Chemically modified antigen preferentially elicits induction of Th1-like cytokine synthesis patterns in vivo.

Abstract

SummaryDifferential activation of CD4 + T cell subsets in vivo leads to the development of qualitatively different effector responses. We identify an approach that allows selective activation of strongly Thl-dominated immune responses to protein antigens. Whereas in vivo administration of ovalbumin (OVA) induces cytokine synthesis that is neither Thl nor Th2 dominated, administration of glutaraldehyde polymerized, high relative molecular weight OVA (OA-POL) leads to 20-fold increase in the ratio of interferon 3' (IFN-3")/IL-4 and IFN-3"/IL-10 synthesis observed after shortterm, antigen-mediated restimulation directly ex vivo. In contrast, concurrent in vivo administration of anti-IFN-3/mAb and OVA or OA-POL results in marked increases in IL-4 and IL-10, and decreased IFN-3' production, reflecting a polarization of the response towards a Th2-1ike pattern of cytokine synthesis. These observations may be useful in clinical settings including hypersensitivity, autoimmune diseases, and vaccine development where the ability to actively select specific patterns of cytokine gene expression would be advantageous.

Endogenous IL-12 synthesis is not required to prevent hyperexpression of type 2 cytokine and antibody responses

“…IL-5 (PharMingen) and IL-13 (R&D Systems) were similarly assessed using cytokine-specific mAb. IFN-+ and IL-10 were determined in ELISA as previously described [51]. Detection limits for these assays were 0.15 U/ml for IFN-+ , 0.2 U/ml for IL-4, 10 pg/ml for IL-5, 0.3 U/ml for IL-10 and 5 pg/ml for IL-13.…”

“…IL-4 levels were determined in an assay using CT.4S cells as previously described [51] and, in most instances, were confirmed in ELISA using 11B11 capture and biotinylated BVD6-24G2 as development mAb (Pharmingen). IL-5 (PharMingen) and IL-13 (R&D Systems) were similarly assessed using cytokine-specific mAb.…”

Vesicle Size Influences the Trafficking, Processing, and Presentation of Antigens in Lipid Vesicles

“…A lthough it is accepted generally that particulate Ags are inherently more immunogenic than soluble Ags (1)(2)(3)(4), it is unclear whether this enhanced immunogenicity is due solely to the increased efficiency of uptake by APCs. This feature of particles makes them attractive adjuvants or delivery systems for vaccine Ags, and consequently, a number of formulation technologies, such as microparticles, liposomes, emulsions, virus-like proteins, and chemically polymerized Ags, have been developed to exploit this phenomenon (4).…”

“…This feature of particles makes them attractive adjuvants or delivery systems for vaccine Ags, and consequently, a number of formulation technologies, such as microparticles, liposomes, emulsions, virus-like proteins, and chemically polymerized Ags, have been developed to exploit this phenomenon (4). Formulation of Ag in particles has also been demonstrated to qualitatively affect the immune response to Ags, with particulate Ags favoring Th1-type responses and soluble Ags promoting Th2 (2,5,6). Despite the significance and widespread application of this observation in vaccine design, it remains unclear exactly how rendering an Ag particulate can affect its immunological profile.…”

Evaluation for synergistic suppression of T cell responses to minor histocompatibility antigens by chloroquine in combination with tacrolimus and a rapamycin derivative, SDZ-RAD

“…19,20 Internal standards of IFN-γ containing Con A-stimulated mouse spleen cell supernatants calibrated against WHO-NIAID international reference reagent Gg02-901-533 (provided by Dr C Laughlin, NIAID, NIH, Bethesda, MD, USA), were included in each ELISA. Each culture supernatant was evaluated using four two-fold dilutions, carried out in duplicate, falling on the linear portion of the standard curve.…”