Acute lymphoblastic leukaemia cells express CCR7 but not higher amounts of IL-10 after CD40 ligation.
OBJECTIVE
Production of cytokines that support T-cell activation and proliferation and migration to lymph nodes is one of the most important terms of cancer vaccine development. In previous studies we and others used CD40 ligation to obtain higher expression of co-stimulatory and adhesion molecules on leukaemic cells from children with acute lymphoblastic leukaemia (ALL). This time we assess the cytokine and chemokine gene expression profile in CD40-stimulated ALL cells.


METHODS
Malignant cells from 25 children with BCP-ALL were stimulated (or not) with huCD40LT and rIL-4 for 96 h. Eleven different molecule, cytokine and chemokine mRNAs levels (CCR7, IL-23, TGF-beta-IP, IFN-gamma, IL-10, CD1a, CD40, CD54, CD80, CD83, CD86) were determined using the real-time PCR technique with TaqMan chemistry using ready-to-use low-density arrays for gene expression by Applied Biosystems.


RESULTS
1) Increases in mRNA levels for CD40, CD54 and CD80 after CD40L and IL-4 stimulation were observed, 2) CCR7 mRNA expression was higher after CD40 ligation than before the culture (p = 0.002), 3) IL-10 mRNA expression was higher after the culture with medium than before the culture (p = 0.01).


CONCLUSIONS
The results show that leukaemia-derived dendritic cells obtained with CD40 ligation express CCR7 - chemokine is involved in migration to lymph nodes and does not produce higher amounts of IL-10, a potent immunosuppressive cytokine. Our preclinical findings could be used in the design of immunotherapy trials for the treatment of children with ALL.
Luczyński
W
W
Department of Pediatric Hematology, Medical University in Białystok, Poland. w.luczynski@wp.pl
Iłendo
E
E
Kovalchuk
O
O
Krawczuk-Rybak
M
M
Malinowska
I
I
Kołtan
A
A
Szczepański
T
T
Wysocka
J
J
Jaworowski
R
R
Olejnik
I
I
Chyczewski
L
L
Matysiak
M
M
Wysocki
M
M
Sońta-Jakimczyk
D
D
Wieczorek
M
M
D019013:CD40 Antigens / Q000494:pharmacology*; D002478:Cells, Cultured; D002648:Child; D003713:Dendritic Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis / Q000235:genetics*; D015847:Interleukin-4 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D054400:Receptors, CCR7; D019707:Receptors, Chemokine / Q000096:biosynthesis / Q000235:genetics*; D015854:Up-Regulation / Q000187:drug effects
C515370:CCR7 protein, human; D019013:CD40 Antigens; D012333:RNA, Messenger; D054400:Receptors, CCR7; D019707:Receptors, Chemokine; D016753:Interleukin-10; D015847:Interleukin-4