Materials and Methods Cont’d. • Patient serum Ig was bound to vaccine T cells using flow cytometry • Results were used in immunoblotting • Also patient serum was used to determine the proliferation of T cells

12. Rationale for Figure 1. • The objective was to test if serum from vaccinated patients influenced the proliferation of T cells • PBMC of vaccinated patients were collected at various time intervals before and after vaccinations • Samples were irradiated with 3H-thymine

13. Results- Proliferative Responses to T cell vaccines (y-axis= stimulation index)

14. Conclusion-Figure 1. • All patients showed proliferative responses after vaccination, indicating an immune response • Proliferative responses often increased after the second and third vaccinations • Some clones were more effective than others

15. Rationale-Figure 2. • Examination of long term proliferative responses using stimulation index • Samples all obtained after the third vaccination • Same irradiation procedures as for figure 1. were used • PHA blasts were used as the control

16. Results-Long Term Proliferative Cellular Responses to T-cell Vaccinaation

17. Conclusion-Figure 2. • SI’s gretaer than 2 were observed for all patiens at 1 and 2 years after vaccination • In some patients the response was greater than others • Indicates a memory T cell system within these patients

18. Rationale-Figure 3. • Determining phenotypes of cytokines which responded to the vaccine clones • Short-term cell line cultures were obtained 3 weeks after vaccination 3 • Results were measured in SI units on the x-axis