• 主页
  • 查找结果
  • Immune response profile elicited by the model antigen ovalbumin expressed in fusion with the bacterial OprI lipoprotein.

Immune response profile elicited by the model antigen ovalbumin expressed in fusion with the bacterial OprI lipoprotein.

Molecular immunology (2014-12-04)
Afonso P Basto, Marina Badenes, Sílvia C P Almeida, Carlos Martins, António Duarte, Dulce M Santos, Alexandre Leitão
摘要

The use of immunogenic formulations targeting pattern recognition receptors towards modulation of immune responses is a promising strategy to develop better vaccines against infectious and malignant diseases. Molecules targeting TLR2 offer interesting properties that are relevant for vaccine development, including the possibility to covalently attach the lipidic ligands to the antigens. However, the type of immune response elicited by these formulations is still controversial. In this work, we used the model antigen ovalbumin (OVA) expressed in fusion with the bacterial lipoprotein OprI in order to characterize the immunomodulatory properties of this TLR ligand. Murine bone marrow-derived dendritic cells stimulated with OprI-OVA fusion lipoprotein produced high levels of the pro-inflammatory cytokines TNF-α and IL-6 and also IL-10, IL-12(p70) and IL-27, while TGF-β and IL-23 were not detected. Using OT-II and OT-I mice, an enhancement of MHC class II and class I antigen presentation was observed for the OVA antigen in fusion with OprI. Mice immunized by intraperitoneal route with this fusion lipoprotein in prime-boost protocols developed strong specific antibody responses including IgG1, IgG2c, IgG2b, IgG3 and IgE. These results, together with data obtained by restimulation of splenocytes from the immunized mice, point to an immune response profile that does not correspond to a strict Th1 or Th2 polarization. Finally, in a challenge experiment using a melanoma syngeneic mouse model (B16-OVA), prophylactic inoculation with OprI fused with the unrelated antigen eGFP increased the tumor growth, while the fusion with the tumor-associated antigen OVA delayed the tumor growth and increased mice survival.

材料
货号
品牌
产品描述

Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, ≥99.5% (titration)
Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
盐酸胍 盐酸盐, for molecular biology, ≥99%
Sigma-Aldrich
尿素, ACS reagent, 99.0-100.5%
Sigma-Aldrich
乙二胺四乙酸 溶液, BioUltra, for molecular biology, pH 8.0, ~0.5 M in H2O
Sigma-Aldrich
乙二胺四乙酸, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
咪唑, ACS reagent, ≥99% (titration)
Sigma-Aldrich
尿素, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
白蛋白 来源于鸡蛋白, lyophilized powder, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
盐酸胍 盐酸盐, ≥99% (titration), organic base and chaeotropic agent
Sigma-Aldrich
乙二胺四乙酸, anhydrous, BioUltra, ≥99% (titration)
Sigma-Aldrich
L-谷氨酰胺, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
HEPES缓冲溶液, 1 M in H2O
Sigma-Aldrich
咪唑, ReagentPlus®, 99%
Sigma-Aldrich
咪唑, for molecular biology, ≥99% (titration)
Sigma-Aldrich
盐酸胍 盐酸盐, ≥98%
Sigma-Aldrich
咪唑, puriss. p.a., ≥99.5% (GC)
Sigma-Aldrich
尿素, ReagentPlus®, ≥99.5%, pellets
Sigma-Aldrich
L-谷氨酰胺, ReagentPlus®, ≥99% (HPLC)
SAFC
盐酸胍 盐酸盐
Sigma-Aldrich
乙二胺四乙酸, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
乙二胺四乙酸 溶液, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
SAFC
羟乙基哌嗪乙硫磺酸
SAFC
L-谷氨酰胺
Sigma-Aldrich
乙二胺四乙酸, purified grade, ≥98.5%, powder
Supelco
尿素, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
咪唑, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
尿素 溶液, BioUltra, ~8 M in H2O
Sigma-Aldrich
尿素, BioXtra, pH 7.5-9.5 (20 °C, 5 M in H2O)