Afuco™ Anti-Human TLR4 ADCC Recombinant Antibody (NI-0101), ADCC Enhanced (CAT#: AFC-591CL)

Figure 1 Interference in toll-like receptor 4 (TLR4) signaling blocks IL-6, TNFα, IL-1β and IL-8 production by monocytes stimulated by synovial fluid from patients with rheumatoid arthritis (RASF).

LR4 signaling was blocked with anti-human TLR4 monoclonal antibody, NI-0101. Representative data shown for monocytes obtained from one of seven patients with rheumatoid arthritis (RA).

Hatterer, E., Shang, L., Simonet, P., Herren, S., Daubeuf, B., Teixeira, S.,... & Sokolove, J. (2016). A specific anti-citrullinated protein antibody profile identifies a group of rheumatoid arthritis patients with a toll-like receptor 4-mediated disease. Arthritis research & therapy, 18(1), 224.

Figure 2 Heterogeneous capacity of rheumatoid arthritis (RA) synovial fluid (RASF) samples to stimulate cytokine production and respond to TLR4 blockade.

RASF samples from patients (Pat) were classified as NI-0101 responders if NI-0101 was able to block (partially or totally) RASF-induced IL-6 production from RA monocytes. Others were classified as NI-0101 non-responders. a, b Representative examples of RASF from non-responders (Pat#13, Pat#35) and from responders (Pat#27, Pat#18). Of the 40 RASF samples tested, 20 were classified as NI-0101 responders (50 %) and 20 as NI-0101 non-responders (50 %). c, d Mean levels of IL-6 in RASF NI-0101 responder and non-responder RASF samples. IL-6 is presented as normalized values with IL-6 levels induced by RASF arbitrarily set as 100 % induction. RASF samples that were unable to induce IL-6 from RA monocytes but were impacted upon by NI-0101 are excluded from the non-responder group.

Hatterer, E., Shang, L., Simonet, P., Herren, S., Daubeuf, B., Teixeira, S.,... & Sokolove, J. (2016). A specific anti-citrullinated protein antibody profile identifies a group of rheumatoid arthritis patients with a toll-like receptor 4-mediated disease. Arthritis research & therapy, 18(1), 224.

Figure 3 Expression of anti-citrullinated protein antibodies (ACPA) and ligands of toll-like receptor 4 in samples of synovial fluid (SF) from patients with rheumatoid arthritis (RA) and correlation between these and NI-0101 response.

a Expression of ACPA, HMGB1 and S100A8/A9 in SF from subjects without RA (non-RA SF) (n = 4 samples) and patients with RA (RASF) (n = 40 samples). b Correlation between ACPA, HMGB1 or S100A8/A9 and NI-0101 response. RASF samples were classified as NI-0101 non-responders (NR) or NI-0101 responders (R) according to Fig. 2.

Hatterer, E., Shang, L., Simonet, P., Herren, S., Daubeuf, B., Teixeira, S.,... & Sokolove, J. (2016). A specific anti-citrullinated protein antibody profile identifies a group of rheumatoid arthritis patients with a toll-like receptor 4-mediated disease. Arthritis research & therapy, 18(1), 224.

Figure 4 Anti-citrullinated protein antibodies (ACPA) fine specificity in the synovial fluid samples of patients with rheumatoid arthritis and their correlation with NI-0101 response.

Antibody reactivity against the citrullinated peptides derived from fibrinogen-α (cFbα − pept), fibrinogen-β (cFbβ-pept) and histone-2A (cH2A-pept) were determined by ELISA and expression in synovial fluids was measured in ACPA-positive rheumatoid arthritis synovial fluid (RASF) samples (a) (n = 22) and both ACPA-negative and ACPA-positive RASF samples (b) (n = 40). RASF samples were classified as NI-0101 non-responders (NR) or NI-0101 responders (R).

Hatterer, E., Shang, L., Simonet, P., Herren, S., Daubeuf, B., Teixeira, S.,... & Sokolove, J. (2016). A specific anti-citrullinated protein antibody profile identifies a group of rheumatoid arthritis patients with a toll-like receptor 4-mediated disease. Arthritis research & therapy, 18(1), 224.

Figure 5 Anti-citrullinated protein antibodies (ACPA) fine specificity in paired sera samples of patients with rheumatoid arthritis (RA) and their correlation with RA synovial fluid (RASF) response to NI-0101.

a Correlation between ACPA in paired samples of sera from patients with RA and synovial fluids (n = 22). b ACPA in paired samples of sera from patients with RA classified according to RASF response to NI-0101 (NI-0101 non-responders (NR) or NI-0101 responders (R)). c, d Antibody reactivity against the citrullinated peptides derived from fibrinogen-α (cFbα-pept), fibrinogen-β (cFbβ-pept) and histone-2A (cH2A-pept) were determined by ELISA in paired samples of sera from patients with RA and correlation with response to NI-0101 was tested. c Paired sera samples from ACPA-positive patients with RA (n = 13). d Paired samples of sera from from ACPA-positive and ACPA-negative patients with RA (n = 22).

Hatterer, E., Shang, L., Simonet, P., Herren, S., Daubeuf, B., Teixeira, S.,... & Sokolove, J. (2016). A specific anti-citrullinated protein antibody profile identifies a group of rheumatoid arthritis patients with a toll-like receptor 4-mediated disease. Arthritis research & therapy, 18(1), 224.

Figure 6 Mean NI-0101 Serum Concentrations (ng/mL) after Single Ascending Dose of NI-0101, (black) Part 1; (red) Part 2

Monnet, E., Lapeyre, G., van Poelgeest, E., Jacqmin, P., de Graaf, K., Reijers, J.,... & de Min, C. (2017). Evidence of NI‐0101 pharmacological activity, an anti‐TLR4 antibody, in a randomized phase I dose escalation study in healthy volunteers receiving LPS. Clinical Pharmacology & Therapeutics, 101(2), 200-208.

Figure 7 Mean IL-6 Percentage Change from Baseline per Genotype (Ex Vivo LPS Challenge) in Part 1 FcγRIIa genotype H/H (a), R/H (b).

Monnet, E., Lapeyre, G., van Poelgeest, E., Jacqmin, P., de Graaf, K., Reijers, J.,... & de Min, C. (2017). Evidence of NI‐0101 pharmacological activity, an anti‐TLR4 antibody, in a randomized phase I dose escalation study in healthy volunteers receiving LPS. Clinical Pharmacology & Therapeutics, 101(2), 200-208.

Figure 8 Change in Pharmacological Parameters after In Vivo LPS Challenge in Part 2 (a) Mean IL-6 serum concentration (pg/mL), (b) Mean CRP serum concentration (mg/L).

Monnet, E., Lapeyre, G., van Poelgeest, E., Jacqmin, P., de Graaf, K., Reijers, J.,... & de Min, C. (2017). Evidence of NI‐0101 pharmacological activity, an anti‐TLR4 antibody, in a randomized phase I dose escalation study in healthy volunteers receiving LPS. Clinical Pharmacology & Therapeutics, 101(2), 200-208.