SEARCH:   Advanced

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Suzuki, K. Articles by Iwamoto, I. Search for Related Content PubMed PubMed Citation Articles by Suzuki, K. Articles by Iwamoto, I. Related Collections Hematopoiesis and Stem Cells Immunobiology Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 15 September 2000, Vol. 96, No. 6, pp. 2172-2180 IMMUNOBIOLOGY Role of common cytokine receptor  chain (c)- and Jak3-dependent signaling in the proliferation and survival of murine mast cells Kotaro Suzuki, Hiroshi Nakajima, Norihiko Watanabe, Shin-ichiro Kagami, Akira Suto, Yasushi Saito, Takashi Saito, and Itsuo Iwamoto From the Department of Internal Medicine II, Chiba University School of Medicine, Chiba; and Department of Molecular Genetics, Chiba University Graduate School of Medicine, Chiba, Japan.     Abstract Top Abstract Introduction Materials and methods Results Discussion References The regulatory roles of the common cytokine receptor  chain (c)- and Jak3-dependent signaling in the proliferation and survival of mast cells were determined using c-deficient (c) and Jak3-deficient (Jak3) mice. Although the mast cells in c and Jak3 mice were morphologically indistinguishable from those in wild-type mice, the number of peritoneal mast cells was decreased in c and Jak3 mice as compared with that in wild-type mice. Among c-related cytokines, interleukin (IL)-4 and IL-9, but not IL-2, IL-7, or IL-15, enhanced the proliferation and survival of bone marrow-derived mast cells (BMMCs) from wild-type mice. However, the effects of IL-4 and IL-9 were absent in BMMCs from c and Jak3 mice. In addition, IL-4R, c, and Jak3, but not IL-2R or IL-7R, were expressed in BMMCs. In contrast, IL-13 did not significantly induce the proliferation and survival of BMMCs even from wild-type mice, and IL-13R1 was not expressed in BMMCs. Furthermore, IL-4 phosphorylated the 65-kd isoform of Stat6 in BMMCs from wild-type mice but not from c and Jak3 mice. These results indicate that c- and Jak3-dependent signaling is essential for IL-4- and IL-9-induced proliferation and survival of murine mast cells, that the effects of IL-4 are mediated by type I IL-4R and that type II IL-4R is absent on mast cells, and that IL-4 phosphorylates the 65-kd isoform of Stat6 in mast cells in a c- and Jak3-dependent manner. (Blood. 2000;96:2172-2180) © 2000 by The American Society of Hematology.     Introduction Top Abstract Introduction Materials and methods Results Discussion References Mast cells are recognized as the major effector cells of the type I hypersensitivity reactions by virtue of their possessing high-affinity receptors for immunoglobulin (Ig) E and are known to play a pivotal role in allergic diseases, such as atopic rhinitis, asthma, and atopic dermatitis.1,2 Mature mast cells are distributed throughout all vascularized tissues, and the development and proliferation of mast cells require proper signaling from several cytokines, among which the c-kit/stem cell factor (SCF) system and interleukin (IL)-3 are the best studied.1-4 The common cytokine receptor  chain (c) is a shared component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15.5,6 Mutations in c result in X-linked severe combined immunodeficiency (XSCID) in humans.7 Like humans with XSCID, mice in which the c gene has been disrupted by homologous recombination exhibit a hypoplastic thymus and are compromised in their ability to respond to antigenic stimuli.8-10 Janus kinase 3 (Jak3) is a tyrosine kinase that is known to transduce c-dependent signals.11 Mutation of the Jak3 gene results in a form of severe combined immunodeficiency (SCID) that is clinically and immunologically indistinguishable from XSCID, except for the autosomal recessive inheritance pattern in Jak3-deficient SCID,12,13 which suggests the essential role of Jak3 in c-dependent signaling for lymphocyte development. IL-7 appears to be the most important cytokine for T-cell and B-cell development,14-16 whereas IL-15 seems to be important for natural killer (NK) cell and NK T-cell development.17-19 Among c-related cytokines, IL-4,20-23 IL-9,24,25 and IL-1526 have been reported to support the proliferation and survival of mast cells. IL-4 exerts a number of biologic activities in the hematopoietic and immune system.27 IL-4 transduces the signals through 2 types of IL-4 receptors (IL-4Rs): Type I IL-4R is a heterodimer of IL-4R and c, and type II IL-4R is a heterodimer of IL-4R and IL-13R1.28,29 Type I IL-4R is preferentially expressed in hematopoietic cells, whereas type II IL-4R is preferentially expressed in nonhematopoietic cells.28,29 However, in some hematopoietic cells, including human B cells30,31 and murine macrophages,32 both types of IL-4R are expressed, and IL-4 can transduce the signals in the absence of c through type II IL-4R. Recently, it has been shown that IL-13, a cytokine that shares many biologic activities with IL-4,33 also uses type II IL-4R for signal transduction.29 Although previous studies have shown that IL-4 is a potent stimulator of mast cell proliferation and survival,20-23 it remains unknown which type of IL-4R is functional on mast cells. Furthermore, the role of IL-13 in mast cell growth also remains unclear. IL-15 is a cytokine that shares many functional properties with IL-2.34 Analysis of IL-15R-deficient mice revealed that IL-15 played important roles in NK cell development and generation of memory CD8+ T cells.19 Recently, it was demonstrated that IL-15 induced mast cell proliferation through an undefined receptor, IL-15R X.26 Interestingly, Tagaya et al found that IL-15 activated Jak2 in mast cells through IL-15R X,26 whereas IL-15 activated Jak1 and Jak3 in T cells and NK cells through a heterodimer of IL-2R and c.34 However, the nature of IL-15R X and IL-15 signaling in mast cell proliferation is still largely unknown. Although analyses of mice lacking c and Jak3 have revealed the importance of c and Jak3 in T-cell and NK cell development in vivo,8-11,35,36 the roles of c and Jak3 in mast cell development remain unclear. We present data that demonstrate an important regulatory role of c- and Jak3-dependent signals in the proliferation and survival of murine mast cells.     Materials and methods Top Abstract Introduction Materials and methods Results Discussion References Mice and genetic analysis c-Deficient (c) mice9 and Jak3-deficient (Jak3) mice35 were back-crossed to BALB/c mice (Jackson Laboratory, Bar Harbor, ME) for at least 6 generations. The mice were genotyped by polymerase chain reaction (PCR) as described previously,36 and littermate wild-type (WT) mice were used as controls. Mice were housed in microisolator cages under pathogen-free conditions. All experiments followed the guidelines of Chiba University. Culture of bone marrow-derived mast cells (BMMCs) Primary culture of IL-3-dependent BMMCs was prepared from 8- to 12-week-old WT, c, or Jak3 mice and maintained as described previously.37 Briefly, the mice were killed and bone marrow was flushed aseptically from femurs and tibias into RPMI 1640 medium containing 10% heat-inactivated fetal calf serum (FCS), 50 µmol/L -mercaptoethanol, 2 mmol/L L-glutamine, 1 mmol/L sodium pyruvate, 0.1 mmol/L nonessential amino acids, antibiotics, and 10% (vol/vol) of murine IL-3 transfectant X63 cell conditioned medium38 (X63-IL-3; kindly provided by Dr H. Karasuyama, Tokyo Metropolitan Institute of Medical Science) as a source of IL-3. The nonadherent bone marrow cells were then maintained at 37°C at a density of 2 to 5 × 105 cells/mL in the same medium, with biweekly replacement of old media with fresh ones. BMMCs obtained after 4 weeks of culture were more than 98% mast cells, as demonstrated by morphologic analysis, and BMMCs were used for the following experiment at 4 to 6 weeks of culture. In preliminary experiments, the maximum proliferation of BMMCs was observed at 2% to 20% of X63-IL-3 conditioned medium. Peritoneal lavage cells Peritoneal lavage was performed by injecting 10 mL of ice-cold phosphate-buffered saline (PBS) into the peritoneal cavity of the mouse. After the cells were centrifuged (400g), resuspended in 1 mL of PBS, and counted using a hemocytometer, differential cell counts were performed on cytospin cell preparations stained with Wright-Giemsa solution. Peritoneal mast cells were identified morphologically according to the criteria described previously.2,39 A fraction of the cells was subjected to flow cytometric analysis as follows. Flow cytometric analysis Cells from the peritoneal cavity and BMMCs were stained and analyzed on a FACScaliber (Becton Dickinson, San Jose, CA) with CELLQuest software. The following antibodies were purchased: anti-CD117 (c-kit) fluorescein isothiocyanate (FITC), phycoerythrin (PE), allophycocyanin (APC) (2B8; PharMingen, San Diego, CA), anti-CD122 (IL-2R) PE (TM-1; PharMingen), anti-c PE (4G2; PharMingen), anti-CDw124 (IL-4R) (1688-01; Genzyme Corp, Cambridge, MA), anti-CD127 (IL-7R) biotin (B12-1; PharMingen), anti-CD16/32 (FcII/III) PE (2.4G2; PharMingen), anti-Gr-1 APC (RB6-8C5; PharMingen), anti-CD4 APC (RM4-5; PharMingen), anti-CD8 APC (53-6.7; PharMingen), anti-CD45R/B220 APC (RA3-6B2; PharMingen), and anti-rat IgG2a FITC (RG7/1.30; PharMingen). Before staining, Fc receptors were blocked with anti-CD16/32 antibody (2.4G2; PharMingen) excepting the detection of anti-CD16/32 PE staining. Negative controls consisted of isotype-matched, directly conjugated, nonspecific antibodies (PharMingen). IgE receptors on mast cells To quantify the levels of IgE receptors expressed on the cell surface, we first incubated the cells with mouse antitrinitrophenol IgE (IgE3; PharMingen) at 4°C for 60 minutes to saturate the IgE receptors, and then labeled them with anti-IgE FITC (R35-72; PharMingen). In some experiments, IgE receptors on BMMCs were visualized directly by FITC-conjugated mouse IgE (IgE3; PharMingen). Cell survival assay BMMCs were washed 3 times with PBS and cultured at 1 × 106 cells/mL in triplicate at 37°C for 1 to 5 days in RPMI 1640 medium without IL-3 in the presence of the indicated cytokines: murine IL-4 (20 ng/mL; Genzyme Corp), murine IL-9 (20 ng/mL; PeproTech Inc, Rocky Hill, NJ), murine IL-13 (20 ng/mL; R&D Systems, Minneapolis, MN), or human IL-15 (20 ng/mL and 129 ng/mL; DIACLONE Research, Besançon Cedex, France). Cells were harvested and viability was determined by fluorescence-activated cell sorting (FACS) with 5 µg/mL of propidium iodide (PI) (Boehringer Mannheim, Indianapolis, IN).40 To examine the survival of peritoneal mast cells, we cultured freshly isolated cells from the peritoneal cavity for 24 hours in RPMI 1640 medium without IL-3 in the presence of murine IL-4 (20 ng/mL) or murine IL-9 (20 ng/mL). The viability of c-kit+ cells was determined by FACS with the use of anti-c-kit FITC and 5 µg/mL of PI. Proliferation assay BMMCs (2 × 105/well) were cultured in triplicate at 37°C for 36 hours in 96-well plates in RPMI 1640 medium containing 10% (vol/vol) of X63-IL-3 conditioned medium with the indicated cytokines: human IL-2 (20 ng/mL; R&D Systems), murine IL-4 (20 ng/mL), murine IL-7 (20 ng/mL; R&D Systems), murine IL-9 (20 ng/mL), murine IL-13 (20 ng/mL), or human IL-15 (20 ng/mL and 129 ng/mL), with 0.5 µCi of [3H] thymidine added for the final 12 hours. Degranulation assay of mast cells BMMCs were cultured at 37°C for 4 days in the presence or absence of murine IL-4 (20 ng/mL) in RPMI 1640 medium containing 10% (vol/vol) of X63-IL-3 conditioned medium. BMMCs were then stimulated with A23187 (200 ng/mL; Sigma, St Louis, MO) at 37°C for 30 minutes. Enzyme activity of -hexosaminidase was evaluated for both the supernatant and the cell lysate using p-nitrophenyl-N-acetyl -D-glucosamine (Sigma) as a substrate.41 The percentage of specific -hexosaminidase release was expressed as: 100 × supernatant activity/(supernatant activity + cell lysate activity). Western blotting After BMMCs were starved for 2 hours from IL-3, the cells were stimulated with IL-4 (20 ng/mL) or IL-13 (20 ng/mL) at 37°C for 15 minutes. As a control, freshly isolated murine splenocytes or human peripheral blood lymphocytes were stimulated with IL-4 or IL-13. The cells were washed with PBS and lysed in lysis buffer (1% Nonidet P-40, 20 mmol/L Tris-HCl [pH 8.0], 50 mmol/L NaCl, 2 mmol/L dithiothreitol, 4 mmol/L EGTA, 10 mmol/L NaF, 1 mmol/L Na3VO4, 5 µg/mL aprotinin, 5 µg/mL leupeptin, 2 µg/mL pepstatin, 0.5 mmol/L phenylmethylsulfonyl fluoride, and 10% glycerol) on ice for 30 minutes, and cell lysates were prepared by centrifugation. A total of 15 µg of cell lysate was separated on 10% sodium dodecyl sulfate (SDS) polyacrylamide gels and transferred to Immobilon-P membranes (Millipore Corp, Bedford, MA). After blocking with PBS containing 0.15% Tween 20 and 3% bovine serum albumin (BSA) for 1 hour at room temperature, the membranes were incubated with antisera to mouse Stat6 (M-20) (Santa Cruz Biotechnology Inc, Santa Cruz, CA), mouse Stat6 (M-200) (Santa Cruz Biotechnology), phospho-Stat6 (New England Biolabs Inc, Beverly, MA), mouse Stat5 (Transduction Laboratories, Lexington, KY), phospho-Stat5 (New England Biolabs), and mouse Jak3 (Upstate Biotechnology, Lake Placid, NY) for 1 hour at room temperature. After washing 3 times with PBS containing 0.15% Tween 20, the membranes were incubated with anti-mouse IgG or anti-rabbit IgG antibodies conjugated with horseradish peroxidase (Amersham Pharmacia Biotech, Little Chalfont, UK) in PBS containing 0.15% Tween 20 and 3% BSA for 1 hour at room temperature. The membranes were then washed with PBS containing 0.15% Tween 20 and developed with an enhanced chemiluminescent substrate (Roche Diagnostics GmbH, Mannheim, Germany). Reverse transcriptase-PCR assay BMMCs and splenocytes were washed twice with PBS, and total RNA was extracted using Isogen reagent (Nippon Gene Co, Tokyo, Japan). The first-strand complementary DNA (cDNA) was then synthesized from total RNA using moloney murine leukemia virus reverse transcriptase (RT) and oligo(dT) primers (Pharmacia Biotech, Buckinghamshire, UK). cDNAs encoding IL-13R142 and -actin (as a control) were amplified by PCR. Data analysis Data are summarized as mean ± SD. Statistical analysis of the results was performed by the unpaired t test. P < .05 was considered significant.     Results Top Abstract Introduction Materials and methods Results Discussion References The number of peritoneal mast cells is decreased in c and Jak3 mice Whereas the importance of SCF and IL-3 in mast cell development is well documented,1-4 the role of c-dependent cytokines in mast cell development is less understood. To determine whether c and Jak3 are required for mast cell development in vivo, we analyzed the number of peritoneal mast cells in c-deficient (c) and Jak3-deficient (Jak3) mice. The total cell numbers recovered from the peritoneal cavity were decreased in c and Jak3 mice, resulting mainly from the diminished number of peritoneal CD5+ B cells (B-1 cells) (Suzuki et al, article in preparation). In contrast, the number of peritoneal macrophages was normal in c and Jak3 mice (data not shown), consistent with a previous report on c mice.32 Although the peritoneal mast cells in c and Jak3 mice were morphologically indistinguishable from those in WT mice, the number of mast cells recovered from the peritoneal cavity was significantly decreased in c and Jak3 mice (WT mice, 5.62 ± 0.98 × 104; c mice, 2.92 ± 0.71 × 104; and Jak3 mice, 2.96 ± 0.79 × 104; mean ± SD, n = 6-8 mice each; P < .01) (Figure 1A). Consistent with the diminished number of peritoneal mast cells in c and Jak3 mice, FACS analysis revealed that c-kit+ cells in the peritoneal cavity were decreased in c and Jak3 mice (data not shown). However, when electrically gated on c-kit+ peritoneal cells, the intensity of IgE receptors was indistinguishable among WT, c, and Jak3 mice (Figure 1B). The expression levels of Bcl-2 as well as the number of apoptotic cells (Annexin V binding-positive cells) were also indistinguishable among c-kit+ peritoneal cells from WT, c, and Jak3 mice (data not shown). In addition, the number of mast cells (c-kit+IgER+Gr-1B220CD4CD8 population) was also decreased in the spleen in c and Jak3 mice (WT mice, 1.36 ± 0.15 × 104; c mice, 0.88 ± 0.09 × 104; and Jak3 mice, 0.81 ± 0.04 × 104; 3-week-old mice, n = 5 each; P < .01). These results suggest that c- and Jak3-dependent signals play an important role in the proliferation of mast cells, but not in the maturation of mast cells in vivo. However, because the numbers of T cells, NK cells, and NK T cells are severely diminished in c and Jak3 mice,8-11,35,36 the decreased cytokine production from these cells may also contribute to the impairment in mast cell development. Therefore, we then studied the role of c-dependent signals in the proliferation and survival of cultured mast cells from c and Jak3 mice. View larger version (31K): [in this window] [in a new window]   Figure 1. The number of peritoneal mast cells is decreased in c and Jak3 mice. (A) Peritoneal mast cells were recovered by lavage and counted in 8- to 12-week-old wild-type (WT), c-deficient (c), and Jak3-deficient (Jak3) mice. Peritoneal mast cells were identified morphologically on cytospin cell preparations stained with Wright-Giemsa solution. Data are means ± SD for 6 to 8 mice in each group. The mean values for c mice and Jak3 mice are significantly different from the mean value for WT mice, *P < .01. (B) Expression of IgE receptors on c-kit+ peritoneal cells. IgE receptors on peritoneal mast cells were quantified by first incubating the cells recovered by peritoneal lavage with mouse IgE, then labeling with anti-IgE FITC, and analyzing by FACS gating on c-kit+ cells. Shown are representative FACS profiles of IgE receptor staining and control staining (dashed lines) on c-kit+ peritoneal cells from WT, c, and Jak3 mice (n = 6-8 mice in each group). Development of IL-3-dependent BMMCs is normal in c and Jak3 mice Murine mast cell lines can be established from bone marrow hematopoietic progenitors in the presence of IL-3.37 To determine the role of c-dependent signals in mast cell development, we prepared primary cultures of IL-3-dependent BMMCs from WT, c, and Jak3 mice. BMMCs obtained after 4 weeks of culture were more than 98% mast cells and were morphologically indistinguishable among these mice (data not shown). The number of BMMCs recovered per mouse was also indistinguishable among these mice (data not shown). Moreover, IL-3-induced proliferation of BMMCs was normal in c and Jak3 mice (Figure 2A). The expression levels of c-kit, FcII/III, and IgE receptor were also indistinguishable among BMMCs from WT, c, and Jak3 mice (Figure 2B). These results indicate that c- and Jak3-dependent signals are not essential for IL-3-induced mast cell development in vitro. View larger version (29K): [in this window] [in a new window]   Figure 2. Development of IL-3-dependent bone marrow-derived mast cells (BMMCs) is normal in c and Jak3 mice. (A) BMMCs (2 × 105/well) were cultured in the presence of IL-3 at 37°C for 36 hours, and the proliferative responses were evaluated by the addition of [3H] thymidine for the final 12 hours. Data are means ± SD for 5 mice in each group. (B) BMMCs from WT, c, and Jak3 mice were stained with anti-c-kit APC, anti-FcII/III PE, and IgE FITC, as described in "Materials and methods." Shown are representative FACS profiles for c-kit, FcII/III, and IgE receptor staining from 5 independent experiments. Dashed lines are FACS profiles of negative controls. Expression of IL-4R, c, and Jak3 in BMMCs We next examined the expression of c, c-related cytokine receptors, and Jak3 in BMMCs. As shown in Figure 3, the expression of IL-2R and IL-7R was absent in BMMCs from WT, c, and Jak3 mice. On the other hand, IL-4R was equally expressed in BMMCs from WT, c, and Jak3 mice (Figure 3). As expected, BMMCs from c mice lacked c expression, confirming the correct recognition by anti-c monoclonal antibody (Figure 3). Interestingly, expression levels of c were significantly increased in BMMCs from Jak3 mice as compared with those from WT mice (WT mice, 20.5 ± 2.6; Jak3 mice, 36.2 ± 3.0; mean fluorescent intensities for c staining, n = 4 each; P < .01) (Figure 3). Increased c expression was also observed in c-kit+ peritoneal cells from Jak3 mice (data not shown). In contrast, the expression of Jak3 in BMMCs was significantly lower in c mice than that in WT mice (Figure 4). As expected, Jak3 was undetectable in Jak3 mice (Figure 4). View larger version (44K): [in this window] [in a new window]   Figure 3. Expression of IL-4R and c in BMMCs. BMMCs from WT, c, and Jak3 mice were analyzed for the expression of IL-2R, IL-4R, IL-7R, and c on c-kit+ cells. Shown are representative FACS profiles from 4 independent experiments. Dashed lines are FACS profiles for the isotype-matched controls. View larger version (18K): [in this window] [in a new window]   Figure 4. Expression of Jak3 in BMMCs. Jak3 expression was analyzed by Western blotting for BMMCs from WT, c, and Jak3 mice as described in "Materials and methods." Shown is a representative anti-Jak3 Western blot from 4 independent experiments. IL-4 enhances the proliferation, survival, and degranulation of BMMCs through c- and Jak3-dependent signaling IL-4 has been shown to function as a mast cell growth factor.20-23 Although it has been shown that IL-4 can transduce the signals from 2 types of receptors and that the types of IL-4Rs expressed on cells differ depending on cell lineage,28,29 it is still unclear which type of IL-4R is functional on mast cells. To address this question, we examined the effect of IL-4 on the proliferation of BMMCs from c and Jak3 mice. Because IL-4 alone did not induce the proliferation of BMMCs from WT mice (data not shown), we examined the synergistic effect of IL-4 on IL-3-induced proliferation of BMMCs. IL-4 significantly enhanced IL-3-induced proliferation of BMMCs from WT mice (IL-3, 14.6 ± 0.8 × 103 cpm; IL-3 + IL-4, 44.8 ± 0.9 × 103 cpm; n = 5 each; P < .005) (Figure 5). IL-4 exhibited no effect on IL-3-induced proliferation of BMMCs from c and Jak3 mice (Figure 5). These results indicate that IL-4 induces the proliferation of BMMCs through c- and Jak3-dependent signals and that the functional IL-4R on BMMCs is type I IL-4R. In contrast to IL-4, IL-13, a cytokine that exhibits IL-4-like functions on B cells43 and endothelial cells44 through type II IL-4R, did not significantly enhance the IL-3-induced proliferation of BMMCs even in WT mice (Figure 5). View larger version (25K): [in this window] [in a new window]   Figure 5. IL-4 induces the proliferation of BMMCs through c- and Jak3-dependent signaling. BMMCs from WT, c, and Jak3 mice were cultured in the presence of IL-3 for 36 hours, with [3H] thymidine added for the final 12 hours. Where indicated, recombinant murine IL-4 or recombinant murine IL-13 was added at 20 ng/mL. Data are means ± SD for 5 mice in each group. The mean value of IL-4-stimulated BMMCs is significantly different from the mean value of control BMMCs (IL-3 alone) in WT mice. *P < .005. We next examined the antiapoptotic effect of IL-4 and IL-13 on BMMCs. IL-3-deprived BMMCs from WT, c, and Jak3 mice were cultured with IL-4 (20 ng/mL) or IL-13 (20 ng/mL) for 1 to 5 days, and the cell viability was then determined by FACS. As shown in Figure 6, the rate of survival after IL-3 withdrawal was similar among BMMCs from WT, c, and Jak3 mice. IL-4 significantly enhanced the survival of BMMCs from WT mice (control, 9.5% ± 0.5%; IL-4, 64.2% ± 7.0% at day 5; n = 5 each; P < .001) (Figure 6). However, IL-4 had no effect on the survival of BMMCs from c and Jak3 mice (Figure 6). IL-13 did not significantly affect the survival of BMMCs from WT, c, or Jak3 mice (Figure 6). These results indicate that IL-4, but not IL-13, enhances the proliferation and survival of BMMCs through c- and Jak3-dependent signaling of type I IL-4R. View larger version (16K): [in this window] [in a new window]   Figure 6. IL-4 induces the survival of BMMCs through c- and Jak3-dependent signaling. BMMCs from WT, c, and Jak3 mice were cultured in the absence of IL-3 for 5 days, and cell viability was determined by FACS using PI (5 µg/mL) at days 0, 1, 3, and 5. Where indicated, recombinant murine IL-4 or recombinant murine IL-13 was added at 20 ng/mL. Data are means ± SD for 5 mice in each group. The mean value of IL-4-stimulated BMMCs is significantly different from the mean value of control BMMCs in WT mice. *P < .001. To determine whether c and Jak3 signaling is involved in IL-4-induced enhancement of mast cell degranulation, we examined the effect of IL-4 on A23187-induced degranulation of BMMCs in WT, c, and Jak3 mice. Consistent with a previous finding on the role of IL-4 in human mast cell degranulation,45 IL-4 significantly enhanced A23187-induced degranulation of BMMCs from WT mice (P < .01) (Figure 7). In contrast, however, IL-4 did not increase the degranulation of BMMCs from c and Jak3 mice (Figure 7). These results indicate that, in addition to its effect on proliferation and survival, c and Jak3 signaling is essential for IL-4-induced enhancement of mast cell degranulation. View larger version (17K): [in this window] [in a new window]   Figure 7. IL-4 induces the degranulation of BMMCs through c- and Jak3-dependent signaling. BMMCs from WT, c, and Jak3 mice were cultured for 4 days in the presence of IL-3 alone or IL-3 plus murine IL-4 (20 ng/mL). BMMCs were then stimulated with A23187 (200 ng/mL) for 30 minutes, and the percentage of specific -hexosaminidase release was determined as described in "Materials and methods." Data are means ± SD for 4 mice in each group. The mean value of IL-4-stimulated BMMCs is significantly different from the mean value of the corresponding BMMCs without IL-4 stimulation in WT mice. *P < .01. IL-4, but not IL-13, phosphorylates the 65-kd Stat6 isoform in BMMCs In the IL-4 signaling pathway, the best-studied signaling molecule is signal transducers and activators of transcription 6 (Stat6).11 Stat6 is also rapidly activated after cellular exposure to IL-13.11 Therefore, we analyzed IL-4- and IL-13-induced Stat6 phosphorylation in BMMCs from WT, c, and Jak3 mice. After BMMCs were deprived of IL-3 for 2 hours, BMMCs were stimulated with either IL-4 or IL-13 for 15 minutes, and tyrosine phosphorylation of Stat6 at tyrosine 641 was detected by anti-phospho Stat6 antibody. Surprisingly, IL-4 phosphorylated the 65-kd isoform of Stat6 in BMMCs from WT mice, whereas IL-4 phosphorylated the 94-kd isoform of Stat6 in WT splenocytes (Figure 8). Reblotting with anti-Stat6 antibody M200, which recognizes the middle part of murine Stat6 (AA280-AA480), revealed that BMMCs expressed the 65-kd isoform of Stat6 but not the 94-kd Stat6 (Figure 8). Interestingly, the 65-kd isoform of Stat6 was not detected by anti-Stat6 antibody M20, which recognizes the c-terminus of murine Stat6, whereas the 94-kd Stat6 in splenocytes was readily detected by M20 (data not shown). These results suggest that the 65-kd isoform of Stat6 lacks the c-terminus. Although the 65-kd isoform of Stat6 was equally expressed in BMMCs among WT, c, and Jak3 mice (Figure 8), IL-4 did not induce the phosphorylation of the 65-kd Stat6 in BMMCs from c or Jak3 mice (Figure 8). In addition, IL-13 did not induce the phosphorylation of 65-kd Stat6 in BMMCs even from WT mice (Figure 8), whereas murine IL-13 did phosphorylate the 94-kd Stat6 in human peripheral blood lymphocytes (data not shown). View larger version (40K): [in this window] [in a new window]   Figure 8. IL-4-induced phosphorylation of 65-kd Stat6 isoform in BMMCs is c- and Jak3-dependent. BMMCs from WT, c, and Jak3 mice were washed with PBS, cultured for 2 hours in the absence of IL-3, and stimulated with IL-4 or IL-13 (20 ng/mL) for 15 minutes. After washing with PBS, cell lysates were prepared, separated on a 10% SDS gel, and blotted with antisera to either phospho-Stat6 or Stat6 (M200). As a control, cell lysates from IL-4-stimulated WT splenocytes were used. Shown is a representative blot from 4 independent experiments. IL-13R1 is not expressed in BMMCs To determine whether IL-13R1 is expressed on BMMCs, we performed RT-PCR analysis for IL-13R1 mRNA expression in BMMCs from WT, c, and Jak3 mice. As shown in Figure 9, IL-13R1 was not detected in BMMCs from WT, c, or Jak3 mice. In contrast, IL-13R1 was expressed in splenocytes from WT mice (Figure 9). The absence of IL-13R1 expression on BMMCs is consistent with our results presented earlier: that IL-4, but not IL-13, functioned on BMMCs and that IL-4-induced effects on BMMCs depended on c and Jak3. View larger version (35K): [in this window] [in a new window]   Figure 9. IL-13R1 is not expressed on murine BMMCs. Total RNA was prepared from BMMCs, and RT-PCR analysis for IL-13R1 and -actin (as a control) was performed as described in "Materials and methods." As a control, splenocytes from WT mice were used. Shown are representative data from 5 independent experiments. IL-9, but not IL-15, enhances the proliferation and survival of BMMCs through c- and Jak3-dependent signaling As described earlier, IL-4 enhanced the proliferation (Figure 5) and survival (Figure 6) of BMMCs through c- and Jak3-dependent signaling. However, because the number of peritoneal mast cells was reported to be normal in IL-4-deficient mice,46 defects of IL-4 signals in c and Jak3 mice may not be responsible for the diminished mast cell numbers in c and Jak3 mice. Therefore, we determined the role of other c-related cytokinesIL-2, IL-7, IL-9, and IL-15in the proliferation of BMMCs. Among these cytokines, only IL-9 significantly enhanced the IL-3-induced proliferation of BMMCs from WT mice (IL-3, 14.6 ± 1.0 × 103 cpm; IL-3 + IL-9, 24.3 ± 1.8 × 103 cpm; n = 5 each; P < .005) (Figure 10). IL-9 had no effect on the proliferation of BMMCs from c and Jak3 mice (Figure 10). Interestingly, and inconsistent with a previous report,26 IL-15 did not significantly enhance the proliferation of BMMCs from WT mice (Figure 10), even when a high concentration of IL-15 (129 ng/mL) was added to the BMMC culture (data not shown). The biologic activity of IL-15 was confirmed by IL-2-dependent CTLL-2 proliferation assay, and the addition of IL-15 achieved a half-maximal proliferation of CTLL-2 cells at approximately 0.01 ng/mL (data not shown). View larger version (41K): [in this window] [in a new window]   Figure 10. IL-9 induces the proliferation of BMMCs through c- and Jak3-dependent signaling. BMMCs from WT, c, and Jak3 mice were cultured in the presence of IL-3 for 36 hours, with [3H] thymidine added for the final 12 hours. Where indicated, recombinant human IL-2, recombinant murine IL-7, recombinant murine IL-9, or recombinant human IL-15 was added at 20 ng/mL. Data are means ± SD for 5 mice in each group. *P < .005. We then determined the effects of IL-9 and IL-15 on the survival of IL-3-deprived BMMCs. Whereas IL-9 enhanced the survival of BMMCs from WT mice (control, 9.7% ± 0.6%; IL-9, 50.1% ± 6.5% at day 5; n = 5; P < .005), IL-9 did not significantly affect the survival of BMMCs from c and Jak3 mice (Figure 11). These results indicate that the antiapoptotic effect of IL-9 on BMMCs depends on c and Jak3. Again, IL-15 did not significantly enhance the survival of BMMCs even in WT mice (Figure 11). View larger version (15K): [in this window] [in a new window]   Figure 11. IL-9 induces the survival of BMMCs through c- and Jak3-dependent signaling. BMMCs from WT, c, and Jak3 mice were cultured in the absence of IL-3 for 5 days, and cell viability was determined by FACS using PI (5 µg/mL) at days 0, 1, 3, and 5. Where indicated, recombinant murine IL-9 or recombinant human IL-15 was added at 20 ng/mL. Data are means ± SD for 5 mice in each group. *P < .05, **P < .005. IL-4 enhances the survival of peritoneal mast cells through c- and Jak3-dependent signaling Finally, we studied the role of c and Jak3 signaling in IL-4- and IL-9-induced survival of freshly isolated peritoneal mast cells. As shown in Figure 12, IL-4 significantly increased the survival of peritoneal mast cells from WT mice (control, 44.4% ± 6.1%; IL-4, 67.3% ± 7.0% at 24 hours; n = 4 each; P < .01). In contrast, IL-9 exhibited only a marginal effect on the survival of peritoneal mast cells from WT mice. Consistent with the in vitro experiments on BMMCs (Figures 6 and 11), IL-4 as well as IL-9 exhibited no effect on the survival of peritoneal mast cells from c and Jak3 mice (Figure 12). These results indicate that IL-4 enhances the survival of peritoneal mast cells through c- and Jak3-dependent signaling. View larger version (27K): [in this window] [in a new window]   Figure 12. IL-4 enhances the survival of peritoneal mast cells through c- and Jak3-dependent signaling. Peritoneal cells from WT, c, and Jak3 mice were cultured in the presence of IL-4 or IL-9 for 24 hours, and cell viability of c-kit+ cells was determined by FACS as described in "Materials and methods." Data are means ± SD for 4 mice in each group. The mean value of IL-4-stimulated peritoneal mast cells is significantly different from the mean value of control peritoneal mast cells in WT mice. *P < .01.     Discussion Top Abstract Introduction Materials and methods Results Discussion References Previous studies have shown that c- and Jak3-dependent signals play the essential regulatory roles in T-cell and NK cell development.8-11,35,36 In this study, we show that c- and Jak3-dependent signaling induces the proliferation and survival of murine mast cells. We found that, among cytokines that use c as a shared receptor component, IL-4 and IL-9, but not IL-2, IL-7, or IL-15, enhanced the proliferation and survival of BMMCs from WT mice and that the effects of IL-4 and IL-9 were absent in BMMCs from c and Jak3 mice (Figures 5, 6, 10, and 11). We also found that IL-4R, c, and Jak3, but not IL-2R, IL-7R, or IL-13R1, were expressed in BMMCs (Figures 3, 4, and 9). Our findings of the decreased number of peritoneal mast cells in c and Jak3 mice also suggest an important role of c- and Jak3-dependent signals in the proliferation and survival of mast cells in vivo (Figure 1). Our results indicate that IL-4-induced proliferation and survival of murine mast cells are mediated through type I IL-4R. It has been shown recently that type I IL-4R is a heterodimer of IL-4R and c, that type II IL-4R is a heterodimer of IL-4R and IL-13R1, and that the usage of IL-4R differs depending on cell lineage.28,29 We found that IL-4 enhanced the proliferation and survival of BMMCs from WT mice but not from c or Jak3 mice (Figures 5 and 6) and that IL-4R and c were expressed in BMMCs (Figure 3). In contrast, although a previous study reported that IL-13 induced the proliferation of a murine mast cell line,47 we found that IL-13 did not significantly enhance the proliferation and survival of BMMCs even from WT mice (Figures 5 and 6) and that IL-13R1 was not expressed in BMMCs (Figure 9). Therefore, although increasing evidence suggests that IL-13 may play an important role in allergic inflammation,48,49 mast cells may not be a target of IL-13. In addition to type I and type II IL-4Rs, the experiments using chimeric proteins that were constructed with the cytoplasmic domain of IL-4R and an extracellular domain of erythropoietin,50,51 CD8,51 or c-kit52 suggested that the homodimerization of IL-4R could transduce IL-4 signals. However, our finding that c-deficient BMMCs, which express IL-4R, did not respond to IL-4 stimulation also indicates that the expression of IL-4R alone is not sufficient to transduce IL-4 signals in mast cells. The best-characterized molecule downstream of c/Jak3 in IL-4 signals is Stat6.11 Interestingly, whereas Stat6 is essential for IL-4-induced proliferation of T cells,53-55 IL-4-induced proliferation of BMMCs is slightly increased in the absence of Stat6.56 In addition, mastocytosis induced by in vivo administration of IL-4 was rather enhanced in Stat6-deficient mice.57 In the present study, we showed that the 65-kd isoform of Stat6, which lacked the c-terminus, was predominantly expressed in mast cells (Figure 8). Moreover, we found that the 65-kd isoform of Stat6 was phosphorylated by IL-4 in a c- and Jak3-dependent manner (Figure 8). Our findings are in agreement with the recent observation by Sherman et al58 that murine mast cells preferentially express 65-kd Stat6. The 65-kd isoform of Stat6 is apparently different from the previously reported Stat6 isoforms in human fibroblast, Stat6b and Stat6c, which encode an NH2-terminal truncation and an SH2-domain deletion, respectively.59 Because deletion mutants of Stat6 that lack the c-terminus still bind to the Stat6-recognition site of DNA but lose transcriptional activity,60 it is possible that the 65-kd isoform of Stat6 may function as a negative regulator of transcription. At present, it is unknown how the 65-kd isoform of Stat6 is generated. In preliminary experiments, we found that when the extract from splenocytes was mixed with the extract from BMMCs and incubated at 37°C, the 94-kd Stat6 in splenocytes was cleaved to 65 kd (our unpublished data), suggesting that the 65-kd Stat6 in mast cells is generated by cleavage of mature protein rather than by alternative splicing of mRNA. In addition to Stat6, it has been reported recently that IL-4 induces the proliferation of murine pro-B-cell line Ba/F3 through activation of Stat5.61 However, we observed no Stat5 phosphorylation in IL-4-stimulated BMMCs (our unpublished data). Thus, it is unlikely that Stat5 activation is responsible for IL-4-induced proliferation and survival of BMMCs. Taken together, our results indicate that IL-4 enhances the proliferation and survival of mast cells in a c- and Jak3-dependent manner, but not in a Stat6- or Stat5-dependent manner. We also show that IL-9 enhances the proliferation and survival of BMMCs through c- and Jak3-dependent signaling (Figures 10 and 11). IL-9 is a Th2-cell-derived cytokine with pleiotropic effects on various cell types, including mast cell growth.24,25,62,63 Because the number of peritoneal mast cells was reported to be normal in IL-4-deficient mice,46 it is conceivable that the diminished peritoneal mast cell numbers in c or Jak3 mice (Figure 1A) may result from the absence of signaling via IL-9 or an undefined c/Jak3-dependent cytokine. IL-9 or IL-9R knockout mice could be valuable in further investigating this issue. IL-15 was also previously shown to function as a mast cell growth factor through an undefined IL-15R X, but not through an IL-2R/c heterodimer.26 However, we found that IL-15 had no significant effect on the proliferation and survival of BMMCs even in WT mice (Figures 10 and 11). In addition, in parallel experiments, we found that IL-15 did induce the proliferation of CTLL-2 cells at 0.01 ng/mL, but IL-15 did not enhance the proliferation of BMMCs even at 129 ng/mL. Because Tagaya et al26 reported that the activity of IL-15 on mast cell proliferation differed between batches of human IL-15, unknown factor(s) possibly contaminated in some batches of IL-15 might induce the expression of IL-15R X, and thereby IL-15 might enhance the proliferation of BMMCs in a c-independent manner. In summary, we have shown that c- and Jak3-dependent signaling is essential for IL-4- and IL-9-induced proliferation and survival of murine mast cells. We have also shown that the effects of IL-4 on mast cells are mediated by type I IL-4R and that IL-13 exhibits no IL-4-like effects on mast cells because of the absence of IL-13R1. In addition, IL-4 phosphorylates the 65-kd isoform of Stat6 in mast cells through c- and Jak3-dependent signaling. These results suggest that c- and Jak3-dependent signaling has an important role in the mast cell expansion of Th2-cell-mediated inflammatory responses.     Acknowledgments We thank Warren J. Leonard for c-deficient mice, Hajime Karasuyama for X63-IL-3 cells, Miki Nishimura and Maki Watanabe for technical help, and Kenji Izuhara and Kazuhiro Kurasawa for valuable discussion.     Footnotes Submitted January 27, 2000; accepted May 23, 2000. Supported in part by grants from the Ministry of Education, Science and Culture, Japan. The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734. Reprints: Hiroshi Nakajima, Department of Internal Medicine II, Chiba University School of Medicine, 1-8-1 Inohana, Chiba City, Chiba 260-8670, Japan; e-mail: nakajimh{at}intmed02.m.chiba-u.ac.jp.     References Top Abstract Introduction Materials and methods Results Discussion References 1. Galli SJ, Hammel I. Mast cell and basophil development. Curr Opin Hematol. 1994;1:33-39[Medline] [Order article via Infotrieve]. 2. Metcalfe DD, Baram D, Mekori YA. Mast cells. Physiol Rev. 1997;77:1033-1079[Abstract/Free Full Text]. 3. Lantz CS, Huff TF. Differential responsiveness of purified mouse c-kit+ mast cells and their progenitors to IL-3 and stem cell factor. J Immunol. 1995;155:4024-4029[Abstract]. 4. Rodewald HR, Dessing M, Dvorak AM, Galli SJ. Identification of a committed precursor for the mast cell lineage. Science. 1996;271:818-822[Abstract]. 5. Sugamura K, Asao H, Kondo M, et al. The interleukin-2 receptor  chain: its role in the multiple cytokine receptor complexes and T cell development in XSCID. Annu Rev Immunol. 1996;14:179-205[Medline] [Order article via Infotrieve]. 6. Leonard WJ. The molecular basis of X-linked severe combined immunodeficiency: defective cytokine receptor signaling. Annu Rev Med. 1996;47:229-239[Medline] [Order article via Infotrieve]. 7. Noguchi M, Yi H, Rosenblatt HM, et al. Interleukin-2 receptor  chain mutation results in X-linked severe combined immunodeficiency in humans. Cell. 1993;73:147-157[Medline] [Order article via Infotrieve]. 8. DiSanto JP, Müller W, Guy-Grand D, Fischer A, Rajewsky K. Lymphoid development in mice with a targeted deletion of the interleukin-2 receptor  chain. Proc Natl Acad Sci U S A. 1995;92:377-381[Abstract/Free Full Text]. 9. Cao X, Shores EW, Hu-Li J, et al. Defective lymphoid development in mice lacking expression of the common cytokine receptor  chain. Immunity. 1995;2:223-238[Medline] [Order article via Infotrieve]. 10. Ohbo K, Suda T, Hashiyama M, et al. Modulation of hematopoiesis in mice with a truncated mutant of the interleukin-2 receptor  chain. Blood. 1996;87:956-969[Abstract/Free Full Text]. 11. Leonard WJ, O'Shea JJ. Jaks and STATs: biological implications. Annu Rev Immunol. 1998;16:293-322[Medline] [Order article via Infotrieve]. 12. Macchi P, Villa A, Gillani S, et al. Mutations of Jak-3 gene in patients with autosomal severe combined immune deficiency (SCID). Nature. 1995;377:65-68[Medline] [Order article via Infotrieve]. 13. Russell SM, Tayebi N, Nakajima H, et al. Mutation of Jak3 in a patient with SCID: essential role of Jak3 in lymphoid development. Science. 1995;270:797-800[Abstract/Free Full Text]. 14. von Freeden-Jeffry U, Vieira P, Lucian LA, McNeil T, Burdach SE, Murray R. Lymphopenia in interleukin (IL)-7 gene-deleted mice identifies IL-7 as a nonredundant cytokine. J Exp Med. 1995;181:1519-1526[Abstract/Free Full Text]. 15. Peschon JJ, Morrissey PJ, Grabstein KH, et al. Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice. J Exp Med. 1994;180:1955-1960[Abstract/Free Full Text]. 16. Puel A, Ziegler SF, Buckley RH, Leonard WJ. Defective IL7R expression in T B+ NK+ severe combined immunodeficiency. Nat Genet. 1998;20:394-397[Medline] [Order article via Infotrieve]. 17. Suzuki H, Duncan GS, Takimoto H, Mak TW. Abnormal development of intestinal intraepithelial lymphocytes and peripheral natural killer cells in mice lacking the IL-2 receptor  chain. J Exp Med. 1997;185:499-505[Abstract/Free Full Text]. 18. Ohteki T, Ho S, Suzuki H, Mak TW, Ohashi PS. Role for IL-15/IL-15 receptor -chain in natural killer 1.1+ T cell receptor-+ cell development. J Immunol. 1997;159:5931-5935[Abstract]. 19. Lodolce JP, Boone DL, Chai S, et al. IL-15 receptor maintains lymphoid homeostasis by supporting lymphocyte homing and proliferation. Immunity. 1998;9:669-676[Medline] [Order article via Infotrieve]. 20. Tepper RI, Levinson DA, Stanger BZ, Campos-Torres J, Abbas AK, Leder P. IL-4 induces allergic-like inflammatory disease and alter T cell development in transgenic mice. Cell. 1990;62:457-467[Medline] [Order article via Infotrieve]. 21. Tsuji K, Nakahata T, Takagi M, et al. Effects of interleukin-3 and interleukin-4 on the development of "connective tissue-type" mast cells: interleukin-3 supports their survival and interleukin-4 triggers and supports their proliferation synergistically with interleukin-3. Blood. 1990;75:421-427[Abstract/Free Full Text]. 22. Madden KB, Urban JF Jr, Ziltener HJ, Schrader JW, Finkelman FD, Katona IM. Antibodies to IL-3 and IL-4 suppress helminth-induced intestinal mastocytosis. J Immunol. 1991;147:1387-1391[Abstract]. 23. Yanagida M, Fukamachi H, Ohgami K, et al. Effects of T-helper 2-type cytokines, IL-3, IL-4, IL-5, and IL-6 on the survival of cultured human mast cells. Blood. 1995;86:3705-3714[Abstract/Free Full Text]. 24. Hultner L, Druez C, Moeller J, et al. Mast cell growth-enhancing activity (MEA) is structurally related and functionally identical to the novel mouse T cell growth factor P40/TCGFIII (interleukin 9). Eur J Immunol. 1990;20:1413-1416[Medline] [Order article via Infotrieve]. 25. Renauld J-C, Kermouni A, Vink A, Louahed J, Van Snick J. Interleukin-9 and its receptor: involvement in mast cell differentiation and T cell oncogenesis. J Leukoc Biol. 1995;57:353-360[Abstract]. 26. Tagaya Y, Burton JD, Miyamoto Y, Waldmann TA. Identification of a novel receptor/signal transduction pathway for IL-15/T in mast cells. EMBO J. 1996;15:4928-4939[Medline] [Order article via Infotrieve]. 27. Paul WE, Seder RA. Lymphocyte responses and cytokines. Cell. 1994;76:241-251[Medline] [Order article via Infotrieve]. 28. Nelms K, Keegan AD, Zamorano J, Ryan JJ, Paul WE. The IL-4 receptor: signaling mechanisms and biologic functions. Annu Rev Immunol. 1999;17:701-738[Medline] [Order article via Infotrieve]. 29. Murata T, Obiri NI, Puri RK. Structure of and signal transduction through interleukin-4 and interleukin-13 receptors. Int J Mol Med. 1998;1:551-557[Medline] [Order article via Infotrieve]. 30. Matthews DJ, Clark PA, Herbert J, et al. Function of the interleukin-2 (IL-2) receptor -chain in biologic responses of X-linked severe combined immunodeficient B cells to IL-2, IL-4, IL-13, and IL-15. Blood. 1995;85:38-42[Abstract/Free Full Text]. 31. Matthews DJ, Hibbert L, Friedrich K, Minty A, Callard RE. X-SCID B cell responses to interleukin-4 and interleukin-13 are mediated by a receptor complex that includes the interleukin-4 receptor  chain (p140) but not the c chain. Eur J Immunol. 1997;27:116-121[Medline] [Order article via Infotrieve]. 32. Andersson A, Grunewald SM, Duschl A, Fischer A, DiSanto JP. Mouse macrophage development in the absence of the common  chain: defining receptor complexes responsible for IL-4 and IL-13 signaling. Eur J Immunol. 1997;27:1762-1768[Medline] [Order article via Infotrieve]. 33. de Vries JE. The role of IL-13 and its receptor in allergy and inflammatory responses. J Allergy Clin Immunol. 1998;102:165-169[Medline] [Order article via Infotrieve]. 34. Waldmann TA, Tagaya Y. The multifaceted regulation of interleukin-15 expression and the role of this cytokine in NK cell differentiation and host response to intracellular pathogens. Annu Rev Immunol. 1999;17:19-49[Medline] [Order article via Infotrieve]. 35. Park SY, Saijo K, Takahashi T, et al. Developmental defects of lymphoid cells in Jak3 kinase-deficient mice. Immunity. 1995;3:771-782[Medline] [Order article via Infotrieve]. 36. Suzuki K, Nakajima H, Saito Y, Saito T, Leonard WJ, Iwamoto I. Jak3 is essential for c-dependent signaling: comparative analysis of c, Jak3, and c and Jak3 double-deficient mice. Int Immunol. 2000;12:123-132[Abstract/Free Full Text]. 37. Ihle JN, Keller J, Oersozlan S, et al. Biological properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth-factor activity, mast cell growth factor activity, P cell-stimulating factor activity and histamine-producing factor activity. J Immunol. 1983;131:282-287[Abstract]. 38. Karasuyama H, Melchers F. Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4, or 5, using modified cDNA expression vectors. Eur J Immunol. 1988;18:97-104[Medline] [Order article via Infotrieve]. 39. Denburg JA. Basophil and mast cell lineages in vitro and in vivo. Blood. 1992;79:846-860[Free Full Text]. 40. Nakajima H, Shores EW, Noguchi M, Leonard WJ. The common cytokine receptor  chain plays an essential role in regulating lymphoid homeostasis. J Exp Med. 1997;185:189-195[Abstract/Free Full Text]. 41. Watanabe N, Akikusa B, Park SY, et al. Mast cells induce autoantibody-mediated vasculitis syn-drome through tumor necrosis factor production upon triggering Fc receptors. Blood. 1999;94:3855-3863[Abstract/Free Full Text]. 42. Schnare M, Blum H, Juttner S, Rollinghoff M, Gessner A. Specific antagonism of type I IL-4 receptor with a mutated form of murine IL-4. J Immunol. 1998;161:3484-3492[Abstract/Free Full Text]. 43. Punnonen J, Aversa G, Cocks BG, et al. Interleukin 13 induces interleukin 4-independent IgG4 and IgE synthesis and CD23 expression by human B cells. Proc Natl Acad Sci U S A. 1993;90:3730-3734[Abstract/Free Full Text]. 44. Schnyder B, Lugli S, Feng N, et al. IL-4 and IL-13 bind to a shared heterodimeric complex on endothelial cells mediating vascular cell adhesion molecule-1 induction in the absence of the common  chain. Blood. 1996;87:4286-4295[Abstract/Free Full Text]. 45. Bischoff SC, Sellge G, Lorentz A, Sebald W, Raab R, Manns MP. IL-4 enhances proliferation and mediator release in mature human mast cells. Proc Natl Acad Sci U S A. 1999;96:8080-8085[Abstract/Free Full Text]. 46. Banks EM, Coleman JW. A comparative study of peritoneal mast cells from mutant IL-4 deficient and normal mice: evidence that IL-4 is not essential for mast cell development but enhances secretion via control of IgE binding and passive sensitization. Cytokine. 1996;8:190-196[Medline] [Order article via Infotrieve]. 47. He Y-W, Malek TR. The IL-2 receptor c chain does not function as a subunit shared by the IL-4 and IL-13 receptors. J Immunol. 1995;155:9-12[Abstract]. 48. Wills-Karp M, Luyimbazi J, Xu X, et al. Interleukin-13: central mediator of allergic asthma. Science. 1998;282:2258-2261[Abstract/Free Full Text]. 49. Grunig G, Warnock M, Wakil AE, et al. Requirement for IL-13 independently of IL-4 in experimental asthma. Science. 1998;282:2261-2263[Abstract/Free Full Text]. 50. Lai SY, Molden J, Liu KD, Puck JM, White MD, Goldsmith MA. Interleukin-4-specific signal transduction events are driven by homotypic interactions of the interleukin-4 receptor  subunit. EMBO J. 1996;15:4506-4514[Medline] [Order article via Infotrieve]. 51. Fujiwara H, Hanissian SH, Tsytsykova A, Geha RS. Homodimerization of the human interleukin 4 receptor  chain induces C germline transcripts in B cells in the absence of the interleukin 2 receptor  chain. Proc Natl Acad Sci U S A. 1997;94:5866-5871[Abstract/Free Full Text]. 52. Reichel M, Nelson BH, Greenberg PD, Rothman PB. The IL-4 receptor -chain cytoplasmic domain is sufficient for activation of JAK-1 andSTAT6 and the induction of IL-4-specific gene expression. J Immunol. 1997;158:5860-5867[Abstract]. 53. Takeda K, Tanaka T, Shi W, et al. Essential role of Stat6 in IL-4 signaling. Nature. 1996;380:627-630[Medline] [Order article via Infotrieve]. 54. Shimoda K, van Deursen J, Sangster MY, et al. Lack of IL-4-induced Th2 response and IgE class switching in mice with disrupted Stat6 gene. Nature. 1996;380:630-633[Medline] [Order article via Infotrieve]. 55. Kaplan MH, Schindler U, Smiley ST, Grusby MJ. Stat6 is required for mediating responses to IL-4 and for development of Th2 cells. Immunity. 1996;4:313-319[Medline] [Order article via Infotrieve]. 56. Ryan JJ, DeSimone S, Klisch G, et al. IL-4 inhibits mouse mast cell FcRI expression through a STAT6-dependent mechanism. J Immunol. 1998;161:6915-6923[Abstract/Free Full Text]. 57. Urban JF Jr, Noben-Trauth N, Donaldson DD, et al. IL-13, IL-4R, and Stat6 are required for the expulsion of the gastrointestinal nematode parasite Nippostrongylus brasiliensis. Immunity. 1998;8:255-264[Medline] [Order article via Infotrieve]. 58. Sherman MA, Secor VH, Brown MA. IL-4 preferentially activates a novel STAT6 isoform in mast cells. J Immunol. 1999;162:2703-2708[Abstract/Free Full Text]. 59. Patel BKR, Pierce JH, LaRochelle WJ. Regulation of interleukin 4-mediated signaling by naturally occurring dominant negative and attenuated forms of human Stat6. Proc Natl Acad Sci U S A. 1998;95:172-177[Abstract/Free Full Text]. 60. Mikita T, Campbell D, Wu P, Williamson K, Schinder U. Requirements for interleukin-4-induced gene expression and functional characterization of Stat6. Mol Cell Biol. 1996;16:5811-5820[Abstract]. 61. Friedrich K, Kammer W, Erhardt I, Brandlein S, Sebald W, Moriggl R. Activation of STAT5 by IL-4 relies on Janus kinase function but not on receptor tyrosine phosphorylation, and can contribute to both cell proliferation and gene regulation. Int Immunol. 1999;11:1283-1294[Abstract/Free Full Text]. 62. Temann UA, Geba GP, Rankin JA, Flavell RA. Expression of interleukin 9 in the lungs of transgenic mice causes airway inflammation, mast cell hyperplasia, and bronchial hyperresponsiveness. J Exp Med. 1998;188:1307-1320[Abstract/Free Full Text]. 63. Godfraind C, Louahed J, Faulkner H, et al. Intraepithelial infiltration by mast cells with both connective tissue-type and mucosal-type characteristics in gut, trachea, and kidneys of IL-9 transgenic mice. J Immunol. 1998;160:3989-3996[Abstract/Free Full Text]. © 2000 by The American Society of Hematology.   CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: F. Martelli, B. Ghinassi, R. Lorenzini, A. M. Vannucchi, R. A. Rana, M. Nishikawa, S. Partamian, G. Migliaccio, and A. R. Migliaccio Thrombopoietin Inhibits Murine Mast Cell Differentiation Stem Cells, April 1, 2008; 26(4): 912 - 919. [Abstract] [Full Text] [PDF] C. Waskow, S. Bartels, S. M. Schlenner, C. Costa, and H.-R. Rodewald Kit is essential for PMA-inflammation-induced mast-cell accumulation in the skin Blood, June 15, 2007; 109(12): 5363 - 5370. [Abstract] [Full Text] [PDF] K. Loesch, L. Deng, J. W. Cowan, X. Wang, K. He, J. Jiang, R. A. Black, and S. J. Frank Janus Kinase 2 Influences Growth Hormone Receptor Metalloproteolysis Endocrinology, June 1, 2006; 147(6): 2839 - 2849. [Abstract] [Full Text] [PDF] K. He, K. Loesch, J. W. Cowan, X. Li, L. Deng, X. Wang, J. Jiang, and S. J. Frank Janus Kinase 2 Enhances the Stability of the Mature Growth Hormone Receptor Endocrinology, November 1, 2005; 146(11): 4755 - 4765. [Abstract] [Full Text] [PDF] A. Lorentz, M. Wilke, G. Sellge, H. Worthmann, J. Klempnauer, M. P. Manns, and S. C. Bischoff IL-4-Induced Priming of Human Intestinal Mast Cells for Enhanced Survival and Th2 Cytokine Generation Is Reversible and Associated with Increased Activity of ERK1/2 and c-Fos J. Immunol., June 1, 2005; 174(11): 6751 - 6756. [Abstract] [Full Text] [PDF] M. Kohno, S. Yamasaki, V. L. J. Tybulewicz, and T. Saito Rapid and large amount of autocrine IL-3 production is responsible for mast cell survival by IgE in the absence of antigen Blood, March 1, 2005; 105(5): 2059 - 2065. [Abstract] [Full Text] [PDF] K. Ikeda, H. Nakajima, K. Suzuki, S.-i. Kagami, K. Hirose, A. Suto, Y. Saito, and I. Iwamoto Mast cells produce interleukin-25 upon Fcepsilon RI-mediated activation Blood, May 1, 2003; 101(9): 3594 - 3596. [Abstract] [Full Text] [PDF] A. R. Migliaccio, R. A. Rana, M. Sanchez, R. Lorenzini, L. Centurione, L. Bianchi, A. M. Vannucchi, G. Migliaccio, and S. H. Orkin GATA-1 as a Regulator of Mast Cell Differentiation Revealed by the Phenotype of the GATA-1low Mouse Mutant J. Exp. Med., February 3, 2003; 197(3): 281 - 296. [Abstract] [Full Text] [PDF] K. B. Madden, L. Whitman, C. Sullivan, W. C. Gause, J. F. Urban Jr., I. M. Katona, F. D. Finkelman, and T. Shea-Donohue Role of STAT6 and Mast Cells in IL-4- and IL-13-Induced Alterations in Murine Intestinal Epithelial Cell Function J. Immunol., October 15, 2002; 169(8): 4417 - 4422. [Abstract] [Full Text] [PDF] M. A. Sherman, D. R. Powell, and M. A. Brown IL-4 Induces the Proteolytic Processing of Mast Cell STAT6 J. Immunol., October 1, 2002; 169(7): 3811 - 3818. [Abstract] [Full Text] [PDF] K. Suzuki, H. Nakajima, S.-i. Kagami, A. Suto, K. Ikeda, K. Hirose, T. Hiwasa, K. Takeda, Y. Saito, S. Akira, et al. Proteolytic Processing of Stat6 Signaling in Mast Cells as a Negative Regulatory Mechanism J. Exp. Med., July 1, 2002; 196(1): 27 - 38. [Abstract] [Full Text] [PDF] R. Malaviya and F. M. Uckun Role of STAT6 in IgE Receptor/Fc{varepsilon}RI-Mediated Late Phase Allergic Responses of Mast Cells J. Immunol., January 1, 2002; 168(1): 421 - 426. [Abstract] [Full Text] [PDF] J. F. Urban Jr., N. Noben-Trauth, L. Schopf, K. B. Madden, and F. D. Finkelman Cutting Edge: IL-4 Receptor Expression by Non-Bone Marrow-Derived Cells Is Required to Expel Gastrointestinal Nematode Parasites J. Immunol., December 1, 2001; 167(11): 6078 - 6081. [Abstract] [Full Text] [PDF] This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Suzuki, K. Articles by Iwamoto, I. Search for Related Content PubMed PubMed Citation Articles by Suzuki, K. Articles by Iwamoto, I. Related Collections Hematopoiesis and Stem Cells Immunobiology Social Bookmarking                   What's this?

	Copyright © 2000 by American Society of Hematology         Online ISSN: 1528-0020