Neupogen

To promote HCMV reactivation, mice were treated with G-CSF and AMD3100 as previously described^{31 (link)}. Briefly, G-CSF (Neupogen, 100 uL at 300 ug/mL, Amgen) was delivered via an osmotic pump (1007D, Azlet) surgically implanted in the subcutaneous space beneath the dorsal skin on the mouse for 7 days. Additionally, on the same day as pump implantation, mice were also given a single IP injection of AMD3100 (1,1′-[1,4-Phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane octahydrochloride at 125 μg).

Human recombinant G-CSF (Neupogen^®) was purchased from Amgen, Inc. (Thousand Oaks, CA, USA). The Neupogen was received in preservative-free vials containing 300 µg/mL. It was diluted to the appropriate concentrations in sterile 5% dextrose solution. The selective CCR-2 antagonist RS 504393 (Chemical Name: 6-Methyl-1′-[2-(5-methyl-2-phenyl-4-oxazolyl)ethyl]-spiro [4H-3,1-benzoxazine-4, 4′-piperidin]-2(1H)-one) was purchased from Tocris Biosci Inc. (Minneapolis, MN, USA). This chemokine receptor CCR-2 and its primary ligand, monocyte chemoattractant protein-1 (MCP-1), represent a critical signaling pathway for the recruitment of peripheral blood monocytes to sites of immune-mediated inflammation, where they become inflammatory macrophages.

Following screening, eligible patients received a mobilization regimen consisting of G-CSF (10 μg/kg/day) for 4 days administered by SC injection (Neupogen^® [Amgen Inc.] or GRAN^® [Kyowa Hakko Kirin Co., Ltd.] only). The baseline peripheral blood CD34+ cell count was determined on the morning of Day 4 and used to assign patients to one of two groups: <10 CD34⁺ cells/μL and ≥10 CD34⁺ cells/μL. Prior to the first dose of plerixafor on the evening of Day 4, patients were randomized in a 1:1 ratio within each CD34⁺ group to receive either plerixafor WB 0.24 mg/kg SC or FD 20 mg SC. Randomization was performed using an automated interactive voice response system incorporating a central randomization and drug supply scheme.
Plerixafor dosing was timed to allow a 10–11-h interval between dosing on Day 4 and the initiation of apheresis on Day 5. Following a dose of G-CSF 10 μg/kg SC 1 h (±15 min) prior, standard apheresis was performed using three blood volumes (±10%). The same regimen was administered (i.e., plerixafor administration in the evening and G-CSF 1 h prior to apheresis the following morning) for a maximum of four aphereses or until ≥5 × 10⁶ CD34⁺ cells/kg had been collected. The decision to continue treatment and apheresis was based on the cumulative cell yield/kg after each apheresis session.