Volumen: 7 # Number : 3
Publication Date : Noviembre - Diciembre Year: 2003
Authors: Dupont J, Fló J, Zorzópulos J, Solimano J, López R, Garay G, Riveros D, Montaner A, Rodriguez J, Fernandez J, Elias F, Cacchione R.
Abstract: Introduction: B-CLL cells are poor stimulators of T-cells because they usually have low expression of costimulatory molecules necessary for T-cells activation (CD40, CD80, CD86), even though they express MHC class I and II molecules. As a consequence, their unique surface Ig idiotype may escape immunological surveillance. ODNs bearing a CpG motif, stimulate B and plasmacytoid dendritic cells. Also, they up-regulate the expression of several co-stimulatory molecules in B-CLL cells and they induce direct apoptosis. Recently, it has been communicated that other ODNs, bearing PyNTTTTGT (Py: pyramidine, N: any nucleotide) motifs are also able to stimulate human B and plasmacytoid dentritic cells. Methods: In this study we investigated the effects of these novel ODNs regardin the expression of co-stimulatory molecules and apoptosis induction in human B-CLL cells. B-CLL cells (CD5, CD19, and CD20 positive) from 20 patients were comparatively incubated with the following 24mer ODNs a)IMT504 bearing PyNTTTTGT motif, b) ODN2006 bearing CpG motif and c) ODN22, a non-stimulatory ODN as control. All incubations were made at the same concentrations of the corresponding ODN. After 24 hrs of incubation, flow cytometric determinatiosn in CD5/CD19 positive cells (Becton Dickinson, San Jose, CA) were made for CD20, CD40, CD80, CD86, expression with fluorescent antisera, (Serotec, Raleigh, NC). Apoptosis induction was evaluated after 40 hrs, using the Annexin V-phosphatidyliserine method (Serotec, Raleigh, NC). Incubation without ODNs determinations were taken as baseline to build an increase index. Mutational status of Ig VH gene and complete clinical data were also obtained. Results: Median increase index for CD40 was 4.84 (1.44-9.20); for CD80 was 1.55 (1.23-2.41); for CD86 was 1.82 (0.9-3.8) in B-CLL cells incubated with IMT504. Median increase index for CD40 was 4.54 (1.40-9.38); for CD80 was 1.38 (1.10-2.36); for CD86 was 1.82 (0.9-3.80) in the B-cells incubated with the ODN2006. Neither ODN2006, nor IMT504 increased the expression index was 1.66 (0.77-2.61) and 2.01 (1.07-2.61) for IMT504 and ODN2006 respectively. ODN22 incubated controls. Mutational satus and clinical correlations were also made. Conclusions: Both, PyNTTTTGT ODNs and CpG ODNs are able to up-regulate co-stimulatory molecules in B-CLL cells and directly stimulate apoptosis in B-CLL cells of some patients. The results here presented strongly suggest the possibility that stimulation of B-CLL cells in vivo with some ODNs, may be an atractive immunotheraphy for B-CLL. Our preclinical (unpublished) in three animal species, including non-human primates. Therefore, phase I/II clinical trials using these ODNs in B-CLL may be warranted.
Key words: B-CLL, Oligodeoxynucleotide, biology Introducción
Pages : 167-175
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