Proliferation Assay

In order to evaluate the in vitro antitumor activity of the prodrugs, compounds 20 and 21 were incubated at varied concentrations with human T-lineage acute lymphoblastic leukemia MOLT-3 cells in the presence or absence of ImM of PGA. The data from the cell proliferation assays are presented in Fig. 5.15. 

Although a number of cell lines were shown to be sensitive to inhibition by PatA (Low et al., 2005), we selected RKO cells (IC50 of 0.4 nM in cell proliferation assay) to prepare lysates for the isolation and identification of target protein(s). We often select RKO cells for target identification of small molecules using biotin-conjugates, because they appear to be particularly suitable for target protein isolation. 

Ex vivo studies have revealed that trichothecenes can both inhibit and stimulate leukocyte function.12 For example, trichothecenes are toxic to alveolar macrophages,13 but drive differentiation of human myeloid leukemic cells.14 Dose-dependent decreases or increases in B- and T-cell mitogen responses are observable in lymphocytes from animals exposed to T-2 toxin, DON, or various macrocyclic trichothecenes these toxins similarly impair or enhance mitogen-induced lymphocyte proliferation in vitro.12 Rank order of inhibitor potency in rodent and human lymphocyte proliferation assays is Type D > Type A group > Type B group and is dependent on degree of acylation as well as of uptake and metabolism. 

Weaver, J. et al., Evaluation of a lymph node proliferation assay for its ability to detect pharmaceuticals with the potential to cause immune mediated drug reactions. J. Immuno-toxicol., 2, 11, 2005. 

Nickel-specific human T-lymphocyte clones have been isolated from blood [373] and inflammatory infiltrates after nickel application [379, 381]. 7-15% of the CD4 1 8 T-lymphocyte clones appeared to be specific for nickel in a proliferation assay [379] this proliferation response required the presence of antigen-presenting cells and was restricted by HLA class II molecules. Nickel-specific T cells from each nickel-allergic patient were extremely heterogenous with respect to their genetic restriction [381], Nickel-specific T lymphocytes sometimes show unusual genetic restrictions and might even respond to nickel without the participation of HLA-II molecules. 

Linear alcohols were studied using MCF7 cells in the proliferation assay and in the expression of estrogen-dependent genes (Table 7.3.4). The AS C12 i4, its ethoxylated derivative (ALFONIC 1214) and linear alcohols from nonanol to dodecanol increase cell proliferation in the E-Screen assay. This effect was seen at 10-100 xM concentrations and,... 

The methods used were acrosin proteolytic activity (APA) assay (with gelatin) and acrosin activity assay with N-a-benzoyl-DL-arginine-p-nitroanihde (BAPNA)-Triton X 100, and BAPNA assay for trypsin activity [9-11]. The antioxidant activity was tested spectrometrical with ABTS+ [12]. Cytotoxicity of extracts was determined by MTT Cell Proliferation Assay [13]. 

Proliferation assays performed with PBMC cultures from a number of healthy donors demonstrated that IRIV indeed elicited cell proliferation in all tested cultures to an extent variable from donor to donor (Fig. 1 A). Further proliferation assays with CD4+ T-cells or CD8-I- T-cells in coculture with CD 14+ cells demonstrated that IRIV induce CD4+ T-cell proliferation, whereas no proliferation of CD8+ T-cells could be observed (Fig. 1C). Dissection of the CD4+ T-cell population into CD4+CD45RO+ and CD4+CD45RA+ cell subsets pointed to CD4+CD45RO+ cells as proliferative responders (Fig. ID). Importantly, culture of cord blood mononuclear cells in presence of IRIV did not result in major cell proliferation (Fig. IB), underlining antigen experience as prerequisite for IRIV-induced cell proliferation. 

Adjuvants enhancing HLA class I-restricted CTL responses are especially needed for treatment or prevention of chronic viral diseases and infections linked to intracellular pathogens, and for cancer immunotherapy. Among the very few adjuvants licensed for human use, we evaluated the capacity of IRIV to enhance HLA class I-restricted CTL responses in vitro. We addressed IRIV-elicited immune responses and the induction of CTL specific to IM58 66 and Melan-A/Mart-127-35 epitopes. Proliferation assays, cytokine expression studies, and phenotypes of CD4+ T-cells demonstrated that IRIV... 

Okubo T, Suzuki T, Yokoyama Y, Kano K, Kano I (2003) Estimation of estrogenic and antiestrogenic activities of some phthalate diesters and monoesters by MCF-7 cell proliferation assay in vitro. Biol Pharm Bull 26 1219-1224... 

The basic principle underlying the LLNA is that sensitizers induce a primary proliferation of lymphocytes in the lymph node draining the site of chemical application. This proliferation is proportional to the dose applied and to the potency of the allergen, and provides a measurement of sensitization. The LLNA assesses this proliferation as a dose-response in which the proliferation in test groups is compared to that in vehicle treated controls. The ratio of the proliferation in treated groups to that in vehicular controls, termed the Stimulation Index , is determined, and must be at least three before a test substance can be further evaluated as a potential skin sensitizer. The test substance, plus vehicle and positive control, is applied for three consecutive days to the ears of test mice, on days 4 and 5 the animals are left alone, and on the 6th day they are prepared for the proliferation assay, sacrificed, and the measurements are done. 

Triciribine, also known as Akt/protein kinase B signaling inhibitor-2 (API-2) compound 52, Fig. 9), was identified by screening the National Cancer Institute Diversity Set chemical library (1992 compounds). The screen was performed using a cell-based proliferation assay with PKB/1 transformed NIH3T3 cells, and triciribine scored positive in this assay with an IC50 value of 50 nM [ 196]. Although the compound blocks the cellular phosphorylation... 

BaF3 cell assays were used to study the effect of AP23464 against the most relevant Abl mutations. In a cell proliferation assay with BaF3 cells expressing... 

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