Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Factor IX administration in the skin primes inhibitor formation and sensitizes hemophilia B mice to systemic factor IX administration

doi: 10.1016/j.rpth.2023.102248

Figure Lengend Snippet: Intradermal factor IX administration does not induce immune tolerance toward FIX. (A) Experimental timeline. C3H/HeJ hemophilia B (HB) mice received FIX intradermally (ID) (0.01-1 IU) twice weekly for 4 weeks before initiation of once weekly intraperitoneal (IP)/intravenous (i.v.) FIX (1 IU) + triprolidine + ABT-491 administration for 5 weeks with intradermal injections continued throughout ( n = 8-9 per group). At 9 weeks, blood was collected for enzyme-linked immunosorbent assay (ELISA) and Bethesda assay. (B) Bethesda assay and anti-FIX immunoglobulin G (IgG) 1 ELISA results in mice that received FIX IP/i.v. only (IP/i.v.) or ID (0.01-1 IU) + IP/i.v. (C) Experimental timeline. C3H/HeJ HB mice received FIX ID (10 −5 -10 −3 IU) twice weekly for 4 weeks before initiation of once weekly IP/i.v. FIX (1 IU) + triprolidine + ABT-491 administration for 6 weeks with intradermal injections continued throughout ( n = 6-8 per group). At 10 weeks, blood was collected for ELISA and Bethesda assay. (D) Bethesda assay and anti-FIX IgG1 ELISA results in mice that received FIX IP/i.v. only (IP/i.v.) or ID (10 −5 -10 −3 IU) + IP/i.v. Shown are means ± SDs and P values from analysis of variance (∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001). BU, Bethesda unit.

Article Snippet: The animals received 10 −5 to 1 IU FIX (Benefix, Pfizer) or 0.01 to 1 IU FIX-Fc (Alprolix, Sanofi) intradermally (ID) in the groin area twice per week for 4 weeks and continued throughout at the same frequency after initiation of intraperitoneal (IP) and i.v. administration of 1 IU FIX (Benefix, Pfizer) ±50 μg triprolidine (antihistamine; Sigma) and 10 μg ABT-491 (platelet-activating factor receptor antagonist; Sigma) in the tail vein once per week for 5 to 6 weeks (1 IP followed by 4 or 5 i.v. injections).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Factor IX administration in the skin primes inhibitor formation and sensitizes hemophilia B mice to systemic factor IX administration

doi: 10.1016/j.rpth.2023.102248

Figure Lengend Snippet: Factor IX administration in the skin alone triggers robust FIX inhibitor formation. (A) Experimental timeline. C3H/HeJ hemophilia B (HB) mice received FIX intradermally (ID) (0.01-1 IU) twice weekly for 4 weeks or once weekly intraperitoneal (IP)/intravenous (i.v.) FIX (1 IU) + triprolidine + ABT-491 administration for 6 weeks ( n = 5-9 per group). At 4 weeks, blood was collected for enzyme-linked immunosorbent assay (ELISA) and Bethesda assay. (B) Bethesda assay and anti-FIX immunoglobulin G (IgG) 1 ELISA results in mice that received FIX IP/i.v. only (IP/i.v.) or ID (0.01-1 IU) only. Shown are means ± SDs and P values from analysis of variance (∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001). BU, Bethesda unit.

Article Snippet: The animals received 10 −5 to 1 IU FIX (Benefix, Pfizer) or 0.01 to 1 IU FIX-Fc (Alprolix, Sanofi) intradermally (ID) in the groin area twice per week for 4 weeks and continued throughout at the same frequency after initiation of intraperitoneal (IP) and i.v. administration of 1 IU FIX (Benefix, Pfizer) ±50 μg triprolidine (antihistamine; Sigma) and 10 μg ABT-491 (platelet-activating factor receptor antagonist; Sigma) in the tail vein once per week for 5 to 6 weeks (1 IP followed by 4 or 5 i.v. injections).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Factor IX administration in the skin primes inhibitor formation and sensitizes hemophilia B mice to systemic factor IX administration

doi: 10.1016/j.rpth.2023.102248

Figure Lengend Snippet: Intradermal factor IX administration sensitizes hemophilia B (HB) mice to systemic FIX administration. (A) Experimental timeline. C3H/HeJ HB mice received FIX intradermally (ID) (1 IU) twice weekly for 4 weeks before initiation of once weekly intraperitoneal (IP)/intravenous (i.v.) FIX (1 IU) + triprolidine + ABT-491 administration for 6 weeks with intradermal injections continued throughout ( n = 4-9 per group). At 10 weeks, blood was collected for enzyme-linked immunosorbent assay (ELISA). (B) Anti-FIX immunoglobulin E (IgE) ELISA results in mice that received FIX IP/i.v. only (IP/i.v.), ID only or ID + IP/i.v. (C) Experimental timeline. C3H/HeJ HB mice received FIX ID (1 IU) twice weekly for 4 weeks before a single i.v. dose of FIX (1 IU) injection, after which the experiment was stopped ( n = 15 per group). (D) Survival of C3H/HeJ HB mice upon receiving 1 IU FIX i.v. with (1 IU ID) or without (1 IU i.v.) intradermal pretreatment. Shown are means ± SDs and P values from analysis of variance or log-rank test (∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001).

Article Snippet: The animals received 10 −5 to 1 IU FIX (Benefix, Pfizer) or 0.01 to 1 IU FIX-Fc (Alprolix, Sanofi) intradermally (ID) in the groin area twice per week for 4 weeks and continued throughout at the same frequency after initiation of intraperitoneal (IP) and i.v. administration of 1 IU FIX (Benefix, Pfizer) ±50 μg triprolidine (antihistamine; Sigma) and 10 μg ABT-491 (platelet-activating factor receptor antagonist; Sigma) in the tail vein once per week for 5 to 6 weeks (1 IP followed by 4 or 5 i.v. injections).

Techniques: Enzyme-linked Immunosorbent Assay, Injection

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Factor IX administration in the skin primes inhibitor formation and sensitizes hemophilia B mice to systemic factor IX administration

doi: 10.1016/j.rpth.2023.102248

Figure Lengend Snippet: Factor IX–Fc does not modulate FIX inhibitor formation. (A) Experimental timeline. C3H/HeJ hemophilia B (HB) mice received FIX-Fc intradermally (ID) (0.01-1 IU) twice weekly for 4 weeks before initiation of once weekly intraperitoneal (IP)/intravenous (i.v.) FIX (1 IU) + triprolidine + ABT-491 administration for 6 weeks with intradermal injections continued throughout ( n = 5-6 per group). At 10 weeks, blood was collected for enzyme-linked immunosorbent assay (ELISA) and Bethesda assay. (B) Bethesda assay and anti-FIX immunoglobulin G (IgG) 1 ELISA results in mice that received FIX IP/i.v. only (IP/i.v.) or FIX-Fc intradermally (0.01-1 IU)+FIX IP/i.v. Shown are means ± SDs and P values from analysis of variance (∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001). BU, Bethesda unit.

Article Snippet: The animals received 10 −5 to 1 IU FIX (Benefix, Pfizer) or 0.01 to 1 IU FIX-Fc (Alprolix, Sanofi) intradermally (ID) in the groin area twice per week for 4 weeks and continued throughout at the same frequency after initiation of intraperitoneal (IP) and i.v. administration of 1 IU FIX (Benefix, Pfizer) ±50 μg triprolidine (antihistamine; Sigma) and 10 μg ABT-491 (platelet-activating factor receptor antagonist; Sigma) in the tail vein once per week for 5 to 6 weeks (1 IP followed by 4 or 5 i.v. injections).

Techniques: Enzyme-linked Immunosorbent Assay