ENHANCED EFFICACY OF LIPOSOMAL ANTISENSE DELIVERY

Abstract: Pharmaceutical compositions for the treatment of pathologic conditions associated with the overexpression of ICAM-1 in a host. These pharmaceutical compositions comprise an effective amount of an ICAM-1 antisense molecule encapsulated in a lipid mixture which is typically a liposome or lipid particle. The lipid mixture will typically comprise at least two members selected from the group consisting of phospholipids, sterols and cationic lipids.

Description: ENHANCED EFFICACY OF LIPOSOMAL ANTISENSE DELIVERY BACKGROUND OF THE INVENTION Research aimed at developing therapeutic drugs based on the inhibition of protein expression by oligonucleotides has encountered a number of limitations. It has been demonstrated in vitro that a number of cell types exhibit low rates of intracellular delivery of antisense molecules. This is of great concern as antisense molecules are designed to bind to mRNA target sites located in the cytoplasm and nucleus of cells. In vivo studies have also indicated that many oligonucleotides are rapidly cleared from circulation; thus a limited time period exits in which the antisense molecule can interact with the target cells. Furthermore, a number of studies have demonstrated that oligonucleotides are quite toxic when administered in vivo. Many of the limitations of free antisense, such as toxicity and nuclease digestion, can potentially be overcome by encapsulating antisense molecules in liposomes. When made as relatively small particles (approximately 100 nm in diameter), liposomes will passively accumulate at sites of inflammation by moving through the restructured vasculature. Numerous studies have determined that liposomes or lipid complexes have the ability to deliver oligonucleotides intracellularly through two mechanisms: cellular uptake of liposomes via endocytosis, and fusion of cationic liposomes with target cell membranes. Comparisons of free and encapsulated oligonucleotides, indicate an enhanced stability for encapsulated oligos in vitro as the liposome prevents nuclease degradation. In contrast, comparison of free and encapsulated phosphorothioate oligonucleotides usually indicate no enhancement as the phosphorothioate oligos are themselves nuclease resistant. Thus the same amount of oligonucleotide can potentially be delivered to the cell whether it is a free phosphorothioate or an encapsulated phosphodiester oligonucleotide. Wang, et al., Proc. Natl. Acad. Sci. 92:3318-3322 (1995). Despite such studies, no evidence has been presented which demonstrates that a conventional neutral liposome (e.g. , phosphatidylcholine/cholesterol) can disrupt the endosome to release its contents into the cytoplasm. SUBSTTTUTE SHEET (RULE 26) One approach to this problem has involved the use of cationic lipid vesicles. These cationic lipid vesicles form "complexes" with DNA, including plasmids and oligonucleotides. These complexes are not liposomes (i.e. an intact bilayer encapsulating an aqueous space) but are aggregates of lipid and DNA held together by electrostatic attraction between the cationic lipid and anionic nucleic acid. Recent literature indicates that complexes of cationic lipids in association with a fusogenic factor such as phosphatidylethanolamine or a fusion protein is required to achieve a significant antisense effect or gene transfection. Farhood, et al., Biochim. Biophys. Acta. 1235:289-295 (1995); Feigner, et al., J. Biol. Chem. 269:2550-2561 (1994); and Wrobel, et al. , Biochim. Biophys. Acta. 1235:296-304 (1995). Still other studies involving intravenous administration of antisense oligonucleotides in monkeys have indicated certain toxicities associated with this practice (see Galbraith, et al., Antisense Res. Dev. 4:201-206 (1994) and Cornish, et al., Antisense Res. Dev. 3:239-247 (1993)). The most likely cause for the observed toxicities is the activation of the complement system, releasing C3a and C5a, vasoactive cleavage products of C3 and C5, respectively. In this regard, both DNA and RNA have been shown to activate complement in various in vitro assays. The complement system is a multi-protein cascade which serves as one of the first lines of defense against foreign particles which have entered the blood. The two basic mechanisms by which complement attacks foreign particles are by opsonization and cell lysis. Opsonization involves the covalent attachment of complement fragments, principally C3b and iC3b, to the surface of a particle or cell, which is then recognized by corresponding receptors present on macrophages. Cell lysis involves the assembly of a multiprotein complex, C5b-9, which perforates cell membranes and generates a pore. Relatively little work has been published concerning the use of neutral phospholipid vesicles (liposomes) as delivery systems for antisense oligonucleotides (see, Juliano, et al., Antisense Research & Development 2:165-176 (1992)). Oligonucleotides are usually 15-25 bases in length and highly charged which means they cannot cross membranes by passive diffusion. The target site for antisense drugs is either the cytoplasm or nucleus and consequently the plasma membrane acts as an effective barrier. Despite this, some free antisense oligonucleotides have been shown active in vitro and in vivo (see, Juliano, et al., Antisense Research & Development 2:165-176 (1992); Bennett, et al., J. Immunol. 152:3530-3540 (1994); Bennett, et al. , Adv. Pharmacol. 28:1-43 SUBSTTTUTE SHEET (RULE 26) (1994); and Stepkowski. et al , J. Immunol. 153:5336-5346 (1994)), furthermore there is some evidence that a receptor mediated uptake mechanism for oligonucleotides may be responsible. Even if a target cell or tissue does exhibit an ability to endocytose antisense molecules the process is not efficient and biological efficacy is normally only observed at very high concentrations. This is most likely because even after endocytosis the nucleotide must still cross the endosomal membrane to reach the cytoplasm before it is degraded by lysosomal enzymes which occurs at a very low frequency. In the case of anti ICAM-1 the free oligonucleotide does not appear to penetrate endothelial cells in culture but when complexed with cationic lipid/phosphatidylethanolamine (PE) complexes extensive uptake is observed. Such cationic lipid/PE complexes are known to disrupt intracellular endosomes and deliver nucleic acids into the cytoplasm. Once in the cytoplasm antisense oligonucleotides rapidly diffuse into the nucleus (Sixou, et al. , Nuc. Acids Res. 22:662-668 (1994)) and this is also observed for anti ICAM-1 delivered by aggregates or complexes of the oligonucleotides and lipids (see Bennett, et al, Mol. Pharmacol. 41:1023-1033 (1992)). As noted above, these latter complexes or aggregates are not liposome formulations and do not use encapsulation methods for construction of the complex. Moreover, the complexes (which are not encapsulated oligonucleotide systems) have not been shown to disrupt endosomes and as a result, will only show intracellular delivery of the oligonucleotide in in vitro studies. Extensive modifications to the structure of oligonucleotides have been made in an attempt to improve stability in vivo and to enhance membrane permeability. For example the phosphorothioates, noted above, in which an oxygen atom is replaced by a sulphur in the phosphate backbone, exhibit increased resistance to nucleases and are more stable in vivo than normal phosphodiester oligonucleotides (Juliano, et al. , Antisense Research & Development 2:165-176 (1992)). Another example are nucleic acid methylphosphonates, which are not only nuclease resistant but also hydrophobic analogues of phosphodiesters and therefore expected to be more membrane permeable (see, Hughes, et al. , J. Pharm. Sci. 83:597-600 (1994) and Tari, et al., Blood 84:601- 607 (1994)). Despite these developments, antisense oligonucleotide therapy is slow to develop mainly because it is difficult to get therapeutic drug levels into cells in target tissue and toxic side effects persist such as complement activation which restrict dosing. Attractive targets for antisense therapy include the nucleic acids which encode intercellular adhesion molecule- 1 (ICAM-1), vascular cell adhesion molecule- 1 SUBSTΓΓUTE SHEET (RULE 26) (VCAM-1), and endothelial leukocyte adhesion molecule-1 (ELAM-1). ICAM-1 , which is a 90- 110 kDa membrane glycoprotein involved in the trafficking of leukocytes out of the vasculature and in antigen presentation to T cells (see Osborn, Cell 56:907-910 (1990) and Springer. Nature (Lond.) 346:425-443 (1990)). ICAM-1 is normally expressed at low levels on the surface of endothelial cells, keratinocytes, fibroblasts and leukocytes. Expression of ICAM-1 is inducible by a number of cytokines, including IL- lj3, tumor necrosis factor-α and interferon-γ. Increased expression of ICAM-1 has been demonstrated in a variety of human diseases and has been shown to correlate with leukocyte infiltration in the diseased tissue. What is needed in the an are new compositions and methods for the delivery of antisense molecules directed toward inhibiting the expression of cellular adhesion molecules. Such compositions should increase the serum stability of the antisense molecules and reduce toxic side effects such as complement activation. Surprisingly, the present invention provides such compositions and methods. SUMMARY OF THE INVENTION The present invention provides pharmaceutical compositions for the treatment of pathologic conditions associated with the overexpression of cellular adhesion molecules, such as ICAM-1 in a host. These pharmaceutical composition comprise an effective amount of an ICAM-1 antisense molecule encapsulated in a lipid mixture which is typically a liposome or lipid particle. The lipid mixture will typically comprise at least two members selected from the group consisting of phosphoiipids, sterols and cationic lipids. In one group of embodiments, the antisense molecule is either a phosphorothioate molecule or a methyl phosphonate molecule, from about 15 to 50 nucleic acids, and is complementary to a portion of the 3 '-untranslated region of ICAM-1. For those embodiments in which the lipid mixture is a liposome composition, the liposome will preferably comprise phosphatidylcholine and cholesterol, more preferably egg phosphatidylcholine and cholesterol. For those embodiments in which the lipid mixture is present as lipid particles, the particles will preferably comprise phosphoiipids and cationic lipids. Additionally, the present invention provides methods for the treatment of pathologic conditions associated with the overexpression of ICAM-1 in a host. Such conditions include Alzheimer's disease, multiple sclerosis, uveitis, Herpes keratitis, renal allograft rejection, glomerulonephritis, liver allograft rejection, viral hepatitis, alcoholic hepatitis, cholangitis, cardiac allograft rejection, atherosclerotic plaques, rheumatoid arthritis, Grave's disease, Hashimoto's thyroiditis, psoriasis, scleroderma, graft v host disease, contact dermatitis, lichen planus, fixed drug eruption, mycosis fungoides, and alopecia areata. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 illustrates a spin column elution profile of encapsulated antisense. 50 μL of encapsulated antisense was applied to a 1 mL Biogel A15m, 200-400 mesh, spin column and separated. Lipid was determined by phosphate analysis (O), and oligonucleotide was detected by measuring A 260 , after Bligh and Dyer extraction (•). Figure 2 illustrates a purified liposomal antisense preparation. Liposome- encapsulated antisense was "purified" on DEAE-sepharose CL-6B columns. Removal of free antisense was assessed by size exclusion chromatography on 1 mL Biogel A15m column. Lipid was determined by phosphate analysis (O), and oligonucleotide was detected by measuring A 260 , after Bligh and Dyer extraction (•). Figure 3 shows a time course for leakage of encapsulated antisense. Leakage of encapsulated antisense was monitored at room temperature for 1 (O), 3 (■) and 5 (•) days. Figure 4 shows the time course for leakage of encapsulated antisense at 4°C. Leakage of encapsulated antisense was monitored for 1 (O), 3 (D), and 5 (•) days, at 4 q C. Figure 5 illustrates complement activation by liposomes. Complement activation was investigated using an EPC/CH liposome preparation. Liposomes were 100 ± 30 nm. Lipid composition is expressed in molar ratios. Figure 6 illustrates the ear swelling characteristics of ICR mice. Measurements refer to increases in ear thickness with respect to baseline measurements (prior to ear challenge). Values are given for two separate experiments. Error bars represent the standard deviation of measurements from 4 mice. Figure 7 illustrates the ear swelling characteristics of BALB/c mice. Measurements refer to increases in ear thickness with respect to baseline measurements (prior to ear challenge). Error bars represent the standard deviation of measurements from 4 mice. Figure 8 shows the liposome accumulation in the ears of ICR mice during various time intervals of inflammation. Liposomes were injected into mice at 0 hr, 24 hr, and 48 hr after initiation of ear inflammation. Liposomes were allowed to circulate for 24 hr at which time ears were recovered, digested, and analyzed for radiolabeled lipid. Error bars represent the standard deviation of measurements from 4 mice. Figure 9 shows the liposome accumulation in the ears of BALB/c mice during various time intervals of inflammation. Liposomes were injected into mice at 0 hr, 24 hr, and 48 hr after initiation of ear inflammation. Liposomes were allowed to circulate for 24 hr at which time ears were recovered, digested, and analyzed for radiolabeled lipid. Error bars represent the standard deviation of measurements from 4 mice. Figure 10 shows the MPO levels in the ears of ICR mice during inflammation. At various times after the initiation of inflammation, inflamed ears (■) and control ears (•) were recovered, homogenized, and assayed for MPO activity. Error bars represent the standard deviation of measurements from 4 mice. Figure 11 shows cell infiltration into the inflamed ear of ICR mice during inflammation. Bone marrow cells and circulating leukocytes were labeled 24 hr prior to the onset of inflammation. At various times after the initiation of inflammation, ears were recovered, digested, and analyzed for radiolabeled cells by liquid scintillation counting. Error bars represent the standard deviation of measurements from 4 mice. Figure 12 shows a typical inflammation experiment involving ICR mice. The following parameters were measured: ear swelling (Δ); liposome accumulation in inflamed (•) and non- inflamed (■) ears; and cell infiltration (O). Figure 13 shows liposome accumulation in the ears of ICR mice during the first 24 hours of inflammation. DSPC:CH liposomes were injected into mice SUBSTrrUTE SHEET (RULE 26) immediately after initiation of ear inflammation. At various times mice were sacrificed and the ears were collected and analyzed (inflamed (•) and non-inflamed (■) ears). Error bars represent the standard deviation of measurements from 4 mice. Figure 14 illustrates the circulation clearance rates of free [ 3 H]-antisense and liposome encapsulated pHJ-antisense. Figure 15 illustrates the tissue biodistribution of free [ 3 H] -antisense (Isis 2302). Figure 16 illustrates the tissue biodistribution profiles for both the lipid and antisense portions of a lipsome encapsulated antisense formulation. Figure 17 illustrates the ability of free antisense to inhibit ear inflammation. Figure 18 illustrates the efficacy of free and encapsulated ICAM-1 antisense formulations in reducing ear inflammation in mice. Figure 19 is a bar graph showing edema formation (based on ear weights) in mice treated with free and encapsulated antisense. DETAILED DESCRflPTION OF THE INVENTION Abbreviations and Definitions Abbreviations used herein have the following meanings: i.v. , intravenous; DC-Choi, 33-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol (see, Gao, et al., Biochem. Biophys. Res. Comm. 179:280-285 (1991)); DDAB, N,N-distearyl-N,N-dimethylammonium bromide; DMRIE, N-(l,2-dimyristyloxyprop-3- yI)-N,N-dimethyl-N-hydroxyethyl ammonium bromide; DODAC, N,N-dioleyl-N,N-dimethylammonium chloride (see commonly owned patent application USSN 08/316,399, incorporated herein by reference); DOGS, dioctadecylamidoglycyl carboxy spermine; DOPE, 1,2-j/t-dioleoyiphoshatidyethanolamine; DOSPA, N-(l-(2,3- dioleyloxy)propyl)-N-(2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate; DOTAP, N-((2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride; DOTMA, N-((2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride; EPC, egg phosphatidylcholine; RT, room temperamre; HEPES, 4-(2-hydroxyethyl)-l- piperazineethanesulfonic acid; HBS, HEPES buffered saline (150 mM NaCl and 20 mM SUBSTTTUTE SHEET (RULE 25) HEPES); PEG-Cer-C 20 , l-O-(2'-(ω-methoxypolyethyleneglycol)succinoyl)-2-N- arachidoyl-sphingosine; PEG-Cer-C 14 , l-O-(2'-(ω-methoxypolyethyleneglycol)succinoyl)- 2-N-myristoyl-sphingosine; PBS, phosphate-buffered saline; EGTA, ethylenebis(oxyethylenenitrilo)-tetraacetic acid; OGP, n-octyl β-D-glycopyranoside (Sigma Chemical Co. , St. Louis, MO); POPC, palmitoyl oleoyl phosphatidylcholine (Northern Lipids, Vancouver, BC); QELS, quasielastic light scattering; TBE, 89 mM Tris-borate with 2 mM EDTA; and EDTA, Ethylenediaminetetraacetic acid (Fisher Scientific, Fair Lawn, NJ); RES, reticuloendothelial system; SPDP, N-succinimidyl 3-(2-pyridyldithio) propionic acid; DTT, dithiothreitol; NEM, N-ethylmaleimide; Choi, cholesterol; DMPC, 1 ,2-s/ι-dimyristoylphosphatidylcholine; DSPC, l,2-5A7-distearoylphosphatidylcholine; EDTA, ethylenediaminetetraacetic acid; MLV, multilammelar vesicles; PEG-DSPE, poly(ethylene glycol)-modified distearoylphosphatidyle anolamine; LUV, large unilamellar vesicles; SUV, small unilamellar vesicles; PEG, poly(ethylene glycol); 3 H-CHE, 3 H-Cholesteryl hexadecyl ether. The term "oligonucleotide" refers to a single or double-stranded polymer of deoxyribonucleotide or ribonucleotide bases read from the 5' to the 3' end. It includes both self-replicating plasmids, infectious polymers of DNA or RNA and non¬ functional DNA or RNA. When used to refer to oligonucleotides, the terms "phosphorothioate" and "methyl phosphonate" refer to those oligonucleotides in which a phosphodiester intemucleotide linkage has been modified by replacing at least one of the non-bridged oxygens of the intemucleotide linkage with sulfur or a methyl group, respectively. Preferably at least 10% of the intemucleotide linkages are modified, more preferably at least 30% of the linkages are so modified. Most preferably, at least 50% of the linkages are modified. The term "complementary" means that one nucleic acid hybridizes selectively to another nucleic acid. Selectivity of hybridization exists when hybridization (or base pairing) occurs that is more selective than total lack of specificity. Typically, selective hybridization will occur when there is at least about 55 % paired bases over a stretch of at least 14-25 nucleotides, preferably at least about 65%, more preferably at least about 75 % , and most preferably at least about 90% . See, M. Kanehisa Nucleic Acids Res. 12:203 (1984), incorporated herein by reference. SUBSTTTUTE SHEET (RULE 26) The term "lipid" refers to any fatty acid derivative which is capable of forming a bilayer such that a hydrophobic portion of the lipid material orients toward the bilayer while a hydrophilic portion orients toward the aqueous phase. Hydrophilic characteristics derive from the presence of phosphato, carboxylic, sulfato. amino, sulfhydryl, nitro, and other like groups. Hydrophobicity could be conferred by the inclusion of groups that include, but are not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups and such groups substituted by one or more aromatic, cycloaliphatic or heterocyclic group(s). Preferred lipids are phosphoglycerides and sphingolipids, representative examples of which include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoyl phosphatidylcholine, lysophosphatidylcholine, lysophosphatidyl- ethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoyl- phosphatidylcholine or dilinoleoylphosphatidylcholine could be used. Other compounds lacking in phosphorus, such as sphingolipid and glycosphingolipid families are also within the group designated as lipid. Additionally, the amphipathic lipids described above may be mixed with other lipids including triglycerides and sterols. The term "cationic lipid" refers to any of a number of lipid species which carry a net positive charge at physiological pH. Such lipids include, but are not limited to, DODAC, DOTMA, DDAB, DOTAP, DC-Choi and DMRIE. Additionally, a number of commercial preparations of cationic lipids are available which can be used in the present invention. These include, for example, LIPOFECTIN ® (commercially available cationic liposomes comprising DOTMA and DOPE, from GIBCO/BRL, Grand Island, New York, USA); LIPOFECTAMINE ® (commercially available cationic liposomes comprising DOSPA and DOPE, from GIBCO/BRL); and TRANSFECTAM ® (commercially available cationic liposomes comprising DOGS from Promega Corp. , Madison, Wisconsin, USA). As used herein, the phrase "pathologic conditions associated with the overexpression of ICAM-1 ", is meant to include diseases of the central nervous system (e.g. Alzheimer's disease and multiple sclerosis), the eye (e.g. uveitis and Herpes keratitis), the kidney (e.g. renal allograft rejection and glomerulonephritis), the liver (e.g. liver allograft rejection, viral hepatitis, alcoholic hepatitis, cholangitis), the heart (e.g. cardiac allograft rejection and atherosclerotic plaques), the bone (e.g. rheumatoid arthritis), the thyroid (e.g. Grave's disease and Hashimoto's thyroiditis), and the skin SUBSTTTUTE SHEET (RULE 26) (e.g. psoriasis, scleroderma, graft v host disease, contact dermatitis, lichen planus. fixed drug eruption, mycosis fungoides, and alopecia areata). As used herein, the term "host" refers to a human, rat, mouse, dog, cow, sheep, horse, cat and goat. Description of the Embodiments The present invention derives from the surprising discovery that antisense molecules which are encapsulated in a liposome or lipid particle composition can be delivered to a site of inflammation in response to overexpression of ICAM-1 and thereby reduce the associated inflammation. It was particularly surprising that liposome formulations which consist essentially of charge neutral lipids and a sterol (e.g., cholesterol) would be effective for antisense delivery in view of the conventional wisdom that cationic liposome formulations or formulations having fusogenic lipids or proteins are necessary for cell or endosome fusion. Accordingly, the present invention provides pharmaceutical compositions for the treatment of conditions associated with the overexpression of cellular adhesion molecules, preferably ICAM-1. These compositions comprise an antisense oligonucleotide encapsulated in a lipid mixture. The lipid mixture can be in either of two forms. The first is a conventional liposome, which is preferably charge neutral, consists essentially of neutral phosphoiipids and a sterol (e.g., cholesterol) and which can be passively loaded with an antisense molecule. The second form is a lipid particle which comprises phosphoiipids, cationic lipids, sterols and combinations thereof. Recently, others have described a cationic liposomal preparations of N- ((2, 3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA) for the delivery of an ICAM-1 antisense molecule (see, Bennett, et al., Mol. Pharmacol. 41: 1023-1033 (1992). The preparation used by Bennett, et al., was a 1 : 1 mixture of DOTMA and dioleoyl phosphatidylethanolamine which formed 250 nm liposomes. These systems provided complexes of the antisense molecules which were not encapsulated. Moreover, these systems are useful only in vitro for intracellular delivery as they do not disrupt endosomes in vivo. SUBSTTTUTE SHEET (RULE 26) ANTISENSE OLIGONUCLEOTIDES The antisense oligonucleotides which are useful in the present invention are those oligonucleotides which are complementary to a portion of a mammalian nucleic acid encoding cellular adhesion molecules such as ELAM-1 (human), VCAM-1 (human) and ICAM-1 (human and mouse) which are provided as Sequence I.Ds. No. 1 , 3, 5 and 7, respectively). ELAM-1 is a 115-kDa membrane glycoprotein which is a member of the selecting family of membrane glycoproteins (see, Bevilacqua, et al., Science, 243: 1160- 1165 (1989)). The amino terminal region of ELAM-1 contains sequences with homologies to members of lectin-like proteins, followed by a domain similar to epidermal growth factor, followed by six tandem 60-amino acid repeats similar to those found in complement receptors 1 and 2. ELAM-1 is encoded by a 3.9-kb mRNA. The 3 '-untranslated region of ELAM-1 mRNA contains sever sequence motifs ATTTA which are responsible for the rapid turnover of cellular mRNA consistent with the transient nature of ELAM-1 expression. ELAM-1 exhibits a limited cellular distribution and has only been identified on vascular endothelial cells. Like ICAM-1, ELAM-1 is inducible by a number of cytokines including tumor necrosis factor, interleukin- 1 and lymphotoxin and bacterial lipopolysaccharide. Unlike ICAM-1, ELAM-1 is not induced by gamma- interferon. The kinetics of ELAM-1 mRNA induction and disappearance in human umbilical endothelial cells precedes the appearance and disappearance of ELAM-1 on the cell surface. VCAM-1 is a 110-kDa membrane glycoprotein encoded by a 3.2-kb mRNA. It appears to be encoded by a single-copy gene which can undergo alternative splicing to yield products with either six or seven immunoglobulin domains (see Osborn, et al., Cell 59:1203-1211 (1989)). The receptor for VCAM-1 is proposed to be CD29 (VLA-4) as demonstrated by monoclonal antibodies which bind to CD29 and block the adherance of Ramos cells to VCAM-1. VCAM-1 is expressed primarily on vascular endothelial cells and is also regulated by treatment with cytokines (see, Rice, et al. , Science 246:1303-1306 (1989) and Rice, et al., J. Exp. Med. 171:1369-1374 (1990)). Increased expression appears to be due to induction of the mRNA. Human ICAM-1 is encoded by a 3.3-kb mRNA resulting in the synthesis of a 55,219 dalton protein. ICAM-1 is heavily glycosylated through N-linked SUBSTTTUTE SHEET (RULE 25) glycosylation sites. The mature protein has an apparent molecular mass of 90 kDa as determined by gel electrophoresis (see Staunton, et al. , Cell 52:925-933 (1988)). ICAM-1 is a member of the immunoglobulin supergene family, containing 5 immunoglobulin-like domains at the amino terminus, followed by a transmembrane domain and a cytoplasmic domain. The primary binding site for LFA-1 and rhinovirus are found in the first immunoglobulin-like domain. However, the binding sites appear to be distinct. ICAM-1 exhibits a broad tissue and cell distribution, and may be found on white blood cells, endothelial cells, fibroblast, keratinocytes and other epithelial cells. The expression of ICAM-1 can be regulated on vascular endothelial cells, fibroblasts, keratinocytes, astrocytes and several cell lines by treatment with bacterial lipopolysaccharide and cytokines such as interleukin- 1 , tumor necrosis factor, gamma- interferon, and lymphotoxin (see, e.g., Frohman, et al., J. Neuroimmunol. 23:117-124 (1989)). In preferred embodiments, the antisense oligonucleotide is complementary to the 3 '-untranslated region of human ICAM-1 mRNA and contains from about 10 to about 50 nucleotides, more preferably about 15 to about 30 nucleotides. In particularly preferred embodiments, the antisense oligonucleotide is a phosphorothioate oligonucleotide or a methyl phosphonate oligonucleotide. Phosphorothioate oligonucleotides (PS-oligos) are those oligonucleotides in which one of the non-bridged oxygens of the intemucleotide linkage has been replaced with sulfur. These PS-oligos are resistant to nuclease degradation, yet retain sequence-specific activity. Similarly, methyl phosphonate oligonucleotides (MeP-oligos) are those oligonucleotides in which one of the non-bridged oxygens of the intemucleotide linkage has been replaced by a methyl group. These MeP-oligos have also proven to be more nuclease resistant than their natural phosphodiester linked derivatives. A number of antisense oligonucleotides which are directed toward inhibiting the production of ICAM-1, as well as VCAM-1 and ELAM-1, and which are useful in the present invention have been described in PCT applications: PCT/US91/05209; PCT/US94/09026; PCT/US94/ 12797 and PCT/US93/08101 , the disclosures of each being incoφorated herein by reference. The antisense oligonucleotides used in the present invention may be synthesized in solid phase or in solution. Generally, solid phase synthesis is preferred. Detailed descriptions of the procedures for solid phase synthesis of oligonucleotides by SUBSTTTUTE SHEET (RULE 26) phosphite-triester, phosphotriester, and H-phosphonate chemistries are widely available. See, for example, Itakura, U.S. Pat. No. 4,401 ,796; Caruthers, et al. , U.S. Pat. Nos. 4,458,066 and 4.500,707; Beaucage, et al. , Tetrahedron Lett. , 22: 1859-1862 (1981); Matteucci, et al , J. Am. Chem. Soc , 103:3185-3191 (1981); Caruthers, et al . Genetic Engineering, 4:1-17 (1982); Jones, chapter 2, Atkinson, et al , chapter 3, and Sproat, et al , chapter 4, in Oligonucleotide Synthesis: A Practical Approach, Gait (ed.), IRL Press, Washington D.C. (1984); Froehler, et al, Tetrahedron Lett. , 27:469-472 (1986); Froehler, et al, Nucleic Acids Res. , 14:5399-5407 (1986); Sinha, et al. Tetrahedron Lett. , 24:5843-5846 (1983); and Sinha, et al , Nucl. Acids Res. , 12:4539-4557 (1984) which are incoφorated herein by reference. Generally, the timing of delivery and concentration of monomeric nucleotides utilized in a coupling cycle will not differ from the protocols typical for commercial phosphoramidites used in commercial DNA synthesizers. In these cases, one may merely add the solution containing the monomers to a receptacle on a port provided for an extra phosphoramidite on a commercial synthesizer (e.g., model 380B, Applied Biosystems, Foster City, California, U.S.A.). However, where the coupling efficiency of a particular monomer is substantially lower than the other phoφhoramidites, it may be necessary to alter the timing of delivery or the concentration of the reagent in order to optimize the synthesis. Means of optimizing oligonucleotide synthesis protocols to correct for low coupling efficiencies are well known to those of skill in the art. Generally one merely increases the concentration of the reagent or the amount of the reagent delivered to achieve a higher coupling efficiency. Methods of deterrriining coupling efficiency are also well known. For example, where the 5'-hydroxyl protecting group is dimethoxytrityl (DMT), coupling efficiency may be determined by measuring the DMT cation concentration during the acidic removal of the DMT group. DMT cation concentration is usually determined by spectrophotometrically monitoring the acid wash. The acid/DMT solution is a bright orange color. Alternatively, since capping prevents further extension of an oligonucleotide where coupling has failed, coupling efficiency may be estimated by comparing the ratio of truncated to full length oligonucleotides utilizing, for example, capillary electrophoresis or HPLC. Solid phase oligonucleotide synthesis may be performed using a number of solid supports. A suitable support is one which provides a functional group for the attachment of a protected monomer which will become the 3' terminal base in the SUBSTTTUTE SHEET (RULE 26) synthesized oligonucleotide. The support must be inert to the reagents utilized in the particular synthesis chemistry. Suitable supports are well known to those of skill in the art. Solid support materials include, but are not limited to polacryloylmoφholide, silica, controlled pore glass (CPG), polystyrene, polystyrene/latex, and carboxyl modified teflon. Preferred supports are amino-functionalized controlled pore glass and carboxyl- functionalized teflon. Solid phase oligonucleotide synthesis requires, as a starting point, a fully protected monomer (e.g., a protected nucleoside) coupled to the solid support. This coupling is typically through the 3'-hydroxyl. Typically, a linker group is covalently bound to the 3 '-hydroxyl on one end and covalently bound to the solid support on the other end. The first synthesis cycle then couples a nucleotide monomer, via its 3 '-phosphate, to the 5 '-hydroxyl of the bound nucleoside through a condensation reaction that forms a 3 '-5' phosphodiester linkage. Subsequent synthesis cycles add nucleotide monomers to the 5 '-hydroxyl of the last bound nucleotide. In this manner an oligonucleotide is synthesized in a 3' to 5' direction producing a "growing" oligonucleotide with its 3' terminus attached to the solid support. Numerous means of linking nucleoside monomers to a solid support are known to those of skill in the art, although monomers covalently linked through a succinate or hemisuccinate to controlled pore glass are generally preferred. Conventional protected nucleosides coupled through a hemisuccinate to controlled pore glass are commercially available from a number of sources (e.g. , Glen Research, Sterling, Vermont, U.S.A.; Applied Biosystems, Foster City, California, U.S.A. ; and Pharmacia LKB, Piscataway, New Jersey, U.S.A.). Once the full length oligonucleotide is synthesized, the oligonucleotide is deprotected and cleaved from the solid support prior to use. Cleavage and deprotection may occur simultaneously or sequentially in any order. The two procedures may be interspersed so that some protecting groups are removed from the oligonucleotide before it is cleaved off the solid support and other groups are deprotected from the cleaved oligonucleotide in solution. The sequence of events depends on the particular blocking groups present, the particular linkage to a solid support, and the preferences of the individuals performing the synthesis. Where deprotection precedes cleavage, the protecting groups may be washed away from me oligonucleotide which remains bound on the solid support. Conversely, where deprotection follows cleavage, the removed SUBSTTTUTE SHEET (RULE 26) protecting groups will remain in solution with the oligonucleotide. Often the oligonucleotide will require isolation from these protecting groups prior to use. In a preferred embodiment, and most commercial DNA syntheses, the protecting group on the 5 '-hydroxyl is removed at the last stage of synthesis. The oligonucleotide is then cleaved off the solid support, and the remaining deprotection occurs in solution. Removal of the 5 '-hydroxyl protecting group typically requires treatment with the same reagent utilized throughout the synthesis to remove the terminal 5 '-hydroxyl protecting groups prior to coupling the next nucleotide monomer. Where the 5'- hydroxyl protecting group is a dimethoxytrityl group, deprotection can be accomplished by treatment with acetic acid, dichloroacetic acid or trichloroacetic acid. Where the oligonucleotide is a ribonucleotide and the 2 '-hydroxyl group is blocked with a tert-butyldimethylsilyl (TBDMS) moiety, the latter group may be removed using tetrabutylammonium fluoride in tetrahydrofuran at the end of synthesis. See Wu, et al , J. Org. Chem. 55:4717-4724 (1990). Phenoxyacetyl protecting groups can be removed with anhydrous ammonia in alcohol (under these conditions the TBDMS groups are stable and the oligonucleotide is not cleaved). The benzoyl protecting group of cytidine is also removed with anhydrous ammonia in alcohol. Cleaved and fully deprotected oligonucleotides may be used directly (after lyophilization or evaporation to remove the deprotection reagent) or they may be purified prior to use. Purification of synthetic oligonucleotides is generally desired to isolate the full lengm oligonucleotide from the protecting groups that were removed in the deprotection step and, more importantly, from the truncated oligonucleotides that were formed when oligonucleotides that failed to couple with the next nucleotide monomer were capped during synthesis. Oligonucleotide purification techniques are well known to those of skill in the art. Methods include, but are not limited to, thin layer chromatography (TLC) on silica plates, gel electrophoresis, size fractionation (e.g. , using a Sephadex column), reverse phase high performance liquid chromatography (HPLC) and anion exchange chromatography (e.g., using the mono-Q column, Pharmacia-LKB, Piscataway, New Jersey, U.S.A.). For a discussion of oligonucleotide purification see McLaughlin, et al , chapter 5, and Wu, et al , chapter 6 in Oligonucleotide Synthesis: A Practical Approach, Gait (ed.), IRL Press, Washington, D.C. , (1984). SUBSTTTUTE SHEET (RULE 26) To test the therapeutic ability of liposomes as delivery vehicles for antisense molecules to ICAM-1 , ISIS Pharmaceuticals donated their antisense molecules ISIS 2302 (human, Sequence I.D. No. 9) and 3082 (murine, Sequence I.D. No. 10) towards the project. These oligonucleotides are phosphorothioate molecules in which each phosphodiester linkage is a phosphorothioate diester linkage. Additionally, each oligonucleotide consists of 20 nucleotides specifically designed to bind to a sequence in the 3' untranslated region of ICAM-1 mRNA. Currently it is theorized that this antisense molecule acts posttranscriptionally to inhibit the expression of ICAM-1 via 2 possible mechanisms: inhibiting the formation of a stabilizing stem-loop structure necessary for mRNA stability, and by increasing the degradation of the target mRNA via increased RNase-H degradation. Preparation of pharmaceutical compositions containing the antisense oligonucleotides will typically involve either encapsulating the oligonucleotide in a liposome or forming a lipid particle in which the oligonucleotide is coated with a lipid mixture. LIPOSOME-ENCAPSULATED ANΗSENSE OLIGONUCLEOTIDES The liposomes which are used in the present invention are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phosphoiipids, preferably neutral phosphoiipids, and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of, e.g. , liposome size and stability of the liposomes in the bloodstream. Typically, the major lipid component in the liposomes is phosphatidylcholine. Phosphatidylcholines having a variety of acyl chain groups of varying chain length and degree of saturation are available or may be isolated or synthesized by well-known techniques. In general, less saturated phosphatidylcholines are more easily sized, particularly when the liposomes must be sized below about 0.3 microns, for puφoses of filter sterilization. Phosphatidylcholines containing saturated fatty acids with carbon chain lengths in the range of C to C 22 are preferred. Phosphatidylcholines with mono or diunsaturated fatty acids and mixtures of saturated and unsaturated fatty acids may also be used. Other suitable lipids include phosphonolipids in which the fatty acids are linked to glycerol via ether linkages rather than ester linkages. Liposomes useful in the present invention may also be composed of SUBSTTTUTE SHEET (RULE 26) sphingomyelin or phosphoiipids with head groups other than choline, such as ethanolamine, serine, glycerol and inositol. Preferred liposomes will include a sterol. preferably cholesterol, at molar ratios of from 0.1 to 1.0 (cholesterol:phospholipid). Most preferred liposome compositions are egg phosphatidylcholine/cholesterol, distearoylphosphatidy Icholine/cholesterol. dipalmitoylphosphatidy Icholine/cholesterol, and sphingomyelin/cholesterol. Methods used in sizing and filter-sterilizing liposomes are discussed below. The liposomes can be prepared by any of the techniques now known or subsequently developed for preparing liposomes. For example, the liposomes can be formed by the conventional technique for preparing multilamellar lipid vesicles (MLVs), that is, by depositing one or more selected lipids on the inside walls of a suitable vessel by dissolving the lipids in chloroform and then evaporating the chloroform, and by then adding the aqueous solution which is to be encapsulated to the vessel, allowing the aqueous solution to hydrate the lipid, and swirling or vortexing the resulting lipid suspension. This process engenders a mixture including the desired liposomes. Alternatively, techniques used for producing large unilamellar lipid vesicles (LUVs), such as reverse-phase evaporation, infusion procedures, and detergent dilution, can be used to produce the liposomes. A review of these and other methods for producing lipid vesicles can be found in the text Liposome Technology, Volume I, Gregory Gregoriadis Ed. , CRC Press, Boca Raton, Florida, (1984), which is incoφorated herein by reference. For example, the lipid-containing particles can be in the form of steroidal lipid vesicles, stable plurilamellar lipid vesicles (SPLVs), monophasic vesicles (MPVs), or lipid matrix carriers (LMCs) of the types disclosed in Lenk, et al. U.S. Patent No. 4,522,803, and Fountain, et al. U.S. Patent Nos. 4,588,578 and 4,610,868, the disclosures of which are incoφorated herein by reference. A particularly preferred method for preparing LUVs is described in U.S. Patent No. 5,008,050. Additionally, in the case of MLVs, if desired, the liposomes can be subjected to multiple (five or more) freeze-thaw cycles to enhance their trapped volumes and trapping efficiencies and to provide a more uniform interlamellar distribution of solute (Mayer, et al, J. Biol. Chem. 260:802-808 (1985)). Following liposome preparation, the liposomes may be sized to achieve a desired size range and relatively narrow distribution of liposome sizes. In preferred SUBSTTTUTE SHEET (RULE 26) embodiments, the liposomes will have diameters of from about 50 to about 150 nm, more preferably from about 75 to about 125 nm. Several techniques are available for sizing liposomes to a desired size. One sizing method is described in U.S. Pat. No. 4,737,323, incoφorated herein by reference. Sonicating a liposome suspension either by bath or probe sonication produces a progressive size reduction down to small unilamellar vesicles (SUVs) less than about 0.05 microns in size. Homogenization is another method which relies on shearing energy to fragment large liposomes into smaller ones. In a typical homogenization procedure, multilamellar vesicles are recirculated through a standard emulsion homogenizer until selected liposome sizes, typically between about 0.1 and 0.5 microns, are observed. In both methods, the particle size distribution can be monitored by conventional laser-beam particle size determination. Extrusion of liposome through a small-pore polycarbonate membrane or an asymmetric ceramic membrane is also an effective method for reducing liposome sizes to a relatively well-defined size distribution (see, U.S. Patent No. 5,008,050 and Hope, et al. , in: Liposome Technology, vol. 1, 2d ed. (G. Gregoriadis, Ed.) CRC Press, pp. 123-139 (1992), the disclosures of which are incoφorated herein by reference). Typically, the suspension is cycled through the membrane one or more times until the desired liposome size distribution is achieved. The liposomes may be extruded through successively smaller-pore membranes, to achieve a gradual reduction in liposome size. For use in the present inventions, liposomes having a size of from about 0.05 microns to about 0.15 microns are preferred. Other useful sizing methods such as sonication, solvent vaporization or reverse phase evaporation are known to those of skill in the art. Liposomes prepared for use in the methods and pharmaceutical compositions of the present invention may be dehydrated for longer storage. The liposomes are preferably dehydrated under reduced pressure using standard freeze-drying equipment or equivalent apparatus. The lipid vesicles and their surrounding medium can also be frozen in liquid nitrogen before being dehydrated or not, and placed under reduced pressure. Dehydration without prior freezing takes longer than dehydration with prior freezing, but the overall process is gentler without the freezing step, and thus there is subsequently less damage to the lipid vesicles and a smaller loss of the internal contents. Dehydration without prior freezing at room temperamre and at a reduced pressure provided by a vacuum pump capable of producing a pressure of about 1 mm Hg SUBSTTTUTE SHEET (RULE 25) typically takes between approximately 24 and 36 hours, while dehydration with prior freezing under the same conditions generally takes between approximately 12 and 24 hours. To ensure that the liposomes will survive the dehydration process without losing a substantial portion of their internal contents, it is important that one or more protective sugars be available to interact with the lipid vesicle membranes and keep them intact as the water in the system is removed. A variety of sugars can be used, including such sugars as trehalose, maltose, sucrose, glucose, lactose, and dextran. In general, disaccharide sugars have been found to work better than monosaccharide sugars, with the disaccharide sugars trehalose and sucrose being most effective. Other more complicated sugars can also be used. For example, aminoglycosides, including streptomycin and dihydrostreptomycin, have been found to protect lipid vesicles during dehydration. Typically, one or more sugars are included as part of either the internal or external media of the lipid vesicles. Most preferably, the sugars are included in both the internal and external media so that they can interact with both the inside and outside surfaces of the liposomes' membranes. Inclusion in the internal medium is accomplished by adding the sugar or sugars to the buffer which becomes encapsulated in the lipid vesicles during the lipid vesicle foπnation process. Since in most cases this buffer also forms the bathing medium for the finished lipid vesicles, inclusion of the sugars in the buffer also makes them part of the external medium. The amount of sugar to be used depends on the type of sugar used and the characteristics of the lipid vesicles to be protected. See, U.S. Patent No. 4,880,635 and Harrigan, et al., Chem. Phys. Lipids 52:139-149 (1990), the disclosures of which are incoφorated herein by reference. Persons skilled in the art can readily test various sugar types and concentrations to determine which combmation works best for a particular lipid vesicle preparation. In general, sugar concentrations on the order of 100 mM and above have been found necessary to achieve the highest levels of protection. In terms of moles of membrane phospholipid, millimolar levels on the order of 100 mM correspond to approximately 5 moles of sugar per mole of phospholipid. In the case of dehydration without prior freezing, if the lipid vesicles being dehydrated are of the type which have multiple lipid layers and if the dehydration is carried to an end point where between about 2 % and about 5 % of the original water in SUBSTTTUTE SHEET (RULE 26) the preparation is left in the preparation, the use of one or more protective sugars may be omitted. Once the lipid vesicles have been dehydrated, they can be stored for extended periods of time until they are to be used. The appropriate temperamre for storage will depend on the make up of the lipid vesicles and the temperamre sensitivity of whatever materials have been encapsulated in the lipid vesicles. For example, as is known in the art, various oligonucleotides are heat labile, and thus dehydrated lipid vesicles containing such oligonucleotides should be stored under refrigerated conditions so that the potency of the agent is not lost. Also, for such agents, the dehydration process is preferably carried out at reduced temperatures, rather than at room temperamre. Methods of loading antisense oligonucleotides into liposomes will typically be carried out using an encapsulation technique in which the antisense oligonucleotide is placed into a buffer and added to a dried film of only lipid components. In this manner, the oligonucleotide will become encapsulated in the aqueous interior of the liposome. The buffer which is used in the formation of the liposomes can be any biologically compatible buffer solution of, for example, isotonic saline, phosphate buffered saline, or other low ionic strength buffers. Generally, the antisense oligonucleotide will be present in an amount of from about 0.01 ng/mL to about 200 mg/mL. The resulting liposomes with the antisense oligonucleotide incoφorated in the aqueous interior or in the membrane are then optionally sized as described above. Alternatively, the antisense oligonucleotide can be formulated in lipid particles such as those described in co-pending U.S. Ser. Nos. 08/484,282 and 08/485,458 each being filed on June 7, 1995 and incoφorated herein by reference. ANTISENSE LIPID PARTICLES Antisense lipid particles can be prepared by combining an antisense oligonucleotide with cationic lipids in a detergent solution to provide a coated antisense- lipid complex. The complex is then contacted with phosphoiipids to provide a solution of detergent, an antisense-lipid complex and phosphoiipids, and the detergent is then removed to provide a solution of serum-stable antisense-lipid particles, in which the SUBSTTTUTE SHEET (RULE 26) antisense oligonucleotide is encapsulated in a lipid bilayer. The particles, thus formed, have a size of about 50-150 nm. Alternatively, serum-stable antisense-lipid particles can be formed by preparing a mixture of cationic lipids and phosphoiipids in an organic solvent; contacting an aqueous solution of the antisense oligonucleotide with the mixture of cationic and phosphoiipids to provide a clear single phase; and removing the organic solvent to provide a suspension of antisense-lipid particles, in which the antisense oligonucleotide is encapsulated in a lipid bilayer, and the particles are stable in serum and have a size of about 50-150 nm. In this aspect of the invention, cationic lipids which are useful will include, for example, DODAC, DOTMA, DDAB, DOTAP, DC-Choi, DORI and DMRIE. These lipids and related analogs, which are also useful in the present invention, have been described in co-pending USSN 08/316,399; U.S. Patent Nos. 5,208,036, 5,264,618, 5,279,833, 5,283,185, and 5,334,761, the disclosures of which are incoφorated herein by reference. Additionally, a number of commercial preparations of cationic lipids are available and can be used in the present invention. These include, for example, LIPOFECTIN ® (commercially available cationic liposomes comprising DOTMA and DOPE, from GIBCO/BRL, Grand Island, New York, USA); LIPOFECTAMINE ® (commercially available cationic liposomes comprising DOSPA and DOPE, from GIBCO/BRL); and TRANSFECTAM ® (commercially available cationic liposomes comprising DOGS from Promega Coφ., Madison, Wisconsin, USA). An initial solution of coated antisense-lipid complexes is formed by combining the antisense oligonucleotides with the cationic lipids in a detergent solution. The detergent solution is preferably an aqueous solution of a neutral detergent having a critical micelle concentration of 15-300 mM, more preferably 20-50 mM. Examples of suitable detergents include, for example, N,N'-((octanoylimino)-bis-(trimethylene))-bis- (D-gluconamide) (BIGCHAP); BRIJ 35; Deoxy-BIGCHAP; dodecylpoly (ethylene glycol) ether; Tween 20; Tween 40; Tween 60; Tween 80; Tween 85; Mega 8; Mega 9; Zwittergent ® 3-08; Zwittergent ® 3-10; Triton X-405; hexyl-, heptyl-, octyl- and nonyl-β- D-glucopyranoside; and heptylthioglucopyranoside; with octyl 0-D-glucopyranoside being the most preferred. The concentration of detergent in the detergent solution is typically about 100 mM to about 2 M, preferably from about 200 mM to about 1.5 M. SUBSTTTUTE SHEET (RULE 26) The cationic lipids and antisense oligonucleotides will typically be combined to produce a charge ratio (+/-) of about 1: 1 to about 20: 1, preferably in a ratio of about 1 : 1 to about 12:1, and more preferably in a ratio of about 2: 1 to about 6: 1. Additionally, the overall concentration of antisense oligos in solution will typically be from about 25 μg/mL to about 150 mg/mL, preferably from about 100 μg/mL to about 50 mg/mL, and more preferably from about 100 μg/mL to about 5 mg/mL. The combination of antisense oligos and cationic lipids in detergent solution is kept, typically at room temperamre, for a period of time which is sufficient for the coated complexes to form. Alternatively, the antisense oligos and cationic lipids can be combined in the detergent solution and warmed to temperatures of up to about 37 °C. For antisense oligos which are particularly sensitive to temperamre, the coated complexes can be formed at lower temperatures, typically down to about 4°C. The detergent solution of the coated antisense-lipid complexes is then contacted with non-cationic lipids to provide a detergent solution of antisense-lipid complexes and non-cationic lipids. The non-cationic lipids which are useful in this step include, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cardiolipin, and cerebrosides. In preferred embodiments, the phosphoiipids are diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide or sphingomyelin. The acyl groups in these lipids are preferably acyl groups derived from fatty acids having C 10 -C 24 carbon chains. More preferably the acyl groups are lauroyl, myristoyl, palmitoyl, stearoyl or oleoyl. In particularly preferred embodiments, the non-cationic lipid will be 1,2-dioleoylphosphatidylethanolamine (DOPE), palmitoyl oleoyl phosphatidylcholine (POPC) or egg phosphatidylcholine (EPC). In the most preferred embodiments, the antisense-lipid particles will be fusogenic particles with enhanced properties in vivo and the phospholipid will be DOPE. In other preferred embodiments, the non-cationic lipids will further comprise polyethylene glycol-based polymers such as PEG 2000, PEG 5000 and polyethylene glycol conjugated to ceramides, as described in co-pending USSN 08/316,429, incoφorated herein by reference. . The amount of phospholipid which is used in the present methods is typically about 2 to about 150 mg/mL of total lipids to about 2 to about 150 mg/mL of antisense oligonucleotide. Preferably the amount of total lipid is from about 50 to about 100 mg/mL to 50 to 100 mg/mL antisense. SUBSTTTUTE SHEET (RULE 26) Following formation of the detergent solution of antisense-lipid complexes and non-cationic lipids, the detergent is removed, preferably by dialysis. The removal of the detergent results in the formation of a lipid-bilayer which surrounds the antisense oligo providing serum-stable antisense-lipid particles which have a size of from about 50 nm to about 150 nm. The particles thus formed do not aggregate and are optionally sized to achieve a uniform particle size. The serum-stable antisense-lipid particles can be sized by any of the methods available for sizing liposomes. The sizing may be conducted in order to achieve a desired size range and relatively narrow distribution of particle sizes. Several techniques are available for sizing the particles to a desired size. One sizing method, used for liposomes and equally applicable to the present particles is described in U.S. Pat. No. 4,737,323, incoφorated herein by reference. Sonicating a particle suspension either by bath or probe sonication produces a progressive size reduction down to particles of less than about 50 nm in size. Homogenization is another method which relies on shearing energy to fragment larger particles into smaller ones. In a typical homogenization procedure, particles are recirculated through a standard emulsion homogenizer until selected particle sizes, typically between about 60 and 80 nm, are observed. In both methods, the particle size distribution can be monitored by conventional laser-beam particle size discrimination, or QELS. Extrusion of the particles through a small-pore polycarbonate membrane or an asymmetric ceramic membrane is also an effective method for reducing particle sizes to a relatively well-defined size distribution. Typically, the suspension is cycled through the membrane one or more times until the desired particle size distribution is achieved. The particles may be extruded through successively smaller-pore membranes, to achieve a gradual reduction in size. As noted above, the serum-stable antisense-lipid particles can also be prepared by combining a mixture of cationic lipids and non-cationic lipids in an organic solvent; contacting an aqueous solution of the antisense oligonucleotide with the mixture of lipids to provide a clear single phase; and removing the organic solvent to provide a suspension of antisense-lipid particles, wherein the antisense oligonucleotide is encapsulated in a lipid bilayer, and the particles are stable in serum and have a size of from about 50 to about 150 nm. SUBSTTTUTE SHEET (RULE 26) The antisense oligos, cationic lipids and phosphoiipids which are useful in this group of embodiments are as described for the detergent dialysis preparative methods described above. The selection of an organic solvent will typically involve consideration of solvent polarity and the ease with which the solvent can be removed at the later stages of particle formation. . The organic solvent, which is also used as a solubilizing agent, is in an amount sufficient to provide a clear single phase mixture of antisense oligonucleotide and lipids. Suitable solvents include chloroform, dichloromethane, diethylether, cyclohexane, cyclopentane, benzene, toluene, methanol, or other aliphatic alcohols such as propanol, isopropanol, butanol, tert-butanol, iso-butanol, pentanol and hexanol. Combinations of two or more solvents may also be used in the present invention. Contacting the antisense oligonucleotide with the organic solution of cationic and phosphoiipids is accomplished by mixing together a first solution of antisense oligonucleotide, which is typically an aqueous solution and a second organic solution of the lipids. One of skill in the art will understand that this mixing can take place by any number of methods, for example by mechanical means such as by using vortex mixers. After the antisense oligonucleotide has been contacted with the organic solution of lipids, the organic solvent is removed, thus forming an aqueous suspension of serum-stable antisense-lipid particles. The methods used to remove the organic solvent will typically involve evaporation at reduced pressures or blowing a stream of inert gas (e.g., nitrogen or argon) across the mixture. The serum-stable antisense-lipid particles thus formed will typically be sized from about 50 nm to 150 nm. To achieve further size reduction or homogeneity of size in the particles, sizing can be conducted as described above. ADMINISTRATION OF THE ANTISENSE FORMULATIONS The present invention further provides methods for the treatment of pathologic conditions associated with the overexpression of ICAM-1 in a host. In these methods a pharmaceutical composition as described above is administered to the host. In preferred embodiments, the host is a mammal, more preferably a mouse, rat, human, horse, dog, cat. cow or pig. Still more preferably, the host is human. Preferred SUBSTTTUTE SHEET (RULE 26) antisense oligonucleotides, lipid mixtures and lipids are as described above for the compounds of the present invention. The antisense oligonucleotide liposomes and lipid particles described above can be administered in any suitable manner, preferably with pharmaceutically acceptable carriers. One skilled in the art will appreciate that suitable methods of administering such compositions in the context of the present invention to an animal are available, and, although more than one route can be used to administer a particular composition, a particular route can provide a more immediate and more effective reaction than another route. Pharmaceutically acceptable carriers are also well-known to those who are skilled in the art. The choice of carrier will be determined in part by the particular composition, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of the pharmaceutical composition of the present invention. Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the antisense oligonucleotide (in a liposome or lipid particle) dissolved in diluents, such as water, saline or PEG 400; (b) suspensions in an appropriate liquid; and (c) suitable emulsions. Formulations suitable for parenteral administration, such as, for example, by intravenous, intramuscular, intradermal, intraperitoneal, and subcutaneous routes, include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The formulations can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water, for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. The dose administered to an animal, particularly a human, in the context of the present invention should be sufficient to effect a therapeutic response in the animal over a reasonable time frame. The dose will be determined by the strength of the particular compound employed and the condition of the animal, as well as the body SUBSTTTUTE SHEET (RULE 26) weight or surface area of the animal to be treated. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects that might accompany the administration of a particular compound in a particular animal. As noted, the antisense oligonucleotides (in liposome or lipid particle formulations) can be administered, for example, by intravenous infusion, orally, topically, intraperitoneally, intravesically or intrathecally. For oral administration, the compositions of the present invention can be administered at the rate up to 3000 mg/m 2 body surface area, which approximates 6 grams/day in the average patient. A preferred rate is from 1 to 300 mg/m 2 body surface area. This can be accomplished via single or divided doses. For intravenous administration, such compounds can be administered at the rate of up to about 2500 mg/m 2 /d, preferably from about 0.1 to about 200 mg/m 2 /d. For intravesicle administration, such compounds can be administered at the rate of up to about 2500 mg/πr/d, preferably from about 0.1 to about 200 mg/m 2 /d. For topical administration, the rate can be up to about 2500 mg/m 2 /d, preferably from about 0.1 to about 200 mg/m 2 /d. The dose for inhalation aerosol administration can be up to about 2500 mg/m 2 /d, preferably from about 0.1 to about 200 mg/m 2 /d. Direct intraperitoneal administration can be performed using up to about 3000 mg/m 2 /d, preferably from about 0.1 to about 100 mg/m 2 /d. The dose for reservoir administration to the brain or spinal fluid can be up to about 2000 mg/m 2 /d, preferably from about 0.1 to about 100 mg/n /d. For slow release intraperitoneal or subcutaneous administration, the dose can be from about 0.1 to about 5000 mg/day in a bolus, preferably from about 1.0 to about 200 mg/day. For intrathecal administration, the dose can be up to about 2000 mg/m 2 /d, preferably from about 0.1 to about 100 mg/m 2 /d. The following examples are offered solely for the puφoses of illustration, and are intended neither to limit nor to define the invention. EXAMPLES Materials l ,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and egg phosphatidylcholine (EPC) was purchased from Avanti Polar Lipids (Alabaster, Alabama, SUBSTTTUTE SHEET (RULE 26) USA). Cholesterol, N-(2-hydroxyethyl) piperazine-N'-(2-ethanesulphonic acid) (HEPES), SEPHAROSE ® CL-4B, SEPHADEX ® G-25, SEPHADEX ® G-50 and dithiothreitol (DTT) were obtained from Sigma Chemical Co. (St. Louis, Missouri, USA). 3 H- Cholesteryl hexadecyl ether ( 3 H-CHE) was obtained from Amersham (Oakville, Ontario, Canada). Female ICR and Balb/c mice were obtained from Harley-Sprague Davis. Buffers 5X Veronal Buffered Saline (VBS). Mix 85.0 g of NaCl, 3.75 g of sodium barbitone, and 5.75 g of barbitone in 1.6 L of distilled water. Heat the mixmre to dissolve the barbitone. Bring the volume up to 2.0 L with distilled water. The pH should be 7.4 - 7.6. Store at 4°C. GVB 2" . Dissolve 0.5 g of gelatin in 100 mL of distilled water and heat to dissolve. Add 100 mL of 5X VBS and make the total volume to 500 mL with distilled water. Store at 4°C. Make this buffer every few days. D5W 2 ". Dissolve 25 g of D-glucose in 500 mL of distilled water. Make this solution every few days. Store at 4°C. DGVB 2+ . Mix 250 mL of D5W with 250 mL of GVB 2 . Add 75 μL of 1.0 M CaCl 2 and 100 μL of 2.5 M MgCl 2 . Store at 4°C. Make this buffer every few days. EDTA»GVB 2 . Mix 1.489 g EDTA, 0.5 g gelatin, 100 mL of 5X VBS, and 200 mL of distilled water. Bring the pH up to 7.4 with 5M NaOH (this will also help to dissolve the EDTA). Make the volume up to 500 mL with distilled water. Store at 4 C C. Make this buffer every few days. EXAMPLE 1 This example illustrates the passive encapsulation of antisense oligonucleotides in liposomes. Additionally, this example illustrates that liposomes and unencapsulated DNA can be separated by their molecular weight difference using size exclusion chromatography . SUBSTTTUTE SHEET (RULE 26) 1.1 Encapsulation of antisense Egg phosphatidylcholine (EPC) (1.78 g) and cholesterol (CH) (0.72 g) were weighed out in a laminar flow hood and combined in chloroform in a 150 mL round bottomed flask. Chloroform was removed by roto-evaporation for 3 hours. The antisense molecule ISIS 2302 (1.5 g) was dissolved in 10 mL of sterile phosphate buffered saline (PBS) and added to the lipid film composed of 2.5 g of EPC:CH (55:45; mol: mol). The resulting multilamellar vesicles (MLV) were transferred to three 5 mL cryovials and subjected to five cycles of freezing in liquid nitrogen and thawing at 40 °C. The freeze-thawed MLV were combined, transferred into a sterile 100 mL capacity extruder (Lipex Biomembranes, Vancouver) and extruded ten times through one 0.1 μm filter. Similar conditions were followed for smaller liposomal antisense preparations, except that 200 mg of oligonucleotide was rehydrated in 250 mg (total) of EPC:CH lipid. 1.2 DEAE-Sepharose Chromatography Non-encapsulated (free) antisense molecule was separated from the entrapped molecule by anion exchange chromatography using DEAE-Sepharose CL-6B. In order to ensure reasonable sterility of the column material, the columns were washed with sterile PBS followed by elution of 300 mL of 100 nm vesicles composed of EPC (24 mg/mL in PBS). Columns consisted of 6 mL syringes with a packed volume of 5 mL. A 400 mL aliquot of the antisense-liposome suspension was eluted on each column and the fractions containing lipid were collected and combined. Each column was used twice and then stored at 4°C for future recovery of the free antisense molecule. The total volume of the column eluant was approximately 110 mL; therefore, it was necessary to concentrate the liposomal antisense. The liposomal antisense was placed in dialysis tubing (MW cutoff, 14000) and concentrated to approximately 28 mL using Aquacide. Results of this purification are shown in Figure 2. 1.3 Measurement of Trapping Efficiency Trapping efficiency was determined by size exclusion chromatography using a 1 mL Biogel A 15m (fine) spin column as described previously (Chonn et al , SUBSTTTUTE SHEET (RULE 26) 1991). Briefly, a 50 μL aliquot of the liposomal antisense was diluted with 70 μL of PBS, and 50 μL was eluted on each of two duplicate columns. Each fraction consisted of the volume eluted ( -40 μL) during centrifugation at 1000 φm for 1 min. The column eluant was collected, 50 μL of PBS was added to the column, and the column was centrifuged again as described above, with the eluant being collected into a new 13 x 100 mm mbe. This was repeated until a total of 36 fractions had been collected. Each fraction was assayed for lipid and DNA after the addition of 250 μL of distilled water. Lipid was assayed by scintillation counting or phosphate analysis (Fiske and Subbarow, 1925) of a 50 μL aliquot from each sample and the remainder (approximately 250 μl) was assayed for DNA by a Bligh-Dyer extraction protocol. The results are shown in Figure 1. 1.4 Extraction Protocol DNA in 250 μL H 2 O was separated from the lipid by the addition of 750 μL of chloroform: methanol (1 :2.1) to form a single phase consisting of chloroform:methanol:H 2 O (1:2.1:1). Additional volumes of H 2 O (250 μL) and chloroform (250 μL) were added to the sample, resulting in a two phase system. The sample was then centrifuged at 3000 φm for 10 min to facilitate rapid separation of the organic and aqueous layers. The upper, aqueous, phase was collected and 400 μL was assayed for DNA by absorbance at 260 nm. The lower, chloroform, phase was washed three times with 300 μL of methanol :H 2 0 (1: 1) and dried under N 2 . The lipid film was hydrated in 1 mL of H 2 0 and an aliquot was taken for phosphate assay. 1.5 Stability of the Encapsulated Liposomal Antisense Since the liposomal antisense preparation was not always used immediately it was necessary to analyze the stability of the preparation over several days. To this end, retention of antisense in liposomes was measured over a 5 day period, at 4°C and at room temperamre. Leakage of entrapped antisense would appear as a peak in the included volume of Biogel A15m spin columns. At various times, 50 μL aliquots of the sample were applied to a 1 mL Biogel A15m column and eluted as described above. Lipid and oligonucleotide were determined as described above. A second peak in the oligonucleotide elution profile was taken to indicate leakage of entrapped oligonucleotide. SUBSTTTUTE SHEET (RULE 26) No leakage was indicated, either at 4°C or room temperamre, even after 5 days (see Figures 3 and 4). EXAMPLE 2 This example illustrates the complement activation by a liposomal antisense formulation. To monitor the effects of various liposome and/or DNA formulations we have established an in vitro complement hemolytic assay using human serum for screening the relative complement activating properties of these formulations. The assay is a two-step procedure. The first step involves consumption of complement by liposomes and/or DNA, while the second step involves the lysis of antibody-sensitized sheep red cells by any residual complement that may not have been activated in the first part of the assay. Complement activation by liposomes and free antisense was investigated using EPC/CH liposome compositions of 100 ± 30 nm diameter. 2.1 Normal Human Serum Pool The first component of the assay that was tested was the activity of the fresh serum pool. This should be tested each time a new serum pool is generated as there will be some differences in complement activity between serum pools which can affect the sensitivity of the assay. A series of serum dilutions was tested to determine what dilution would give both maximal red cell lysis and minimal interference in absorbance readings (the more concentrated the serum dilution the more background absorbance is observed). Anything less than a 100-fold serum dilution gave reasonable levels of red cell lysis. EDTA-GVB 2' was added at the end of the assay to inhibit complement activity. The volume of EDTA-GVB 2" can be modified to increase or decrease the absorbance range of the assay depending on the activity of the semm pool. For all subsequent assays a 50-fold serum dilution was used (25-fold dilution in step 1 and 2-fold dilution in step 2). Furthermore, the amount of EDTA-GVB 2' used to stop the assay was 1.0 mL, giving an absorbance range (A 4U) ) of 0 - 0.8. SUBSTTTUTE SHEET (RULE 26) Blood from seven healthy males and six healthy females was gathered into chilled serum mbes and immediately placed in an ice/ water bath. Thirty mL of blood was collected per individual. Tubes were centrifuged at 2500 φm for 10 min at 4°C, every six mbes (to avoid clotting). Plasma was removed from all mbes and pooled into a 250 mL beaker, on ice. The pooled plasma was then incubated at 37°C for 30 min, in the presence of several cloning sticks (to help recess the clot). The clot was removed and recessed, generating approximately 100 mL of serum. The semm was aliquoted (1.0 mL) into 1.5 mL Eppendorf mbes and stored at -65°C until use. 2.2 Preparation of Sensitized Sheep Red Cells (EA cells) An aliquot of whole sheep blood (a 50% solution in Alsever's; Cedarlane) was withdrawn from the stock solution and centrifuged for 10 min at 1500 φm. The cells were then washed three times with 10 volumes of EDTA-GVB 2" . The washed cells were resuspended in a volume of EDTA-GVB 2 ' that is approximately 5-times that of the initial aliquot. An aliquot (100 μL) of the resuspended cells was mixed with 2.9 mL of distilled water in a cuvette and the absorbance at 541 nm was measured. The concentration of the cells was adjusted to 1 x 10 9 cells/mL with EDTA-GVB 2 according to the following information: [RBC] O.D. λ (cells/mL) (nm) 1 x 10 9 0.385 541 5 10 8 0.192 541 2 10 8 0.654 414 l x 10 8 0.327 414 The cell suspension was warmed to 37 °C in a shaking bath and rabbit anti-sheep red blood cell antibody (hemolysin) was added to give a final antibody dilution of 1/500 (i.e. 20 μL of antibody into 10 mL of cells). This mixmre was incubated for 30 min at 37°C. Following the. incubation, the cells were centrifuged at 1500 φm for 5 min at 4°C, the supernatant removed, and the cells washed with EDTA-GVB 2' . The cells were then washed 2 times with DGVB 2+ in order to further remove any free antibody and to introduce cations into the cell suspension. Finally, the cell concentration was adjusted to SUBSTTTUTE SHEET (RULE 26) 2 x 10 a cells/mL with DGVB 2+ using the information given above. Cells were maintained at 4°C at all times, after preparation, and were used on the same day. 2.3 Complement Hemolytic Assay All samples were serially diluted over a concentration range covering several orders of magnitude. Diluted liposome or oligonucleotide samples (100 μL) were added to 100 μL of normal human semm (NHS) that had been diluted 5-times in DGVB 2+ . Samples were incubated for 30 min at 37 °C and subsequently placed on ice. Ice-cold DGVB 2+ (300 μL) was added to each mbe to dilute any complement not already consumed. Samples (100 μL) of the diluted incubation mixmre were added to 100 μL of EA cells or 100 μL of DGVB 2+ (for color blanks - these are usually required at liposome concentrations > 1 mM). The mixmre was incubated for 30 min at 37 °C and subsequently placed on ice. EDTA-GVB 2 (1.0 mL) was added to the sample to inhibit complement activity and the mixmre was centrifuged for 5 min at 4°C and 1500 φm. Aliquots of the supernatant (250 μL) were transferred to a microtiter plate, in triplicate, with care not to disturb the pelleted red cells. The absorbance of the supernatant was measured at 410 nm on an electronic plate reader. Figure 5 depicts the complement activating ability of the liposome composition. As can be seen, the neutral EPC:CH liposomes showed no observable complement activation over the concentration ranges studied. EXAMPLE 3 This example illustrates the treatment of mice with liposomal antisense compositions to reduce inflammation due to delayed type hypersensitivity (DTH). A murine model of contact sensitivity, a form of delayed type hypersensitivity (DTH), has been established. This model involves the sensitization of the abdominal region of mice with a strong contact sensitizing agent, dinitrofluorobenzene (DNFB). Inflammation can then be induced at a later time (5 days after initial sensitization) by challenging the ear with a dilute solution DNFB. This model has been characterized with respect to several common features of inflammation; specifically, SUBSTTTUTE SHEET (RULE 26) edema (ear thickness measurements), vascular leak (liposome accumulation in inflamed ears), and cell infiltration (myeloperoxidase assays for neutrophils/monocytes or by prelabeling bone marrow cells and circulating leukocytes with [ 3 H] -thymidine). Furthermore, both inbred (BALB/c) and outbred (ICR) mice have been tested in this model, with similar patterns of inflammation being observed for both strains of mice. 3.1 Mice Female BALB/c and ICR mice were obtained from Harley and Sprague Davis. BALB/c mice were used at 6-9 weeks of age, while ICR mice were used at 8-10 weeks of age. Each experimental group consists of four mice and the experiments were repeated at least twice. 3.2 Sensitization and Elicitation of Contact Sensitivity Mice were sensitized by applying 25 μL of 0.5% 2,4-dinitro-l- fluorobenzene (DNFB) in acetone: olive oil (4: 1) to the shaved abdominal wall for two consecutive days. Four days after the second application, mice were challenged on the dorsal surface of the left ear with 10 μL of 0.2% DNFB in acetone:olive oil (4: 1). Mice received no treatment on the contralateral (right) ear. In some cases, control mice received 10 μL of vehicle on the dorsal surface of the left ear. 3.3 Evaluation of Ear Swelling Ear thickness was measured immediately prior to ear challenge, and at various time intervals after DNFB challenge, using an engineer's micrometer (Mitutoyo, Tokyo, Japan). Increases in ear thickness measurements were determined by subtracting the pre-challenge from post-challenge measurements. The progression of ear inflammation over a 3 day period for ICR (outbred) and BALB/c (inbred) mice is indicated in Figures 6 and 7, respectively. Erythema was evident almost immediately after ear challenge and gradually declined in intensity over the remainder of the study. ICR mice exhibited peak ear thickness at 24 hours after the induction of ear inflammation. Maximal ear thickness measurements were found to be 170 x 10" inches, corresponding to a 70% increase in ear thickness. Although ear SUBSTTTUTE SHEET (RULE 26) swelling gradually declines at 48 and 72 hours after inflammation initiation, ear measurements still have not returned to baseline thickness levels (90-100 x 10" inches). BALB/c mice demonstrate peak ear thickness measurements between 24-48 hours after ear challenge. The maximal ear thickness measurements exhibited by these mice were 130 x 10" inches, which corresponds to an increase of -50% over baseline values (75-85 x 10" inches). 3.4 Evaluation of Liposome Accumulation Large unilamellar vesicles composed of distearylphosphatidylcholine (DSPC) and cholesterol (CH), at a 55:45 molar ratio, were prepared in HEPES-buffered saline (20 mM HEPES, 145 mM NaCl, pH 7.4) by extrusion through 2 stacked 100 nm polycarbonate filters. Liposomes contained a non-exchangeable radioactive lipid marker, [ 3 H]cholesterylhexadecylether (CHE). LUVs were administered at a dose of 100 mg/kg (200 μL; - 2 μCi of CHE/mouse) via the dorsal tail vein at 0, 24, 48, 72 hr after initiation of ear inflammation. Liposomes were allowed to circulate for 24 hr after taking ear measurements. Mice were then terminated and the ears were collected for analysis of liposome accumulation and cell infiltration. Analyses of liposome accumulation were performed so as to give a relative indication of the degeneration of the vasculature during various stages of inflammation. Consequently, liposome accumulation was measured over the following time intervals: 0-24 hr, 24-48 hr, 48-72 hr (and in some cases 72-96 hr). For both strains of mice maximal liposome accumulation occurred during the first 24 hr (Figures 8 and 9). Thus, the most prominent changes in the vasculature likely occurred during the 0-24 hr time period. 3.5 Evaluation of Cell Infiltration Cellular infiltration in inflamed and non-inflamed ears was assessed by an enzymatic assay for neutrophil (and monocyte) myeloperoxidase (MPO) activity, or by pre-labeling bone marrow cells and circulating leukocytes with [ 3 H] -methyl thymidine. In order to extract MPO from the azurophilic granules of the cells rather harsh conditions were employed. Ears were finely minced with scissors and added to 2.0 mL of phosphate buffer containing 0.5% hexadecyltrimethylammonium bromide (HTAB) (50 mM potassium phosphate, 0.5 % HTAB, pH 6.0). The sample was then homogenized using a polytron homogenizer for 1 min (high output), and sonicated for 30 sec (power output, 4; 40% pulse). The sample was then divided into two 1.0 mL portions. One portion was digested, using Solvable, and analyzed for CHE by standard liquid scintillation counting. The second portion was freeze-thawed five times in liquid nitrogen and again sonicated for 30 sec (power output, 4; 40% pulse). The sample was then centrifuged for 10 min at 18 000 x g to remove cellular debris. The supernatant was removed and assayed for MPO activity by incubating 0.1 mL aliquots with 2.9 mL of substrate buffer (50 mM potassium phosphate, pH 6.0 containing 0.167 mg/mL o- dianisidine dihydrochloride and 0.0005% hydrogen peroxide). Absorbance at 460 nm was monitored for several minutes and, in most cases, was taken from the absorbances at 30 and 90 seconds. All solutions were maintained on ice as much as possible through the procedure. One unit of MPO activity is defined as the amount of enzyme that degrades 1 μmol of peroxide per minute at 25 °C. For experiments in which blood cells were prelabeled with pH] -methyl thymidine, all mice received a 500 μL i.p. injection pH] -methyl thymidine in sterile saline (1 μCi/g of body weight) 24 hours prior to challenging the ear with DNFB. Ears were removed after taking ear thickness measurements, digested with Solvable, and analyzed for labeled cells by standard liquid scintillation analysis. A relative estimate of cell infiltration was made by expressing the ratio of the radioactivity observed in the inflamed versus the non-inflamed ear. Figure 10 shows a time course MPO accumulation in inflamed versus non- inflamed ears. Peak MPO activity occurs at approximately 48 hr and returns to baseline levels by 96 hr. Based on the accumulation of radiolabeled blood cells (predominantly neutrophils, monocytes, and T-lymphocytes), cell accumulation peaks by 24 hr and remains relatively high over 72 hours (see Figure 11). The major difference in these two procedures is that the MPO assay primarily measures neutrophil accumulation (neutrophils have three times more MPO than monocytes), whereas the [ 3 H] -methyl thymidine procedure measures the influx of all cells equally. Neutrophil accumulation at sites of inflammation has been demonstrated to rise rapidly over the first 24 hours and to decrease almost as rapidly. From 24-48 hours, increased levels of monocytes and T-cells are observed at the inflammation site. SUBSTTTUTE SHEET (RULE 26) 3.6 A Typical Inflammation Experiment Figure 12 details a typical inflammation experiment involving ICR mice. Ear swelling begins to show increases at approximately 12 hr after ear challenge. Lipid accumulation begins almost immediately and consistently increases over 24 hr (the 6 hr, 12 hr, and 24 hr lipid accumulation time points in this figure were injected at t=0 hr). Finally, a lag period can be observed before the onset of cell infiltration, which begins to increase consistently after 6 hr, peaks at 24 hr, and then slowly declines over 72 hr. EXAMPLE 4 This example illustrates the passive targeting of large unilamellar vesicles to sites of inflammation using a murine ear inflammation model. Liposome accumulation appeared to be maximal during the 0-24 and 24-48 hr time periods after the onset of inflammation, corresponding to peak inflammatory events. After this, liposome accumulation decreased dramatically, corresponding to remodeling and repair of the "leaky" vasculature. Of primary importance to the development of inflainmation are the moφhological and functional alterations that occur in dermal microvascular cells. When activated by cytokines, endothelial cells vasodilate, resulting in increased vascular blood flow to the region of inflammation. In addition, the blood vessels are stimulated to structurally remodel, thus enabling immune cells to extravasate from the vasculamre and access the inflammation site. After such alterations, the endothelium is optimized for the infiltration of leukocytes and macromolecules to the site of inflammation. Consequently, it would be expected that relatively small vesicles, such as liposomes (100 nm diameter in our smdies), would avidly move through the "leaky" vasculamre and passively accumulate at sites of inflammation. -- 4.1 Methods The mice used, as well as the sensitization and elicitation of contact sensitivity were carried out as described above in Example 3. Liposome accumulation was monitored over the first 24 hours of inflammation. LUVs were administered at a dose of 100 mg/kg (200 μL; - 2 μCi of CHE/mouse) via the dorsal tail vein immediately SUBSTTTUTE SHEET (RULE 26) after ear challenge with DNFB. At 6, 12, and 24 hr after ear challenge, mice were terminated. Ears were collected and analyzed as described above. 4.2 Liposome Accumulation at a Site of Inflammation Liposome accumulation was examined during various stages of murine ear inflammation so as to give a relative indication of the ability of these vesicles to extravasate through the "inflamed" vasculamre. This is of interest for the passive targeting of liposomal dmgs, such as anti-ICAM-1 oligonucleotides and corticosteroids, to sites of inflammation. Liposome accumulation was measured over the following time intervals: 0-24 hr, 24-48 hr, 48-72 hr. For both ICR and BALB/c mice, maximal liposome accumulation occurred during the first 24 hr of inflammation. Thus, it appears that the most prominent stmcmral changes to the vasculamre occurred during the 0-24 hr time period. This is consistent with previous reports detailing vascular leakage of relatively small molecules and proteins, such as BSA. 4.3 Liposome Accumulation over the first 24 hr of Inflammation Figure 13 depicts the accumulation of DSPC:CH liposomes, administered immediately after ear challenge (t=0 hr), over the initial 24 hr of inflammation. As might be expected, lipid steadily accumulated in the inflamed ear during the first 24 hr, corresponding to the remodeling of the vascular endothelium in the inflamed region. No such increases in lipid accumulation were observed for control ears (non- inflamed), since no remodeling of the endothelium has occurred. As Figure 13 illustrates, passive accumulation of LUVs occurs at sites of inflammation. Liposome accumulation appears to be maximal during the 0-24 hr time periods (after the onset of inflammation). After this, accumulation decreases dramatically, corresponding to remodeling and repair of the "leaky" vasculamre. EXAMPLE 5 This example illustrates the plasma clearance and biodistribution of free and encapsulated oligonucleotides. SUBSTTTUTE SHEET (RULE 26) Mice were sensitized and challenged as described in Example 3. Fifteen minutes after ear challenge, various antisense formulations were administered by the lateral tail (200 μL) at an oligonucleotide dose of 50 mg/kg. Control mice were injected with PBS or saline. At various timepoints blood was withdrawn from the mice by cardiac puncture and collected into plasma mbes containing EDTA. An aliquot of whole blood was removed for analysis. The blood was centrifuged at 3000 φm for 10 min and an aliquot of the plasma was counted by standard liquid scintillation analysis. Organs were collected, homogenized, digested and analyzed for the presence of radiolabeled antisense and radiolabeled lipid by standard liquid scintillation counting techniques. The relative rates of clearance for free and encapsulated oligonucleotide are shown in Figure 14. The circulation half-life for free oligonucleotide (ISIS 2302) was very short (about 2.5 minutes). However, the formulation with antisense encapsulated in EPC:CH (55:45) liposomes had a much slower rate of clearance from the circulation. Instead of a circulation half-life of minutes, the encapsulated oligonucleotide had a half- life of about 8 hours. Thus, encapsulation increases the circulation half-life approximately 100-fold. Figure 15 shows the tissue distribution of pH] -antisense in the liver, spleen, lung, and kidney after intravenous injection of oligonucleotide (50 mg/kg dose). The antisense molecule was rapidly cleared from the plasma with the majority of the dose distributing primarily to the liver and kidney. Only minor accumulation was observed in the spleen and lung. On the basis of organ weight, however, the kidney was the most efficient organ for antisense removal. This result is consistent with reports that indicate that the major route of elimination of free antisense from the body is by urinary excretion. Figure 16 demonstrates the biodistribution profiles for both the lipid and antisense portions of the encapsulated formulation. The liver was found to be the primary organ of accumulation for both antisense and lipid. Some accumulation was noted in the spleen with trace amounts being detected in the lungs and kidneys. This pattem of biodistribution is very similar to standard liposome biodistribution profiles but is significantly different from the kidney and liver tissue distribution exhibited by free oligonucleotide. Thus, the biodistribution of the lipid component determines the distribution of the antisense molecule as would be expected if the antisense molecule does not leak out of the liposomes. SUBSTTTUTE SHEET (RULE 26) EXAMPLE 6 This example illustrates the efficacy of mouse anti-ICAM oligonucleotide. The efficacy of antisense oligonucleotide against mouse ICAM-1 mRNA was tested using the ear inflammation model described above (Example 3). The test oligonucleotide was developed by Isis Pharmaceuticals and is referred to as Isis 3082. The antisense is a 20 base (20 mer) phosphorothioate against a sequence in the untranslated 5 '-region of murine ICAM-1 mRNA (see, Bennett, et al., J. Immunol. 152:3530-3540 (1994); Bennett, et al., Adv. Pharmacol 28:1-43 (1994); and Chiang, et al, J. Biol. Chem. 266:18162-18171 (1991)). This oligonucleotide has recently been shown to exhibit activity in reducing heart allograft rejection in mice (Stepkowski, et al, J. Immunol. 153:5336-5346 (1994) and Stepkowski, et al. Transplant. Proc. 27:113 (1995) but had not been found effective in the DTH model (personal communication, F. Bennett, Isis). Ear thickness in the mice was measured immediately prior to ear challenge and 24 hr after DNFB challenge using an engineer's micrometer (Mitutoyo, Tokyo, Japan). Increases in ear thickness measurements were determined by subtracting the pre- challenge measurements from the post-challenge measurements. Fifteen minutes after the initiation of murine ear inflammation, various antisense formulations were administered at an oligonucleotide dose of 50 mg/kg (in 200 μL) via the dorsal tail vein. Inflammation control mice were injected with PBS. Ear measurements were taken 24 hours later to assess edema formation (ear swelling). The effect of free anti-ICAM- 1 oligonucleotides on the development of murine ear inflammation is indicated in Figure 17. As can be seen, each of the sets of mice (treated with either free ISIS 2302 or 3082) exhibited increased ear thickness measurements equivalent to the inflammation control mice which had been injected with PBS. Thus, a single dose (50 mg/kg) of free oligonucleotide was insufficient for preventing the development of ear edema. Encapsulated antisense oligonucleotides were injected as described above and the results are shown in Figure 18. Increases in ear thickness were observed in mice SUBSTTTUTE SHEET (RULE 26) that received an injection of PBS. Similarly, mice treated with a single dose of free ISIS 3082 antisense, empty EPC:CH liposomes, or an encapsulated human ICAM-1 specific oligonucleotide (ISIS 2302) showed significant increases ( > 100%) in ear thickness. However, a murine ICAM-1 specific antisense molecule (ISI 3082) encapsulated in EPC:CH liposomes was able to significantly reduce ear edema, whether injected 30 min prior to or immediately after initiating inflammation. This result is comparable to the control in which mice were treated topically with corticosteroid (HBP). In a separate experiment, the ear weights of the DNFB-challenged and untreated ear were compared for mice treated with a single dose of free ISIS 3082, encapsulated ISIS 3082, and PBS (see Figure 19). An approximate 2-fold increase in the weight of the challenged ear was observed for mice treated with either the free antisense molecule or PBS. However, little difference was observed between left and right ear weights of mice treated with encapsulated ISIS 3082, indicating a lack of edema. All publications, patents and patent applications mentioned in this specification are herein incoφorated by reference into the specification to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incoφorated herein by reference. Although the foregoing invention has been described in some detail by way of illustration and example for puφoses of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims. SUBSTTTUTE SHEET (RULE 26) SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: University of British Columbia (B) STREET: 2194 Health Sciences Mall, Room IRC 331 (C) CITY: Vancouver (D) STATE (PROVINCE) : British Columbia (E) COUNTRY: Canada (F) POSTAL CODE: V6T 1Z3 (G) TELEPHONE: (604) 822-8580 (H) TELEFAX: (604) 822-8589 (I) TELEX: (ii) TITLE OF INVENTION: Enhanced Efficacy of Liposomal Antisense (iii) NUMBER OF SEQUENCES: 10 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Townsend and Townsend and Crew LLP (B) STREET: Two Embarcadero Center, Eighth Floor (C) CITY: San Francisco (D) STATE: California (E) COUNTRY: USA (F) ZIP: 94111-3834 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: PCT/CA97/00347 (B) FILING DATE: 22-MAY-1997 (C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 08/657,753 (B) FILING DATE: 30-MAY-1996 (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Kezer, William B. (B) REGISTRATION NUMBER: 37,369 (C) REFERENCE/DOCKET NUMBER: 16303-003600PC (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (415) 576-0200 (B) TELEFAX: (415) 576-0300 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2490 base pairs (B) TYPE: nucleic acid . (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA SUBSTTTUTE SHEET (RULE 26) ( ix) FEATURE : (A) NAME/KEY: CDS (B) LOCATION: 117..1949 (D) OTHER INFORMATION: /product= "human endothelial leukocyte adhesion molecule l (ELAM-1)" (xi) SEQUENCE DESCRIPTION: SEQ ID N0:1: CCTGAGACAG AGGCAGCAGT GATACCCACC TGAGAGATCC TGTGTTTGAA CAACTGCTTC 60 CCAAAACGGA AAGTATTTCA AGCCTAAACC TTTGGGTGAA AAGAACTCTT GAAGTC 116 ATG ATT GCT TCA CAG TTT CTC TCA GCT CTC ACT TTG GTG CTT CTC ATT 164 Met He Ala Ser Gin Phe Leu Ser Ala Leu Thr Leu Val Leu Leu He 1 5 10 15 AAA GAG AGT GGA GCC TGG TCT TAC AAC ACC TCC ACG GAA GCT ATG ACT 212 Lys Glu Ser Gly Ala Trp Ser Tyr Asn Thr Ser Thr Glu Ala Met Thr 20 25 30 TAT GAT GAG GCC AGT GCT TAT TGT CAG CAA AGG TAC ACA CAC CTG GTT 260 Tyr Asp Glu Ala Ser Ala Tyr Cys Gin Gin Arg Tyr Thr His Leu Val 35 40 45 GCA ATT CAA AAC AAA GAA GAG ATT GAG TAC CTA AAC TCC ATA TTG AGC 308 Ala He Gin Asn Lys Glu Glu He Glu Tyr Leu Asn Ser He Leu Ser 50 55 60 TAT TCA CCA AGT TAT TAC TGG ATT GGA ATC AGA AAA GTC AAC AAT GTG 356 Tyr Ser Pro Ser Tyr Tyr Trp He Gly He Arg Lys Val Asn Asn Val 65 70 75 80 TGG GTC TGG GTA GGA ACC CAG AAA CCT CTG ACA GAA GAA GCC AAG AAC 404 Trp Val Trp Val Gly Thr Gin Lys Pro Leu Thr Glu Glu Ala Lys Asn 85 90 95 TGG GCT CCA GGT GAA CCC AAC AAT AGG CAA AAA GAT GAG GAC TGC GTG 452 Trp Ala Pro Gly Glu Pro Asn Asn Arg Gin Lys Asp Glu Asp Cys Val 100 105 110 GAG ATC TAC ATC AAG AGA GAA AAA GAT GTG GGC ATG TGG AAT GAT GAG 500 Glu He Tyr He Lys Arg Glu Lys Asp Val Gly Met Trp Asn Asp Glu 115 120 125 AGG TGC AGC AAG AAG AAG CTT GCC CTA TGC TAC ACA GCT GCC TGT ACC 548 Arg Cys Ser Lys Lys Lys Leu Ala Leu Cys Tyr Thr Ala Ala Cys Thr 130 135 140 AAT ACA TCC TGC AGT GGC CAC GGT GAA TGT GTA GAG ACC ATC AAT AAT 596 Asn Thr Ser Cys Ser Gly His Gly Glu Cys Val Glu Thr He Asn Asn 145 150 155 160 TAC ACT TGC AAG TGT GAC CCT GGC TTC AGT GGA CTC AAG TGT GAG CAA 644 Tyr Thr Cys Lys Cys Asp Pro Gly Phe Ser Gly Leu Lys Cys Glu Gin 165 170 175 ATT GTG AAC TGT ACA GCC CTG GAA TCC CCT GAG CAT GGA AGC CTG GTT 692 He Val Asn Cys Thr Ala Leu Glu Ser Pro Glu His Gly Ser Leu Val 180 185 190 TGC AGT CAC CCA CTG GGA AAC TTC AGC TAC AAT TCT TCC TGC TCT ATC 740 Cys Ser His Pro Leu Gly Asn Phe Ser Tyr Asn Ser Ser Cys Ser He 195 200 205 AGC TGT GAT AGG GGT TAC CTG CCA AGC AGC ATG GAG ACC ATG CAG TGT 788 Ser Cys Asp Arg Gly Tyr Leu Pro Ser Ser Met Glu Thr Met Gin Cys 210 215 220 SUBSTTTUTE SHEET (RULE 26) ATG TCC TCT GGA GAA TGG AGT GCT CCT ATT CCA GCC TGC AAT GTG GTT 836 Met Ser Ser Gly Glu Trp Ser Ala Pro He Pro Ala Cys Asn Val Val 225 230 235 240 GAG TGT GAT GCT GTG ACA AAT CCA GCC AAT GGG TTC GTG GAA TGT TTC 884 Glu Cys Asp Ala Val Thr Asn Pro Ala Asn Gly Phe Val Glu Cys Phe 245 250 255 CAA AAC CCT GGA AGC TTC CCA TGG AAC ACA ACC TGT ACA TTT GAC TGT 932 Gin Asn Pro Gly Ser Phe Pro Trp Asn Thr Thr Cys Thr Phe Asp Cys 260 265 270 GAA GAA GGA TTT GAA CTA ATG GGA GCC CAG AGC CTT CAG TGT ACC TCA 980 Glu Glu Gly Phe Glu Leu Met Gly Ala Gin Ser Leu Gin Cys Thr Ser 275 280 285 TCT GGG AAT TGG GAC AAC GAG AAG CCA ACG TGT AAA GCT GTG ACA TGC 1028 Ser Gly Asn Trp Asp Asn Glu Lys Pro Thr Cys Lys Ala Val Thr Cys 290 295 300 AGG GCC GTC CGC CAG CCT CAG AAT GGC TCT GTG AGG TGC AGC CAT TCC 1076 Arg Ala Val Arg Gin Pro Gin Asn Gly Ser Val Arg Cys Ser His Ser 305 310 315 320 CCT GCT GGA GAG TTC ACC TTC AAA TCA TCC TGC AAC TTC ACC TGT GAG 1124 Pro Ala Gly Glu Phe Thr Phe Lys Ser Ser Cys Asn Phe Thr Cys Glu 325 330 335 GAA GGC TTC ATG TTG CAG GGA CCA GCC CAG GTT GAA TGC ACC ACT CAA 1172 Glu Gly Phe Met Leu Gin Gly Pro Ala Gin Val Glu Cys Thr Thr Gin 340 345 350 GGG CAG TGG ACA CAG CAA ATC CCA GTT TGT GAA GCT TTC CAG TGC ACA 1220 Gly Gin Trp Thr Gin Gin He Pro Val Cys Glu Ala Phe Gin Cys Thr 355 360 365 GCC TTG TCC AAC CCC GAG CGA GGC TAC ATG AAT TGT CTT CCT AGT GCT 1268 Ala Leu Ser Asn Pro Glu Arg Gly Tyr Met Asn Cys Leu Pro Ser Ala 370 375 380 TCT GGC AGT TTC CGT TAT GGG TCC AGC TGT GAG TTC TCC TGT GAG CAG 1316 Ser Gly Ser Phe Arg Tyr Gly Ser Ser Cys Glu Phe Ser Cys Glu Gin 385 390 395 400 GGT TTT GTG TTG AAG GGA TCC AAA AGG CTC CAA TGT GGC CCC ACA GGG 1364 Gly Phe Val Leu Lys Gly Ser Lys Arg Leu Gin Cys Gly Pro Thr Gly 405 410 415 GAG TGG GAC AAC GAG AAG CCC ACA TGT GAA GCT GTG AGA TGC GAT GCT 1412 Glu Trp Asp Asn Glu Lys Pro Thr Cys Glu Ala Val Arg Cys Asp Ala 420 425 430 GTC CAC CAG CCC CCG AAG GGT TTG GTG AGG TGT GCT CAT TCC CCT ATT 1460 Val His Gin Pro Pro Lys Gly Leu Val Arg Cys Ala His Ser Pro He 435 440 445 GGA GAA TTC ACC TAC AAG TCC TCT TGT GCC TTC AGC TGT GAG GAG GGA 1508 Gly Glu Phe Thr Tyr Lys Ser Ser Cys Ala Phe Ser Cys Glu Glu Gly 450 455 460 TTT GAA TTA TAT GGA TCA ACT CAA CTT GAG TGC ACA TCT CAG GGA CAA 1556 Phe Glu Leu Tyr Gly Ser Thr Gin Leu Glu Cys Thr Ser Gin Gly Gin 465 470 475 480 TGG ACA GAA GAG GTT CCT TCC TGC CAA GTG GTA AAA TGT TCA AGC CTG 1604 Trp Thr Glu Glu Val Pro Ser Cys Gin Val Val Lys Cys Ser Ser Leu 485 490 495 SUBSTTTUTE SHEET (RULE 26) GCA GTT CCG GGA AAG ATC AAC ATG AGC TGC AGT GGG GAG CCC GTG TTT 1652 Ala Val Pro Gly Lys He Asn Met Ser Cys Ser Gly Glu Pro Val Phe 500 505 510 GGC ACT GTG TGC AAG TTC GCC TGT CCT GAA GGA TGG ACG CTC AAT GGC 1700 Gly Thr Val Cys Lys Phe Ala Cys Pro Glu Gly Trp Thr Leu Asn Gly 515 520 525 TCT GCA GCT CGG ACA TGT GGA GCC ACA GGA CAC TGG TCT GGC CTG CTA 1748 Ser Ala Ala Arg Thr Cys Gly Ala Thr Gly His Trp Ser Gly Leu Leu 530 535 540 CCT ACC TGT GAA GCT CCC ACT GAG TCC AAC ATT CCC TTG GTA GCT GGA 1796 Pro Thr Cys Glu Ala Pro Thr Glu Ser Asn He Pro Leu Val Ala Gly 545 550 555 560 CTT TCT GCT GCT GGA CTC TCC CTC CTG ACA TTA GCA CCA TTT CTC CTC 1844 Leu Ser Ala Ala Gly Leu Ser Leu Leu Thr Leu Ala Pro Phe Leu Leu 565 570 575 TGG CTT CGG AAA TGC TTA CGG AAA GCA AAG AAA TTT GTT CCT GCC AGC 1892 Trp Leu Arg Lys Cys Leu Arg Lys Ala Lys Lys Phe Val Pro Ala Ser 580 585 590 AGC TGC CAA AGC CTT GAA TCA GAC GGA AGC TAC CAA AAG CCT TCT TAC 1940 Ser Cys Gin Ser Leu Glu Ser Asp Gly Ser Tyr Gin Lys Pro Ser Tyr 595 600 605 ATC CTT TAAGTTCAAA AGAATCAGAA ACAGGTGCAT CTGGGGAACT AGAGGGATAC 1996 He Leu 610 ACTGAAGTTA ACAGAGACAG ATAACTCTCC TCGGGTCTCT GGCCCTTCTT GCCTACTATG 2056 CCAGATGCCT TTATGGCTGA AACCGCAACA CCCATCACCA CTTCAATAGA TCAAAGTCCA 2116 GCAGGCAAGG ACGGCCTTCA ACTGAAAAGA CTCAGTGTTC CCTTTCCTAC TCTCAGGATC 2176 AAGAAAGTGT TGGCTAATGA AGGGAAAGGA TATTTTCTTC CAAGCAAAGG TGAAGAGACC 2236 AAGACTCTGA AATCTCAGAA TTCCTTTTCT AACTCTCCCT TGCTCGCTGT AAAATCTTGG 2296 CACAGAAACA CAATATTTTG TGGCTTTCTT TCTTTTGCCC TTCACAGTGT TTCGACAGCT 2356 GATTACACAG TTGCTGTCAT AAGAATGAAT AATAATTATC CAGAGTTTAG AGGAAAAAAA 2416 TGACTAAAAA TATTATAACT TAAAAAAATG ACAGATGTTG AATGCCCACA GGCAAATGCA 2476 TGGAGGGTTG TTAA 2490 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 610 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2: Met He Ala Ser Gin Phe Leu Ser Ala Leu Thr Leu Val Leu Leu He 1 5 10 15 Lys Glu Ser Gly Ala Trp Ser Tyr Asn Thr Ser Thr Glu Ala Met Thr 20 25 30 SUBSTTTUTE SHEET (RULE 26) Tyr Asp Glu Ala Ser Ala Tyr Cys Gin Gin Arg Tyr Thr His Leu Val 35 40 45 Ala He Gin Asn Lys Glu Glu He Glu Tyr Leu Asn Ser He Leu Ser 50 55 60 Tyr Ser Pro Ser Tyr Tyr Trp He Gly He Arg Lys Val Asn Asn Val 65 70 75 80 Trp Val Trp Val Gly Thr Gin Lys Pro Leu Thr Glu Glu Ala Lys Asn 85 90 95 Trp Ala Pro Gly Glu Pro Asn Asn Arg Gin Lys Asp Glu Asp Cys Val 100 105 110 Glu He Tyr He Lys Arg Glu Lys Asp Val Gly Met Trp Asn Asp Glu 115 120 125 Arg Cys Ser Lys Lys Lys Leu Ala Leu Cys Tyr Thr Ala Ala Cys Thr 130 135 140 Asn Thr Ser Cys Ser Gly His Gly Glu Cys Val Glu Thr He Asn Asn 145 150 155 160 Tyr Thr Cys Lys Cys Asp Pro Gly Phe Ser Gly Leu Lys Cys Glu Gin 165 170 175 He Val Asn Cys Thr Ala Leu Glu Ser Pro Glu His Gly Ser Leu Val 180 185 190 Cys Ser His Pro Leu Gly Asn Phe Ser Tyr Asn Ser Ser Cys Ser He 195 200 205 Ser Cys Asp Arg Gly Tyr Leu Pro Ser Ser Met Glu Thr Met Gin Cys 210 215 220 Met Ser Ser Gly Glu Trp Ser Ala Pro He Pro Ala Cys Asn Val Val 225 230 235 240 Glu Cys Asp Ala Val Thr Asn Pro Ala Asn Gly Phe Val Glu Cys Phe 245 250 255 Gin Asn Pro Gly Ser Phe Pro Trp Asn Thr Thr Cys Thr Phe Asp Cys 260 265 270 Glu Glu Gly Phe Glu Leu Met Gly Ala Gin Ser Leu Gin Cys Thr Ser 275 280 285 Ser Gly Asn Trp Asp Asn Glu Lys Pro Thr Cys Lys Ala Val Thr Cys 290 295 300 Arg Ala Val Arg Gin Pro Gin Asn Gly Ser Val Arg Cys Ser His Ser 305 310 315 320 Pro Ala Gly Glu Phe Thr Phe Lys Ser Ser Cys Asn Phe Thr Cys Glu 325 330 335 Glu Gly Phe Met Leu Gin Gly Pro Ala Gin Val Glu Cys Thr Thr Gin 340 345 350 Gly Gin Trp Thr Gin Gin He Pro Val Cys Glu Ala Phe Gin Cys Thr 355 360 365 Ala Leu Ser Asn Pro Glu Arg Gly Tyr Met Asn Cys Leu Pro Ser Ala 370 375 380 SUBSTTTUTE SHEET (RULE 25) Ser Gly Ser Phe Arg Tyr Gly Ser Ser Cys Glu Phe Ser Cys Glu Gin 385 390 395 400 Gly Phe Val Leu Lys Gly Ser Lys Arg Leu Gin Cys Gly Pro Thr Gly 405 410 415 Glu Trp Asp Asn Glu Lys Pro Thr Cys Glu Ala Val Arg Cys Asp Ala 420 425 430 Val His Gin Pro Pro Lys Gly Leu Val Arg Cys Ala His Ser Pro He 435 440 445 Gly Glu Phe Thr Tyr Lys Ser Ser Cys Ala Phe Ser Cys Glu Glu Gly 450 455 460 Phe Glu Leu Tyr Gly Ser Thr Gin Leu Glu Cys Thr Ser Gin Gly Gin 465 470 475 480 Trp Thr Glu Glu Val Pro Ser Cys Gin Val Val Lys Cys Ser Ser Leu 485 490 495 Ala Val Pro Gly Lys He Asn Met Ser Cys Ser Gly Glu Pro Val Phe 500 505 510 Gly Thr Val Cys Lys Phe Ala Cys Pro Glu Gly Trp Thr Leu Asn Gly 515 520 525 Ser Ala Ala Arg Thr Cys Gly Ala Thr Gly His Trp Ser Gly Leu Leu 530 535 540 Pro Thr Cys Glu Ala Pro Thr Glu Ser Asn He Pro Leu Val Ala Gly 545 550 555 560 Leu Ser Ala Ala Gly Leu Ser Leu Leu Thr Leu Ala Pro Phe Leu Leu 565 570 575 Trp Leu Arg Lys Cys Leu Arg Lys Ala Lys Lys Phe Val Pro Ala Ser 580 585 590 Ser Cys Gin Ser Leu Glu Ser Asp Gly Ser Tyr Gin Lys Pro Ser Tyr 595 600 605 He Leu 610 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2220 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS . (B) LOCATION: 1..2220 (D) OTHER INFORMATION: /product= "human vascular cell adhesion molecule 1 (VCAM-1) " SUBSTTTUTE SHEET (RULE 26) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: ATG CCT GGG AAG ATG GTC GTG ATC CTT GGA GCC TCA AAT ATA CTT TGG 48 Met Pro Gly Lys Met Val Val He Leu Gly Ala Ser Asn He Leu Trp 1 5 10 15 ATA ATG TTT GCA GCT TCT CAA GCT TTT AAA ATC GAG ACC ACC CCA GAA 96 He Met Phe Ala Ala Ser Gin Ala Phe Lys He Glu Thr Thr Pro Glu 20 25 30 TCT AGA TAT CTT GCT CAG ATT GGT GAC TCC GTC TCA TTG ACT TGC AGC 144 Ser Arg Tyr Leu Ala Gin He Gly Asp Ser Val Ser Leu Thr Cys Ser 35 40 45 ACC ACA GGC TGT GAG TCC CCA TTT TTC TCT TGG AGA ACC CAG ATA GAT 192 Thr Thr Gly Cys Glu Ser Pro Phe Phe Ser Trp Arg Thr Gin He Asp 50 55 60 AGT CCA CTG AAT GGG AAG GTG ACG AAT GAG GGG ACC ACA TCT ACG CTG 240 Ser Pro Leu Asn Gly Lys Val Thr Asn Glu Gly Thr Thr Ser Thr Leu 65 70 75 80 ACA ATG AAT CCT GTT AGT TTT GGG AAC GAA CAC TCT TAC CTG TGC ACA 288 Thr Met Asn Pro Val Ser Phe Gly Asn Glu His Ser Tyr Leu Cys Thr 85 90 95 GCA ACT TGT GAA TCT AGG AAA TTG GAA AAA GGA ATC CAG GTG GAG ATC 336 Ala Thr Cys Glu Ser Arg Lys Leu Glu Lys Gly He Gin Val Glu He 100 105 110 TAC TCT TTT CCT AAG GAT CCA GAG ATT CAT TTG AGT GGC CCT CTG GAG 384 Tyr Ser Phe Pro Lys Asp Pro Glu He His Leu Ser Gly Pro Leu Glu 115 120 125 GCT GGG AAG CCG ATC ACA GTC AAG TGT TCA GTT GCT GAT GTA TAC CCA 432 Ala Gly Lys Pro He Thr Val Lys Cys Ser Val Ala Asp Val Tyr Pro 130 135 140 TTT GAC AGG CTG GAG ATA GAC TTA CTG AAA GGA GAT CAT CTC ATG AAG 480 Phe Asp Arg Leu Glu He Asp Leu Leu Lys Gly Asp His Leu Met Lys 145 150 155 160 AGT CAG GAA TTT CTG GAG GAT GCA GAC AGG AAG TCC CTG GAA ACC AAG 528 Ser Gin Glu Phe Leu Glu Asp Ala Asp Arg Lys Ser Leu Glu Thr Lys 165 170 175 AGT TTG GAA GTA ACC TTT ACT CCT GTC ATT GAG GAT ATT GGA AAA GTT 576 Ser Leu Glu Val Thr Phe Thr Pro Val He Glu Asp He Gly Lys Val 180 185 190 CTT GTT TGC CGA GCT AAA TTA CAC ATT GAT GAA ATG GAT TCT GTG CCC 624 Leu Val Cys Arg Ala Lys Leu His He Asp Glu Met Asp Ser Val Pro 195 200 205 ACA GTA AGG CAG GCT GTA AAA GAA TTG CAA GTC TAC ATA TCA CCC AAG 672 Thr Val Arg Gin Ala Val Lys Glu Leu Gin Val Tyr He Ser Pro Lys 210 215 220 AAT ACA GTT ATT TCT GTG AAT CCA TCC ACA AAG CTG CAA GAA GGT GGC 720 ' Asn Thr -Val He Ser Val Asn Pro Ser Thr Lys Leu Gin Glu Gly Gly 225 230 235 240 TCT GTG ACC ATG ACC TGT TCC AGC GAG GGT CTA CCA GCT CCA GAG ATT 768 Ser Val Thr Met Thr Cys Ser Ser Glu Gly Leu Pro Ala Pro Glu He 245 250 255 SUBSTTTUTE SHEET (RULE 26) TTC TGG AGT AAG AAA TTA GAT AAT GGG AAT CTA CAG CAC CTT TCT GGA 816 Phe Trp Ser Lys Lys Leu Asp Asn Gly Asn Leu Gin His Leu Ser Gly 260 265 270 AAT GCA ACT CTC ACC TTA ATT GCT ATG AGG ATG GAA GAT TCT GGA ATT 864 Asn Ala Thr Leu Thr Leu He Ala Met Arg Met Glu Asp Ser Gly He 275 280 285 TAT GTG TGT GAA GGA GTT AAT TTG ATT GGG AAA AAC AGA AAA GAG GTG 912 Tyr Val Cys Glu Gly Val Asn Leu He Gly Lys Asn Arg Lys Glu Val 290 295 300 GAA TTA ATT GTT CAA GAG AAA CCA TTT ACT GTT GAG ATC TCC CCT GGA 960 Glu Leu He Val Gin Glu Lys Pro Phe Thr Val Glu He Ser Pro Gly 305 310 315 320 CCC CGG ATT GCT GCT CAG ATT GGA GAC TCA GTC ATG TTG ACA TGT AGT 1008 Pro Arg He Ala Ala Gin He Gly Asp Ser Val Met Leu Thr Cys Ser 325 330 335 GTC ATG GGC TGT GAA TCC CCA TCT TTC TCC TGG AGA ACC CAG ATA GAC 1056 Val Met Gly Cys Glu Ser Pro Ser Phe Ser Trp Arg Thr Gin He Asp 340 345 350 AGC CCT CTG AGC GGG AAG GTG AGG AGT GAG GGG ACC AAT TCC ACG CTG 1104 Ser Pro Leu Ser Gly Lys Val Arg Ser Glu Gly Thr Asn Ser Thr Leu 355 360 365 ACC CTG AGC CCT GTG AGT TTT GAG AAC GAA CAC TCT TAT CTG TGC ACA 1152 Thr Leu Ser Pro Val Ser Phe Glu Asn Glu His Ser Tyr Leu Cys Thr 370 375 380 GTG ACT TGT GGA CAT AAG AAA CTG GAA AAG GGA ATC CAG GTG GAG CTC 1200 Val Thr Cys Gly His Lys Lys Leu Glu Lys Gly He Gin Val Glu Leu 385 390 395 400 TAC TCA TTC CCT AGA GAT CCA GAA ATC GAG ATG AGT GGT GGC CTC GTG 1248 Tyr Ser Phe Pro Arg Asp Pro Glu He Glu Met Ser Gly Gly Leu Val 405 410 415 AAT GGG AGC TCT GTC ACT GTA AGC TGC AAG GTT CCT AGC GTG TAC CCC 1296 Asn Gly Ser Ser Val Thr Val Ser Cys Lys Val Pro Ser Val Tyr Pro 420 425 430 CTT GAC CGG CTG GAG ATT GAA TTA CTT AAG GGG GAG ACT ATT CTG GAG 1344 Leu Asp Arg Leu Glu He Glu Leu Leu Lys Gly Glu Thr He Leu Glu 435 440 445 AAT ATA GAG TTT TTG GAG GAT ACG GAT ATG AAA TCT CTA GAG AAC AAA 1392 Asn He Glu Phe Leu Glu Asp Thr Asp Met Lys Ser Leu Glu Asn Lys 450 455 460 AGT TTG GAA ATG ACC TTC ATC CCT ACC ATT GAA GAT ACT GGA AAA GCT 1440 Ser Leu Glu Met Thr Phe He Pro Thr He Glu Asp Thr Gly Lys Ala 465 470 475 480 CTT GTT TGT CAG GCT AAG TTA CAT ATT GAT GAC ATG GAA TTC GAA CCC 1488 Leu Val Cys Gin Ala Lys Leu His He Asp Asp Met Glu Phe Glu Pro 485 490 495 AAA CAA AGG CAG AGT ACG CAA ACA CTT TAT GTC AAT GTT GCC CCC AGA 1536 Lys Gin Arg Gin Ser Thr Gin Thr Leu Tyr Val Asn Val Ala Pro Arg 500 505 510 GAT ACA ACC GTC TTG GTC AGC CCT TCC TCC ATC CTG GAG GAA GGC AGT 1584 Asp Thr Thr Val Leu Val Ser Pro Ser Ser He Leu Glu Glu Gly Ser 515 520 525 SUBSTTTUTE SHEET (RULE 26) TCT GTG AAT ATG ACA TGC TTG AGC CAG GGC TTT CCT GCT CCG AAA ATC 1632 Ser Val Asn Met Thr Cys Leu Ser Gin Gly Phe Pro Ala Pro Lys He 530 535 540 CTG TGG AGC AGG CAG CTC CCT AAC GGG GAG CTA CAG CCT CTT TCT GAG 1680 Leu Trp Ser Arg Gin Leu Pro Asn Gly Glu Leu Gin Pro Leu Ser Glu 545 550 555 560 AAT GCA ACT CTC ACC TTA ATT TCT ACA AAA ATG GAA GAT TCT GGG GTT 1728 Asn Ala Thr Leu Thr Leu He Ser Thr Lys Met Glu Asp Ser Gly Val 565 570 575 TAT TTA TGT GAA GGA ATT AAC CAG GCT GGA AGA AGC AGA AAG GAA GTG 1776 Tyr Leu Cys Glu Gly He Asn Gin Ala Gly Arg Ser Arg Lys Glu Val 580 585 590 GAA TTA ATT ATC CAA GTT ACT CCA AAA GAC ATA AAA CTT ACA GCT TTT 1824 Glu Leu He He Gin Val Thr Pro Lys Asp He Lys Leu Thr Ala Phe 595 600 605 CCT TCT GAG AGT GTC AAA GAA GGA GAC ACT GTC ATC ATC TCT TGT ACA 1872 Pro Ser Glu Ser Val Lys Glu Gly Asp Thr Val He He Ser Cys Thr 610 615 620 TGT GGA AAT GTT CCA GAA ACA TGG ATA ATC CTG AAG AAA AAA GCG GAG 1920 Cys Gly Asn Val Pro Glu Thr Trp He He Leu Lys Lys Lys Ala Glu 625 630 635 640 ACA GGA GAC ACA GTA CTA AAA TCT ATA GAT GGC GCC TAT ACC ATC CGA 1968 Thr Gly Asp Thr Val Leu Lys Ser He Asp Gly Ala Tyr Thr He Arg 645 650 655 AAG GCC CAG TTG AAG GAT GCG GGA GTA TAT GAA TGT GAA TCT AAA AAC 2016 Lys Ala Gin Leu Lys Asp Ala Gly Val Tyr Glu Cys Glu Ser Lys Asn 660 665 670 AAA GTT GGC TCA CAA TTA AGA AGT TTA ACA CTT GAT GTT CAA GGA AGA 2064 Lys Val Gly Ser Gin Leu Arg Ser Leu Thr Leu Asp Val Gin Gly Arg 675 680 685 GAA AAC AAC AAA GAC TAT TTT TCT CCT GAG CTT CTC GTG CTC TAT TTT 2112 Glu Asn Asn Lys Asp Tyr Phe Ser Pro Glu Leu Leu Val Leu Tyr Phe 690 695 700 GCA TCC TCC TTA ATA ATA CCT GCC ATT GGA ATG ATA ATT TAC TTT GCA 2160 Ala Ser Ser Leu He He Pro Ala He Gly Met He He Tyr Phe Ala 705 710 715 720 AGA AAA GCC AAC ATG AAG GGG TCA TAT AGT CTT GTA GAA GCA CAG AAA 2208 Arg Lys Ala Asn Met Lys Gly Ser Tyr Ser Leu Val Glu Ala Gin Lys 725 730 735 TCA AAA GTG TAG 2220 Ser Lys Val (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 739 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein SUBSTTTUTE SHEET (RULE 26) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4: Met Pro Gly Lys Met Val Val He Leu Gly Ala Ser Asn He Leu Trp 1 5 10 15 He Met Phe Ala Ala Ser Gin Ala Phe Lys He Glu Thr Thr Pro Glu 20 25 30 Ser Arg Tyr Leu Ala Gin He Gly Asp Ser Val Ser Leu Thr Cys Ser 35 40 45 Thr Thr Gly Cys Glu Ser Pro Phe Phe Ser Trp Arg Thr Gin He Asp 50 55 60 Ser Pro Leu Asn Gly Lys Val Thr Asn Glu Gly Thr Thr Ser Thr Leu 65 70 75 80 Thr Met Asn Pro Val Ser Phe Gly Asn Glu His Ser Tyr Leu Cys Thr 85 90 95 Ala Thr Cys Glu Ser Arg Lys Leu Glu Lys Gly He Gin Val Glu He 100 105 110 Tyr Ser Phe Pro Lys Asp Pro Glu He His Leu Ser Gly Pro Leu Glu 115 120 125 Ala Gly Lys Pro He Thr Val Lys Cys Ser Val Ala Asp Val Tyr Pro 130 135 140 Phe Asp Arg Leu Glu He Asp Leu Leu Lys Gly Asp His Leu Met Lys 145 150 155 160 Ser Gin Glu Phe Leu Glu Asp Ala Asp Arg Lys Ser Leu Glu Thr Lys 165 170 175 Ser Leu Glu Val Thr Phe Thr Pro Val He Glu Asp He Gly Lys Val 180 185 190 Leu Val Cys Arg Ala Lys Leu His He Asp Glu Met Asp Ser Val Pro 195 200 205 Thr Val Arg Gin Ala Val Lys Glu Leu Gin Val Tyr He Ser Pro Lys 210 215 220 Asn Thr Val He Ser Val Asn Pro Ser Thr Lys Leu Gin Glu Gly Gly 225 230 235 240 Ser Val Thr Met Thr Cys Ser Ser Glu Gly Leu Pro Ala Pro Glu He 245 250 255 Phe Trp Ser Lys Lys Leu Asp Asn Gly Asn Leu Gin His Leu Ser Gly 260 265 270 Aβn Ala Thr Leu Thr Leu He Ala Met Arg Met Glu Asp Ser Gly He 275 280 285 Tyr Val Cys Glu Gly Val Asn Leu He Gly Lys Asn Arg Lys Glu Val 290 295 300 " Glu Leu ' He Val Gin Glu Lys Pro Phe Thr Val Glu He Ser Pro Gly 305 310 315 320 Pro Arg He Ala Ala Gin He Gly Asp Ser Val Met Leu Thr Cys Ser 325 330 335 Val Met Gly Cys Glu Ser Pro Ser Phe Ser Trp Arg Thr Gin He Asp 340 345 350 SUBSTTTUTE SHEET (RULE 25) Ser Pro Leu Ser Gly Lys Val Arg Ser Glu Gly Thr Asn Ser Thr Leu 355 360 365 Thr Leu Ser Pro Val Ser Phe Glu Asn Glu His Ser Tyr Leu Cys Thr 370 375 380 Val Thr Cys Gly His Lys Lys Leu Glu Lys Gly He Gin Val Glu Leu 385 390 395 400 Tyr Ser Phe Pro Arg Asp Pro Glu He Glu Met Ser Gly Gly Leu Val 405 410 415 Asn Gly Ser Ser Val Thr Val Ser Cys Lys Val Pro Ser Val Tyr Pro 420 425 430 Leu Asp Arg Leu Glu He Glu Leu Leu Lys Gly Glu Thr He Leu Glu 435 440 445 Asn He Glu Phe Leu Glu Asp Thr Asp Met Lys Ser Leu Glu Asn Lys 450 455 460 Ser Leu Glu Met Thr Phe He Pro Thr He Glu Asp Thr Gly Lys Ala 465 470 475 480 Leu Val Cys Gin Ala Lys Leu His He Asp Asp Met Glu Phe Glu Pro 485 490 495 Lys Gin Arg Gin Ser Thr Gin Thr Leu Tyr Val Asn Val Ala Pro Arg 500 505 510 Asp Thr Thr Val Leu Val Ser Pro Ser Ser He Leu Glu Glu Gly Ser 515 520 525 Ser Val Asn Met Thr Cys Leu Ser Gin Gly Phe Pro Ala Pro Lys He 530 535 540 Leu Trp Ser Arg Gin Leu Pro Asn Gly Glu Leu Gin Pro Leu Ser Glu 545 550 555 560 Asn Ala Thr Leu Thr Leu He Ser Thr Lys Met Glu Asp Ser Gly Val 565 570 575 Tyr Leu Cys Glu Gly He Asn Gin Ala Gly Arg Ser Arg Lys Glu Val 580 585 590 Glu Leu He He Gin Val Thr Pro Lys Asp He Lys Leu Thr Ala Phe 595 600 605 Pro Ser Glu Ser Val Lys Glu Gly Asp Thr Val He He Ser Cys Thr 610 615 620 Cys Gly Asn Val Pro Glu Thr Trp He He Leu Lys Lys Lys Ala Glu 625 630 635 640 Thr Gly Asp Thr Val Leu Lys Ser He Asp Gly Ala Tyr Thr He Arg 645 650 655 Lys Ala Gin Leu Lys Asp Ala Gly Val Tyr Glu Cys Glu Ser Lys Asn 660 665 670 Lys Val Gly Ser Gin Leu Arg Ser Leu Thr Leu Asp Val Gin Gly Arg 675 680 685 Glu Asn Asn Lys Asp Tyr Phe Ser Pro Glu Leu Leu Val Leu Tyr Phe 690 695 700 SUBSTTTUTE SHEET (RULE 26) Ala Ser Ser Leu He He Pro Ala He Gly Met He He Tyr Phe Ala 705 710 715 720 Arg Lys Ala Asn Met Lys Gly Ser Tyr Ser Leu Val Glu Ala Gin Lys 725 730 735 Ser Lys Val (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1846 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 13..1611 (D) OTHER INFORMATION: /product= "human intercellular adhesion molecule 1 (ICAM-1)" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: CTCAGCCTCG CT ATG GCT CCC AGC AGC CCC CGG CCC GCG CTG CCC GCA 48 Met Ala Pro Ser Ser Pro Arg Pro Ala Leu Pro Ala 1 5 10 CTC CTG GTC CTG CTC GGG GCT CTG TTC CCA GGA CCT GGC AAT GCC CAG 96 Leu Leu Val Leu Leu Gly Ala Leu Phe Pro Gly Pro Gly Asn Ala Gin 15 20 25 ACA TCT GTG TCC CCC TCA AAA GTC ATC CTG CCC CGG GGA GGC TCC GTG 144 Thr Ser Val Ser Pro Ser Lys Val He Leu Pro Arg Gly Gly Ser Val 30 35 40 CTG GTG ACA TGC AGC ACC TCC TGT GAC CAG CCC AAG TTG TTG GGC ATA 192 Leu Val Thr Cys Ser Thr Ser Cys Asp Gin Pro Lys Leu Leu Gly He 45 50 55 60 GAG ACC CCG TTG CCT AAA AAG GAG TTG CTC CTG CCT GGG AAC AAC CGG 240 Glu Thr Pro Leu Pro Lys Lys Glu Leu Leu Leu Pro Gly Asn Asn Arg 65 70 75 AAG GTG TAT GAA CTG AGC AAT GTG CAA GAA GAT AGC CAA CCA ATG TGC 288 Lys Val Tyr Glu Leu Ser Asn Val Gin Glu Asp Ser Gin Pro Met Cys 80 85 90 TAT TCA AAC TGC CCT GAT GGG CAG TCA ACA GCT AAA ACC TTC CTC ACC 336 Tyr Ser Asn Cys Pro Asp Gly Gin Ser Thr Ala Lys Thr Phe Leu Thr 95 100 105 GTG TAC TGG ACT CCA GAA CGG GTG GAA CTG GCA CCC CTC CCC TCT TGG 384 Val Tyr Trp Thr Pro Glu Arg Val Glu Leu Ala Pro Leu Pro Ser Trp 110 115 120 CAG CCA GTG GGC AAG AAC CTT ACC CTA CGC TGC CAG GTG GAG GGT GGG 432 Gin Pro Val Gly Lys Asn Leu Thr Leu Arg Cys Gin Val Glu Gly Gly 125 130 135 140 SUBSTTTUTE SHEET (RULE 26) GCA CCC CGG GCC AAC CTC ACC GTG GTG CTG CTC CGT GGG GAG AAG GAG 480 Ala Pro Arg Ala Asn Leu Thr Val Val Leu Leu Arg Gly Glu Lys Glu 145 150 155 CTG AAA CGG GAG CCA GCT GTG GGG GAG CCC GCT GAG GTC ACG ACC ACG 528 Leu Lys Arg Glu Pro Ala Val Gly Glu Pro Ala Glu Val Thr Thr Thr 160 165 170 GTG CTG GTG AGG AGA GAT CAC CAT GGA GCC AAT TTC TCG TGC CGC ACT 576 Val Leu Val Arg Arg Asp His His Gly Ala Asn Phe Ser Cys Arg Thr 175 180 185 GAA CTG GAC CTG CGG CCC CAA GGG CTG GAG CTG TTT GAG AAC ACC TCG 624 Glu Leu Asp Leu Arg Pro Gin Gly Leu Glu Leu Phe Glu Asn Thr Ser 190 195 200 GCC CCC TAC CAG CTC CAG ACC TTT GTC CTG CCA GCG ACT CCC CCA CAA 672 Ala Pro Tyr Gin Leu Gin Thr Phe Val Leu Pro Ala Thr Pro Pro Gin 205 210 215 220 CTT GTC AGC CCC CGG GTC CTA GAG GTG GAC ACG CAG GGG ACC GTG GTC 720 Leu Val Ser Pro Arg Val Leu Glu Val Asp Thr Gin Gly Thr Val Val 225 230 235 TGT TCC CTG GAC GGG CTG TTC CCA GTC TCG GAG GCC CAG GTC CAC CTG 768 Cys Ser Leu Asp Gly Leu Phe Pro Val Ser Glu Ala Gin Val His Leu 240 245 250 GCA CTG GGG GAC CAG AGG TTG AAC CCC ACA GTC ACC TAT GGC AAC GAC 816 Ala Leu Gly Asp Gin Arg Leu Asn Pro Thr Val Thr Tyr Gly Asn Asp 255 260 265 TCC TTC TCG GCC AAG GCC TCA GTC AGT GTG ACC GCA GAG GAC GAG GGC 864 Ser Phe Ser Ala Lys Ala Ser Val Ser Val Thr Ala Glu Asp Glu Gly 270 275 280 ACC CAG CGG CTG ACG TGT GCA GTA ATA CTG GGG AAC CAG AGC CAG GAG 912 Thr Gin Arg Leu Thr Cys Ala Val He Leu Gly Asn Gin Ser Gin Glu 285 290 295 300 ACA CTG CAG ACA GTG ACC ATC TAC AGC TTT CCG GCG CCC AAC GTG ATT 960 Thr Leu Gin Thr Val Thr He Tyr Ser Phe Pro Ala Pro Asn Val He 305 310 315 CTG ACG AAG CCA GAG GTC TCA GAA GGG ACC GAG GTG ACA GTG AAG TGT 1008 Leu Thr Lys Pro Glu Val Ser Glu Gly Thr Glu Val Thr Val Lys Cys 320 325 330 GAG GCC CAC CCT AGA GCC AAG GTG ACG CTG AAT GGG GTT CCA GCC CAG 1056 Glu Ala His Pro Arg Ala Lys Val Thr Leu Asn Gly Val Pro Ala Gin 335 340 345 CCA CTG GGC CCG AGG GCC CAG CTC CTG CTG AAG GCC ACC CCA GAG GAC 1104 Pro Leu Gly Pro Arg Ala Gin Leu Leu Leu Lys Ala Thr Pro Glu Asp 350 355 360 AAC GGG CGC AGC TTC TCC TGC TCT GCA ACC CTG GAG GTG GCC GGC CAG 1152 Asn Gly Arg Ser Phe Ser Cys Ser Ala Thr Leu Glu Val Ala Gly Gin 365 370 375 380 CTT ATA CAC AAG AAC CAG ACC CGG GAG CTT CGT GTC CTG TAT GGC CCC 1200 Leu He His Lys Asn Gin Thr Arg Glu Leu Arg Val Leu Tyr Gly Pro 385 390 395 CGA CTG GAC GAG AGG GAT TGT CCG GGA AAC TGG ACG TGG CCA GAA AAT 1248 Arg Leu Asp Glu Arg Asp Cys Pro Gly Asn Trp Thr Trp Pro Glu Asn 400 405 410 SUBSTTTUTE SHEET (RULE 26) TCC CAG CAG ACT CCA ATG TGC CAG GCT TGG GGG AAC CCA TTG CCC GAG 1296 Ser Gin Gin Thr Pro Met Cys Gin Ala Trp Gly Asn Pro Leu Pro Glu 415 420 425 CTC AAG TGT CTA AAG GAT GGC ACT TTC CCA CTG CCC ATC GGG GAA TCA 1344 Leu Lys Cys Leu Lys Asp Gly Thr Phe Pro Leu Pro He Gly Glu Ser 430 435 440 GTG ACT GTC ACT CGA GAT CTT GAG GGC ACC TAC CTC TGT CGG GCC AGG 1392 Val Thr Val Thr Arg Asp Leu Glu Gly Thr Tyr Leu Cys Arg Ala Arg 445 450 455 460 AGC ACT CAA GGG GAG GTC ACC CGC GAG GTG ACC GTG AAT GTG CTC TCC 1440 Ser Thr Gin Gly Glu Val Thr Arg Glu Val Thr Val Asn Val Leu Ser 465 470 475 CCC CGG TAT GAG ATT GTC ATC ATC ACT GTG GTA GCA GCC GCA GTC ATA 1488 Pro Arg Tyr Glu He Val He He Thr Val Val Ala Ala Ala Val He 480 485 490 ATG GGC ACT GCA GGC CTC AGC ACG TAC CTC TAT AAC CGC CAG CGG AAG 1536 Met Gly Thr Ala Gly Leu Ser Thr Tyr Leu Tyr Asn Arg Gin Arg Lys 495 500 505 ATC AAG AAA TAC AGA CTA CAA CAG GCC CAA AAA GGG ACC CCC ATG AAA 1584 He Lys Lys Tyr Arg Leu Gin Gin Ala Gin Lys Gly Thr Pro Met Lys 510 515 520 CCG AAC ACA CAA GCC ACG CCT CCC TGAACCTATC CCGGGACAGG GCCTCTTCCT 1638 Pro Asn Thr Gin Ala Thr Pro Pro 525 530 CGGCCTTCCC ATATTGGTGG CAGTGGTGCC ACACTGAACA GAGTGGAAGA CATATGCCAT 1698 GCAGCTACAC CTACCGGCCC TGGGACGCCG GAGGACAGGG CATTGTCCTC AGTCAGATAC 1758 AACAGCATTT GGGGCCATGG TACCTGCACA CCTAAAACAC TAGGCCACGC ATCTGATCTG 1818 TAGTCACATG ACTAAGCCAA GAGGAAGG 1846 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 532 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Met Ala Pro Ser Ser Pro Arg Pro Ala Leu Pro Ala Leu Leu Val Leu 1 5 10 15 Leu Gly Ala Leu Phe Pro Gly Pro Gly Asn Ala Gin Thr Ser Val Ser 20 25 30 Pro Ser Lys Val He Leu Pro Arg Gly Gly Ser Val Leu Val Thr Cys ' 35 40 45 Ser Thr Ser Cys Asp Gin Pro Lys Leu Leu Gly He Glu Thr Pro Leu 50 55 60 Pro Lys Lys Glu Leu Leu Leu Pro Gly Asn Asn Arg Lys Val Tyr Glu 65 70 75 80 SUBSTTTUTE SHEET (RULE 26) Leu Ser Asn Val Gin Glu Asp Ser Gin Pro Met Cys Tyr Ser Asn Cys 85 90 95 Pro Asp Gly Gin Ser Thr Ala Lys Thr Phe Leu Thr Val Tyr Trp Thr 100 105 110 Pro Glu Arg Val Glu Leu Ala Pro Leu Pro Ser Trp Gin Pro Val Gly 115 120 125 Lys Asn Leu Thr Leu Arg Cys Gin Val Glu Gly Gly Ala Pro Arg Ala 130 135 140 Asn Leu Thr Val Val Leu Leu Arg Gly Glu Lys Glu Leu Lys Arg Glu 145 150 155 160 Pro Ala Val Gly Glu Pro Ala Glu Val Thr Thr Thr Val Leu Val Arg 165 170 175 Arg Asp His His Gly Ala Asn Phe Ser Cys Arg Thr Glu Leu Asp Leu 180 185 190 Arg Pro Gin Gly Leu Glu Leu Phe Glu Asn Thr Ser Ala Pro Tyr Gin 195 200 205 Leu Gin Thr Phe Val Leu Pro Ala Thr Pro Pro Gin Leu Val Ser Pro 210 215 220 Arg Val Leu Glu Val Asp Thr Gin Gly Thr Val Val Cys Ser Leu Asp 225 230 235 240 Gly Leu Phe Pro Val Ser Glu Ala Gin Val His Leu Ala Leu Gly Asp 245 250 255 Gin Arg Leu Asn Pro Thr Val Thr Tyr Gly Asn Asp Ser Phe Ser Ala 260 265 270 Lys Ala Ser Val Ser Val Thr Ala Glu Asp Glu Gly Thr Gin Arg Leu 275 280 285 Thr Cys Ala Val He Leu Gly Asn Gin Ser Gin Glu Thr Leu Gin Thr 290 295 300 Val Thr He Tyr Ser Phe Pro Ala Pro Asn Val He Leu Thr Lys Pro 305 310 315 320 Glu Val Ser Glu Gly Thr Glu Val Thr Val Lys Cys Glu Ala His Pro 325 330 335 Arg Ala Lys Val Thr Leu Asn Gly Val Pro Ala Gin Pro Leu Gly Pro 340 345 350 Arg Ala Gin Leu Leu Leu Lys Ala Thr Pro Glu Asp Asn Gly Arg Ser 355 360 365 Phe Ser Cys Ser Ala Thr Leu Glu Val Ala Gly Gin Leu He His Lys 370 375 380 Asn Gin Thr Arg Glu Leu Arg Val Leu Tyr Gly Pro Arg Leu Asp Glu 385 390 395 400 Arg Asp Cys Pro Gly Asn Trp Thr Trp Pro Glu Asn Ser Gin Gin Thr 405 410 415 Pro Met Cys Gin Ala Trp Gly Asn Pro Leu Pro Glu Leu Lys Cys Leu 420 425 430 SUBSTTTUTE SHEET (RULE 26) Lys Asp Gly Thr Phe Pro Leu Pro He Gly Glu Ser Val Thr Val Thr 435 440 445 Arg Asp Leu Glu Gly Thr Tyr Leu Cys Arg Ala Arg Ser Thr Gin Gly 450 455 460 Glu Val Thr Arg Glu Val Thr Val Asn Val Leu Ser Pro Arg Tyr Glu 465 470 475 480 He Val He He Thr Val Val Ala Ala Ala Val He Met Gly Thr Ala 485 490 495 Gly Leu Ser Thr Tyr Leu Tyr Asn Arg Gin Arg Lys He Lys Lys Tyr 500 505 510 Arg Leu Gin Gin Ala Gin Lys Gly Thr Pro Met Lys Pro Asn Thr Gin 515 520 525 Ala Thr Pro Pro 530 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2522 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 23..1636 (D) OTHER INFORMATION: /product- "mouse intercellular adhesion molecule 1 (ICAM-l) " (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: AGCACTCTGC CCTGGCCCTG CA ATG GCT TCA ACC CGT GCC AAG CCC ACG CTA 52 Met Ala Ser Thr Arg Ala Lys Pro Thr Leu 1 5 10 CCT CTG CTC CTG GCC CTG GTC ACC GTT GTG ATC CCT GGG CCT GGT GAT 100 Pro Leu Leu Leu Ala Leu Val Thr Val Val He Pro Gly Pro Gly Asp 15 20 25 GCT CAG GTA TCC ATC CAT CCC AGA GAA GCC TTC CTG CCC CAG GGT GGG 148 Ala Gin Val Ser He His Pro Arg Glu Ala Phe Leu Pro Gin Gly Gly 30 35 40 TCC GTG CAG GTG AAC TGT TCT TCC TCA TGC AAG GAG GAC CTC AGC CTG 196 Ser Val Gin Val Asn Cys Ser Ser Ser Cys Lys Glu Asp Leu Ser Leu 45 50 55 GGC TTG GAG ACT CAG TGG CTG AAA GAT GAG CTC GAG AGT GGA CCC AAC 244 -Gly Leu Glu Thr Gin Trp Leu Lys Asp Glu Leu Glu Ser Gly Pro Asn 60 65 70 TGG AAG CTG TTT GAG CTG AGC GAG ATC GGG GAG GAC AGC AGT CCG CTG 292 Trp Lys Leu Phe Glu Leu Ser Glu He Gly Glu Asp Ser Ser Pro Leu 75 80 85 90 SUBSTTTUTE SHEET (RULE 26) TGC TTT GAG AAC TGT GGC ACC GTG CAG TCG TCC GCT TCC GCT ACC ATC 340 Cys Phe Glu Asn Cys Gly Thr Val Gin Ser Ser Ala Ser Ala Thr He 95 100 105 ACC GTG TAT TCG TTT CCG GAG AGT GTG GAG CTG AGA CCT CTA CCA GCC 388 Thr Val Tyr Ser Phe Pro Glu Ser Val Glu Leu Arg Pro Leu Pro Ala 110 115 120 TGG CAG CAA GTA GGC AAG GAC CTC ACC CTG CGC TGC CAC GTG GAT GGT 436 Trp Gin Gin Val Gly Lys Asp Leu Thr Leu Arg Cys His Val Asp Gly 125 130 135 GGA GCA CCG CGG ACC CAG CTC TCA GCA GTG CTG CTC CGT GGG GAG GAG 484 Gly Ala Pro Arg Thr Gin Leu Ser Ala Val Leu Leu Arg Gly Glu Glu 140 145 150 ATA CTG AGC CGC CAG CCA GTG GGT GGG CAC CCC AAG GAC CCC AAG GAG 532 He Leu Ser Arg Gin Pro Val Gly Gly His Pro Lys Asp Pro Lys Glu 155 160 165 170 ATC ACA TTC ACG GTG CTG GCT AGC AGA GGG GAC CAC GGA GCC AAT TTC 580 He Thr Phe Thr Val Leu Ala Ser Arg Gly Asp His Gly Ala Asn Phe 175 180 185 TCA TGC CGC ACA GAA CTG GAT CTC AGG CCG CAA GGG CTG GCA TTG TTC 628 Ser Cys Arg Thr Glu Leu Asp Leu Arg Pro Gin Gly Leu Ala Leu Phe 190 195 200 TCT AAT GTC TCC GAG GCC AGG AGC CTC CGG ACT TTC GAT CTT CCA GCT 676 Ser Asn Val Ser Glu Ala Arg Ser Leu Arg Thr Phe Asp Leu Pro Ala 205 210 215 ACC ATC CCA AAG CTC GAC ACC CCT GAC CTC CTG GAG GTG GGC ACC CAG 724 Thr He Pro Lys Leu Asp Thr Pro Asp Leu Leu Glu Val Gly Thr Gin 220 225 230 CAG AAG TTG TTT TGC TCC CTG GAA GGC CTG TTT CCT GCC TCT GAA GCT 772 Gin Lys Leu Phe Cys Ser Leu Glu Gly Leu Phe Pro Ala Ser Glu Ala 235 240 245 250 CGG ATA TAC CTG GAG CTG GGA GGC CAG ATG CCG ACC CAG GAG AGC ACA 820 Arg He Tyr Leu Glu Leu Gly Gly Gin Met Pro Thr Gin Glu Ser Thr 255 260 265 AAC AGC AGT GAC TCT GTG TCA GCC ACT GCC TTG GTA GAG GTG ACT GAG 868 Asn Ser Ser Asp Ser Val Ser Ala Thr Ala Leu Val Glu Val Thr Glu 270 275 280 GAG TTC GAC AGA ACC CTG CCG CTG CGC TGC GTT TTG GAG CTA GCG GAC 916 Glu Phe Asp Arg Thr Leu Pro Leu Arg Cys Val Leu Glu Leu Ala Asp 285 290 295 CAG ATC CTG GAG ACG CAG AGG ACC TTA ACA GTC TAC AAC TTT TCA GCT 964 Gin He Leu Glu Thr Gin Arg Thr Leu Thr Val Tyr Asn Phe Ser Ala 300 305 310 CCG GTC CTG ACC CTG AGC CAG CTG GAG GTC TCG GAA GGG AGC CAA GTA 1012 Pro Val Leu Thr Leu Ser Gin Leu Glu Val Ser Glu Gly Ser Gin Val 315 320 325 330 ACT GTG AAG TGT GAA GCC CAC AGT GGG TCG AAG GTG GTT CTT CTG AGC 1060 Thr Val Lys Cys Glu Ala His Ser Gly Ser Lys Val Val Leu Leu Ser 335 340 345 GGC GTC GAG CCT AGG CCA CCC ACC CCG CAG GTC CAA TTC ACA CTG AAT 1108 Gly Val Glu Pro Arg Pro Pro Thr Pro Gin Val Gin Phe Thr Leu Asn 350 355 360 SUBSTTTUTE SHEET (RULE 26) GCC AGC TCG GAG GAT CAC AAA CGA AGC TTC TTT TGC TCT GCC GCT CTG 1156 Ala Ser Ser Glu Asp His Lys Arg Ser Phe Phe Cys Ser Ala Ala Leu 365 370 375 GAG GTG GCG GGA AAG TTC CTG TTT AAA AAC CAG ACC CTG GAA CTG CAC 1204 Glu Val Ala Gly Lys Phe Leu Phe Lys Asn Gin Thr Leu Glu Leu His 380 385 390 GTG CTG TAT GGT CCT CGG CTG GAC GAG ACG GAC TGC TTG GGG AAC TGG 1252 Val Leu Tyr Gly Pro Arg Leu Asp Glu Thr Asp Cys Leu Gly Asn Trp 395 400 405 410 ACC TGG CAA GAG GGG TCT CAG CAG ACT CTG AAA TGC CAG GCC TGG GGG 1300 Thr Trp Gin Glu Gly Ser Gin Gin Thr Leu Lys Cys Gin Ala Trp Gly 415 420 425 AAC CCA TCT CCT AAA ATG ACC TGC AGA CGG AAG GCA GAT GGT GCC CTG 1348 Asn Pro Ser Pro Lys Met Thr Cys Arg Arg Lys Ala Asp Gly Ala Leu 430 435 440 CTG CCC ATC GGG GTG GTG AAG TCT GTC AAA CAG GAG ATG AAT GGT ACA 1396 Leu Pro He Gly Val Val Lys Ser Val Lys Gin Glu Met Asn Gly Thr 445 450 455 TAC GTG TGC CAT GCC TTT AGC TCC CAT GGG AAT GTC ACC AGG AAT GTG 1444 Tyr Val Cys His Ala Phe Ser Ser His Gly Asn Val Thr Arg Asn Val 460 465 470 TAC CTG ACA GTA CTG TAC CAC TCT CAA AAT AAC TGG ACT ATA ATC ATT 1492 Tyr Leu Thr Val Leu Tyr His Ser Gin Asn Asn Trp Thr He He He 475 480 485 490 CTG GTG CCA GTA CTG CTG GTC ATT GTG GGC CTC GTG ATG GCA GCC TCT 1540 Leu Val Pro Val Leu Leu Val He Val Gly Leu Val Met Ala Ala Ser 495 500 505 TAT GTT TAT AAC CGC CAG AGA AAG ATC AGG ATA TAC AAG TTA CAG AAG 1588 Tyr Val Tyr Asn Arg Gin Arg Lys He Arg He Tyr Lys Leu Gin Lys 510 515 520 GCT CAG GAG GAG GCC ATA AAA CTC AAG GGA CAA GCC CCA CCT CCC 1633 Ala Gin Glu Glu Ala He Lys Leu Lys Gly Gin Ala Pro Pro Pro 525 530 535 TGAGCCTGCT GGATGAGACT CCTGCCTGGA CCCCCTGCAG GGCAACAGCT GCTGCTGCTT 1693 TTGAACAGAA TGGTAGACAG CATTTACCCT CAGCCACTTC CTCTGGCTGT CACAGAACAG 1753 GATGGTGGCC TGGGGGATGC ACACTTGTAG CCTCAGAGCT AAGAGGACTC GGTGGATGGA 1813 GCAAGACTGT GAACACGTGT GACCCGGACC CACCTACAGC CCGGTGGACC TTCAGCCAAG 1873 AAACGCTGAC TTCATTCTCT ATTGCCCCTG CTGAGGGGCT CCTGCCTAAG GAAGACATGA 1933 TATCCAGTAG ACACAAGCAA GAAGACCACA CTTCCCCCCC GACACAGGAA AGCTGAGACA 1993 TTGTCCCCAA CTCTTCTTGA TGTATTTATT AATTTAGAGT TTTACCAGCT ATTTATTGAG 2053 TACCCTGTAT ATAGTAGATC AGTGAGGAGG TGAATGTATA AGTTATGGCC TGGACCCTGC 2113 TGCAGATGCT GTGAGAGTCT GGGGAAAGAT CACATGGGTC GAGGGTTTCT CTACTGGTCA 2173 GGATGCTTTT CTCATAAGGG TCGACTTTTT TCACCAGTCA CATAAACACT ATGTGGACTA 2233 GCAGTGGTTC TCTGCTCCTC CACATCCTGG AGCGTCCCAG CACCTCCCCA CCTACTTTTG 2293 TTCCCAATGT CAGCCACCAT GCCTTAGCAG CTGAACAATC GAGCCTCATG CTCATGAAAT 2353 SUBSTTTUTE SHEET (RULE 26) CATGGTCCCA GGCGGCTCCA CCTCAAAGAG AAAGCCTGGA AGGAAATGTT CCAACTCCTT 2 13 AGAAGGGTCG TGCAAGCTGC TGTGGGAGGG TAAGCACCCC TCCCAGCAGC AGAAACCTTT 2473 CCTTTGAATC AATAAAGTTT TATGTCGGCC TGAAAAAAAA AAAAAAAAA 2522 (2) INFORMATION FOR SEQ ID NO:8 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 537 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8 : Met Ala Ser Thr Arg Ala Lys Pro Thr Leu Pro Leu Leu Leu Ala Leu 1 5 10 15 Val Thr Val Val He Pro Gly Pro Gly Asp Ala Gin Val Ser He His 20 25 30 Pro Arg Glu Ala Phe Leu Pro Gin Gly Gly Ser Val Gin Val Asn Cys 35 40 45 Ser Ser Ser Cys Lys Glu Asp Leu Ser Leu Gly Leu Glu Thr Gin Trp 50 55 60 Leu Lys Asp Glu Leu Glu Ser Gly Pro Asn Trp Lys Leu Phe Glu Leu 65 70 75 80 Ser Glu He Gly Glu Asp Ser Ser Pro Leu Cys Phe Glu Asn Cys Gly 85 90 95 Thr Val Gin Ser Ser Ala Ser Ala Thr He Thr Val Tyr Ser Phe Pro 100 105 110 Glu Ser Val Glu Leu Arg Pro Leu Pro Ala Trp Gin Gin Val Gly Lys 115 120 125 Asp Leu Thr Leu Arg Cys His Val Asp Gly Gly Ala Pro Arg Thr Gin 130 135 140 Leu Ser Ala Val Leu Leu Arg Gly Glu Glu He Leu Ser Arg Gin Pro 145 150 155 160 Val Gly Gly His Pro Lys Asp Pro Lys Glu He Thr Phe Thr Val Leu 165 170 175 Ala Ser Arg Gly Asp His Gly Ala Asn Phe Ser Cys Arg Thr Glu Leu 180 185 190 Asp Leu Arg Pro Gin Gly Leu Ala Leu Phe Ser Asn Val Ser Glu Ala 195 200 205 Arg Ser Leu Arg Thr Phe Asp Leu Pro Ala Thr He Pro Lys Leu Asp 210 215 220 Thr Pro Asp Leu Leu Glu Val Gly Thr Gin Gin Lys Leu Phe Cys Ser 225 230 235 240 Leu Glu Gly Leu Phe Pro Ala Ser Glu Ala Arg He Tyr Leu Glu Leu 245 250 255 SUBSTTTUTE SHEET (RULE 26) Gly Gly Gin Met Pro Thr Gin Glu Ser Thr Asn Ser Ser Asp Ser Val 260 265 270 Ser Ala Thr Ala Leu Val Glu Val Thr Glu Glu Phe Asp Arg Thr Leu 275 280 285 Pro Leu Arg Cys Val Leu Glu Leu Ala Asp Gin He Leu Glu Thr Gin 290 295 300 Arg Thr Leu Thr Val Tyr Asn Phe Ser Ala Pro Val Leu Thr Leu Ser 305 310 315 320 Gin Leu Glu Val Ser Glu Gly Ser Gin Val Thr Val Lys Cys Glu Ala 325 330 335 His Ser Gly Ser Lys Val Val Leu Leu Ser Gly Val Glu Pro Arg Pro 340 345 350 Pro Thr Pro Gin Val Gin Phe Thr Leu Asn Ala Ser Ser Glu Asp His 355 360 365 Lys Arg Ser Phe Phe Cys Ser Ala Ala Leu Glu Val Ala Gly Lys Phe 370 375 380 Leu Phe Lys Asn Gin Thr Leu Glu Leu His Val Leu Tyr Gly Pro Arg 385 390 395 400 Leu Asp Glu Thr Asp Cys Leu Gly Asn Trp Thr Trp Gin Glu Gly Ser 405 410 415 Gin Gin Thr Leu Lys Cys Gin Ala Trp Gly Asn Pro Ser Pro Lys Met 420 425 430 Thr Cys Arg Arg Lys Ala Asp Gly Ala Leu Leu Pro He Gly Val Val 435 440 445 Lys Ser Val Lys Gin Glu Met Asn Gly Thr Tyr Val Cys His Ala Phe 450 455 460 Ser Ser His Gly Asn Val Thr Arg Asn Val Tyr Leu Thr Val Leu Tyr 465 470 475 480 His Ser Gin Asn Asn Trp Thr He He He Leu Val Pro Val Leu Leu 485 490 495 Val He Val Gly Leu Val Met Ala Ala Ser Tyr Val Tyr Asn Arg Gin 500 505 510 Arg Lys He Arg He Tyr Lys Leu Gin Lys Ala Gin Glu Glu Ala He 515 520 525 Lys Leu Lys Gly Gin Ala Pro Pro Pro 530 535 (2) INFORMATION FOR SEQ ID NO:9 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single- (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA SUBSTTTUTE SHEET (RULE 26) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GCCCAAGCTG GCATCCGTCA 20 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: TGCATCCCCC AGGCCACCAT 20 SUBSTTTUTE SHEET (RULE 26)

Claims: WHAT IS CLAIMED IS: 1. A pharmaceutical composition for the treatment of pathologic conditions associated with the overexpression of ICAM-1 in a host, said pharmaceutical composition comprising an effective amount of an ICAM-1 antisense molecule encapsulated in a lipid mixture, said lipid mixmre comprising at least two members selected from the group consisting of phosphoiipids, sterols and cationic lipids. 2. A pharmaceutical composition in accordance with claim 1 , wherein said lipid mixmre is in the form of liposomes and consists essentially of neutral phosphoiipids and a sterol. 3. A pharmaceutical composition in accordance with claim 1, wherein said lipid mixmre is in the form of lipid particles. 4. A pharmaceutical composition in accordance with claim 1, wherein said ICAM-1 antisense molecule comprises from about 15 to about 50 nucleic acids and is complementary to a portion of the 3' -untranslated region of ICAM-1. 5. A pharmaceutical composition in accordance with claim 4, wherein said ICAM-1 antisense molecule is a phosphorothioate molecule. 6. A pharmaceutical composition in accordance with claim 4, wherein said ICAM-1 antisense molecule is a methyl phosphonate molecule. 7. A pharmaceutical composition in accordance with claim 1 , wherein said lipid mixmre is in the form of liposomes consisting essentially of phosphatidylcholine and a sterol. 8. A pharmaceutical composition in accordance with claim 1, wherein said lipid mixmre is in the form of liposomes consisting essentially of egg phosphatidylcholine and cholesterol. SUBSTTTUTE SHEET (RULE 26) 9. A pharmaceutical composition in accordance with claim 1 , wherein said lipid mixmre is in the form of liposomes consisting essentially of neutral phosphoiipids and cholesterol, said liposomes having diameters of from about 50 to about 150 nm. 10. A pharmaceutical composition in accordance with claim 9, wherein said liposomes have diameters of from about 75 to about 125 nm. 11. A pharmaceutical composition in accordance with claim 1, wherein said lipid mixmre is in the form of lipid particles, said lipid particles being from about 50 to about 90 nm in diameter. 12. A pharmaceutical composition in accordance with claim 1, wherein said lipid mixmre is in the form of lipid particles, said lipid particles being from about 60 to about 80 nm in diameter. 13. A pharmaceutical composition in accordance with claim 12, wherein said lipid particles comprise phosphoiipids and cationic lipids, wherein said cationic lipids are members selected from the group consisting of DODAC, DDAB, DOTAP, DOTMA, DOSPA, DOGS, DC-Chol and combinations thereof, and said phosphoiipids are members selected from the group consisting of DOPE, POPC, EPC and combinations thereof. 14. A method for the treatment of pathologic conditions associated with the overexpression of ICAM-1 in a host, said method comprising delivering to said host a pharmaceutical composition in accordance with claim 1. 15. A method in accordance with claim 14, wherein said composition is a liposomal composition. 16. A method in accordance with claim 14, wherein said composition comprises lipid particles. SUBSTTTUTE SHEET (RULE 26) 17. A method in accordance with claim 14, wherein said ICAM-1 antisense molecule comprises from about 15 to about 50 nucleic acids and is complementary to a portion of the 3 '-untranslated region of ICAM-1. 18. A method in accordance with claim 14, wherein said delivering comprises administering intravenously. 19. A method in accordance with claim 14, wherein said delivering comprises administering topically. 20. A method in accordance with claim 16, wherein said lipid particles comprise phosphoiipids and cationic lipids, said cationic lipids being members selected from the group consisting of DODAC, DDAB, DOTAP, DOTMA, DOSPA, DOGS, DC- Chol and combinations thereof, and said phosphoiipids being members selected from the group consisting of DOPE, POPC, EPC and combinations thereof. 21. A method in accordance with claim 14, wherein said condition is a member selected from the group consisting of Alzheimer's disease, multiple sclerosis, uveitis, Herpes keratitis, renal allograft rejection, glomerulonephritis, liver allograft rejection, viral hepatitis, alcoholic hepatitis, cholangitis, cardiac allograft rejection, atherosclerotic plaques, rheumatoid arthritis, Grave's disease, Hashimoto's thyroiditis, psoriasis, scleroderma, graft v host disease, contact dermatitis, lichen planus, fixed drug eruption, mycosis fungoides, and alopecia areata. 22. A method in accordance with claim 14, wherein said condition is contact dermatitis. SUBSTTTUTE SHEET (RULE 26)

Inventors: HOPE MICHAEL J, KLIMUK SANDRA K, SCHERRER PETER, SEMPLE SEAN C, HOPE, MICHAEL, J., KLIMUK, SANDRA, K., SCHERRER, PETER, SEMPLE, SEAN, C.
Applicants: UNIV BRITISH COLUMBIA, UNIVERSITY OF BRITISH COLUMBIA
Application year: 1997
Main classification: C12N15
IPC codes: A61K9, A61K31, A61K38, A61K47, A61K48, A61P1, A61P5, A61P9, A61P17, A61P19, A61P25, A61P27, A61P31, A61P37, A61P43, C12N15