Serum-free generation of antigen presenting cells from acute myeloid leukaemic blasts for active specific immunisation.
OBJECTIVE
Immunotherapy holds promise as a new strategy for the eradication of residual cells in acute myeloid leukaemia (AML). Leukaemic antigen presenting cells (APCs) combining optimal antigen presentation and tumour antigenicity could be used as potent T cell activators. For clinical purposes it is desirable to culture APCs under serum-free conditions. Therefore, we compared morphological, immunophenotypical and functional outcome of the serum-free culture of AML-APCs to their serum-enriched culture.


METHODS
AML blasts (n=19) were cultured in the presence of either a cytokine mix or calcium ionophore (CI) for 14 and 2 days, respectively, in FCS-containing medium (FCS), StemSpan serum-free medium (SP) and CellGro serum-free medium (CG). After culture relative yields were calculated and immunophenotypic analysis of APC markers was performed. The mixed leukocyte reaction (MLR) was used to determine T cell stimulating capacity.


RESULTS
Serum-free culture of AML-APCs resulted in comparable morphology, relative yields and immunophenotype to serum-enriched culture. By comparing both serum-free media we observed a trend towards a more mature phenotype of CI-cultured AML-APCs in SP. MLR showed that serum-free cultured cells have equal T cell stimulatory capacity in comparison with serum-enriched culture.


CONCLUSIONS
These data show that the serum-free culture of AML-APCs is feasible and that these APCs are comparable to serum-enriched cultured AML-APCs with regard to morphological, immunophenotypical and functional characteristics. These AML-APCs are suitable for the development of active specific immunisation protocols which meet the criteria for good clinical practise (GCP).
Houtenbos
Ilse
I
Department of Haematology BR 250, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
Westers
Theresia M
TM
Stam
Anita G M
AG
de Gruijl
Tanja D
TD
Scheper
Rik J
RJ
Ossenkoppele
Gert J
GJ
van de Loosdrecht
Arjan A
AA
D000208:Acute Disease; D000328:Adult; D000368:Aged; D000938:Antigen-Presenting Cells / Q000276:immunology*; D015703:Antigens, CD / Q000378:metabolism*; D000951:Antigens, Neoplasm / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000276:immunology / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D016895:Culture Media, Serum-Free; D005260:Female; D006801:Humans; D007114:Immunization; D016130:Immunophenotyping; D007951:Leukemia, Myeloid / Q000276:immunology*; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D014611:Vaccination
D015703:Antigens, CD; D000951:Antigens, Neoplasm; D016895:Culture Media, Serum-Free