PBMC , human

Human PBMC Isolation and Stimulation with DIMS... 

IFN production of PBMCs cultured in the presence of selected DIMS is shown from six healthy individuals (ELISA mean data of IFNa and IFNfl) and CBA mean data (IFNy) from four healthy individuals (Table 2). Different DIMS are capable of inducing IFN with different efficiency in human PBMCs (see Notes 1, 2, 5, 7, 8, 10-15 for comments on results). 

The three levels of DIMS structures and their correlation with the induction of IFNs in human PBMCs in vitro... 

CpG context in DIMS9051, but otherwise maintaining the same set and pattern of motifs as in DIMS9054, will totally abolish the DIMS ability to induce IFNs in cultured healthy human PBMCs (Table 2). 

DIMS capability to induce IFNs (a, P, y) in cultured healthy human PBMCs in vitro well correlates with its ability to form parallel tetramer of G-quadruplex (parallel G4-tetramer) (Table 2 Fig. 1). CD spectra demonstrated appearance of the characteristic peaks in the range of 252-264 nm, typical of a parallel G-4 tetramer structure, for DIMS9052, DIMS9054, DIMS9058, and DIMS9059 but not for DIMS9051 (Fig. 3). 

This may be complemented with ex vivo gene profiling of PBMC from patients on immunosuppressive drugs. Finally, instead of using fresh human PBMC and rodent splenocytes, cell lines such as the human Jurkat T-cell line and the mouse EL4 thymoma cell line can be used. 

Four different amounts of the four gangliosides were added to microtiter wells containing human PBMC s and 18 micrograms of concanavalin-A. All four gangliosides markedly inhibited lymphoblastic transformation when 12.6 nanomoles was added to the cultures with a total volume of 0.2 ml (Table II). As little as 1.6-nanomoles of GDlb and GTlb resulted in substantial inhibition, while much less inhibition occurred with equivalent amounts of GM1 and GDla. 

Engraftment of NOD-scid IL2rynu" Mice with Human PBMC... 

Previous attempts to measure immune function of human cells following PBMC engraftment of immunodeficient mice have been met with limited success (34). The lack of a vigorous primary immune response mediated by human PBMC engrafted into earlier humanized mouse models was associated with host murine NK cell activity and other innate immune function as well as an overwhelming xenogeneic GVHD response of the... 

Hauk, P.J., et al., Induction of corticosteroid insensitivity in human PBMCs by microbial superantigens. J. Allergy Clin. Immunol. 105, 782-787, 2000. 

In vitro T-cell immunogenicity data obtained with this conjugate using T cell hybridomas, lymph node cells from immunized mice, and human PBMC cultures from PPD-positive individuals indicated that both epitopes were efficiently recognized (10). 

There are striking parallels between the effects of opioids and cannabinoids on cytokine production. A -THC has also been shown to suppress proinflammatory cytokines crucial in host defense, such as TNF-a (Fischer-Stenger et al., 1993). Further studies showed a biphasic dose response to cytokine production by A -THC on human PBMCs, but it was the opposite of that observed with morphine, with low doses inhibiting TNF-a, lL-6 and lL-8, and high doses increasing these cytokines (Berdyshev et al., 1997). In a mouse model of Legionnaires Disease, A -THC blocked IFN-y, lL-12, and lL-12 receptor expression and increased lL-4 levels, reflecting a polarization of the immune response towards a Th2 phenotype (Perez-Castrillon et al, 1992 Klein and Cabral, 2(X)6). Similar... 

M. Jemal, S. Rao, M. Gatz, D. Whigan, LC-AdS-AdS quantitative determination of the HIV protease inhibitor atazanavir in human PBMC practical approaches to PBMC preparation and PBMC assay design for high-throughput analysis, J. Chromatogr. B, 795 (2003) 273. 

F. Becher, A. Pruvost, C. Goujard, C. Guerreiro, J.-F. Delfraissy, J. Grassi, H. Benech, Improved method for the simultaneous determination of d4T, 3TC and ddl intracellular phosphorylated anabolites in human PBMC using LC-MS-MS, Rapid Commun. Mass Spectrom., 16 (2002) 555. 

F. Becher, D. Schlemmer, A. Pmvost, M.-C. Nevers, C. Goujard, S. Jorajuria, T. Brossette, L. Lebeau, C. Creminon, J. Grassi, H. Benech, Development of a direct assay for measuring intracellular AZT triphosphate in human PBMC, Anal. Chem., 74(2002)4220. 

In addition to these simpler cell-based assays, more complex assays can be employed to measure the induction of specific T cell responses to haptens. It is possible to stimulate naive T cells to proliferate in vitro in the presence of antigen-presenting cells such as LCs.27-28 In addition, effector T cells (T cell clones) can be used to investigate T cell stimulation induced by haptens in the presence of the appropriate antigen-presenting cells.29 30 Another complex, but easy to use assay is based on the culture of purified human PBMC in the presence of potential reactive chemicals, and the measurement of proliferation or cytokine production.31-33... 

IL-2R ji- and y-chains are both required for binding IL-15 and for signal transduction the a-chain is not required. Like IL-2, IL-15 causes the release of sIL-2Ra fi om PHA-activated human PBMCs in a concentration and time-dependent manner. This effect of IL-15 is largely direct and independent of IL-2. Blocking of the JL 2R(5-chain prevents the release of sIL-2Ra by IL-15 but not by IL-2. IL-7, another cytokine using the y-chain of the IL-2R, drives the release of sIL-2Ra as well. Several clinical condi-... 

IL-18 augments NK-cell activity in cultures of human PBMCs similar to that of the structurally unrelated cytokine IL-12. In addition, it has been found to enhance the production of IFNy and GM-CSF while decreasing the production of IL-10 by mitogen-stimulated PBMCs but had no effect on IL-4 production by concanavalin A-stimulated PBMC. lL-18 enhances T-celi proliferation apparently through an IL-2-dependent pathway. It also enhances Thl cytokine pro-duction in vitro and exhibits synergism when combined with IL-12 in terms of enhanced IFNy production but not IL-2 and GM-CSF production. ... 

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