Expression of high-affinity IL-4 receptors on human melanoma, ovarian and breast carcinoma cells (original) (raw)
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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.
Cancer Research
We have reported that human malignant glioma cell lines express high levels of plasma membrane interleukin-4 receptors (IL-4R). We have also reported that biopsy/surgical samples or primary explant cell cultures from brain tumors express mRNA and protein for the IL-4R␣ chain, a primary IL-4-binding protein. However, whether IL-4R are expressed in brain tumors in situ has not been resolved. In addition, expression of IL-4R on the cell surface of various normal brain tissues is not known. We examined the expression of IL-4R by using a monoclonal antibody to the IL-4R␣ chain (also known as IL-4R ) in surgical/biopsy samples of brain tumor tissues by immunohistochemistry. Our data indicate that 15 of 18 glioblastoma multiforme (GBMs) tumors obtained from two different institutions and 12 other brain tumor samples are moderately to intensely positive for IL-4R␣. In contrast, although IL-4R␣ mRNA was expressed, no IL-4R protein was detectable in two adult and one pediatric brain tissue specimens. In addition, a commercially available human neural tissue grid containing fixed tissues from various areas of brain showed no positive staining for the IL-4R␣ chain. IL-4R␣ expression was also demonstrated on astrocytoma grades I, II, and III. Because IL-4 cytotoxin comprised of a circularly permutated IL-4 and a mutated form of Pseudomonas exotoxin [IL4(38-37)-PE38KDEL] is cytotoxic to IL-4R-expressing cells, we tested whether primary GBM explant cell cultures are sensitive to IL-4 cytotoxin. Our data indicate that 13 of 15 GBM cell cultures were 25-74 times more sensitive to IL-4 cytotoxin compared with normal human astrocytes or the NT2 neuronal cell line. These observations indicate that human brain tumors in situ overexpress IL-4R compared with normal brain tissues, thus confirming our previous conclusions that IL-4R in brain tumors may serve as an attractive target for anticancer therapy.
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
Interleukin 4 receptor expression on human lung tumors and normal lung
Cancer research, 1991
Interleukin 4 (IL-4) receptors were detected by a monoclonal antibody on tumor cells of 10 of 29 squamous cell carcinomas and 6 of 17 adenocarcinomas of the lung. None of the small cell carcinomas or carcinoid tumors stained. Parallel sections stained for epidermal growth factor receptors showed that all but 2 of the IL-4 receptor-positive tumors also expressed epidermal growth factor receptors. Positive labeling for IL-4 receptors was also obtained on nonneoplastic bronchial epithelium and on lymphocytes and macrophages infiltrating the tumor stroma. The role of IL-4 and its receptor in normal human lung is unknown, but the expression of IL-4 receptors on particular subtypes of lung tumors suggests that they may have a role in differentiation or proliferation of squamous and adenocarcinomas.
Cancer research, 1995
Interleukin 4, a T cell-derived 20-kDa glycoprotein, plays an important role in regulating the immune response of B cells, T cells, and macrophages against infections and malignant cells. For this reason recombinant human interleukin 4 (rhIL-4) has entered early clinical trials in cancer patients. In the present study we report that rhIL-4 has an antiproliferative effect on five of nine cell lines derived from human colon tumors, head and neck tumors, and glioblastomas as measured by a decrease of colony formation in human tumor cloning assays. All of the cell lines with in vitro responsiveness express at least 100 high-affinity receptors for human interleukin 4 per cell on their cell surface, whereas the nonresponsive tumor cell lines lack expression of high-affinity receptors for human interleukin 4 on their cell surface. In the next series of experiments we have xenotransplanted some of the responsive cell lines into athymic nude mice. Subsequently, the animals were treated s.c. ...
Interleukin 4 Receptor Expression on Human Lung Tumors and Normal Lung1
Interleukin 4 (11-4) receptors were detected by a monoclonal antibody on tumor cells of 10 of 29 squamous cell carcinomas and 6 of 17 adenocarcinomas of the lung. None of the small cell carcinomas or carcinoid tumors stained. Parallel sections stained for epidermal growth factor receptors showed that all but 2 of the 11-4 receptor-positive tumors also expressed epidermal growth factor receptors. Positive labeling for IL-4 receptors was also obtained on nonneoplastic bronchial epithelium and on lymphocytes and macrophages infiltrating the tumor stroma. The role of IL-4 and its receptor in normal human lung is unknown, but the expression of IL-4 receptors on particular subtypes of lung tumors suggests that they may have a role in differentiation or proliferation of squamous and adenocarcinomas.
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.
British Journal of Cancer, 1996
The recent use of interleukin 2 (IL-2) and interleukin 4 (IL-4) single cytokine modified tumour cells in rodent models has demonstrated a potential use of these cytokines to produce autologous cancer cell vaccines. Here we compare the potential therapeutic benefit of transduction with IL-2 or IL-4 alone, and combined IL-2 + IL-4 in B16F1O cells, a murine malignant melanoma of poor immunogenicity. Transduction of B16F1O cells (MHC class I and II negative) to express either IL-2 or IL-4 alone delays the formation of tumours, IL-4 being more effective than IL-2. However, combined expression of IL-2 + IL4 reduces tumorigenicity more than either cytokine alone. The eventual formation of tumours may result from loss of gene expression, and preliminary results suggest methylation of the retroviral long terminal repeat (LTR), rather than loss of the transduced DNA sequences. Histological examination of tumours expressing either IL-2 or IL-4 alone shows a non-specific inflammatory reaction with an increased tissue infiltrate of immune effectors (monocytes/macrophages, lymphocytes, granulocytes) localised around the tumour. In comparison, when cells expressing combined IL-2 + IL-4 were injected there were more granulocytes present, and perhaps more importantly, these were mainly localised within the tumour. The benefit of combined IL-2 + IL-4 expression results from a local rather than systemic effect as the growth of tumours from cells expressing IL-2 or IL-4 alone injected at distant sites was comparable with a single inoculation of cells expressing either cytokine alone. However, when cells expressing single cytokines IL-2 or IL-4 were mixed and injected at the same site, in comparison with the clonal population of cells expressing combined IL-2 + IL-4, tumour growth was characteristic of IL-4 alone rather than IL-2 + IL-4. Treatment of established tumours with a single injection of lethally irradiated tumour cells expressing IL-2 + IL-4 was sufficient to either reject tumours, or at least delay further tumour development. Furthermore, treatment stimulated an initial non-specific immune reaction that lead to a systemic immunity. Lethally irradiated wild-type cells were also successful in treating some established tumours, although this did not induce any systemic immunity. However, although successful in treatment studies, neither wild-type nor combined IL-2 + IL-4 expressing cells were able to vaccinate animals against a subsequent challenge with live wild-type tumour. These results indicate a potential therapeutic benefit with the use of combination IL-2 + IL-4 transduction of autologous cancer cells.
Journal of Experimental Medicine, 1988
Addition of IL-4 (1,000 U/ml) to either high or low concentrations of IL-2 augmented tumor-infiltrating lymphocytes (TIL) growth from human melanoma. Weekly restimulation with irradiated tumor cells in conjunction with IL-4 allowed enhanced growth of TIL. With low-dose IL-2 (10 U/ml) and IL-4, expanded TIL had little cytolytic activity against Daudi or allogeneic tumors. Further, IL-4 augmented the total lytic activity against autologous tumors in most cases. With high-dose IL-2 (1,000 U/ml), IL-4 addition decreased nonspecific killing activity against Daudi or allogeneic melanomas in many cases, and reciprocally augmented cytolytic activity against the autologous melanoma in many cases. This suggests the possible use of IL-4 in cancer therapy, especially in adoptive cellular immunotherapy using TIL or in conjunction with IL-2 administration.