Cell cycle controlling the silencing and functioning of mammalian activators.

Naïve CD4(+) helper T (T(H)) cells respond to stimulation by terminally differentiating into two mature classes, T(H)1 cells, which express interferon gamma (IFN-gamma), and T(H)2 cells, which express interleukin 4 (IL-4). The transcriptional activators T-bet and Gata-3 mediate commitment to the T(H)1 and T(H)2 fates, respectively, including chromatin remodeling of signature genes. The cytokine IL-12 fosters growth of committed T(H)1 cells, while IL-4 fosters growth of committed T(H)2 cells. IL-12 and IL-4 also play critical roles in commitment by promoting transcriptional silencing of Gata-3 and T-bet, respectively. We now show that both T-bet and Gata-3 are induced in a cell cycle-independent manner in bipotent progenitor cells. In contrast, both lineage-restricted gene induction by the activator proteins and heritable silencing of the transcription of each activator, the hallmarks of terminal differentiation, are cell cycle dependent. We found that cells that cannot cycle remain uncommitted and bipotent in response to the most polarizing signals for maturation. These results provide mechanistic insight into a mammalian model of terminal differentiation by illustrating that cell cycle-coupled epigenetic effects, as originally described in yeast, may represent an evolutionarily conserved strategy for organizing signaling and cell fate.

Induction of cytotoxic T lymphocyte antigen 4 (CTLA-4) restricts clonal expansion of helper T cells.

Cytotoxic T lymphocyte antigen (CTLA)-4 plays an essential role in immunologic homeostasis. How this negative regulator of T cell activation executes its functions has remained controversial. We now provide evidence that CTLA-4 mediates a cell-intrinsic counterbalance to restrict the clonal expansion of proliferating CD4(+) T cells. The regulation of CTLA-4 expression and function ensures that, after approximately 3 cell divisions of expansion, most progeny will succumb to either proliferative arrest or death over the ensuing three cell divisions. The quantitative precision of the counterbalance hinges on the graded, time-independent induction of CTLA-4 expression during the first three cell divisions. In contrast to the limits imposed on unpolarized cells, T helper type 1 (Th1) and Th2 effector progeny may be rescued from proliferative arrest by interleukin (IL)-12 and IL-4 signaling, respectively, allowing appropriately stimulated progeny to proceed to the stage of tissue homing. These results suggest that the cell-autonomous regulation of CTLA-4 induction may be a central checkpoint of clonal expansion of CD4(+) T cells, allowing temporally and spatially restricted growth of progeny to be dictated by the nature of the threat posed to the host.

Role of T-bet in commitment of TH1 cells before IL-12-dependent selection.

How cytokines control differentiation of helper T (TH) cells is controversial. We show that T-bet, without apparent assistance from interleukin 12 (IL-12)/STAT4, specifies TH1 effector fate by targeting chromatin remodeling to individual interferon-gamma (IFN-gamma) alleles and by inducing IL-12 receptor beta2 expression. Subsequently, it appears that IL-12/STAT4 serves two essential functions in the development of TH1 cells: as growth signal, inducing survival and cell division; and as trans-activator, prolonging IFN-gamma synthesis through a genetic interaction with the coactivator, CREB-binding protein. These results suggest that a cytokine does not simply induce TH fate choice but instead may act as an essential secondary stimulus that mediates selective survival of a lineage.

A streptomycete antagonist to Phellinus weirii, Fomes annosus, and Phytophthora cinnamomi.

An actinomycete isolated from the rhizoplane of nitrogen-fixing nodules of Ceanothus velutinus was identified as a variety of Streptomyces griseoloalbus. Streptomyces griseoloalbus is a strong antagonist to three destructive root pathogens, Phellinus weirii, Fomes annosus, and Phytophthora cinnamomi, inhibiting all three on several culture media and preventing establishment of F. annosus on hemlock wood disks. The stability and longevity of the antimicrobial substance produced by it, its consistent effect on the pathogens on all substrates, its ability to colonize wood, and its ability to grow at 10 degrees C suggest biological control possibilities for this organism in the Pacific Northwest.

In vitro inhibition of root-rot pathogensPhellinus weirii, Armillariella mellea, Fomes annosus, andPhytophthora cinnamomi by a newly isolatedBacillus sp.

A Gram-positive bacterium that inhibits several root-rot pathogens was isolated from alder forest soil on the Oregon coast. This organism, similar toBacillus cereus, produces in culture media a heat-stable, broad-spectrum antibiotic which inhibits growth of four important Northwest conifer root disease fungi:Phellinus weirii (Murr.) Gilbertson,Fomes annosus (Fr.) Cke.,Armillariella mellea (Fr.) Karst., andPhytophthora cinnamomi Rands.