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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

Inactive Publication Date: 2005-07-13
坎莫森特里克斯公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Also, there is a long time lag between immunization and when the immune system provides protection to the subject

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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...

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PUM

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Abstract

The inventors have discovered that a CKbeta8-1 truncation variant, CKbeta8-1 (25-116), is a bifunctional ligand for two distinct GPCRs, chemokine receptor CCR1 and formyl peptide receptor like 1 (FPRL1). Hence, the inventors have discovered that, in addition to its functional activity on CCR1, CKbeta8-1(25-116) is also a functional ligand for the GPCR receptor FPRL1 that is involved in inflammatory reactions and innate immunity by recruiting monocytes and neutrophils. In addition, the inventors have discovered an alternatively spliced exon of CKbeta8-1, named SHAAGtide. SHAAGtide, along with its parent chemokine CKbeta8-1 (25-116), is fully functional on both monocytes and neutrophils that are known to express FPRL1. This application relates generally to enhancing immune responses. Such immune responses may be elicited by vaccine administration. Compositions and methods for inducing or enhancing an immune response to an antigen are provided. The compositions and methods are useful for vaccine formulations for therapeutic and prophylactic use (immunization) and for production of antibodies.

Description

field of invention [0001] The present invention relates to compositions and methods for enhancing or modulating an immune response, such as that elicited by vaccination. The compositions and methods of the invention are useful, among other uses, in the formulation of vaccines (immunization) for prophylactic and therapeutic use and for the production of useful antibodies (eg, monoclonal antibodies for therapeutic or diagnostic use). Background of the invention [0002] In 1979, almost 200 years after giving the first smallpox vaccine (from a cowpox-infected milkmaid) to a young boy named Jim Phipp, the World Health Organization declared smallpox conquered. The boy survived smallpox because Edward Jenner discovered that cowpox-exposed milkmaids were rarely infected. This accident occurred because vaccinia and smallpox had a similar molecular structure. When smallpox invaded, Philip's immune system could quickly generate a specific response and quickly eliminate the invader. ...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K9/127A61K35/12A61K38/00A61K39/00A61K39/02A61K39/145A61K39/29A61K39/39A61K47/04A61K47/10A61K47/34A61K47/36A61K47/46A61K47/48A61K48/00A61P31/04A61P31/14A61P31/16A61P35/00A61P37/04A61P43/00C07K7/00C07K14/00
CPCA61K39/39A61K2039/55522C07K14/523A61K38/00A61P31/04A61P31/14A61P31/16A61P35/00A61P37/00A61P37/04A61P43/00Y02A50/30A61K38/16A61K39/02A61K39/09A61K39/385
Inventor T·J·沙尔缪振华R·贝拉霍维奇Z·魏M·霍瓦德B·普雷麦克
Owner 坎莫森特里克斯公司
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