Background: This study aimed to investigate if treatment response could retrospectively be related to inflammatory or axonal patho logy as measured by plasma surrogate markers. Methods: In this 1-year observational study 30 multiple sclerosis (MS) patients with relapsing-remitting disease were treated with intramuscular IFNb-1a or subcutaneous IFNb-1b. Responders and nonresponders were defined according to clinical and magnetic resonance imaging criteria. The control group consisted of 14 healthy subjects. Plasma levels of surrogate markers for inflammation (nitric oxide metabolites (NO x)), astrocytic activation (S100B) and axonal damage (NfHSMI35) were measured using standard assays. Results: There were 11 nonresponders and 19 responders to IFNb treatment. Median S100B levels were elevated in a higher proportion of treatment responders (63%, 42.9 pg/mL) compared to nonresponders (18%, 11.7 pg/mL, PB- 0.05, Fisher’s exact test) and controls (0%, 2 pg/mL, PB- 0.001). Levels of NO x were found to be more frequently elevated in nonresponders (72%, 39 mM) compared to healthy controls (0%, 37 mM, PB- 0.05). Levels of NfHSMI35 were more frequently elevated in responders (58%, 300 pg/mL, PB- 0.001) and nonresponders (72%, 500 pg/mL, PB- 0.001) compared to controls (0%, 4.5 pg/mL). Conclusion: Patients with relapsing-remitting MS who had surrogate marker supported evidence for astrocytic activation responded more frequently to treatment with IFNb.