Adju phos

Animals were anesthetized as previously described [32 (link)] with isoflurane inhalation and intranasally inoculated with a fluid volume of 100μL. For bronchoalveolar lavage studies, cotton rats were inoculated with either 5μg of purified G protein, or 5μg or 20μg recombinant mouse CX3CL1 (458-MF, R&D Systems, Minneapolis, MN, USA). To determine the best serotype of AAV for studies in the cotton rat respiratory tract, cotton rats were inoculated with 10⁹⁻¹⁰ DNAase resistant particles (DRP) with different serotypes of AAV expressing GFP for flow cytometry studies. Blood samples were obtained by retro-orbital bleed in isofluorane narcosis. For the studies of inflammation and immunogenicity, cotton rats were inoculated with 2x10¹⁰ DRP of various AAV constructs and challenged with 10⁵ TCID₅₀ of RSV A2. 100μg house dust mite (HDM, Dermatophagoides pteronyssinus) antigen was absorbed to aluminum phosphate (AdjuPhos, Brenntag, Ballerup, Denmark) at a 1:1 ratio for 30 minutes at room temperature and injected into cotton rats intraperitoneally (IP). Sensitization was followed 8 days later with intranasal administration of 100μg HDM in a volume of 100μL PBS. Four days after HDM administration, cotton rats were euthanized through CO₂ inhalation and lungs were collected for histologic examination.

Hartley guinea pigs were obtained from Charles River Laboratories. Purified CHIKV VLPs derived from infection of SfBasic with AcMNPV-CHIKV37997 and VLP standard derived from transient transfection of HEK293 cells were adjuvanted onto Adju-Phos aluminum based adjuvant (Brenntag Biosector). Guinea pigs (4 animals per group) were vaccinated intramuscularly with doses of 0.01, 0.1, 1, or 10 μg of CHIKV VLPs. Animals were vaccinated at Day 0 and Day 14, and serum was sampled on Day 14 (prior to dosing) and on Day 21 (at study completion). A pre-vaccination serum sample was taken prior to the first vaccination for the purpose of establishing the serum IgG ELISA background.

100μg house dust mite (HDM, Dermatophagoides pteronyssinus) antigen was absorbed in aluminum phosphate (AdjuPhos, Brenntag, Ballerup, Denmark) at 1:1 ratio for 30 minutes at room temperature. HDM was administered intraperitoneally (IP). Sensitization was followed 8 days later with intranasal (IN) administration of 100μg HDM in a volume of 100μL PBS. Four days after IN HDM administration, cotton rats were anesthetized and forced oscillation technique was performed or lungs were collected for histologic examination.

The aluminium adjuvant preparations used in this study were Alhydrogel; AlO(OH) and Adju-Phos, Al(OH)x(PO₄)y, purchased from Brenntag Biosector (Frederikssund, Denmark). Dealuminated zeolite Y (USY) was purchased from Tosoh Corporation, Japan.
Lumogallion (CAS 4386–25-8) was purchased from TCI Europe N.V., Antwerp, Belgium, and lipopolysaccharide (LPS, from Escherichia coli O111:B4) was purchased from Sigma-Aldrich, St. Louis, MO, USA.

Six- to ten-week-old C57BL/6, B6.129S2-H2^dlAb1-Ea/J (major histocompatibility complex class II [MHC-II] knockout [KO]), B6.129S2-Cd40lg^tm1Imx/J (CD40L KO), and B6.129P2-Cd40^tm1Kik/J (CD40 KO) mice were purchased from the Jackson Laboratory (Bar Harbor, ME). Thymectomized C57BL/6 mice underwent adult thymectomy at the Jackson Laboratory (Bar Harbor, ME). Mice were immunized intramuscularly (i.m.) with a volume of 100 μl divided between the two quadriceps. The previously described E1/E3-deleted Ad26-SIV_mac239 Env or Ad5-SIV_mac239 Env was used at a dose of 10⁹ or 10¹⁰ viral particles (vp) (24 (link)). Simian immunodeficiency virus (SIV) Env 32H gp140 was used at 50 μg with 100 μg Adju-Phos (Brenntag) (25 (link)). All animal experiments were performed in accordance with Institutional Animal Care and Use Committee guidelines of the Beth Israel Deaconess Medical Center.