A novel gene coding for the pre-B-cell colony-enhancing factor (PBEF) has been isolated from a human peripheral blood lymphocyte cDNA library. The expression of this gene is induced by pokeweed mitogen and superinduced by cycloheximide. It is also induced in the T-lymphoblastoid cell line HUT 78 after phorbol ester (phorbol myristate acetate) treatment. The predominant mRNA for PBEF is approximately 2.4 kb long and codes for a 52-kDa secreted protein. The 3' untranslated region of the mRNA has multiple TATT motifs, usually found in cytokine and oncogene messages. The PBEF gene is mainly transcribed in human bone marrow, liver tissue, and muscle. We have expressed PBEF in COS 7 and PA317 cells and have tested the biological activities of the conditioned medium as well as the antibody-purified protein in different in vitro assays. PBEF itself had no activity but synergized the pre-B-cell colony formation activity of stem cell factor and interleukin 7. In the presence of PBEF, the number of pre-B-cell colonies was increased by at least 70% above the amount stimulated by stem cell factor plus interleukin 7. No effect of PBEF was found with cells of myeloid or erythroid lineages. These data define PBEF as a novel cytokine which acts on early B-lineage precursor cells.

Cloning and characterization of the cDNA encoding a novel human pre-B- cell colony-enhancing factor

Hematopoiesis: A Human Perspective

Cloning andCharacterization ofthecDNA Encoding a Novel HumanPre-B-Cell Colony-Enhancing Factor

Generation and proliferation of early B-cell progenitors have been known to require stromal cell-derived molecules. A stromal cell line, PA6, was found to produce a soluble mediator, which was distinct from interleukin 7 (IL-7) and stem cell factor and supported the proliferation of a stromal cell-dependent pre-B-cell clone, DW34. A cDNA clone encoding this DW34 growth-stimulating factor was isolated by expression cloning. The nucleotide sequence contained a single substantial open reading frame of 267 nucleotides encoding an 89-amino acid polypeptide. The amino acid sequence of this cytokine, designated pre-B-cell growth-stimulating factor (PBSF), revealed that it is a member of intercrine alpha subfamily. Recombinant PBSF stimulated the proliferation of DW34 cells for itself and, furthermore, synergistically augmented the growth of DW34 as well as bone marrow B-cell progenitors in the presence of IL-7.

https://www.pnas.org/content/pnas/91/6/2305.full.pdf

Molecular cloning and structure of a pre-B-cell growth-stimulating factor

The biology of stem cell factor and its receptor C-kit.

Bone marrow stromal cell regulation of B lymphopoiesis: interleukin-1 (IL-1) and IL-4 regulate stromal cell support of pre-B cell production in vitro.

In mammals, B lymphocytes are continuously generated in the bone marrow along with other hematopoietic lineages. Differentiation along the B cell lineage results in clones of immunocompetent cells that express identical immunoglobulin (Ig) variable region specificity. Although the molecular details of the Ig gene rearrangements that result in clonotype specificity have been well documented, only during the last few years have we begun to appreciate that the tempo of B cell generation in the marrow is regulated by a hierarchy of positive and negative signals. We and others have described roles for several regulatory cell types that act to maintain B lymphopoietic differentiation; the essential regulatory component in vitro appears to be a population of fibroblastic stromal cells. These stromal elements of the marrow produce hematopoietic and lymphopoietic cytokines'-' and are capable of supporting both Ig gene rearrangement and B lymphoid cell development when maintained under defined culture condition^.^*^ It appears that these same stromal cells function to potentiate myeloid differentiation2v6.' and that, to some extent, their capacity to support the differentiation of hematopoietic lineages is regulated by environmental levels of ~ytokines.~**.~ Our studies of the role of stromal cells in regulating B cell development have demonstrated that several cytokines known to accelerate myeloid differentiation in v i m have negative effects on B cell genesis. Although some of these negative regulatory interactions appear to act directly at the level of lymphoid precursor cells, the studies presented here establish that a number of cytokines inhibit B cell generation indirectly, by altering stromal cell function. The studies presented here demonstrate that either interleukin-1 (IL-1) or IL-4 can alter stromal cell function and act as a functional negative regulator of B cell development in vitro. We have also found that elevated levels of colony-stimulating factors (CSF) can also act as negative regulators of B lymphopoiesis. Taken together our data suggest that both macrophages and T lymphocytes play a role in regulating stromal cell function and that B lymphocyte and myeloid cell production in the marrow are developmentally linked and coordinately regulated.

Suppression of bone marrow stromal cell function.

Differential roles of stromal cells, interleukin-7, and kit-ligand in the regulation of B lymphopoiesis

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Bone marrow stromal cell regulation of B lymphopoiesis: interleukin-1 (IL-1) and IL-4 regulate stromal cell support of pre-B cell production in vitro.

Suppression of bone marrow stromal cell function.

Differential roles of stromal cells, interleukin-7, and kit-ligand in the regulation of B lymphopoiesis