Phenotypic and molecular analysis of six human cell lines derived from patients with plasma cell dyscrasia.
Cell lines RPMI 8226, JJN3, U266 B1, NCI-H929 (all EBV-) and ARH77 and HS-Sultan (both EBV+) have been extensively characterized in this study. EBV- lines expressed the phenotype (CD138-, CD19+, CD20+) whereas EBV+ were (CD138+, CD19-, CD20-). CD56 expression was restricted to EBV- cell lines, with the exception of U266 B1, whereas PCA-1 was strongly expressed on five of the six cell lines. Only EBV+ cell lines bound peanut-agglutinin (PNA). However, all cell lines bound the lectin Jacalin that binds the same receptor as PNA, irrespective of the receptors sialylation status. By RT-PCR and direct sequencing of their IgH V/D/J domains, ARH77 was demonstrated to use the germline sequence VH4-34/dm1/JH6b, whereas no arrangement was demonstrated for RPMI 8226, suggesting IgH gene deletion or mutation. HLA class I and II antigens were detected using HLA typing on all cell lines warranting their use as suitable targets for HLA-restricted cytotoxic T cells. By sensitive RT-PCR, mRNA for IL-6, IL-6R and TNFbeta was found expressed in all cell lines. IL-1 mRNA expression was predominantly associated with the EBV+ phenotype. Although mRNA for IL-3 and GM-CSF was never detected, transcripts for c-kit ligand and, more commonly, its receptor were. Likewise GM-CSF, M-CSF and erythropoietin mRNA transcripts were detected in the majority of cell lines.
Gooding
R P
RP
Myeloma Unit, Department of Haematology, Imperial College School of Medicine, Hammersmith Hospital, London. rgooding@rpms.ac.uk
Bybee
A
A
Cooke
F
F
Little
A
A
Marsh
S G
SG
Coelho
E
E
Gupta
D
D
Samson
D
D
Apperley
J F
JF
D001483:Base Sequence; D015139:Blotting, Southern; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D015321:Gene Rearrangement; D006680:HLA Antigens / Q000378:metabolism; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D016130:Immunophenotyping; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000235:genetics; D010265:Paraproteinemias / Q000235:genetics*; D010641:Phenotype; D012313:RNA / Q000032:analysis; D018121:Receptors, Cytokine / Q000378:metabolism
D016207:Cytokines; D006680:HLA Antigens; D007143:Immunoglobulin Heavy Chains; D018121:Receptors, Cytokine; D012313:RNA