Methods and compositions for inducing an immune response
A technology of immune response and composition, which is applied in the field of composition for enhancing or regulating immune response, and can solve problems such as long time intervals
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment
[0157] Example 1 method
[0158] Reagents were from Sigma Chemical Company (St. Louis, MO) unless otherwise noted.
[0159] Preparation of SHAAYtide Polypeptide (SEQ ID NO: 4): The polypeptide of SEQ ID NO: 4, "SHAAYtide" (Phoenix Pharmaceuticals; Belmont, CA), was chemically synthesized and purified. The material was suspended in phosphate-buffered saline (PBS) at a concentration of about 1 mg / ml and stored at -20°C.
[0160] Enzyme-linked immunosorbent assays (ELISAs): First, 96-well U-bottom plastic plates were covered with 1 μg ovalbumin (OVA) in 100 μl PBS per well, overnight. The next day, the plate was rinsed with PBS, blocked with PBS containing 5% fetal bovine serum (FBS), and rinsed with PBS again. Plasma samples from experimental animals (see below) were diluted 10 2 -10 5 times, and then added to the plate to react for 2 hours, and then rinse the plate with PBS again. The plate was then incubated with biotinylated goat anti-monkey IgG detection antibody, rinsed ...
Embodiment 2
[0164] Example 2 SHAAGtide variant (SEQ ID NO: 2) attracts dendritic cells
[0165] This example describes the in vivo assay of the ability of several chemokines and SHAAGtide (SEQ ID NO: 2) to attract dendritic cells.
[0166] The following chemokines were from R&D Systems (Minneapolis, MN): vMCK-2, mClO and GM-CSF. The following peptides were synthesized at Phoenix Pharmaceuticals (San Carlos, CA): SHAAYtide (SEQ ID NO: 4), several structurally altered peptides of SHAAGtide variants (SEQ ID NO: ) (i.e., cyclization with MPR-Cys linkage) cyclization), control peptide (SEQ ID NO: 17, Gly Ala Ala His Ser Leu Thr Met Gln Pro GlyIle Lys Arg Arg Trp Leu Met), randomly linked to OVA in a ratio of 1:1 or 1:4 (by MBS binding method), linked to OVA at the C-terminus (C-terminus, prepared by addition of cysteine), and a SHAAGtide variant (SEQ ID NO: 2). Chemokines and peptides (2 μg or 20 μg in PBS) were injected intradermally into BALB / c or C57B1 / 6 mice (Jackson Laboratory; Bar Harb...
Embodiment 3
[0171] Example 3 SHAAYtide (SEQ ID NO: 4) was administered to rhesus monkeys Rhesus monkeys were administered with different polypeptides (see Table 10) in different amounts (8, 20 or 60 μg in 100 μl PBS) under anesthesia. Injections are given intradermally. After 24 and 48 hours, 6 mm skin punch biopsies were taken using aseptic technique and divided. A portion of the biopsy was embedded in OCT compound, quickly frozen in liquid nitrogen and stored at -70°C. Other biopsies were soaked in formalin and then embedded in paraffin, and the microtome-cut thin sections were stained with hematoxylin and eosin, and the infiltration of cells into the dermis was observed under a microscope (Table 10). Monkeys were injected with PBS without polypeptide as a negative control.
[0172] The infiltration of mononuclear cells is recorded as 0-5; 0 means that there is very slight inflammatory infiltration of extravascular mononuclear cells in the dermis; 1 means that there is a slight inflam...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap