Immune cell regulators in mouse mammary development and involution.

Dramatic changes in cell composition and function occur in the mammary gland during a pregnancy-lactation-involution cycle. We investigated the transcriptional changes associated with these biological events by using microarray analysis and identified the critical genes involved by using genetically modified mice. Two surprising findings arose from these studies. First, the microarray data showed that postlactational regression was associated with an acute phase inflammatory response, in addition to cell death. Conditional deletion of signal transducer and activator of transcription (Stat)3 or the nuclear factor-kappaB regulatory kinase inhibitor of kappa B kinase beta resulted in a failure of cell death induction during involution, an indication that these signaling pathways are essential mediators of the involution process. Both Stat3 and nuclear factor-kappaB have been shown to regulate acute phase gene expression in addition to apoptosis regulators. Four distinct transcriptional profiles are present in the first 4 d of involution, whereas there are 3 in lactation. At the peak of lactation (i.e., d 10 in mouse), more than 400 genes reach their maximum expression before declining dramatically in the first 12 h of involution. A reciprocal pattern was observed for more than 500 genes that were specifically upregulated within the first 12 h of forced involution. We are now investigating the role of a subset of these genes in involution. We also uncovered a role for genes normally associated with immune cell signaling in the differentiation of luminal mammary epithelial cells during pregnancy. Genetic deletion of the transcription factor Stat6 resulted in delayed development during pregnancy, and this phenotype was recapitulated in mammary tissue from IL-4 and IL-13 doubly deficient mice. Furthermore, we showed that mammary epithelial cells secrete T-cell regulatory cytokines. T-helper type 1 cytokines, such as interferon-gamma and IL-12a, are secreted by undifferentiated mammary epithelial cells, whereas T-helper type 2 cytokines, including IL-4 and IL-13, are secreted by differentiated cells. This unexpected finding demonstrates a role for immune cell signaling in mammary epithelial cell fate and function.