©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Receptor for Interleukin 13
INTERACTION WITH INTERLEUKIN 4 BY A MECHANISM THAT DOES NOT INVOLVE THE COMMON CHAIN SHARED BY RECEPTORS FOR INTERLEUKINS 2, 4, 7, 9, AND 15 (*)

Nicholas I. Obiri (1), Waldemar Debinski (2), Warren J. Leonard (3), Raj K. Puri (1)(§)

From the (1) Laboratory of Molecular Tumor Biology, Division of Cellular and Gene Therapies, Food and Drug Administration, Center for Biologics Evaluation and Research, Bethesda, Maryland 20892, the (2) Department of Surgery, Division of Neurosurgery, and the Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Milton S. Hershey Medical Center, Hershey, Pennsylvania 17033, and the (3) Laboratory of Molecular Immunology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892

ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES

ABSTRACT

Interleukin 13 (IL-13) shares many biological properties with IL-4, and although the receptor for IL-4 (IL-4R) has been characterized, the expression and structure of IL-13 receptor are unknown. We report here that human renal cell carcinoma (RCC) cells express large numbers of functional IL-13R. Human B lymphocytes and monocytes expressed a very small number of IL-13R, while resting or activated human T cells expressed little or no IL-13R. IL-4 did not compete for IL-13 binding, while IL-13 competed for IL-4 binding, even though IL-4R and IL-13R are structurally distinct on human RCC cells. IL-13 cross-linked with one major protein that is similar in size to the subunit of IL-2, -4, -7, -9, and -15 receptors but was not recognized by anti-or anti-IL-4R antibodies. IL-4, on the other hand, cross-linked with two major proteins, the smaller of which appears to be similar in size to IL-13R and , but (like the IL-13R) it did not react with anti-antibody. Although as shown in this study and in previous studies, is a functional component of IL-4R in lymphoid cells, it does not appear to be associated with IL-4R on RCC cells. Even in the absence of common chain IL-4 and IL-13 were able to up-regulate intracellular adhesion molecule-1 antigen on RCC cells. These data suggest that the interaction of IL-13 with IL-4R does not involve and IL-13R itself may be a novel subunit of the IL-4R.


INTRODUCTION

IL-13() is a 12-kDa lymphokine that has been cloned from activated T cells (1, 2) . The gene for IL-13 is closely linked to the IL-4 gene (3) , and IL-13 protein has been shown to have a 30% identity in the amino acid sequence to IL-4 protein (4) . IL-13 shares many biological properties with IL-4. Like IL-4, it inhibits the production of inflammatory cytokines (2, 5) and up-regulates major histocompatibility complex class II and CD23 expression on monocytes (6) . Similarly, on B cells, IL-13 enhances proliferative responses to anti-IgM and anti-CD40 antibodies, major histocompatibility complex class II, and CD23 expression, induces anti-CD40-dependent IgE class switch, and induces IgG and IgM synthesis (2, 7, 8, 9, 10) . In sharp contrast to IL-4, IL-13 has been shown not to affect resting or activated T cells (9, 11) .

Despite the knowledge of many effects of IL-13, the receptor for IL-13 (IL-13R) has not been reported on. It was hypothesized that the subunit of the IL-2 receptor (IL-2R) (12, 13) , which is associated with IL-4, IL-7, IL-9, and IL-15 (14, 15, 16, 17, 18) receptors, may also be associated with IL-13R (12, 13, 14, 15, 31) . In addition, others have suggested that IL-4R and IL-13R may share a common component that appears to function in signal transduction (4) .

We have previously demonstrated that human renal cell carcinoma (RCC) cells express high affinity IL-4R (19) . Since many of the observed effects of IL-13 are similar to those of IL-4, we have examined RCC cells for the expression of IL-13R. We demonstrate that human RCC cells express large numbers of functional IL-13R and that IL-4 and IL-13 interact with each other's receptors. We provide evidence that IL-13R and IL-4R on RCC cells are composed of different subunits and that IL-13R is mainly composed of a single protein similar in size to the smaller of the two IL-4R subunits. The IL-13R protein does not cross-react with anti-or anti-IL-4R antibodies. Similarly, to our surprise, the smaller of the two IL-4R proteins does not react with anti-antibody. These results suggest that chain is not associated with IL-13R or IL-4R on human RCC cells and further suggest that IL-13R is a monomeric or dimeric protein, which itself may be a novel subunit of IL-4R.


EXPERIMENTAL PROCEDURES

Cytokines and Reagents

Recombinant human IL-13 (2) was kindly provided by Dr. A. Minty (Sanofi Elf Bio Recherches, Laberge, France). For some experiments, we used recombinant human IL-13 expressed in Escherichia coli and produced by one of us (W. D.). Prior to use, we compared IL-13 prepared from the two sources and found that they were identical in biological activity ( i.e. support of TF-1 cell proliferation) as well as in their ability to displace I-IL-13 from the cell surface. Recombinant human IL-4 was provided by Paul Trotta (Schering-Plough Research Institute, Kenilworth, NJ). A polyclonal rabbit antibody to the high affinity human IL-4R (P7) was a kind gift from Immunex Corporation (Seattle, WA). The antibodies to human IL-4 and intercellular adhesion molecule (ICAM-1) were purchased from Genzyme Corp. (Boston, MA) and AMAC, Inc. (Westbrook, ME), respectively. The ICAM-1 cDNA probe, a 1.9-kilobase cDNA cloned from stimulated HL-60 cells (20) , was a gift from Dr. Steve Shaw (NCI, Bethesda, MD).

Cells

The primary cultures of renal cell carcinoma cell lines WS-RCC, HL-RCC, and MA-RCC were established in our laboratory and are maintained as described previously (19) . TF-1 cell line, a human premyeloid erythroleukemic cell line that responds to various cytokines including IL-13 (4) , and other cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum glutamine (2 m M), sodium pyruvate (1 m M), essential amino acids (1 m M), penicillin (100 units/ml), and streptomycin (100 µg/ml).

Radioreceptor Binding Assay

Recombinant human IL-4 and recombinant human IL-13 were labeled with I (Amersham Corp.) by using the IODO-GEN reagent (Pierce) according to the manufacturer's instructions. The specific activity of the radiolabeled cytokines was estimated to range from 20 to 100 µCi/µg of protein.

For binding experiments, typically 1 10RCC tumor cells were incubated at 4 °C for 2 h with I-IL-13 (100 p M) with or without increasing concentrations (up to 500 n M) of unlabeled IL-13. In some experiments, IL-13R expression was examined as described previously (19) . The data were analyzed with the LIGAND program (21) to determine receptor number and binding affinity.

Affinity Cross-linking Studies

Cells (5 10) were labeled with I-IL-13 or I-IL-4 in the presence or absence of excess IL-13 or IL-4 for 2 h at 4 °C. The bound ligand was cross-linked to its receptor with disuccinimidyl suberate (Pierce) at a final concentration of 2 m M for 30 min. Cells were lysed in a buffer containing 1% Triton X-100, 1 m M phenylmethylsulfonyl fluoride, 0.02 m M leupeptin, 5.0 µ M trypsin inhibitor, 10 m M benzamidine HCl, 1 m M phenanthroline iodoacetamide, 50 m M aminocaproic acid, 10 µg/ml pepstatin, and 10 µg/ml aprotinin. The cell lysates were cleared by boiling in sample buffer containing 2-mercaptoethanol and analyzed by electrophoresis through 8% SDS-polyacrylamide gel. The gel was subsequently dried and autoradiographed. In some experiments, the receptor-ligand complex was immunoprecipitated from the lysate overnight at 4 °C by incubating with protein A-Sepharose beads that had been preincubated with P7 anti-human IL-4R or anti-antibody and analyzed as above.

Flow Cytometric Analysis

RCC cells were cultured with IL-13 (10 ng/ml) over a 48-h period, washed, and incubated at 4 °C for 60 min with anti-human ICAM-1 antibody (84H10). Control cells were incubated in staining buffer alone or with isotype control (IgG) antibody. Cells were then washed and stained for 60 min with a secondary antibody consisting of F(ab`)fragment of mouse IgG conjugated to fluorescein isothiocyanate. After washing, cells were analyzed on FACScan/C32 equipment (Becton Dickinson, San Jose, CA) using a LYSIS II program. Fluorescence intensity was expressed as mean channel number on a 256-channel/10log scale.

Northern Blot Analysis

Equal amounts of total RNA were examined by Northern blot analysis (19, 22) . The membrane was probed with P-labeled cDNA for or ICAM-1 at 42 °C for 18 h. In some experiments, the membrane was stripped and probed with a cDNA probe for glyceraldehyde-3-phosphate dehydrogenase.


RESULTS AND DISCUSSION

Expression of IL-13R

Four human RCC lines (WS-RCC, HL-RCC, PM-RCC, and MA-RCC) that were examined bound I-IL-13 specifically, and the density of IL-13R varied from 2100 sites/cell in WS-RCC cells to 150,000 sites/cell in HL-RCC cells (Fig. 1 and ). Scatchard analyses (23) revealed that only one affinity class of receptors was expressed on each cell line. The binding affinities ( K) ranged between 100 p M and 400 p M in three RCC cell lines, while HL-RCC cells expressed lower affinity receptors ( K 3 n M).

Although IL-13 responsiveness has previously been reported in human monocytes, B cells, and premyeloid (TF-1) cells (2, 3, 4, 5, 6, 7, 8, 9, 10, 11) , little is known about IL-13R structure or its binding char-acteristics in these cells. Our results show that freshly isolated human monocytes, Epstein-Barr virus-transformed B cell line, and TF-1 cell line express very few IL-13 binding sites (100-300 binding sites/cell) compared with human RCC cells (). On the other hand, no binding of I-IL-13 was observed on H9 T cells, LAK cells, and resting or phytohemagglutinin-activated peripheral blood lymphocyte. This is compatible with the fact that IL-13 responsiveness has not been observed in T lymphocytes (9, 11) .

Interaction of IL-13 and IL-4 with IL-13R and IL-4R

Recently, it was proposed that IL-2R is associated with IL-13R (14, 15, 17, 18) and that IL-13R may share a common component with IL-4R (4, 24) . To directly address these possibilities, we first performed radioligand binding experiments on HL-RCC and WS-RCC cells using I-IL-4 or I-IL-13 in the presence or absence of excess of either cytokine. Our results (Fig. 2) show that unlabeled IL-4 more efficiently inhibited I-IL-4 from binding to RCC cells (84 and 72% displacement of total binding in WS-RCC and HL-RCC, respectively) than IL-13, which also displaced I-IL-4 binding to these cells (61% of total binding in WS-RCC and 51% in HL-RCC) under similar conditions. On the other hand, while I-IL-13 binding was effectively displaced by IL-13 (about 85% of the total in both cell types), it was only minimally displaced by IL-4 (12% of total displacement in WS-RCC and 7% in HL-RCC). These results indicate that IL-4 and IL-13 both interact with each other's receptors; however, the interaction is not identical, since IL-4 inhibition of I-IL-13 binding was weak and IL-13 inhibition of I-IL-4 binding was not complete. Our results agree with previous observations in which IL-13 was found to compete with IL-4 binding on TF-1 cells (4) . However, in that report the converse experiment was not done. Here we provide results showing that even though IL-13 competed for IL-4 binding, IL-4 did not compete for IL-13 binding.

We also investigated whether IL-13 competes for the IL-4 binding on lymphoid MLA 144 cells or RAJI cell lines. These cells were incubated with radiolabeled IL-4 with or without excess unlabeled IL-4 or IL-13. As shown in Fig. 3, A and B, I-IL-4 bound to MLA 144 and RAJI cells, and excess unlabeled IL-4 effectively displaced radiolabeled IL-4, while excess IL-13 could not compete this binding. This observation is at variance to that seen with RCC cells in which IL-13 competed for IL-4 binding. The inability of IL-13 to compete for I-IL-4 binding to MLA 144 is consistent with our observation that IL-13 did not bind to peripheral blood T (or MLA 144) cells. However, the inability of IL-13 to compete for IL-4 binding on the RAJI (B cell lineage) cell line is not clear. As discussed below, these cells express common chain, and it is possible that common chain interferes with the binding of IL-13. Studies are under way to address this point.


Figure 3: Lack of competition of I-IL-4 binding by IL-13 on MLA 144 and RAJI cells. MLA 144 cells ( A) or RAJI (Burkitt's lymphoma) cells ( B) were incubated with 0.5 n M of I-IL-4 in the absence or presence of 200-fold molar excess (RAJI) or various concentrations (up to 1000-fold molar excess) of IL-13 (MLA 144). Total I-IL-4 bound was 4569 ± 36 cpm on MLA 144 cells and 9975 ± 283 cpm on RAJI cells.



Subunit Structure of IL-13R and IL-4R

We next investigated the subunit structure of IL-13R on RCC cells by cross-linking studies. Our results indicate that I-IL-13 cross-linked to one major protein on all four RCC cells and that the complex migrated as a single broad band ranging between 68 and 80 kDa (Fig. 4, A and B). A single band was also observed on human premyeloid TF-1.J61 cells (Fig. 4 B) only after much longer exposure of the gel. After subtracting the molecular mass of IL-13 (12 kDa), the size of the IL-13 binding protein was estimated at 56-68 kDa. The I-IL-13 cross-linked band was not observed when the cross-linking was performed in the presence of a 200-fold molar excess of IL-13 (Fig. 4, A, lanes 2 for each tumor cell line, and B, lane 2). In addition to the major band, a faint band of approximately 45 kDa was also observed in HL-RCC and PM-RCC but not on MA-RCC cells (Fig. 4 A). This band appeared to be specifically associated with IL-13R, because unlabeled IL-13 competed for the binding of I-IL-13. This band could represent an IL-13R-associated protein or a proteolytic fragment of the larger band. In contrast to the displacement of I-IL-13 binding by unlabeled IL-13 (Fig. 2), an excess of unlabeled IL-4 did not prevent the appearance of IL-13R band in RCC cell lines (Fig. 4 A, lanes 3 for each tumor). IL-13 on the other hand competed for I-IL-4 binding to both major proteins on WS-RCC cells (Fig. 4 C, lane 3). It is of interest that I-IL-13 cross-linked protein was slightly larger in size in TF-1.J61, WS-RCC, PM-RCC, and HL-RCC cell lines compared with that seen in MA-RCC (Figs. 4, A and B, and 5, A and B). Post-translational modifications, such as glycosylation or phosphorylation, may account for this difference.


Figure 4: SDS-polyacrylamide gel electrophoresis analysis of I-IL-13IL-13R or I-IL-4IL-4R complexes. Five million RCC cells ( A) or nine million TF1.J61 cells ( B) were labeled with I-IL-13 in the absence or presence of 200-fold molar excess of unlabeled IL-13 or IL-4. MA-RCC, HL-RCC, and PM-RCC cells were incubated with I-IL-13 alone ( A, lane 1 of each tumor type) or in the presence of excess IL-13 ( A, lane 2 of each tumor type, or B, lane 2) or IL-4 ( A, lane 3 of each tumor type). WS-RCC cells were incubated with 1 n M I-IL-4 alone ( C, lane 1) or in the presence of excess IL-4 ( C, lane 2) or IL-13 ( C, lane 3). The radiolabeled ligand was cross-linked to its receptor and analyzed by SDS-polyacrylamide gel electrophoresis and autoradiographed for 2 days ( A), 6 days ( B), or 4-19 days ( C).




Figure 2: Competition of I-IL-4 or I-IL-13 binding. RCC cells (1 10) were incubated with I-IL-4 (0.64 n M) or I-IL-13 (0.64 n M) with or without excess IL-4 (128 n M) or IL-13 (128 n M). Competition for radioligand binding is expressed as percentage of total binding. Total I-IL-4 bound to two RCC cell cultures was 17,832 ± 1099 and 14,631 ± 780 cpm ± S.D. for HL-RCC and WS-RCC, respectively, and total binding of I-IL-13 was 49,945 ± 3164 and 8119 ± 122 cpm ± S.D. to HL-RCC and WS-RCC cells, respectively.



Immunoprecipitation of IL-13R and IL-4R

To characterize the relationship between IL-4R and IL-13R, we immunoprecipitated I-IL-13IL-13R and I-IL-4IL-4R complexes with the polyclonal anti-IL-4R antibody, P7, which has been used to immunoprecipitate IL-4R subunits (14) . Our results show that anti-IL-4R did not co-immunoprecipitate IL-13R on MA-RCC cells (Fig. 5 A, lane 3) or WS-RCC cells (Fig. 5 B, lane 3). Thus, IL-13R is not immunoreactive with the anti-IL-4R antibody. However, as expected, anti-IL-4R antibody immunoprecipitated IL-4R (140 and 75 kDa) on WS-RCC cells (Fig. 5 C, lane 1), which express IL-4R (19) .


Figure 5: Immunoprecipitation of IL-13R and IL-4R. RCC or MLA 144 cells were labeled with 1 n M either I-IL-13 or I-IL-4 in the absence or presence of unlabeled IL-13 or IL-4. The cell lysates were incubated with the anti-IL-4R (P7) or anti-antibody overnight at 4 °C to immunoprecipitate IL-4R or and analyzed on SDS-polyacrylamide gel electrophoresis. A, radiolabeled IL-13 bound to MA-RCC cells in the absence ( lane 1) or presence ( lane 2) of excess unlabeled IL-13 or immunoprecipitated with anti-IL-4R ( lane 3) or anti-antibody ( lane 4). B, I-IL-13 cross-linked to WS-RCC cells in the absence ( lane 1) or presence ( lane 2) of excess unlabeled IL-13 or immunoprecipitated with IL-4R ( lane 3) or antibody ( lane 4). C, I-IL-4 cross-linked to WS-RCC cells in the absence ( lane 1) or presence ( lane 2) of excess IL-4 or immunoprecipitated with antibody to IL-4R ( lanes 1 and 2) or to ( lane 3). Lane 4 represents immunoprecipitation of I-IL-4-labeled receptors on MLA-144 cells by anti-antibody. D, Northern analysis of mRNA expression. Total cellular RNA was extracted from three RCC (WS-RCC, MA-RCC, and CAKI-1), two T cell lymphoma, one B-cell Burkitt's lymphoma cell line, or a murine fibroblast cell line (L cells), and 20 µg was electrophoresed through 1% agarose/formaldehyde denaturing gel. These were then probed with a P-labeled cDNA (30) and exposed to autoradiographic film for 2 days.



We next immunoprecipitated IL-13R and IL-4R on RCC cells with anti-antibody. Antibody to the IL-2R has been shown to co-immunoprecipitate IL-4R and IL-7R (14, 16) . However, this antibody did not immunoprecipitate the IL-13R on MA-RCC cells (Fig. 5 A, lane 4) or on WS-RCC (Fig. 5 B, lane 4). As a control, anti-did immunoprecipitate the IL-4R bands on lymphoid MLA 144 cells (14) (Fig. 5 C, lane 4). Although has been shown to be associated with IL-4R on lymphoid cells (14, 15) , it does not appear to be associated with the IL-4R on WS-RCC tumor cells because anti-did not immunoprecipitate IL-4R on these cells (Fig. 5 C, lane 3). The absence of expression on RCC tumor cells was substantiated by Northern analysis. All three RCC cell lines examined were negative for mRNA, while lymphoid cells were strongly positive (Fig. 5 D).

The lack of association of chain with IL-4R in RCC cells is a novel and surprising observation. But interestingly, in the absence of chain, IL-4Rs were functional on these cells as IL-4 inhibited their proliferation in vitro (19) and modulated ICAM-1 antigen expression() (shown below). Similarly, we previously demonstrated that murine L cells, which constitutively express IL-4R, do not express common chain (14) . However, the introduction of chain into these cells caused the phosphorylation of IRS-1, which was not observed in untransfected cells, indicating that chain is necessary at least for IRS-1 phosphorylation. These data suggest that IL-4 may involve at least two independent signaling pathways. The involvement of two independent signaling pathways has also been previously suggested (25, 26) . In addition, in a recent study utilizing monocytic cell lines (Mono Mac 6 and THP-1) that have many properties in common with normal human monocytes, it was demonstrated that IL-4 caused signaling even though these cells did not express common chain by polymerase chain reaction (12, 27) . These data indicate that IL-4R is comprised of different receptor components in different cell types and that they are able to signal by different pathways.

Our data indicate that IL-13R do not associate with chain of IL-2, IL-4, IL-7, IL-9, and IL-15R in RCC cells. Since RCC cells did not express this chain, it is possible that chain associates with IL-13R in other cells. The possible association of IL-13R with chain was considered in cells that expressed this subunit. Human B cells and T cells, including MLA 144 cells, have been shown to express chain (14, 15) . Since these cells expressed no or only about 100-300 IL-13 binding sites/cell and very few bound ligand cross-links to the receptor, it is difficult to analyze receptor structure by cross-linking studies. However, the fact that IL-13 did not compete for IL-4 binding on these cells suggests that common chain does not associate with IL-13R. In addition, the minimal binding of I-IL-13 to cells expressing chain also suggests that chain does not interact with IL-13R.

Functional Significance of IL-13R Expression on Human RCC Cells

We have previously shown that RCC cells express high levels of ICAM-1 antigen on their cell surface, which is up-regulated by IL-4.As shown in Fig. 6, A and B, WS-RCC and PM-RCC expressed significant basal levels of ICAM-1, and treatment with IL-13 significantly increased this expression. The IL-13 modulation of ICAM-1 expression was also evident at the mRNA level as shown in Fig. 6 C. When these data were normalized to glyceraldehyde-3-phosphate dehydrogenase, we observed that IL-13 induced a 44% increase in ICAM-1 mRNA levels compared with 32% by IL-4 and 224% by a positive control, IFN-. These data demonstrate that IL-13 on RCC cells is functional.


Figure 6: Effect of IL-13 on ICAM-1 expression on RCC cells. WS-RCC ( A) or PM-RCC cells ( B) were incubated in medium alone or in medium containing IL-13 (10 ng/ml) for 48 h. Cells were then analyzed for ICAM-1 expression by flow cytometry. Cells from control and IL-13-treated groups showed identical patterns when analyzed using isotype control antibody. Data from single experiments are shown. Experiments were repeated 2-6 times with a similar result. C, WS-RCC cells were cultured with or without IL-13, IL-4, or interferon- ( IFN) for 48 h. Total RNA from these cells was electrophoresed (15.6 µg/lane) and hybridized with a P-labeled cDNA probe for ICAM-1. For densitometric scanning, the blot was stripped and reprobed with a P-labeled cDNA for glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) as an internal standard. We quantitated the bands on a densitometer and compared the ICAM-1 bands after normalizing them with their corresponding glyceraldehyde-3-phosphate dehydrogenase bands. Results shown are representative of three experiments.



Conclusion

We have demonstrated that human RCC cells express between 15- and 500-fold higher numbers of functional high or intermediate affinity IL-13R than normal immune cells. Although IL-13 appears to interact with IL-4R, IL-4 has minimal interaction with IL-13R. The subunit structure of IL-13R appears to be different from that of IL-4R. I-IL-13 cross-links to a major protein of 56-68 kDa, while I-IL-4 cross-links to two different proteins of 140 and 75 kDa. Even though the sizes of IL-13R and one of the two proteins of the IL-4R are similar to those of the subunit of IL-2R, IL-4R, IL-7R, IL-9R, and IL-15R, anti-antibody did not cross-react with IL-13R or with IL-4R on RCC cells. The absence of was confirmed by the lack of mRNA for in RCC cells. These data suggest that the composition of IL-4R on RCC cells is different from that on immune cells that do express chain. Because of the similarity in the molecular size of IL-13R and one subunit of IL-4R, it is possible that IL-13R is a component of the IL-4R, which may be one explanation for how IL-13 can interact with IL-4R in RCC cells as shown in this study and in TF-1 cells as shown by Zurawski et al. (4) . Further studies are needed to support or refute this hypothesis.

Finally, the expression of IL-13R on human RCC cells may provide a unique antigen that can be used for many diagnostic or therapeutic purposes. For example, radionuclides conjugated to IL-13 or to an antibody to IL-13R may be useful for nuclear medicine scanning diagnosis of RCC. Similarly, IL-13R may be targets for many receptor-directed therapeutics. We have observed that other human solid tumors also express IL-13R that are internalized following binding to a ligand consisting of IL-13 and Pseudomonas exotoxin.() The large numbers of IL-13R on RCC cells and the apparent paucity of IL-13R on normal cells should make RCC or other solid tumor cells preferential targets for the cytotoxic effects of these and similar tumoricidal chimeric molecules.

  
Table: Expression of IL-13R on human cells



FOOTNOTES

*
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked `` advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§
To whom correspondence should be addressed. Tel.: 301-827-0471; Fax: 301-827-0449; E-mail: ``PURI@A1.CBER.FDA.GOV''.

The abbreviations used are: IL, interleukin; IL-13R and IL-4R, interleukin 13 and interleukin 4 receptor(s), respectively; RCC, renal cell carcinoma; ICAM-1, intercellular adhesion molecule; LAK, lymphokine-activated killer.

N. I. Obiri, N. Tandon, and R. K. Puri (1995) Int. J. Cancer (in press).

W. Debinski, N. I. Obiri, I. Pastan, and R. K. Puri, submitted for publication.


ACKNOWLEDGEMENTS

We thank P. Leland and H. Mostowski for expert technical assistance and flow cytometric analysis, respectively; Dr. P. Munson (NIH) for help in designing binding experiments and statistical interpretation of the IL-13 binding data using the LIGAND program; Dr. G. Haas (Wayne State University, Detroit, MI) for PM-RCC cells; Dr. P. Burd (Center for Biologics Evaluation and Research (CBER)) for TF1.J6 cells; G. Tosato (CBER) for DH cells; Chiron Corp. (Emerryville, CA) for IL-2 and Dr. I. Pastan (National Cancer Institute) for helpful discussions and critical reading of this manuscript.


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