Anti-LINGO-1 has no detectable immunomodulatory effects in preclinical and phase 1 studies

Abstract

Objective: To evaluate whether the anti-LINGO-1 antibody has immunomodulatory effects.

Methods: Human peripheral blood mononuclear cells (hPBMCs), rat splenocytes, and rat CD4⁺ T cells were assessed to determine whether LINGO-1 was expressed and was inducible. Anti-LINGO-1 Li81 (0.1-30 μg/mL) effect on proliferation/cytokine production was assessed in purified rat CD4⁺ T cells and hPBMCs stimulated with antibodies to CD3 +/- CD28. In humans, the effect of 2 opicinumab (anti-LINGO-1/BIIB033; 30, 60, and 100 mg/kg) or placebo IV administrations was evaluated in RNA from blood and CSF samples taken before and after administration in phase 1 clinical trials; paired samples were assessed for differentially expressed genes by microarray. RNA from human CSF cell pellets was analyzed by quantitative real-time PCR for changes in transcripts representative of cell types, activation markers, and soluble proteins of the adaptive/innate immune systems. ELISA quantitated the levels of CXCL13 protein in human CSF supernatants.

Results: LINGO-1 is not expressed in hPBMCs, rat splenocytes, or rat CD4⁺ T cells; LINGO-1 blockade with Li81 did not affect T-cell proliferation or cytokine production from purified rat CD4⁺ T cells or hPBMCs. LINGO-1 blockade with opicinumab resulted in neither significant changes in immune system gene expression in blood and CSF, nor changes in CXCL13 CSF protein levels (clinical studies).

Conclusions: These data support the hypothesis that LINGO-1 blockade does not affect immune function.

Classification of evidence: This study provides Class II evidence that in patients with MS, opicinumab does not have immunomodulatory effects detected by changes in immune gene transcript expression.

Figure 1. Anti-LINGO-1 (Li81) has no effect on activated T-cell proliferation

(A) Western blot analysis with anti-LINGO-1 (3C11) shows that it was readily detected in neurons but not in activated human peripheral blood lymphocytes, rat splenocytes, or rat CD4⁺ T cells. (B and C) Stimulation with anti-CD3 antibody activated ³H thymidine incorporation into rat CD4⁺ T cells (B, left panel) and hPBMC (C, left panel), and in both cases, this was unaffected by the addition of anti-LINGO-1 (Li81) antibody at concentrations up to 30 μg/mL (B and C, right panels). CPM = counts per minute; hPBMC = human peripheral blood mononuclear cell; IgG = immunoglobulin G; NS = no stimulation.

Figure 2. Anti-LINGO-1 (Li81) does not affect cytokine production

Anti-LINGO-1 (Li81) does not affect cytokine production in the hPBMC, as determined by ELISA. Ab = antibody; hPBMC = human peripheral blood mononuclear cell; IFN = interferon; IgG = immunoglobulin G; IL = interleukin; TNF = tumor necrosis factor.

Figure 3. Gene expression in CSF cell pellets

Gene expression in CSF cell pellets reflecting (A.a–A.b) T cells, (B) B cells, (C.a–C.e) plasma cells, (D.a–D.b) myeloid cells, (E) T-cell activation, and (F) type I IFN response before and after opicinumab or placebo treatment. Each line connects the 2 time points of an individual patient. Patients from the 30-, 60-, and 100-mg/kg groups have been combined. Healthy range (shaded area) was determined by the minimum and maximum values of 5 healthy control patients evaluated. Ig = immunoglobulin; TCRα = T-cell receptor alpha.

Figure 4. Opicinumab does not affect the level of CXCL13 in CSF

Opicinumab does not affect the level of CXCL13 protein in CSF as measured by the CXCL13-Imperacer assay. ^aCorresponding day 0 (before dose) sample above the limit of quantification. ^bCorresponding day 0 sample not done. ^cDay 0 sample below the limit of quantification.