H9 cells

Au(CN)2] is taken up into H9 cells, a T-cell line susceptible to HIV infection and retards the proliferation in these cells. At low concentrations, as low as 20 ppb, it is well tolerated in arthritic patients and may serve well as an adjunct alongside existing HIV treatments [146]. 

Brinkmann T, Schaefers J, Guertler L, Kido H, Niwa Y, Katunuma N, et al. Inhibition of tryptase TL2 from human T4+ lymphocytes and inhibition of HIV-1 replication in H9 cells by recombinant aprotinin and bikunin homologs. J Protein Chem 1997 16 651-660. 

Concentration that inhibits uninfected H9 cell growth by 50%. Concentration that inhibits viral replication by 50%. " Therapeutic Index, TI = IC50/EC50. 

Concentration that inhibits uninfected H9 cell growth by 50%. 

AzddU) (92) for further clinical evaluation has been discussed previously. Introducing an iodo or bromo group resulted in a considerable loss of potency compared with AZT, but these derivatives still retained some antiviral activity. Indeed, very recent studies at the NCI indicate that 3 -azido-2, 3 -dideoxy-5-iodouridine (166) and 3 -azido-2, 3 -dideoxy-5-bro-mouridine (167) are excellent antiretroviral agents in vitro, combining minimal toxicity to uninfected human lymphoid H9 cells, with efficient phosphorylation, potent inhibition of RT (A = 0.028 and 0.043 /tM, respectively) and relatively poor activity against host cell DNA polymerase a (ATj = 42.0 and 47.0 /rM, respectively) [224]. 

Split the target lymphoblastoid cells (e.g., C8166 or H9 cells) on d 0 into two or four parts depending on density and resuspend in 40 mL fresh medium (RPMI-1640 plus 10% bovine calf serum) in a 250-mL flask. 

Fig. 3. Effect of immunotoxin treatment on cell morphology. Scanning electron micrographs (80,000x) were made of uninfected H9 cells (A), persistently infected H9/NL4-3 cells (B), H9/NL4-3 cells following treatment with antibody (C), and H9/NL4-3 following immunotoxin treatment (D). Budding virions are plainly evident on H9/NL4-3 cells. Antibody has no effect on the cells immunotoxin results in massive disruption of the cell membrane. Interestingly, remains of budding virions can be seen following immunotoxin treatment, but these are no longer infectious. From ref. 7, courtesy of Mary Ann Liebert Publishers.

A chain and either human anti-gp41 or CD4 kill H9 cells infected with different isolates of HIV, but do not inhibit normal T or B cell function../. AIDS 3,609-614. 

Briggs CJ, Ott DE, Coren LV, Oroszlan S, Tozser J. Comparison of the effect of FK506 and cyclosporin A on vims production in H9 cells chronically and newly infected by HIV-1. Arch Virol 144(11) 2151-2160, 1999. 

They are generally active at the two steps of the integration but are not selective for IN and the practical utility of those catechol-containing inhibitors is severely reduced by cytotoxicity even though they have been found to inhibit HIV-1 integrase in vitro. Nevertheless, lithospermic acids, Fig. (19) were found to be active on purified IN as well as on HIV-1 infected H9 cells, and showed no cytotoxicity to H9 cells [80]. 

Tepperman and colleagues have found that A CN) - is taken up into H9 cells, a continuous T-cell line that is susceptible to HIV infection. At concentrations as low as 20ppb, it retards the proliferation of HIV in these cells. The concentration used is well tolerated in arthritis patients, which suggests that Au(CN)2 may have promise as a complement for existing HIV treatments. ... 

In the course of the search for novel potent anti-AIDS agents, the ethanolic extract of the roots of T. wilfordii was found to show significant anti-HIV activity. Bioassay-directed fractionation of the active extract has led to the isolation and characterization of a new anti-HIV principle, a kaurane-type diterpene lactone, tripterifordin (90), Fig (27). This compound inhibited HIV replication in H9 lymphoc e cells with an EC50 of 1 pg/ml (6 (M) but it did not inhibit uninfected H9 cell growth at 15 xM [180]. From the same species a new kaurene type diterpene, neotripterifordin (91) was also isolated, which showed potent anti-HIV replication activity in H9 lymphocyte cells with an EC50 of 25 nM and a... 

Le Dinh T, Freneaux E, Labbe G, Letteron P, Degott C, Geneve J, Berson A, Larrey D, Pessayre D (1988) Amineptine, a tricyclic antidepressant, inhibits the mitochondrial oxidation of fatty acids and produces microvesicular steatosis of the liver in mice. J Pharmacol Exp Ther 247 745-750 Le Roy F, Bisbal C, Silhol M, Martinand C, Lebleu B, Salehzada T (2001) The 2-5A/RNase L/RNase inhibitor (RLl) pathway regulates mitochondrial mRNAs stability in interferon-a-treated H9 cells. J Biol Chem 276 48473 8482 Le Roy F, Silhol M, Salehzada T, Bisbal C (2007) Regulation of mitochondrial mRNA stability by RNase L is translation-dependent and controls IFNalpha-induced apoptosis. Cell Death Differ 14 1406-1413... 

Assay of Protein Synthesis. For radiolabeling, H9 cells (3 x 10 cells per well in microtiter plates) were grown in the presence of ascorbate at 0, 75, 100, and 150 ig/ml as described earlier. On days 1, 2, and 4, cells were harvested, washed, and resuspended in methionine- and cysteine-free medium and then incubated at 37°C for 30 min in 0.5 ml of the same medium supplemented with 50 juCi of Tran S-label (specific activity, 1013 Ci/mmol ICN), Labeled cells were pelleted, washed in isotonic phosphate-buffered saline, resuspended in lysis buffer containing 1% Nonidet P-40, and... 

Fig. 1. Analysis of cytotoxicity of ascorbate (AA) for HTLV-IIlB-infecled H9 cells, as determine by trypan blue dye exclusion. Each point is the mean of four cell counts.

Effect of Ascorbate on Cell Metabolism. We addressed the question of whether ascorbate-induced suppression of RT and p24 production in H9/HTLV-IIIb cells was a virus-specific effect or an indirect effect due to inhibition of cellular metabolism or protein synthesis. The metabolic activity of uninfected H9 cells in the presence and absence of ascorbate was determined by using a quantitative colorimetric assay that utilizes the tetrazolium salt MTT (18). MTT is used to measure the activity of various dehydrogenases in viable cells (18, 19). H9 cells grown in the presence of various concentrations of ascorbate (0-150 /tg/ml) showed an increase in cellular metabolic activity on day 1 (Fig. 4). This correlated with stimulation of cell proliferation by ascorbate. On days 2 and 4, no significant change in metabolic activity was noted between control cultures and those exposed to ascorbate at 75,100, and 150 /xg/ml. 

Effect of Ascorbate on Cellular Protein Synthesis. The effect of ascorbate on cellular protein synthesis was determined by growing uninfected H9 cells for 4 days with ascorbate at 0,75, 100, and 150 M8/tnl (17). On day 1, ascorbate stimulated protein synthesis, consistent with stimulation of metabolic activity and cell growth. On days 2 and 4, the difference in the apparent rates of cellular protein synthesis between ascorbate-treated and control cultures was less than a factor of 2 (Fig. 5). Thus the suppressive effects on HIV production could not be ascribed to a general inhibition of cellular metabolism or protein synthesis. 

Concentration that is toxic to 50% of mock-infected H9 cells. 

Inhibition of HIV in virto (IC50) are cytoxicity (TC50) measured in H9 cells (compound (21) and (25-28)) or C8166 cells (compounds (29)-(31)) infected with HlV-ljg and HIV-l p,... 

W. E. Muller, M. Bachmann, B. E. Weiler, H. C. Schroder, G. Uhlenbruck, T. Shinoda, H. Shimizu, H. Ushijima, Antibodies against defined carbohydrate structures of Candida albicans protect H9 cells against infection with human immunodeficiency virus-1 in vitro. J. Acquir. Immun. Defic. Syndr. 1991, 4, 694-703. 

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