Features of hpdODN B consist in a stretch of pyrimidines spanning nucleotides 1

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Features of hpdODN B consist in a stretch of pyrimidines spanning nucleotides 1

Post  huwan123456 on Thu May 22, 2014 5:42 am

The percentages of amount of ganglioside GD2 of all gangliosides isolated from cell lines EL 4, IMR 32 and mS were 60%, 45% and 35%, respectively, Ganglioside GD2 was not detected in ganglioside extracts of Jurkat, Neuro 2A and A375 cell lines. Thus, we confirmed biochemically that we chose appropriate GD2 positive and GD2 negative cell lines to study physiological [You must be registered and logged in to see this link.] effects of anti GD2 mAbs. Cytotoxic effects of two types of anti GD2 mAbs 14G2a and ME361 on GD2 positive and GD2 negative tumor cell lines The cytotoxic effects of anti GD2 mAbs on selected GD2 positive and GD2 negative cell lines were further investigated using two different monoclonal antibodies 14G2a and ME361.

We found that after 24 h of incuba tion of tumor cells with anti GD2 mAbs at concentra tion of 5 ug ml GD2 positive cells underwent significant morphological changes, shrinkage and [You must be registered and logged in to see this link.] rounding of the cells, their detachment from plates, and formation of cell aggregates. All of these morphological changes were the most dramatic for GD2 positive EL 4 and mS cell lines, These anti GD2 mAbs had no effect on morphology of all examined GD2 negative cell lines, Next, we investigated DNA fragmentation in the popu lation of the cells treated with anti GD2 mAbs. After incubation with anti GD2 antibodies, the cells were fixed, permeabilized and stained with DNA binding dye propidium iodide, The percentage of the cells in hypodiploid peak of three tested GD2 positive tumor cell lines EL 4, mS and IMR 32 was increased after anti GD2 treatment when compared to untreated cells.

After incubation with two different anti GD2 antibodies 14G2a and ME361 at concentrations of 5 ug ml the percentage of EL 4 cells with fragmented DNA increased 5. 0 0. 7 and 3. 1 0. 9 fold above baseline level, respect ively, When compared [You must be registered and logged in to see this link.] to EL 4 cells, an in crease in percentage of the cells with fragmented DNA for IMR 32 and mS cell lines was slightly lower, but still sta tistically significant After incubation with 14G2a and ME361 mAbs, the proportion of IMR 32 cells with frag mented DNA increased 2. 5 0. 5 and 1. 7 0. 4 fold, re spectively. For mS cells treated with 14G2a and ME361 antibodies, these values were 3. 2 0. 4 and 2. 3 0. 5, re spectively, Anti GD2 mAbs did not affect GD2 negative tumor cell lines, We further investigated the viability of tumor cells incubated with various concentrations of anti GD2 mAbs using MTT assay.

As shown in Figure 4, anti GD2 anti bodies substantially decreased viability of GD2 positive EL 4, mS and IMR 32 cell lines, without a significant influence on GD2 negative cell lines Neuro 2A, A375, and Jurkat. Note that the anti GD2 antibodies 14G2a were more cytotoxic for GD2 positive cell lines when compared to ME361 antibodies, After 72 h of incubation of the cells with the highest concentra tion of 14G2a antibodies, the strongest effect was observed for EL 4 lymphoma cells, which express the highest level of GD2. While the viability of the EL 4 cells was reduced by more than 80%, the viability of mS and IMR 32 cells decreased by 60 70%. The cytotoxic effect of ME361 antibodies was weaker, but still substantial, and the differences in viability of GD2 positive and GD2 negative cell lines were statistically significant for concen trations of antibodies higher than 2.


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