CD40L and IL-4 stimulation of acute lymphoblastic leukemia cells results in upregulation of mRNA level of FLICE--an important component of apoptosis.
The use of cancer vaccines based on dendritic cells (DC) presenting tumor antigens can be a promising tool in the treatment of leukemia. The functional characteristics of leukemia derived DC is still to be elucidated. CD40 promotes survival, proliferation and differentiation of normal B cells. CD40 triggering was used to enhance the poor antigen-presenting capacity of leukemic B-cells. Since it is still unclear whether CD40 ligation drives neoplastic B-cells to apoptosis or not, we assessed the mRNA expression of FLICE, FAS, FADD and TRADD - important components of apoptosis machinery, using real-time PCR in acute lymphoblastic leukemia cells before and after CD40 and IL-4 stimulation. ALL cells stimulated with CD40L/IL-4 expressed dendritic cell phenotype at mRNA and protein levels (upregulation of main costimulatory and adhesion molecules noted in real-time RT PCR and flow cytometry); they also expressed higher amounts of mRNA for FLICE, TRADD and FADD after CD40L/IL-4 stimulation. However differences statistically significant comparing cells cultured with CD40L/IL-4 and medium alone regarded only FLICE. Concluding, we showed upregulation of important elements of apoptosis at mRNA level in ALL cells after CD40 ligation.
łuczyński
Włodzimierz
W
Departments of Pediatric Hematology and Oncology, Medical University in Białystok, Poland. w.luczynski@wp.pl
Kowalczuk
Oksana
O
Iłendo
Elibieta
E
Stasiak-Barmuta
Anna
A
Krawczuk-Rybak
Maryna
M
Malinowska
Iwona
I
Kołtan
Andrzej
A
Szczepalński
Tomasz
T
Olejnik
Igor
I
Jaworowski
Radosław
R
Chyczewski
Lech
L
Matysiak
Michał
M
Wysocki
Mariusz
M
Sońta-Jakimczyk
Danuta
D
Wieczorek
Maria
M
D017209:Apoptosis; D023201:CD40 Ligand / Q000494:pharmacology*; D053181:Caspase 8 / Q000235:genetics* / Q000378:metabolism; D003713:Dendritic Cells / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology*; D008297:Male; D010641:Phenotype; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism; D053199:TNF Receptor-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism
C505533:FADD protein, human; C491901:FAS protein, human; D053200:Fas-Associated Death Domain Protein; D012333:RNA, Messenger; D053199:TNF Receptor-Associated Death Domain Protein; D019014:fas Receptor; D023201:CD40 Ligand; D015847:Interleukin-4; D053181:Caspase 8