| Abstract |
Background: Most cell lines derived from primary effusion lymphomas (PEL)
are both HHV-8+ and EBV+. A new cell line (ISI-1) which is only HHV-8+ was
established from the lymphomatous pleural effusion of a HIV-infected
patient.
Cell line characteristics: The morphology and phenotype of the original
lymphomatous cells as well as of the cells in culture were characteristic
of PEL: large/anaplastic cells, CD2-, CD3-, CD19-, CD20-, CD38+, CD71+,
CD45+, CD30+, MHC class I +, class II -. A clonal rearrangement of IgH
gene was demonstrated by Southern blot analysis. The cytogenetic study
revealed multiple chromosome abnormalities. Presence of HHV-8 and absence
of EBV DNA sequences were demonstrated using PCR and Southern blot
analysis. These characteristics have remained stable after 20 months.
Cytokines production: The cell line was maintained in RPMI-1640 and FCS
(15%) and the spontaneous cytokines production was evaluated in the
culture supernatant using ELISA assays. Large amounts of IL10 (26218 pg/ml
[106 cells]) and low concentrations of IL6 (49 pg/ml [106 cells]) were
detected. Intracytoplasmic IL10 production and the expression of the IL10
receptor were demonstrated using flow cytometry.
Animal model: 5x10^6 cells were infused intra peritoneously in 6 NOD/SCID
mice. All mice developed lymphomatous ascitis between day 27 and 96. Nodal
and splenic involvement were present in 3/3. New cell lines (ISI-11, ISI-12)
could be established from the ascitis and spleen of two mice and
exhibited the same phenotype and clonal IgH gene rearrangement than ISI-1.
At day 14 ex vivo, ISI-11 produces IL10 (3179 pg/ml [106 cells]) and no
detectable IL6.
Conclusions: A new HHV-8+ EBV -B cell line (ISI-1) produces large amounts
of IL10 and induces tumors in NOD/SCID mice.
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