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Establishment and characterization of a permanent human IgA2/kappa myeloma cell line
J Lohmeyer1, M Hadam, S Santoso
1Department for Internal Medicine, University of Giessen, F.R.G.
Abstract:
A human myeloma cell line designated LOPRA-1 has been established from ascites fluid containing malignant plasma cells of a patient with IgA2/kappa multiple myeloma. The cultured cells which are Epstein-Barr virus (EBV) negative have retained the morphological, cytochemical, ultrastructural and immunophenotypical features of well-differentiated plasma cells. They express the plasma cell antigen PCA-1, the antigens CD28 (Kolt-2) and CD38 (OKT10), the transferrin-receptor (OKT9), and some epitopes of the CD24 antigen (HB8, VIB E3), but are negative for surface immunoglobulins. HLA class II antigens (HLA-DP, -DQ, -DR) and other B-cell markers such as CD10 (CALLA), CD19 (B4), CD20 (B1), CD21 (B2), CD22 (HD39), CD23 (MHM6), CD37 (BL14) and CD39 (G28-8) as analysed by both flow cytometry and immunocytochemistry (PAP/APAAP). With respect to immunoglobulin synthesis, two stable clones were selected by single cell cloning: clone LOPRA-1/5 synthesizes large amounts of alpha 2 heavy and kappa light chains, but secretes only small amounts of these molecules, whereas clone LOPRA-1/4 is clearly devoid of intracellular immunoglobulin heavy and light chains and thus appears to be a chain loss variant. Cytogenetic analysis revealed a pseudotriploid phenotype with several structurally abnormal marker chromosomes: 3n + -, 70, XX, -X, -1, -4, -6, -8, -8, -13, -16, +7, +18, +21, +i(1q), +i(1q), +6q-, +3mar.
Insights
A new human myeloma cell line, LOPRA-1, was established from a patient with IgA2/kappa multiple myeloma. This cell line retains plasma cell features and offers insights into immunoglobulin synthesis and chain loss variants in myeloma.
Area of Science:
- Hematology
- Cell Biology
- Immunology
Background:
- Multiple myeloma is a hematologic malignancy characterized by the proliferation of malignant plasma cells.
- Establishing and characterizing novel cell lines is crucial for understanding disease mechanisms and developing therapeutic strategies.
- The IgA2/kappa subtype of multiple myeloma presents unique research challenges.
Observation:
- A human myeloma cell line, LOPRA-1, was successfully established from ascites fluid.
- The LOPRA-1 cells are Epstein-Barr virus (EBV) negative and exhibit stable morphological, cytochemical, and ultrastructural features of well-differentiated plasma cells.
- Immunophenotypic analysis revealed expression of plasma cell-specific antigens (PCA-1, CD28, CD38, CD24 epitopes) while lacking surface immunoglobulins and other B-cell markers.
Findings:
- LOPRA-1 cells express specific plasma cell antigens and transferrin receptor but are negative for common B-cell markers and surface immunoglobulins.
- Single-cell cloning yielded two distinct variants: LOPRA-1/5, synthesizing but poorly secreting IgA2/kappa, and LOPRA-1/4, a chain loss variant devoid of intracellular immunoglobulin chains.
- Cytogenetic analysis showed a pseudotriploid karyotype with multiple abnormal marker chromosomes, indicating significant genomic instability.
Implications:
- The LOPRA-1 cell line provides a valuable in vitro model for studying IgA2/kappa multiple myeloma.
- The characterized clones offer unique tools for investigating immunoglobulin synthesis regulation and B-cell differentiation pathways.
- Understanding the cytogenetic abnormalities in LOPRA-1 may contribute to identifying novel therapeutic targets in multiple myeloma.
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