Hematopoietic prostaglandin D synthase suppresses intestinal adenomas in ApcMin/+ mice.

Aspirin and other nonsteroidal anti-inflammatory drugs prevent some cases of colon cancer by inhibiting prostaglandin (PG) synthesis. PGE(2) promotes colon neoplasia, as shown by knockout mouse studies on enzymes and receptors in the PG cascade. A few experiments 20 to 30 years ago suggested that PGD(2) may suppress tumors, but a role for biosynthetic enzymes for PGD(2) in tumor development has not been studied. We report here that disruption of the gene for hematopoietic PGD synthase in Apc(Min/+) mice led to approximately 50% more intestinal adenomas compared with controls. Tumor size was not affected. By immunohistochemistry, we detected hematopoietic PGD synthase mainly in macrophages and monocytes of the gut mucosa. The mean number of tumors did not increase with knockout of the gene for the lipocalin type of the enzyme, which is not produced in the intestine. On the other hand, Apc(Min/+) mice with transgenic human hematopoietic PGD synthase tended to have 80% fewer intestinal adenomas. The transgene produced high mRNA levels (375-fold over endogenous). There was a suggestion of higher urinary excretion of 11beta-PGF(2alpha) and a lower excretion of a PGE(2) metabolite in transgenic mice, but differences (30-40%) were not statistically significant. The results support an interpretation that hematopoietic PGD synthase controls an inhibitory effect on intestinal tumors. Further studies will be needed to prove possible mechanisms, such as routing of PG production away from protumorigenic PGE(2) or inhibition of the nuclear factor-kappaB cascade by PGD(2) metabolites.

c-Kit--a hematopoietic cell essential receptor tyrosine kinase.

The receptor tyrosine kinase c-Kit is expressed in hematopoietic stem and progenitor cells and in several non-hematopoietic tissues. In the hematopoietic system, c-Kit is critical for proliferation, survival and differentiation. During recent years exploration of the signalling pathways downstream of this receptor has yielded significant new insights in the field. In this review, we will summarise the c-Kit background, structure, downstream signalling and medical significance with particular focus on its role in hematopoietic progenitor cells and mast cells.

Expression of multiple Na+,K+-adenosine triphosphatase isoform genes in human hematopoietic cells. Behavior of the novel A3 isoform during induced maturation of HL60 cells.

Multiple isoenzymes of the Na+,K+-ATPase (alpha, alpha+, and alpha 3) have been identified by molecular cloning (Shull, G. E., J. Greeb, and J. B. Lingrel. 1986. Biochemistry. 25:8125-8132; and Schneider, J. W., R. W. Mercer, and E. J. Benz, Jr. 1987. Clin. Res. 35:585A. [Abstr.]). At least one of these, the alpha 3 chain, represents a novel form for which protein products and enzymatic activities are just beginning to be defined in rodents. We have recently demonstrated that expression of alpha 3 is largely confined to neuromuscular tissues of fetal and adult rats (Schneider, J. W., R. W. Mercer, M. Gilmore-Hebert, M. F. Utset, C. Lai, A. Greene, and E. J. Benz, Jr. 1988. Proc. Natl. Acad. Sci. USA. 85:284-288). We now report that certain human leukemia cell lines including HL60, HEL, and Molt 4 express mRNA for both alpha and alpha 3 isoforms of Na+,K+-ATPase; mRNA was not detected in several other cell lines, including K562 and U937; no cell lines expressed alpha+ mRNA. In uninduced HL60 cells, alpha 3 mRNA comprised 20-30% of total Na+,K+-ATPase mRNA. Furthermore, in HL60 and HEL cells, both alpha and alpha 3 mRNA declined after induction of maturation by DMSO, retinoic acid, or hemin. However, the reduction in alpha 3 mRNA was far more dramatic. alpha 3 mRNA virtually disappeared, but alpha mRNA declined by only approximately 50%. In contrast, when maturation of HL60 cells along the monocyte/macrophage lineage was induced by exposure to phorbol esters, alpha 3 mRNA remained abundant. Moreover, mRNA for the beta subunit of the Na+,K+-ATPase increased dramatically. Our results demonstrate that the alpha 3 isoform, formerly thought to be confined to neuromuscular tissues, is expressed in restricted lineages of hematopoietic origin. These leukemia cell lines should provide a useful model for analyzing regulation of the alpha 3 isoform gene and characterization of alpha 3 isoform activities.

Selective up-regulation of type II inosine 5'-monophosphate dehydrogenase messenger RNA expression in human leukemias.

The discovery of isozymes (types I and II) of IMP dehydrogenase (IMPDH; EC 1.1.1.205), the rate-limiting enzyme of de novo GTP biosynthesis, has attracted attention as a possible novel approach to cancer diagnosis and selective tumor cell chemotherapy. To elucidate differences in expression and regulation of the two IMPDH isozymes, we examined the steady-state levels of these mRNAs in various types of leukemic cells from patients. Northern blot analysis revealed that type II IMPDH was more active transcriptionally (1.5- to 5.1-fold) in all the leukemic cells examined than in normal lymphocytes, whereas type I expression was similar. The increased expression of type II mRNA in leukemic cells was closely linked with the increase in total IMPDH activity (r = 0.92). When leukemic cells from a patient with chronic granulocytic leukemia in blast crisis were separated into blast-rich and mature leukocyte-rich fractions, the expression of type II mRNA correlated positively with the population of immature leukemic cells, whereas type I expression was unchanged. Treatment of leukemic blasts with 12-O-tetradecanoyl-phorbol-13-acetate for 5 days resulted in a 90% decrease in the expression of type II mRNA with macrophage-like differentiation, while the expression of type I mRNA was relatively stable. These observations suggest that expression of type II IMPDH is stringently linked with immature characteristics of leukemic cells; thus, it should be a selective target for antileukemic chemotherapy.

bcr/abl expression in 32D cl3(G) cells inhibits apoptosis induced by protein tyrosine kinase inhibitors.

Eight protein tyrosine kinase inhibitors with in vitro epidermal growth factor receptor kinase 50% inhibitory concentration values ranging from 0.043 to 22 microM were studied for their ability to inhibit the growth of the murine interleukin-3 (IL-3) dependent myeloid 32D cl3(G) cell line and, a subclone (LG7) transformed to IL-3 independent growth by retroviral transduction and expression of the chronic myelogenous leukemia-associated protein tyrosine kinase p210bcr/abl. Cell proliferation 50% inhibitory concentration values ranged from 4 to 250 microM, and one compound was not inhibitory at 500 microM. The dose-cell proliferation curves were remarkably similar for parental 32D cl3(G) cells + IL-3 and LG7 +/- IL-3, and reversion of LG7 cells to IL-3 dependence was not observed, suggesting that none of the compounds tested could selectively inhibit p210bcr/abl. However, 6 compounds induced the appearance of a 200-base pair nucleosomal DNA ladder characteristic of apoptosis at 24 h in parental 32D cl3(G) cells + IL-3, which mimicked the effects of IL-3 withdrawal alone, but not in similarly growth arrested LG7 cells that eventually developed a necrotic pattern of DNA fragmentation. These studies suggest that the expression of p210bcr/abl can suppress apoptotic signal transduction and that this may contribute to the development of the myeloid hyperplasia that occurs in chronic phase chronic myelogenous leukemia.

C-RAF-1 serine/threonine kinase is required in BCR/ABL-dependent and normal hematopoiesis.

BCR/ABL oncogenic tyrosine kinase is responsible for initiating and maintaining the leukeic phenotype of Philadelphia chromosome-positive cells. c-RAF-1 serine/threonine kinase is known to be activated by receptor and nonreceptor tyrosine kinases. To determine whether c-RAF-1 plays a role in the growth of BCR/ABL-dependent cells, we examined whether c-RAF-1 associates with and/or is regulated by BCR/ABL and, if so, whether this interaction is functionally significant for BCR/ABL-dependent growth of chronic myelogenous leukemia cells and for growth factor-dependent proliferation of normal bone marrow cells. We show that c-RAF-1 enzymatic activity is regulated by BCR/ABL, although the protein does not associate with BCR/ABL. Downregulation of c-RAF-1 expression with antisense oligodeoxynucleotides or cDNA constructs, and inhibition of c-RAF-1 activity by its dominant negative mutants, inhibited both BCR/ABL-dependent growth of chronic myelogenous leukemia cells and growth factor-dependent proliferation of normal hematopoietic progenitors and the MO7 cell line without affecting the BCR/ABL-and growth factor-independent proliferation of HL-60 cells. These results indicate that c-RAF-1 plays an important role in Philadelphia chromosome-positive and normal hematopoiesis.

ErbB-4 ribozymes abolish neuregulin-induced mitogenesis.

The epidermal growth factor-like receptor tyrosine kinase (ErbB) family is frequently overexpressed in a variety of human carcinomas, including breast cancer. To assist in characterizing the role of ErbB-4 in breast cancer, we generated three specific hammerhead ribozymes targeted to the ErbB-4 mRNA. These ribozymes, Rz6, Rz21, and Rz29, efficiently catalyzed the specific cleavage of ErbB-4 message in a cell-free system. We demonstrated that the neuregulin-induced mitogenic effect was abolished in ribozyme Rz29- and Rz6-transfected 32D/ErbB-4 cells. Inhibition of mitogenesis was characterized by ribozyme-mediated down-regulation of ErbB-4 expression. In addition, we provide the first evidence that different threshold levels of ErbB-4 expression and activation correlate with different responses to neuregulin stimulation. High levels of ErbB-4 expression, phosphorylation, and homodimerization are necessary for neuregulin-stimulated, interleukin 3-independent cell proliferation in the 32D/E4 cells. In the case of Rz29-transfected 32D/E4 cells, low levels of ErbB-4 expression allowed neuregulin-induced phosphorylation but were insufficient to couple the activated receptor to cellular signaling. Furthermore, expression of the functional ErbB-4 ribozyme in T47D human breast carcinoma cells led to a down-regulation of endogenous ErbB-4 expression and a reduction of anchorage-independent colony formation. These studies support the use of ErbB-4 ribozymes to define the role of ErbB-4 receptors in human cancers.

Expression of a novel form of Tec kinase in hematopoietic cells and mapping of the gene to chromosome 5 near Kit.

The Tec kinase was initially identified as a novel cytoplasmic protein tyrosine kinase that is preferentially expressed in the liver and is highly homologous to the Drosophila Dsrc28C src-related tyrosine kinase. In screening of interleukin 3 (IL-3)-dependent myeloid leukemia cells for protein tyrosine kinases, we observed that all cell lines examined expressed high levels of Tec transcripts. However, characterization of Tec cDNAs indicated that they differed significantly from the published sequence. Most strikingly, an insertion of 41 bp in the 5' region affects the initiation codon and results in replacing the published 13 amino acid amino-terminal sequences with 94 amino acids. Using polymerase chain reaction (PCR) analysis, only the form containing the insertion was detected in hematopoietic cells. In addition, we found an in-frame insertion of 66 bp that introduces an additional 22 amino acids into the SH3 domain. This insertion restores conserved SH3 sequences that are found in the src gene family and in the Dsrc28C gene. By PCR analysis, approximately equal levels of Tec transcripts containing the intact SH3 domain and containing the 22 amino acid deletion were found in hematopoietic cells. Lastly, by interspecies backcross analysis, we show that the Tec gene is tightly linked to the c-Kit gene on mouse chromosome 5.

Heart-specific activation of LTK results in cardiac hypertrophy, cardiomyocyte degeneration and gene reprogramming in transgenic mice.

Leukocyte tyrosine kinase (LTK) is a receptor-type protein tyrosine kinase belonging to the insulin receptor superfamily. To elucidate its biological role, we generated transgenic mice expressing LTK under the control of cytomegarovirus enhancer and beta-actin promoter. The transgenic mice exhibited growth retardation and most of the transgenic mice died within several months after birth. Interestingly, although LTK was expressed in several major organs, the activation (tyrosine-phosphorylation, kinase activity, and multimerization) of LTK was observed selectively in the heart, where LTK was localized on intracellular membrane, presumably on endoplasmic reticulum. Echocardiography showed that the transgenic heart underwent severe concentric hypertrophy, which resulted in reduced cardiac output, low blood pressure, and increased heart rate. Histological examination of the heart exhibited focal degeneration of cardiomyocytes. These histological changes were considered to be due to apoptosis, based on the finding that the sarcolemmas of the degenerative cardiomyocytes were well preserved. In addition, expression of fetal genes, such as atrial natriuretic peptide and skeletal alpha-actin, was markedly induced in the transgenic heart. These results indicate that a certain tissue-specific mechanism of activating LTK exists in the heart and that the activated LTK resulted in cardiac hypertrophy, cardiomyocyte degeneration and gene reprogramming. These findings will provide novel insights into the activating mechanism and biological role of LTK in vivo.

Prostaglandin D2 and sleep regulation.

Prostaglandin (PG) D2 is recognized as the most potent endogenous sleep-promoting substance whose action mechanism is the best characterized among the various sleep-substances thus far reported. The PGD2 concentration in rat cerebrospinal fluid (CSF) shows a circadian change coupled to the sleep-wake cycle and elevates with an increase in sleep propensity during sleep deprivation. Lipocalin-type PGD synthase is dominantly produced in the arachnoid membrane and choroid plexus of the brain, and is secreted into the CSF to become beta-trace, a major protein component of the CSF. The PGD synthase as well as the PGD2 thus produced circulates in the ventricular system, subarachnoidal space, and extracellular space in the brain system. PGD2 then interacts with DP receptors in the chemosensory region of the ventro-medial surface of the rostral basal forebrain to initiate the signal to promote sleep probably via the activation of adenosine A2A receptive neurons. The activation of DP receptors in the PGD2-sensitive chemosensory region results in activation of a cluster of neurons within the ventrolateral preoptic area, which may promote sleep by inhibiting tuberomammillary nucleus, the source of the ascending histaminergic arousal system.

Rho GTPases in hematopoietic cells.

The ubiquitous Rho GTPases are instrumental in the organization of the actin cytoskeleton, but also for the control of gene expression. Here we review the role of the major members of this family, i.e., RhoA, Rac1, Rac2, and Cdc42, and their intracellular signaling in hematopoietic cells. Although these proteins have been classically implicated in chemotaxis, there are now clear indications on how differential signaling toward other, more specific functions, such as phagocytosis or the production of reactive oxygen species, is regulated by relatively small differences in primary sequence. The identification of mutations in these GTPases or their regulators has provided novel insights in their function as well as their relevance for the development of hematological diseases.

A hematopoietic protein tyrosine phosphatase (HePTP) gene that is amplified and overexpressed in myeloid malignancies maps to chromosome 1q32.1.

Tyrosine phosphorylation is an important regulator of cell growth and differentiation reflecting the interaction of protein tyrosine kinases (PTK) and protein tyrosine phosphatases (PTP). Although excessive PTK activity can result in hematopoietic cell transformation, perturbation of either of these two modulators may result in uncontrolled cell growth. Myeloid cells are responsive to growth factors and cytokines that induce tyrosine phosphorylation and can become ligand independent when endogenous PTKs become dysregulated. Specific PTPs, through mutation or altered expression, may enhance PTK activities and also cause myeloid ligand independence, though this has not yet been demonstrated. We have previously reported the isolation of a hematopoietic specific cytoplasmic PTP (HePTP). We now report that this gene maps to chromosome 1q32.1 utilizing fluorescent in situ chromosomal hybridization (FISH). This site is frequently amplified in preleukemic myeloproliferative diseases. FISH analysis of a patient with myelodysplastic syndrome characterized by myeloid hypoplasia and monocytosis reveals triplication of the HePTP gene on one allele with elevated protein expression in neoplastic myelomonocytic cells. Elevated expression is also identified in blasts from some patients with acute leukemia. These observations prompted us to examine the experimental effects on cell growth of HePTP overexpression. Though normal myeloid cells show minimal HePTP expression, all hematopoietic cell lines tested show high expression of HePTP. Gene transfer of HePTP into NIH 3T3 cells was therefore performed, which caused altered cell morphology, disorganized growth, anchorage independent colony formation and subtle differences in the pattern of tyrosine phosphoproteins compared to control cell lines. We conclude that amplification and overexpression of HePTP may be an important cofactor contributing to abnormal myeloid cell growth.

The Dtk receptor tyrosine kinase, which binds protein S, is expressed during hematopoiesis.

Dtk (Tyro 3/Sky/Rse/Brt/Tif) belongs to a recently recognized subfamily of receptor tyrosine kinases that also includes Ufo (Axl/Ark) and Mer (Eyk). Ligands for Dtk and Ufo have been identified as protein S and the related molecule Gas6, respectively. This study examined expression of Dtk during ontogeny of the hematopoietic system and compared the pattern of expression with that of Ufo. Both receptors were abundantly expressed in differentiating embryonic stem cells, yolk sac blood islands, para-aortic splanchnopleural mesoderm, fractionated AA4+ fetal liver cells, and fetal thymus from day 14 until birth. Although Ufo was expressed at moderate levels in adult bone marrow, expression of Dtk in this tissue was barely detectable. In adult bone marrow subpopulations fractionated using counterflow centrifugal elutriation, immunomagnetic bead selection for lineage-depletion and FACS sorting for c-kit expression, very low levels of Dtk and/or Ufo were detected in some cell fractions. These results suggest that Dtk and Ufo are likely to be involved in the regulation of hematopoiesis, particularly during the embryonic stages of blood cell development.

Tissue specific expression of Yrk kinase: implications for differentiation and inflammation.

The Src family of proto-oncogenes is a highly conserved group of non-receptor tyrosine kinases with very similar, but not identical, tissue distributions and functions. Yrk is a recently discovered new member of this family. Here we report the patterns of expression of this kinase in a variety of chicken tissues during development and after hatching, and experiments that correlate some of the observed patterns of expression with potential functions. The results show that the Yrk protein is primarily found in neuronal and epithelial cells and in monocyte/macrophages. In neuronal tissues of hatched chicks, Yrk is expressed in Purkinje cells, in the gigantocellularis of the brain-stem, and in retinal ganglion cells. In addition, staining for this kinase is also seen as thread-like and punctate patterns suggesting staining in neurites and growth cones. Epithelial cells express Yrk in the stomach during late developmental stages and after hatching but, in other epithelia such as in the peridermis, intestine and kidney, expression is high during development but low (skin) or undetectable (intestine and kidney) after hatching. These results suggest that Yrk may have several functional roles, specifically in cell migration and or differentiation during neuronal and epithelial cell development and in maintenance of the differentiated phenotype. In this study we also show that significant levels of Yrk are detected in monocytes of the blood and in tissue macrophages. Analysis of chicken hematopoietic cell lines confirmed the expression of Yrk in cells of monocyte/macrophage lineage and show for the first time in experimentally-induced inflammation that Yrk kinase activity is high during the period of monocyte infiltration, raising the possibility that this kinase plays a role in inflammation and/or response to injury.

Isolation of tyrosine kinase related genes expressed in the early hematopoietic system.

Transmembrane tyrosine kinase receptors are involved in cellular interactions which promote proliferation and differentiation of many cell types. To identify receptor tyrosine kinases important in embryonic hematopoietic cell development we have utilized the polymerase chain reaction (PCR) and degenerate oligonucleotides for isolation of such genes from mouse yolk sac and fetal liver. Sequence analysis of PCR amplified cDNAs from these hematopoietic sites of day 8 and 14 embryos, resulted in the isolation of nine tyrosine kinase and three serine/threonine kinase related clones. Two of these receptors, tek and flk-1, are expressed in both yolk sac and fetal liver and have been shown previously to be important for endothelial cell development. Two other clones, 9B4 and 9A2 appeared novel upon isolation but have been recently described as ryk and SK2 (rat homologue). Here we describe the twelve isolated kinases, the specific expression patterns of flk-1, tek and ryk kinases and their potential relationship to the development of the hematopoietic system.

Evaluating function of transmembrane protein tyrosine phosphatase CD148 in lymphocyte biology.

The transmembrane protein tyrosine phosphatase CD148 is expressed on numerous cell types, including most cells of the hematopoietic lineage. CD148 has been shown to regulate density-dependent inhibition of cell growth as well as cellular differentiation in nonhematopoietic cells and has been shown to regulate signal transduction processes in several nonlymphoid hematopoietic cell types. Analysis of CD148 expression on lymphoid cells has demonstrated that CD148 is expressed at low levels on T cells and that it is upregulated in response to activation. Several groups have observed that CD148 negatively regulates T cell activation in response to crosslinking of the T cell antigen receptor, suggesting that it may play a role in feedback inhibition of the T cell immune response. In the B cell compartment, CD 148 expression appears to be restricted to the memory subpopulation, raising the possibility that it serves a unique function in these cells, which has yet to be determined. Recent studies have shown that CD148 interacts with the PDZ domain-containing protein syntenin, raising the possibility that its function or its localization with substrates in T and B cells may be controlled through this or a related interaction with another PDZ domain protein.

Hematopoietic tissues, as a playground of receptor tyrosine kinases of the PDGF-receptor family.

Three receptor tyrosine kinases of the PDGF receptor family (RTKP) that clustered within 1000 Kb of the mouse chromosome 5 constitute an interesting unit that are expressed in three distinct cell lineages essential for constructing hematopoietic tissues. Namely, the c-kit gene that is expressed in hematopoietic stem cells is flanked by pdgfr alpha and flk genes expressed respectively in stromal cells and vascular endothelial cells. In this article, we review our results on their expression in the embryonic hematopoietic tissues. We found that co-expression of Flkl and c-Kit was frequently detected either in vascular endothelial cells or hematopoietic cells in the early hematopoietic tissues. On the other hand, the three RTKPs are expressed in different cell lineages in the fetal liver. On the basis of this finding, we propose two modes of embryonic hematopoiesis; hematogenic angiopoiesis and hematopoiesis.

First determination of the inhibitor complex structure of human hematopoietic prostaglandin D synthase.

Hematopoietic prostaglandin (PG) D synthase (H-PGDS) is responsible for the production of PGD(2) as an allergy or inflammation mediator in mast and Th2 cells. We determined the X-ray structure of human H-PGDS complexed with an inhibitor, 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT) at 1.9 A resolution in the presence of Mg(2+). The styryl group of the inhibitor penetrated to the bottom of the active site cleft, and the tetrazole ring was stabilized by the stacking interaction with Trp104, inducing large movement around the alpha5-helix, which caused the space group of the complex crystal to change from P2(1) to P1 upon binding of BSPT. The phthalhydrazidyl group of BSPT exhibited steric hindrance due to the cofactor, glutathione (GSH), increasing the IC(50) value of BSPT for human H-PGDS from 36.2 micro M to 98.1 micro M upon binding of Mg(2+), because the K(m) value of GSH for human H-PGDS was decreased from 0.60 micro M in the presence of EDTA to 0.14 micro M in the presence of Mg(2+). We have to avoid steric hindrance of the GSH molecule that was stabilized by intracellular Mg(2+) in the mM range in the cytosol for further development of structure-based anti-allergic drugs.

Hematopoietic prostaglandin D synthase.

The biological actions of prostaglandin (PG) D(2) include vasodilatation, bronchoconstriction, inhibition of platelet aggregation, and recruitment of inflammatory cells. Characterization of DP receptor null mice in which antigen-induced airway and inflammatory responses are attenuated and identification of CRTH2 as a novel PGD(2) receptor have shed light on the role of PGD(2) in the immune and inflammatory responses. Hematopoietic PGD synthase (H-PGDS) is a cytosolic enzyme that isomerizes PGH(2), a common precursor for all PGs and thromboxanes, to PGD(2) in a glutathione-dependent manner. H-PGDS is expressed in mast cells, antigen-presenting cells, and Th2 cells, and is the only mammalian member of the Sigma class of cytosolic glutathione S-transferases. In this review, we focus on the molecular biology of H-PGDS, the determination of its three-dimensional structure, characterization of the regulation of its gene expression, and information gleaned from transgenic animals.

Sharing of receptor subunits and signal transduction pathway between the IL-4 and IL-13 receptor system.

In this review, we summarize the subunit structure of the interleukin (IL)-4 and IL-13 receptor system and the molecular mechanism of signals through the cytokine receptor systems. We have demonstrated that two different forms of IL-4R exist, classical and alternative. Classical IL-4R is predominantly expressed in hematopoietic cells and consists of IL-4R p140 (beta) and IL-2R gamma (gamma c) chains. The alternative form of IL-4R is predominantly expressed in nonhematopoietic cells and consists of IL-4R beta and IL-13R alpha' chains. Moreover, the alternative form of IL-4R is also used as a functional component in the IL-13R complex. For signal transduction through IL-4R and IL-13R, we have demonstrated that in nonhematopoietic cells, Janus protein tyrosine kinase (JAK) 2 is phosphorylated and activated instead of JAK3 tyrosine kinase. While JAK3 is required for signal transducer and activator of transcription-6 (STAT6) activation in hematopoietic cells, we recently demonstrated that in nonhematopoietic cells JAK2 is required for STAT6 activation for the alternative form of IL-4R. Thus, a major difference exists between hematopoietic and nonhematopoietic cells with regard to structure and signal transduction through the IL-4R and IL-13R systems.