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Molecular investigation of the cytokines produced by normal and malignant B lymphocytes
M Schena1, G Gaidano, D Gottardi
1Dipartimento di Scienze Biomediche e Oncologia Umana, Sezione Clinica, Torino, Italy.
Insights
Normal and malignant B-cells produce various cytokines. Resting B-cells are key cytokine producers, and B-cell chronic lymphocytic leukemia (B-CLL) cells mimic this resting profile.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- B-cells play crucial roles in immunity and are implicated in various malignancies.
- Understanding cytokine production by normal and malignant B-cells is vital for comprehending immune responses and disease pathogenesis.
Purpose of the Study:
- To investigate in vivo cytokine production by different normal and malignant human B-cell populations.
- To determine the relationship between cytokine expression and the proliferative state of B-cells.
Main Methods:
- Purification of normal B-cell subsets (germinal center B-blasts, mantle B-cells) and malignant B-cells (B-chronic lymphocytic leukemia).
- Analysis of gene expression for key interleukins (IL-1α, IL-1β, IL-2, IL-4, IL-6), tumor necrosis factor alpha (TNF-α), and transforming growth factor beta (TGF-β) using Northern and Western blotting.
Main Results:
- Tumor necrosis factor alpha (TNF-α) mRNA was detected in both resting (mantle B-cells) and proliferating (germinal center B-blasts) normal B-cells.
- Transforming growth factor beta (TGF-β) mRNA was highly expressed in resting mantle B-cells but lower in proliferating germinal center B-blasts and activated CD5+ B-cells.
- Interleukin-2 (IL-2) production was specific to actively proliferating germinal center B-blasts, while IL-1β and IL-6 were produced by resting mantle B-cells.
- Malignant CD5+ B-chronic lymphocytic leukemia (B-CLL) cells exhibited a cytokine production profile similar to resting mantle B-cells, producing IL-1β, TNF-α, TGF-β, and IL-6.
Conclusions:
- Resting mantle B-lymphocytes are the most significant cytokine producers among normal B-cell populations.
- Malignant B-cells in B-chronic lymphocytic leukemia (B-CLL) reflect the cytokine production pattern of normal resting B-lymphocytes.
Abstract:
Different normal and malignant human B-cell populations were studied with a twofold aim: to define which cytokines are produced in vivo, and to assess the relationship between cytokine production and kinetic state. To analyse normal B-cells representative of different stages of activation and proliferation in vivo, we purified germinal centre (GC)-B blasts and mantle B (M-B) cells from tonsils. To compare malignant B lymphocytes with their closest normal equivalent cells, we separated malignant CD5+B lymphocytes from the peripheral blood of patients with B-chronic lymphocytic leukemia (B-CLL) and normal CD5+B lymphocytes from cord blood. The expression of interleukins (IL) IL-1 alpha, IL-1 beta, tumour necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), IL-2, IL-4, and IL-6 genes was analysed using Northern and Western blotting techniques. TNF-alpha mRNA is produced by resting (M-B) and actively proliferating (GC-B) normal B lymphocytes. TGF-beta mRNA is present at high levels in resting normal M-B cells, while the transcript levels are lower in proliferating GC-B and in activated CD5+B lymphocytes. IL-2 production is limited to the actively proliferating GC-B blasts, IL-1 beta and IL-6 to resting M-B cells. The cytokine production profile of CD5+ malignant B-CLL cells differs from that of their putative normal counterparts and is more like the profile of M-B cells, since B-CLL cells produce IL-1 beta, TNF-alpha, TGF-beta, and IL-6. These observations lead to the following conclusions: among normal B lymphocyte populations, resting M-B lymphocytes are the most active cytokine producers, and B-CLL malignant B cells reflect the production pattern of normal resting B lymphocytes.
