Human PBMC peripheral blood

Abbreviations used in this chapter AZT, 3 -azidothymidine CD, cluster of differentiation CHS, Chediak-Higashi syndrome Con A, concanavalin-A EAE, experimental allergic encephalomyelitis HIV, human immunodeficiency virus HTLV-1, human T-cell leukemia virus-1 IL, interleukin INF, interferon NAC, N-acetyl cysteine NF-kB, neurotrophic factor-k beta PBMC, peripheral blood mononuclear cells PM A, phorbol 12-myristate 13-acetate PMN, polymorphonuclear neutrophil RT, reverse transcriptase TNF-2 tumor necrosis factor alpha. 

Recently, it has been found that NO donors inhibit HIV-1 replication in acutely infected human peripheral blood mononuclear cells (PBMCs), and have an additive inhibitory effect on HIV-1 replication in combination with 3 -azido-3 -deoxythymisylate (AZT) [139, 140]. S-nitrosothiols (RSNOs) inhibit HIV-1 replication at a step in the viral replicative cycle after reverse transcription, but before or during viral protein expression through a cGMP-independent mechanism. In the latently infected U1 cell line, NO donors and intracellular NO production stimulate HIV-1 reactivation. These studies suggest that NO both inhibits HIV-1 replication in acutely infected cells and stimulates HIV-1 reactivation in chronically infected cells. Thus, NO donors may be useful in the treatment of HIV-1 disease by inhibiting acute infection, or reactivating a latent virus. 

For other efflux transporters such as BCRP (ABCG2), human pharmacokinetic and pharmacodynamic data are currently rare. However, an investigation of the influence of polymorphisms in ABC-transporter genes on the accumulation of nelfinavir in peripheral blood mononuclear cells (PBMCs) revealed no associations between the polymorphisms in the transporters analyzed and the accumulation of nelfinavir in the PBMCs [151], A second study in patients clearly demonstrated an increase in the AUC of the orally and intravenously administered BCRP substrate topotecan when it is given with GF120918, an inhibitor of P-glycoprotein and BCRP [152],... 

Although cell lines are often used in preclinical studies, it is also important to test a compound on primary cells from the target tissue itself. Because this tissue must be taken from healthy, human volunteers, such tissue is often difficult to obtain in sufficient quantities for many types of assays that require large numbers of cells. In Figure 7.9, normal peripheral blood mononuclear cells (PBMCs) from four unrelated, healthy donors were treated... 

Several benzothiadiazepines, as potent and selective TACE inhibitors, have been synthesized with variation in P1 and PI and evaluated versus porcine TACE, and the initial selectivity was assessed with counterscreens of MMP-1, MMP-2, and MMP-9. Several potent and selective inhibitors were discovered, 440 being the most active against porcine TACE. Most compounds were assessed in the human peripheral blood mononuclear cell (PBMC) assay and the human whole blood assay (WBA) to determine their ability to suppress tumor necrosis factor alpha (TNF-a). Compound 441 was found to be the most potent in the PBMC assay (IC50 = 0.35 pM), while 442 was the most active in the WBA (IC50 = 1.4 pM) <2003JME1811>. 

Recruit volunteers who met entrance requirements 2. Screen botanical ingredients for inhibition of IL-1 p production 3. Pilot clinical evaluation of IL-1 inhibitory lead candidate botanicals from activity 2 Establishing a clinical genotyped database Human monocyte cell lines stimulated in vitro with LPS Clinical + laboratory assay of peripheral blood mononuclear cells (PBMCs), and plasma Adequate number of healthy subjects with CRP = 2-10 mg/L and stratified by IL1 composite genotype Select lead candidate botanicals based on IL-1 protein inhibitory dose compared to untreated cultures IL1 gene expression in PBMCs and ex vivo IL-1 production in plasma from test subjects after 2-week dosing of candidate botanicals... 

D Avolio A et al (2012) HPLC-MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human peripheral blood mononuclear cell (PBMC). J Pharm Biomed Anal 59 109-116... 

Organic and aqueous feverfew powdered leaf extracts were found to inhibit IL-1-induced prostaglandin E2 release from synovial cells, IL-2-induced thymidine uptake by lymphoblasts, and mitogen-induced uptake of thymidine by peripheral blood mononuclear cells (PBMCs) (23). Parthenolide also inhibited thymidine uptake by PBMCs. Both parthenolide and the extracts were cytotoxic to the PBMCs and synovial cells thus, the anti-inflammatory effects of feverfew may be secondary to cytotoxicity. These results reflect those of previous researchers who found parthenolide and other sesquiterpene lactones to be cytotoxic to cultures of human fibroblasts, human laryngeal carcinoma cells, and human cells transformed with simian virus 40 (24). 

Lymphotoxicity (cytotoxicity or functional impairment, e.g., proliferation upon anti-CD3 or anti-CD28 stimulation) [7] Human peripheral blood lymphocytes (whole blood or PBMC) or rodent splenocytes are typically used. Polyclonal stimuli include anti-CD3, anti-CD28, plant lectins such as concanavalin A (ConA) and phytohemagglutinin (PHA), superantigens etc. Mixed lymphocyte reactions possible but less used for immunotoxicity... 

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