Recombinant human interleukin 4 has antiproliferative activity on human tumor cell lines derived from epithelial and nonepithelial histologies (original) (raw)
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Effects of Interleukin 4 Upon Human Tumoricidal Cells Obtained From Patients Bearing Solid Tumors
Journal of Leukocyte Biology, 1991
The use of human interleukin 4 (1L4) in the generation of tumoricidal eftector cells derived from cancer patients undergoing either interleukin 2/lymphokine activated killer cells (lL2/LAK) therapy or tumor derived activated cell (TDAC) therapy was examined in the present study. Human lL4 alone did not generate LAK cells from patients undergoing IL2/LAK therapy when cultured in media containing 2% AB serum (five out of six patients). However, when the serum free medium, Aim V, was used, lL4 did generate LAK cells (eight out of nine patients), although not as cytotoxic as those activated with lL2. When cells were cultured in both lL2 and IL4 during the generation of LAK cells (3-7 days), better cell recoveries were frequently observed. The cytolytic activity of these cells against Daudi target cells was slightly reduced when compared to that response induced by lL2. This inhibition of LAK activity by 1L4 was dose dependent with 1,000 U/mI lL4 producing maximal effects. This form of inhibition did not correlate with any phenotypic differences between those cells cultured in lL2 and those cells cultured in 1L2 plus IL4. The lL4 mediated inhibition was also observed when the cells were cultured in Aim V medium which contains indomethacin. This inhibition induced by IL4 could not be overcome by using supra-optimal lL2. In addition to its effect during the generation of 1L2 induced LAK cells, IL4 also appeared to reduce the cytolytic activity of pre-activated mature LAK cells. These results suggest that 1L4 has a complex role in regulating the actions of LAK cells induced by lymphokines. When IL4 was used with IL2 during the first 5 weeks of growth of TDAC, an enhanced growth was observed when compared to the growth of TDAC when only one lymphokine was used. Besides the growth enhancement of TDAC, a better cytolytic response was observed when both lymphokines were used together. Thus, for the best growth of TDAC both lL2 and lL4 are required.
Tumor cells expressing membrane-bound form of IL-4 induce antitumor immunity
Gene Therapy, 2000
Local cytokine concentrations are required for inhibition of tumor growth with less toxic side-effects. However, genetically engineered tumor cells secreting cytokines still induce toxicity and activate bystander cells. To circumvent such problems, membrane-bound forms of IL-4 (IL-4m) were expressed on MethA fibrosarcoma tumor cells. Chimeric forms of IL-4 with the type I transmembrane protein CD4 or type II transmembrane protein TNF were designed to express IL-4 in opposite orientations on the tumor cell surface. The IL-4m on tumor clones was able to support cell growth of the IL-4 dependent cytotoxic cell line (CT.4S) and
Cancer Research, 2006
An interleukin (IL)-4-containing tumor environment is reported to be beneficial for immune clearance of tumor cells in vivo; however, the effect of IL-4 on the effector CD8 + T cells contributing to tumor clearance is not well defined. We have used the immunogenic HLA-CW3-expressing P815 (P.CW3) mastocytoma and investigated whether IL-4 expression by the tumor affects tumor clearance and, if so, whether it alters the tumor-induced VB10 + CD8 + T-cell response. P.CW3 were stably transfected with IL-4 or the empty control vector, and independent cell lines were injected i.p. into syngeneic DBA/2 mice. After apparent clearance of primary tumors over 12 to 15 days, secondary tumors arose that lacked surface expression and H-2-restricted antigen presentation of CW3 in part due to the loss of the HLA-CW3 expression cassette. Surprisingly, mice that received IL-4-producing tumor cells showed delayed primary tumor clearance and were significantly more prone to develop secondary tumors compared with mice receiving control tumor cells. Tumor clearance was dependent on CD8 + T cells. The IL-4-secreting P.CW3 tumor cells led to markedly higher mRNA expression of IL-4 and granzyme A and B but no differences in IFN-; and IL-2 production, cell proliferation, or ex vivo CTL activity in primary VB10 + CD8 + T cells when compared with the control tumor cells. We concluded that tumor-derived IL-4 selectively changed the quality of the tumor-induced CD8 + T-cell response and resulted in unexpected negative effects on tumor clearance. These data bring into question the delivery of IL-4 to the tumor environment for improving tumor immunotherapy. (Cancer Res 2006; 66(1): 571-80) Requests for reprints: Norbert Kienzle,
Journal of Clinical Investigation, 1993
Previously, Puri et . 1991. Cancer Res. 51:3011-3017) have demonstrated that murine sarcoma and colon adenocarcinoma cells express high affinity interleukin4 receptors (IL-4R) which are internalized after binding to a chimeric ligand consisting of IL-4 and Pseudomonas exotoxin. In the present study, we have tested primary cultures of human renal cell carcinoma (RCC) cells, generated from tumor specimens obtained after nephrectomy, for the expression of IL4R and their modulation by IL4. By using iodinated IL4 in a receptor binding assay, we observed that renal cell carcinoma cells expressed a single class of high affinity IL4R ranging from 1,425±207 (mean±SEM) to 3,831±299 (mean±SEM) IL4R molecules/cell with a Kd ranging from 112±11 pM to 283±71 pM. Northern blot analysis for IL4R gene expression, performed with a cDNA probe to IL4R, revealed that all RCC cells exhibited a single mRNA species of 4 kb. IL4 downregulated the surface expression of IL4R on one RCC tumor cell line. The function of IL-4R expression on RCC tumor cells was further determined by investigating the effect of IL4 on tumor cell growth in vitro and comparing it with IL-4 effect on growth of normal fibroblast and endothelial cell lines. Tumor cell growth, as measured by [3H]thymidine incorporation, was inhibited by IL4 from 20 to 68% in a dose-dependent manner. A neutralizing antibody to human IL-4 was able to reverse the growth inhibitory effect of IL4. Normal human fibroblast and endothelial cell lines also expressed high affinity IL4R, however, IL4 did not inhibit their growth in vitro. In fact, IL-4 caused modest stimulation of their growth. Taken together, our findings can help develop strategies for the treatment of RCC in which IL-4R may be used as a target for IL4 itself, for IL4 toxin therapy or, alternatively, in gene therapy. (J. Clin. Invest. 1993. 91:88-93.) Key words: IL4 receptors on renal cell carcinoma * modulation of IL4 receptors * tumor cell growth inhibition experiments. The human Burkitt lymphoma-B cell lines, Molt-4 and Daudi, were kindly provided by Dr. J. A. Hank (University ofWisconsin, Madison, WI). Normal human skin fibroblast cell lines (39-Sk and 969-Sk) and human umbilical vein endothelial (HUVE) cells were obtained from American Type Culture Collection, Rockville, MD.
An antiproliferative bioassay for interleukin-4
Journal of Immunological Methods, 1996
Inrerleukin-4 (IL41 is cwremly being used for therapeutic inlervention in a wide range of malignan direares as an antitumwr ageor Altbagb bioassays have been developed mar measure tie pmlifemrive capaciry of IL-4. none measure the antiproliferative tiviry of this mokcuk. We have developed a simple. sensitive bioassay for human IL-4 based on the ability of this ~ytokinc to inhibit the proliferation of the humzm lune, carcinoma line, CCL-l85 M easy to maintain. cymkine i&&&m. cell line. It is rapid. ~repmducible and sensitive. able to detect 2 pg/ml IL-4 The assay is completely muqwnsive to atl aher imertcuki~ from n-2 to IL-12, to ti colony stimulating factors and hansforming growth factor baa and is 10%fold less sensitive to imaferon-o olmour wrosir factor-a. IL-Ip and IL-13. The assay can be made completely spexitii for n-4 by incmding specific neutralizing antibodies for IL-4 a-4 in suitable fo: tire estimation of IL-4 m boyl plasma ard serum sampler
Expression of high-affinity IL-4 receptors on human melanoma, ovarian and breast carcinoma cells
Clinical & Experimental Immunology, 2008
It has previously been shown that murine sarcoma cells express high-affinity IL-4 receptors (IL-4R) which are internalized after binding to the ligand (Puri et al., Cancer Res 1991; 51:3011-7). We have also reported that human renal cell carcinoma cells express high-affinity IL-4R, and IL-4 inhibits tumour growth in vitro (Obiri et al., J Clin Invest 1993; 91:88). In this study we investigated the expression and function of IL-4R on other human solid tumours. Human melanoma, ovarian carcinoma and breast carcinoma cell lines were assessed for the cell surface expression of IL-4R by radio-ligand receptor binding and for IL-4R gene expression by Northern blot analysis. Primary cultures of mesothelioma and neurofibrosarcoma cells were similarly investigated. Human melanoma, ovarian carcinoma and breast carcinoma cell lines expressed IL-4R on their cell surface with a dissociation constant (Kd) of 140-549 pM. These tumour lines expressed a single 4 kb species of mRNA for IL-4R. Similarly, primary cultures of mesothelioma and neurofibrosarcoma cells were positive for the IL-4R mRNA by Northern blot analysis. Fresh, non-cultured mesothelioma and neurofibrosarcoma tumour sections were also positive for the presence of IL-4R as determined by immunohistochemistry of frozen sections using anti-IL-4R antibody. In order to study possible functions of IL-4R, we evaluated the effects of IL-4 on cell growth and its effect on MHC antigen expression in the presence or absence of interferon-gamma (IFN-y). In tissue culture, IL-4 reduced the growth of tumour cell lines and primary cell cultures studied. IL-4 had very little effect on MHC class I antigen expression on ovarian, breast and melanoma cell lines; however, MHC class II (HLA-DR) expression was enhanced on melanoma and breast carcinoma cells. IL-4 also enhanced the IFN-y-induced class II expression on melanoma and breast carcinoma cells. Taken together, our observations indicate that IL-4R are expressed on a variety of human solid tumours and these receptors may be functional. IL-4 alone and in combination with IFN-y may play a role in host immune response against cancers.
International Journal of Cancer, 1994
of 7 human colorectal-carcinoma cell lines. In 5 cell lines (HT29, WiDr, LS41 IN, LS5 13, LS1034) a dosedependent reduction of proliferation was documented. At I00 U/ml, IL-4 inhibited thymidine incorporation between 45 and 75% and M l T conversion (26 to 4 I YO). The ability of LS5 I 3 and WiDr cells to form colonies after IL-4 treatment was reduced by 85 and 62% respectively. LS5 I 3 was the most sensitive cell line, with I L 4 inducing half-maximal inhibition at 5 to 6 U/ml. The inhibitory effect of 11-4 was completely neutralized by anti-IL-4 antibodies. Northern-blot analysis revealed the presence of IL-4-receptor (IL-4R) mRNA in all cell lines. The membrane expression of the 130-kDa IL-4R was assessed by FACS, utilizing an anti-IL4R monoclonal antibody and was confirmed by biotinylated IL-4 binding. Our results attribute an important role for IL-4 as a negative regulator of colorectal-carcinoma cell growth, thus indicating a possible avenue for intervention in this disease. o 1994 Wiley-Liss, Inc.
Interleukin 4 receptor expression and growth inhibition of gastric carcinoma cells by interleukin 4
Cancer research, 1992
The expression of the interleukin 4 (IL-4) receptor (IL-4R) and ef facts of human recombinant IL-4 on human gastric carcinoma cell lines were studied. We demonstrated that 11-4 inhibited the growth of gastric carcinoma cells in a dose dependent manner (0.1â€"100anita/mi) in a I3Hlthymldine incorporation proliferation assay. The gastric carcinoma cells varied in sensitivity to treatment with low dose IL-4. Treatment of calls with IL-4 altered the morphology of the calls to a â€oeflattened― morphological shape resembling differentiation. The IL-4-mediated growth inhibition was significantly abrogated by neutralization of 11-4 with specific anti-IL-4 antibody. IL-4R expression on the call surface was determined by assessing biotin-labeled IL-4 binding to cells using flow cytometry. IL-4R expression ranged from 5 to 85% of total cell population in the gastric carcinoma cell lines assessed. There was a positive correlation between the sensitivity to IL-4-mediated growth inhibition and IL-4R expression. By Northern blot analysis, we dem onstrated that mRNA of IL-4R was expressed in the gastric carcinoma cells. Using in situ hybridization, we confirmed that IL-4R mRNA was expressed in the gastric carcinoma cell at the single cell level. By using a sensitive polymerase chain reaction technique, we demonstrated that gastric carcinoma cells expressed IL-4 mRNA, suggesting a possible autocrine loop. These studies indicate that IL-4 can significantly mod ulate gastric carcinoma cells that possess IL-4R. IL-4R on gastric car cinema cells may be a potential therapeutic target site for IL-4-directed therapy.
Apoptosis resistance in epithelial tumors is mediated by tumor-cell-derived interleukin-4
Cell Death and Differentiation, 2008
We investigated the mechanisms involved in the resistance to cell death observed in epithelial cancers. Here, we identify that primary epithelial cancer cells from colon, breast and lung carcinomas express high levels of the antiapoptotic proteins PED, cFLIP, Bcl-xL and Bcl-2. These cancer cells produced interleukin-4 (IL-4), which amplified the expression levels of these antiapoptotic proteins and prevented cell death induced upon exposure to TRAIL or other drug agents. IL-4 blockade resulted in a significant decrease in the growth rate of epithelial cancer cells and sensitized them, both in vitro and in vivo, to apoptosis induction by TRAIL and chemotherapy via downregulation of the antiapoptotic factors PED, cFLIP, Bcl-xL and Bcl-2. Furthermore, we provide evidence that exogenous IL-4 was able to upregulate the expression levels of these antiapoptotic proteins and potently stabilized the growth of normal epithelial cells rendering them apoptosis resistant. In conclusion, IL-4 acts as an autocrine survival factor in epithelial cells. Our results indicate that inhibition of IL-4/IL-4R signaling may serve as a novel treatment for epithelial cancers.