GENOMICS

9,

44-50

( 1991)

Chromosomal

Location

of Murine

and Human IL-I Receptor

Genes

NEAL G. COPELAND,* COLLEEN M. SILAN,” DAVID M. KINGSLEY,* NANCY A. JENKINS,* LINDA A. CANNIZZARO,~ CARLO M. CROCE,~ KAY HUEBNER,~ AND JOHN E. SIMS* *Mammalian Genetics Laboratory, ABL-Basic Research Program, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland 21707; tFe/s Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 79 740; and Slmmunex Corporation, Seattle, Washington 987 0 1 Received

June

15,

199O,revisedAugust6,

In all cell types so far examined, both IL-la and IL-ID are able to bind to the same receptor molecules, albeit sometimes with significantly different affinities (Bird and Saklatvala, 1986; Dower et al., 1986; Kilian et al., 1986; Chin et al., 1987). The current evidence suggests that at least two types of IL-l receptor exist (Bomsztyk et al., 1989; Chizzonite et al., 1989). Recently cDNA clones for the IL-l receptor (IL-1R) expressed in T cells and fibroblasts (the type I receptor) have been isolated from both murine (Sims et al., 1988) and human (Sims et al., 1989) cell lines. We report here that in the human, the gene for this receptor (IL-lR1) maps to chromosome 2, near the genes for IL-la and IL-l/l. The murine IL-1R gene (Ilklrl) maps to chromosome 1, in a region of synteny with human chromosome 2, but separate from the IL-l genes, which map on murine chromosome 2.

The gene for the type I interleukin-1 (IL-l) receptor has been mapped in both mouse and human. In the human genome, a combination of segregation analysis of rodent-human hybrid cells and chromosomal in situ hybridization has placed the gene on the long arm of chromosome 2, at band 2q12. This is near the reported map position of the loci for IL-la and IL-l/3 (2q13+2q21). The murine gene has been mapped by analysis of restriction fragment length polymorphisms in interspecific backcrosses to the centromerit end of chromosome 1, in a region that is syntenic to a portion of human chromosome 2. The murine 11-lrl gene has thus been separated from the IL-l genes, which lie on IIIUriUe chromosome 2. e! 1991 Academic PRSS, k.

INTRODUCTION

IL-la and IL-10 (collectively IL-l)’ are cytokines that play a major role in host defense mechanisms. They exert inductive or regulatory influences on immune responses, inflammatory reactions, and hematopoiesis (Oppenheim et al., 1986; Martin and Resch, 1988). The two IL-1s are only distantly related in primary sequence, despite having broadly overlapping, if not identical, spectra of biological activities; in the human they share 26% amino acid identity (March et al., 1985). There is great conservation of each form across species, however. Murine and human IL-la share 62% amino acid identity; murine and human IL-1B show 67% identity (March et al., 1985; Gray et al., 1986). The genes for IL-la and IL-10 are closely linked on chromosome 2 in both mouse and human (Webb et al., 1986; D’Eustachio et al., 1987; Modi et al., 1988; Boultwood et al., 1989; Lafage et al., 1989), suggesting that the two forms may have arisen by gene duplication.

MATERIALS

METHODS

Mice. Interspecific backcross progeny were generated by mating (C57BL/6J X Mus spretus)F, females and C57BL/6J males as previously described (Buchberg et al., 1988). One hundred ninety-six backcross progeny were analyzed in these studies. Eighty-four of the progeny were typed for all seven probes (see Results). DNA isolation and hybridization analysis. Mouse genomic DNA isolation, restriction enzyme digestion, agarose gel electrophoresis, Southern blot transfer, and probe hybridization were performed essentially as described (Jenkins et al., 1982). All blots were prepared with Zetabind nylon membrane (AMF-Cuno). The 11-lrl, Ctla-4, Acrg, Bcl-2, En-l, and Ren-2 probes were prepared by nick translation. The DlFcrl probe was prepared by random-hexanucleotide priming (Feinberg and Vogelstein, 1984). Washing was done to a final stringency of 1X SSCP, 0.1% SDS, 65°C for 44

Inc. reserved.

AND

Murine Gene

1 Abbreviations used: IL-l, interleukin-1: II,-lR, IL-l receptor; Il-lrl. gene for the human type I IL-l receptor: Zl-lrl, gene for the murine type I IL-l receptor. 0888-7543/91 $3.00 Copyright iC1 1991 by Academic Press. All rights of reproduction in any form

1990

INTERLEUKIN-1

RECEPTOR

all probes except I>1 Fcrl, which was washed at a final stringency of 0.2X SSCP, 0.1% SDS, 65°C. Probes. The Illlrl probe was a 529-bp HindIII/ BamHI mouse cDNA fragment from the cytoplasmic region (Sims et al., 1988). The DlFcrl probe was a I-kb PuuII fragment of a 4.4-kb mouse genomic clone. The Ctla-4 probe was a 1.9-kb HindIII/EcoRI mouse cDNA fragment. The Acrg probe was a 1.7-kb EcoRI fragment of mouse cDNA. The Bcl-2 probe was a 6.0kb SalI/BamHI mouse genomic fragment containing the 3’untranslated region of the Bcl-2 locus. The En-l probe was a 3.6-kb EcoRIISstI fragment of mouse genomic DNA. The Ren-2 probe was a full-length cDNA corresponding to the Ren-2 locus of DBA/BJ mice. Statistical analysis. Mouse gene order was determined by minimizing the number of recombination events required to explain the allele distribution patterns (“pedigree analysis,” Avner et al., 1988) using the computer program “Spretus Madness” developed by D. Dave (Data Management Services, Inc., Frederick, MD) and A. M. Buchberg (BRI-Basic Research Program, Frederick, MD). All reported gene orders were confirmed by maximum likelihood analysis (Bishop, 1985). Human

Gene

Probes. The IL-l R cDNA probe used for Southern blots was a 471-bp Hind111 to Asp718 fragment from the cytoplasmic region of human IL-1R (Sims et al., 1989). Additionally a 1.3-kb genomic EcoRI fragment containing an exon encoding amino acids 202-224 of the extracellular region (J.S., unpublished) was used for some Southern analyses and chromosomal in situ hybridization. Molecular probes used as chromosomal markers for regional localization of the IL-1R gene were a CD8 clone that maps to 2~12 (Sukhatme et al., 1985), a COL3Al clone that maps to 2q31-+q32.3 (Emanuel et al., 1985), and a HOX4 clone that maps to 2q31+q37 (Cannizzaro et al., 1987). For filter hybridization, total plasmid DNA was radiolabeled by nick-translation with [a-32P]dNTPs to a specific activity of -1 X 10’ cpm/O.l pg, and 1 X 10’ cpm was used for each filter hybridization. For in situ hybridization, total plasmid DNA was nick-translated with [3H]dNTPs to a specific activity of -4 X lo7 cpm/pg. Cells. Isolation, propagation, and characterization of most parental cells and somatic cell hybrids used in this study have been described (Cannizzaro et al., 1987; Durst et al., 1987; Finger et al., 1988). The presence of specific human chromosomes or regions of chromosomes has been confirmed by DNA hybridization using probes for genes assigned to specific human chromosome regions. A hybrid retaining an 8q+ chromosome (8pter*8q24: : 2p12+2pter) from a

CHROMOSOMAL

45

LOCATION

Burk.itt lymphoma with a t(2; 8) (p12; q24) translocation (Erikson et al., 1983) and hybrids PB5 and CSK12, retaining partial chromosomes 2 (defined by presence or absence of chromosome 2-linked probes) (Finger et al., 1988), have allowed a regional localization of the IL-1R locus. Southern blot analysis. DNAs from human peripheral blood lymphocytes or human cell lines, mouse cell lines, and rodent-human hybrid cells were prepared by cell lysis, proteinase K digestion, phenol extraction, and ethanol precipitation. Cellular DNAs were digested with an excess of appropriate restriction enzymes, sized in 0.8% agarose gels, transferred to nylon filters, and hybridized under conditions recommended by the manufacturer. Chromosomul in situ hybridization. Metaphase chromosome preparations were obtained by culturing peripheral blood lymphocytes from a normal male subject (46,XY) for 72 h in RPMI-1640 medium supplemented with 15% fetal bovine serum. In situ hybridizat.ion for the mapping of the IL-IR probes was performed by a modified protocol as described (Emanuel et al., 1985; Finger et al., 1988). Slides were aged lo-14 days at 4°C and treated with RNase A (Sigma) for 1 h at 37°C. Chromosomal DNA was denatured at 70°C for 2 min in 70% formamide/ 2~ SSC (1X SSC is 0.15 M NaC1/0.015 sodium citrate, pH 7.0). Probe DNA was denatured in hybridization mixture (25% formamide/2x SSC/lO% dextran sulfate). The final concentration of probe DNA placed on each slide was 0.07 pg/ml. Hybridization was carried out at 37°C for 18 h. Slides were then rinsed at 39”C, dehydrated in an ethanol series, airdried, dipped in nuclear track emulsion (Kodak NTB-2), fan-dried, and stored in lightt,ight boxes at 4°C. At different time intervals, slides were developed and fixed at 15”C, air-dried, and then stained with a modified Wright-Giemsa staining protocol (Cannizzaro et al., 1987; Cannizzaro and Emanuel, 1984). Each slide was dipped for lo-16 s in a pH 9.2 borate buffer (50 mM Na,S0,/2.5 mM Na,B,O,) at 35°C and then stained for 2-3 min in a staining dish containing 3 parts pH 9.2 buffer and 1 part Wright-Giemsa stain (Baker). Grains situated on nonoverlapping chromosome regions were counted and scored. RESULTS

The Murine 11-lrl Chromosome 1

Gene Maps to Mouse

The murine chromosomal location of the type I IL1 receptor gene (Il-lrl) was determined by interspecific backcross analysis. Interspecific backcross progeny were derived from matings of (C57BL/6J X M. spretus)F1 X C57BL/6J mice (Buchberg et al., 1988).

46

COPELAND

ET

TABLE Chromosome

Locus Il-lrl DlFcrl Cal-4

Acrg RCl-2

Gene

name

Interleukin-1 receptor Anonymous mouse genomic fragment Cytotoxic Tlymphocyte-associated protein 4 Acetylcholine receptor, subunit B-cell leukemia-2

1 Loci

Probe

my

Mapped

Enzyme

AL.

1

in Interspecific C57BL/6J fragment size (kb)

a Restriction

Engrailed-1 Renin-2

fragment

Mice

M. spretus fragment size (kb)”

Reference

pG78.Int pZR-4.4~

EcoRI TayI

8.8, 7.2 5.8, 4.5

5.8 5.0

F41F4

MspI

8.5

3.5

PA-g

EcoRI

10.1, 9.4, 1.1

10.1, 9.4, 4.8

(24)

pMBCL-3-4

MspI

6.6,

4.8, 1.9, 1.6, 0.7, 0.5 2.9, 2.0, 0.8 10.4, 7.1

(37)

4.8,

1.9,

0.7,

0.5 En-l Ren-2

Backcross

w Id-2

sizes set in boldface

PstI BamHI

indicate

the restriction

This interspecific backcross mapping panel has been typed for over 500 loci distributed among all of the autosomes as well as the X chromosome. Using this mapping panel, we currently estimate that we have a nearly 100% probability of mapping any new gene. C57BL/6J and M. spretus DNAs were digested with several different restriction enzymes and analyzed by Southern blot hybridization for Il-lrl restriction fragment length polymorphisms (RFLPs). EcoRI digestion produced two fragments of 8.8 and 7.2 kb in C57BL/6J DNA and a single 58kb fragment in M. spretus DNA (Table 1). This RFLP was subsequently typed in the interspecific backcross progeny. As expected, all backcross progeny were either homozygous for the C57BL/6J allele or heterozygous for the C57BL/6J and M. spretus alleles. The results of this analysis indicated that Il-lrl maps to the proximal region of mouse chromosome 1. Table 2 summarizes the recombination frequencies observed between Il-lrl and other proximal chromosome 1 loci typed in this panel, including DlFcrl, Ctla-4, Acrg, Bcl-2, En-l, and Ren-2 (Table 1). Gene order was determined by minimizing the number of double or multiple recombination events required to explain the allele distribution [“pedigree analysis” (Avner et al., 1988)]. The position of loci with respect to the centromere was determined using Ctla-4, Acrg, Bcl-2, En-2, and Ren-2 that had previously been mapped by other laboratories to the proximal region of chromosome 1 using conventional intraspecific crosses (Davisson et al., 1989), interspecific crosses (Heidman et al., 1986; Barnum et al., 1989; Mock et al., 1989), and in situ hybridization (Brunet et al., 1987). DlFcrl is an anonymous mouse genomic frag-

2.2, 2.0, 0.8 10.4,

5.8

fragments

typed

(4’) D. M. Kingsley, unpublished observations

(8)

6.8,

in the backcross

(28) D. Dickinson and K. W. Gross, unpublished observations analysis.

ment that we fortuitously mapped to chromosome 1. Figure 1 shows the different chromosome classesobserved for 84 backcross progeny typed for all proximal chromosome 1 loci. The following are the gene order and estimated distances (+- SE) between Il-lrl and other proximal chromosome 1 loci: centromere-Il-lrl (6.2 f 2.5 cM)-DlFcrl (7.6 k 2.3 cM)-Ctla-4 (13.4 & 3.0 cM)-Acrg (3.6 f 2.0 CM)-Bcl-2 (3.5 +- 2.0 CM)En-l (4.6 +- 1.5 cM)-Ren-2 (Table 2, Figure 2). The Human IL-lR1 Gene Maps to the Long Arm of Chromosome 2, Centromeric to the COLSAI Locus The chromosomal location of the human type I IL1 receptor gene (IL-l Rl) was determined by correlating the presence of IL-1R sequenceswith the presence of specific chromosomes in a panel of somatic cell hybrid DNAs. Parental and hybrid DNAs were digested with an appropriate restriction enzyme, fractionated in agarose gels, transferred to filters, and hybridized to the IL-1R probe, either cDNA or genomic clone, to detect the presence of the cognate restriction fragments. An example of the results of this analysis is shown in Fig. 3 where lane 2 (human control) and lanes 11 and 13 are positive for presence of the 2.8-kb EcoRI fragment of the human IL-lR1 gene. The hybrids represented in lanes 11 and 13 retain a partial and an entire human chromosome 2, respectively (see legend to Fig. 3), while hybrids in lanes 3-10 and lane 12 do not retain human chromosome 2. The results of the hybrid analysis are summarized in Fig. 4, which demonstrates that the presence of the IL-l Rl gene in the hybrids correlates with the presence of a portion of human chromosome 2. Hybrid 514-2 retains the

INTERLEIJKJN-I

RECEPTOR

CHROMOSOMAL

TABLE Recombination

Frequencies of IZ-lrl in C57BL/6J-M.

Ctln-4

2r

Locus II-Irl

h/97 (6.2

(125

--

IIlFcrl

with Other Proximal spretus Backcross

(35.0 -+ 4.4) 42/180 (25.6 -+ 3.3) 18/134 (13.4 i 3.0)

10/1X2 Ik 2.3)

Ctln-4

Chromosome Mice

Hcl-2

24196

t 3.41

(7.6

2

AUg

12/95

_+ 2.5)

47

LOCATION

1 Markers

En-l

RCV-2

31/98

:19/98 (39.8 + 4.91

27186 f 5.0) 21/86

(31.6 i 4.71 56/182

i 4.6) 14/84

(30.8 k 3.4) %6/136

IL 4.1) 3/84 (3.6 It 2.0)

(19.1 -c 3.4) U/194

(31.4 (24.4 (16.7

(6.2

i 1.7) 3186 (3.5 i 2.0)

66/185 (35.7 2 3.5) 34/136 (25.0 2 3.7)

z/194 (10.8 +- 2.2) S/86 c9.a + 3.1)

9/196 (4.6 + 1.51 ” Number

uf recombinant

animals/total

number

of animals

analyzed

region 2p12-+2pter and is negative for IL-lR1; hybrid CSK-12 retains the short arm of chromosome 2 and is negative for the IL-lR1 gene; hybrid PB5 retains most of chromosome 2 but is missing the distal part of the long arm (i.e., is negative for COL3Al and HOX4 probes, data not shown) and retains the IL-tRf gene (see Fig. 4). Thus somatic cell hybrid analysis places the IL-lR1 gene between 2cen and 2q32.

Chromosomal in Situ Hybridization Confirms Localization of the IL-lR1 Gene to 2q Chromosomal in situ hybridization was performed using the 3H-labeled genomic probe for IL-lR1. After hybridization, a total of 558 chromosomally localized grains were counted over 200 metaphases; 32% (177/ 558) were localized over the long arm of chromosome 2, with 53% (93/177) of the 2q grains localized over region 2qI2 (see Fig. 5). Significant clustering of grains above background was not observed for other chromosome regions. Thus, in situ chromosomal hy-

E,lbl

~Zm~~BlImrIImO~Ell’m

/irn-2 n T-1n 25 1.5

[I_I W II7 W Ll 3 3 ? 4 In I

W 17 12

n I?

0

W ~ I 23

FIG. 1. Summary of the results of interspecific hackcross mapping. The seven genes mapped in these studies are shown at the left. Each column represents a particular type of chromosome transmitted from the fC57BL/6J X M. spretus)F, parents to the hackcross offspring. The black boxes represent the presence of a C57BL/6J allele and the white boxes represent the presence of a M. spretus allele. The number of offspring inheriting each type of chromosome is listed at the bottom. Only the 84 chromosomes that were typed for all seven markers are shown.

and (recombination

frequencies

? SE) are listed

for each pair of loci.

bridization confirms the localization of the human IL-lR1 gene to the long arm of chromosome 2 and defines a narrower localization at 2q12. DISCUSSION

The murine and human type I IL-l receptor genes have been mapped to chromosomes 1 and 2, respectively, in an area of conservation between the two species. It is intriguing that the genes for IL-la and IL-l/3 also map to human chromosome 2 (Webb et al., 1986; Modi et al., 1988; Lafage et al., 1989), in a position near that proposed here for the IL-lR1 gene. In the mouse, however, the linkage between the IL-1 and IL-l R genes has not been conserved during evolution, since the mouse IL-l genes are located on mouse chromosome 2 (D’Eustachio et al., 1987; Boultwood et al., 1989). Only in a limited number of caseshave the genes for both a hormone and its corresponding receptor been mapped. Among this small sample, the genes for CSF-1 (M-CSF) and its receptor (CSF-1R; the c-fms protein) are both located on the long arm of human chromosome 5 (Pettenati et al., 1987; Roberts et al., 1988). In all other cases, however, t,he hormone and receptor genes map to separate chromosomes, as do the murine Csfm and Csfmr genes, which are located on chromosomes 3 (Buchberg et al., 1989) and 18 (Hoggan et al., 1988), respectively. The murine Il-lrl gene was mapped to the centromerit region of mouse chromosome 1 based upon its segregation in an interspecific backcross mapping panel with other chromosome 1 markers, including DlFcrl, Ctla-4, Acrg, Bcl-2, En-l, and Ren-2 (Fig. 2). An important consideration when using interspecific backcrosses in mapping is whether recombination distances obtained from these crosses are comparable to those reported using conventional intraspecific

48

COPELAND

ET

AL. 12

3

4

5 6

7

2.8 kbp-

6?

FIG. 2. Linkage maps of mouse chromosome 1. The chromosome on the left shows the location of the seven genes mapped in the current studies, with distances between adjacent genes shown in centimorgans. The chromosome on the right shows the May 1990 version of the composite chromosome 1 linkage map compiled by T. H. Roderick, M. T. Davisson, A. I,. Hillyard, and D. P. Doolittle (personal communication). With the exception of Ken-l”. only genes that have been mapped in humans are included. This map is based largely on genetic crosses between conventional M. muscuLus/M. domvsticus laboratory mouse strains. The map positions for A4~1f, Vi!, An-d, and Acrg have only been determined in two point crosses. Their positions on the composite map have been inferred from their placement on the interspecific map. Boxed loci represent genes placed on both maps. Mouse genes that have not been mapped in humans are underlined. Locations of these genes on human chromosomes are shown between the two maps; arrowheads point to the locus that has been mapped in humans. The two locus. mouse linkage maps were aligned at the Rcn-l’

crosses. Three of the loci mapped in this study-&l2, En-l, and Ren-2-had previously been mapped to chromosome 1 using intraspecific crosses. The gene order and map distances obtained in these crosses are in good agreement with those obtained in the interspecific backcross (Fig. 2). Acrg was previously mapped to chromosome 1 by interspecific backcross analysis (Heidman et al., 1986); however, its proximal or distal relationship to two other chromosome 1 markers used in the mapping study, Idh-1 (isocitrate dehydrogenase 1) and Mylf (myosin light chain, alkali, fast skeletal muscle), could not be determined. Our interspecific backcross mapping data are most consistent with the following gene order: centromere-Idh-1 -Mylf-Acrg-telomere, since placement of Mylf proximal to Idh-1 would result in Acrg being proximal, not distal, to Ctla-4 (Fig. 2). Ok-4 has also been previously positioned on chromosome 1 by interspecific backcross analysis. Again, our map position for Ctla-4 does not differ with the

8

9 101l1213 c*“iLe) -d;;“~

gene segregates with human chromosome 2 FIG. 3. The IL-IR in rodent-human hybrids. DNA C-10 &lane) from mouse (lane 1); human (lane 2); hybrid cl21 retaining human chromosome 7 (lane 3); hybrid GB31 retaining 17 (lane 4); hybrid M442S9 retaining partial 4,8, 12. 13, and 14 (lane 5): hybrid EF3 retaining partial 8. 19. 20, 21, partial 22, and X (lane 6); hybrid Ph1124 retaining chromosomes 1 and X (lane 7); hybrid S5 retaining 3,5, partial 6,7. 9. 11, 13.15, 17, 18, partial 22, and X (lane 8); hybrid (:I,5 retaining 4. 6--8, 14, 15, 17-20, and X (lane 9): hybrid 77-31 retaining 1, 3, partial 4. 5-9, partial IO, 13, 14. 17. Z&22, and X (lane 10); hybrid PBS retaining partial 1. Bpter to approximately Zq:12,5q, 8. llq, 17, and 20 (lane 11); hybrid .?a retaining 4q. 6p, 12, 14, 17 ‘11. and 22 (lane 12); and hybrid BD3 retaining I-8. 9-16. 18-22, iid X (lane 13) was digested with restriction enzyme Ecr~R1, fractionated, transferred to nylon membrane, and hybridized to the radiolabeled human II,- I R cDNA probe. Approximate molrclllar weights of mouse and human bands detected are shown on the left. The difference in intensity of the hybridizing band in the positive hybrids (lanes 11 and 13) is due to variability in t.he percentage of cells in the hybrid clone that retain human chromosome 2.

previous placement of Ctla-4 at the 95% confidence interval. Many of the genes that map in the proximal region of mouse chromosome 1 map to the long arm of hu-

Human

Chromosomes

Hybrid Cl21 GE31 Cl77 AA3 N9 GL3 c/2 77~37 77-30 S5 3c9 C65 Cl1

803 CSK-12 PE JW2 Jl6-5

FIG. 4. Presence of the IL-IR gene in a panel of 18 rodeni-human somatic cell hybrids. [B] Hybrid named in the left column contains the chromosome indicated in the upper row; [O], presence of the long arm (or a part of the long arm, indicated by a smaller fraction of stippling) of the chromosome shown above the column; [Cl], presence of the short arm (or partial short arm) of the chromosome listed above the column; (01, absence of the chromosome listed above the column. The column for chromosome 2 is boldly outlined and stippled to highlight correlation of the presence of this chromosome (or region of this chromosome) with the presence of the IL-1R probes. The pattern of retention of the ILIR gene in the panel is shown in the column at the right, where the presence of the probe in the hybrid is indicated by a stippled box with a plus sign and the absence of the probe is indicated by an open box enclosing a minus sign.

INTERLEUKIN-1

RECEPTOR

120

CHROMOSOMAL

49

LOCATION

and Eastman Kodak. D.M.K. is a Lucille P. Markey Scholar in Biomedical Science. The NCIIFrederick Cancer Research and Development Center is fully accredited by the American Association for Accreditation of Laboratory Animal Care.

100 80 60 F 40 t

REFERENCES

FIG. 5. Chromosomal in situ hybridization localizes the human IL-lR1 locus to 2~12. The diagram shows the distribution of 558 autoradiographic grains on 200 metaphases after probing with the genomic IL-1R probe. The abscissa represents the chromosome banding pattern of each human chromosome in relative size proportion; the ordinate shows the number of silver grains. Thirtytwo percent of grains were localized over chromosome Zq, with 53% of the 2y grains localized to 2ql2.

man chromosome 2 (Fig. 2) (Nadeau, 1989). Ourplacement of Il-lrl on mouse chromosome 1 considerably lengthens this region of homology, which now extends from En-l to II-lrl and covers approximately 34 CM (Fig. 2). This region of homology is, however, interrupted at one end in the mouse by Bcl-2, which has been mapped to human chromosome 18 band q21 (Tsujimoto et al., 19851, and at the other end in the human by the IL-1 genes, which lie on murine chromosome 2 (D’Eustachio et al., 1987; Boultwood et al., 1989). The IL-l receptor gene mapped in this paper encodes the receptor expressed in T cells and fibroblasts (the type I receptor) (Sims et al., 1988, 1989; Chizzonite et al., 1989). There is strong evidence to suggest the existence of another form of IL-l receptor (the type II receptor) (Bomszytk et al., 1989; Chizzonite et al., 1989). This second form of the receptor has been best characterized in B cells (Bomsztyk et al., 1989; Chizzonite et al., 1989; Matsushima et al., 1986; Horuk and McCubrey, 1989). It will be of great interest to determine whether the type II receptor gene is closely linked to the type I receptor gene and the IL-l genes in mouse and/or human.

1.

AVNER, (1988). crosses.

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BIRCH, I).. AND Cox, of murine interleukin-la

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J.-F., DENIZOT, F., LUCIANI, M. F., ROUX-DOSSETO, M., SUZAN, M., MATTEI, M.-G., AND GOLSTEIN, P. (1987). new member of the immunoglobulin superfamily-CTLA-4. Nature (London) 328: 267-270.

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BUCHBERG, A. M., BEDIGIAN, H. G., TAYI,OR, B. A., BROWNELL, E., IHLE, J. N., NAGATA, S., JENKINS, N. A., AND COPELAND, N. G. (19881. Localization of Eui-P to chromosome 11: Linkage to other proto-oncogene and growth factor loci using interspecific backcross mice. Oncogene Rex 2: 1499165.

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BUCHBERG, A. M., JENKINS, N. A., AND COPELAND, N. G. (1989). Localization of the murine macrophage colony-stimulating factor gene to chromosome 3 using interspecific hackcross analysis. Genomics 5: 363-367.

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CANNIZZARO, I,. A., AND EMANUEL, B. S. t 19841. An improved method for G-banding chromosomes after in situ hybridization. (Ivtogenet. Cell Genet.38: 308-309. CANNIZZARO, L. A., CROCE, C. M., GRIFFIN, C. A., SIMEONE, A., BONCINELLI, E., AND HUEBNER, K. (1987). Human homeobox-containing genes located at chromosome regions 2q31+2y37 and 12q12+12y13. Amer. J. Hum &net. 41: I-15.

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CHIN, J., CAMERON, P. M., RUPP, E.. AND SCHMIDT, J. A. (19871. Identification of a high-affinity receptor for native human interleukin 16 and interleukin la on normal human lung fibroblasts. J. Exp. Med. 165: 70-86.

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ACKNOWLEDGMENTS We thank Beverly Mock. Pierre Golstein, John Merlie, Alexandra .Joyner, Gail Martin, Douglass Dickinson, and Kenneth Gross for providing probes and Mary Beth Cybulski, Brian Cho, Teresa Druck, and Cathy McMahan for expert technical assistance. This research was supported by the National Cancer Institute (NCI), DHHS, under Contract NOl-CO-74101 with ABL; by NIH Grant CA-25875; and by a joint venture between Immunex Corporation

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