IL190101A - Anti sr-b1 antibodies and compositions comprising same for diagnosing and treating an inflammation - Google Patents

Description

n iin ^wm n ^ ηιυπί/ι D' WDn COMPOSITIONS AND MBTI IODS FOR DIAGNOSING AND TREATING AN INFLAMMATION FIELD AND BACKGROUND OF THE INVENTION The present invention relates to antibodies, compositions and methods for diagnosing and treating inflammation. More particularly, the present invention relates to the use of and scavenger receptor antibodies in treatment of an inflammatory response.

Inflammation is a physiological condition characterized in the acute form by the classical signs of pain, heat, redness, swelling and loss of function. Inflammation often accompanies diseases such as Multiple Sclerosis (MS), osteoarthritis, Inflammatory Bowl Disease (IBD) including Crohn's disease and ulcerative colitis, Rheumatoid Arthritis (RA), SEE, type I diabetes (IDDM), atherosclerosis, encephalomyelitis, Alzheimer's disease, stroke, traumatic brain injury, Parkinson's disease, septic shock and others. In most eases, there is no effective cure for inflammation associated with such disease and existing treatments are palliative and largely fail to control the underlying causes of tissue degradation.

Scavenger receptors (SRs) are cell surface proteins, which are typically found on macrophages and bind various types of chemically modified lipoproteins (1-3), such as low-density lipoprotein (LDL). This family of trans-membrane receptors which are highly varied in structure are involved in receptor-mediated endocytosis, phagocytosis of apoptolic cells and bacteria, as well as in cell adhesion [Peiser L. el al., Curr. Opin. Immun. 14(1 ): 123- 128, 20U2J. Since the massive receptor-mediated uptake of cholesterol from modified LDL can convert cultured macrophages into cholesteryl ester— filled foam cetls, similar to those found in atherosclerotic plaques, it has been postulated that these receptors also function in deposition of LDL cholesterol of macrophages in artery walls during the initial stages of atherosclerotic plaque formation 111.

Scavenger receptors (SRs) are termed as such since they mediate the binding of remarkably wide variety of polyanionic ligands [e.g., modified proteins, sulfated polysaccharides and certain polynucleotides [ I, 3, 4|. This property led to the hypothesis that these receptors form a part of an in innate immune response by serving pattern recognition receptors that bind a wide variety of pathogen components [2-5].

SK-BI (also referred to as S -BI or CLA-I) is a macrophage scavenger molecule and a receptor for high-density lipoprotein (IIDL) [2, 3, 6, 7] that mediates cholesterol uptake from cells [Rigolti A. et al., Curr. Opiii. Lipidol., 8:181-8, 1997; Rigotli A. et al., Proc. Natl. Acad. Sci., 94:12610-5, 1 97]. SR- I can also serve as a receptor for non-1-1 DL lipoproteins and appears to play an important role in reverse cholesterol transport. In vivo experiments showed that this receptor is important for HDL metabolism in mice, and lor the metabolism of LDL and H L cholesterol in humans [Slang H. el ah, J. Biol. Chem.274:32692-8., 1999; Swarnakar S. el al., J. Biol. Chem.274:29733-9., 1999]. Studies involving the manipulation of SR- I gene expression in mice, indicate that its expression protects against atherosclerosis [Kozarsky K. F., and Krieger M., Curr. Opin. Lipidol.10:491-7., 1 99; Ueda Y. et al., J. Biol. Chem.275:20368-73., 2000; Acton S. L. el al., Mol. Med. Today 5:518-24., 1999]. It vvas also suggested that HDL and particularly its protein fraction Apo-AI affect the in vitro production of pro-inflammatory mediators by macrophages (8). Among mediators derived from macrophages that propagate inflammation are interleukin 12 (IL-12), TNF-a and possibly IL-6 whereas, TGF-|3 and IL-10 have predominantly anti-inflammatory effects [KJefer R. et al., Frog. Neurobiol.64(2): 109-27,2001].

PCT Publication No. WO 2004/041179 teaches targeting of scavenger receptor SR-BI (Cla-I) for the treatment of infectious diseases associated with invasion of foreign antigens such as bacterial or viral antigens (e.g., infection, sepsis and associated inflammation). This is based on the discovery that peptides with an amphipalhie helical motif block cellular uptake of LPS (lipopolysaccharide) and proinflammatory responses induced by LPS, LTA (lipoteichoic acid) and bacterial cpn60 (Chaperonin 60) and amyloid peptides in vitro. Thus the inventors of PCT Publication No. WO 2004/041179 conclude that agents with an amphipathic motif targeting SR-BI (scavenger receptor class B type 1) could potentially be used to treat sepsis, bacterial and viral infections and inflammatory diseases where LPS, LTA, viral envelope proteins, and/or heat shock proteins contribute to pathogenesis.

WO 2004/041179 docs not suggest the above-described agents for the favourable treatment of autoimmune diseases (which arc not associated with foreign pathogenic agents such as LPS, nor with amyloid) such as IBD. Nor does the art teach the use of SR-B I specific antibody sequences having an anti-inflammatory activity activity for the treatment of inllammatory diseases in general and autoimmune diseases in particular, such as multiple sclerosis.

There is thus, a witlely recognized need for and it would be highly advantageous to have novel agents and methods using same for targeting SR-B I and treating autoimmune diseases.

SUMMARY OF THE INVENTION According to otie aspect of the present invention there is provided an isolated polypeptide comprising an antigen recognition domain capable of speci fically binding a human scavenger receptor, wherein the antigen recognition domain comprises at least three CDR amino acid sequences selected from the group consisting of SEQ I D NO: I I , 1 5, 19, 23, 27 and 3 1 .

According to another aspect of the present invention there is provided an isolated polypeptide comprising an antigen recognition domain capable of speci fically binding a human scavenger receptor, wherein the antigen recognition domain comprises CDR amino acid sequences as set forth in SEQ I D NO: NO: I I , 1 5, 1 9, 23, 27 and 3 1 .

According to yet another aspect of the present invention there is provided an isolated polynucleotide comprising a nucleic acid sequence encoding the polypeptide.

According to sti ll another aspect of the present invention there is provided a pharmaceutical composition comprising as an active ingredient the polypeptide.

According to an additional aspect of the present invention there is provided a method of reducing mJlummalion in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the polypeptide, thereby reducing the inflammation in the subject.

According to yet an additional aspect of the present invention there is provided use of the polypeptide for the manufacture of a medicament identi fied for treating IBD.

According to stil l an additional aspect of the present invention there is provided use of the polypeptide for the manufacture of a medicament ideiitilled for treating multiple sclerosis.

According to a f urther aspect of the present invention there is provided use of the polypeptide for the manufacture of a medicament identi fied for treating an autoimmune disease.

The present invention successfully addresses the shortcomings of the presently known configurations by providing novel compositions and methods containing same for diagnosing and treating an inflammatory response.

Unless otherwise defined, all technical and scienti fic terms used herein have the same meaning as commonly understood by one of ordinary ski ll in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In case of conllict, the patent specification, including dellnilions, wil l control. In addition, the materials, methods, and examples are i llustrative only and not intended to be l imiting.

BRI EF DESCRI PTION OF THE DRAWINGS The invention is herein described, by way of example only, with reference to the accompanying drawings. With speci fic reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of ill ustrative discussion o f the preferred embodiments of the present invention only, and arc presented in the cause of providing what is believed to be the most useful and readi ly understood description of the principles and conceptual aspects of the invention. I n this regard, no attempt is made to show structural details of the invention in more detai l than is necessary for a fundamental understanding of the invention, the description taken with the drawings making apparent to those skilled in the art how the several forms of the invention may be embodied in practice.

In the drawings: FIG. 1 is a photograph depicting cross-reactivity of monoclonal anti SR- I antibody, E l 2, with human and mouse orthologs. Recombinant proteins were resolved on SDS-FAGE and transferred to nitrocellulose membrane. The membrane was subjected to l 2; .

FIG. 2 is a graph depicting dose-dependent induction of lL- 10 secretion from cultured peritoneal macrophages treated with E 12; FIG. 3 is a bar graph depicting dose-dependent suppression of NO levels in cultured peritoneal macrophages treated with 1212; FIG. 4 is a graph depicting the effect of E 12 (closed squares), control isotype matching antibody (circles) or no treatment on ongoing EA E in mice induced with such, as determined by reduction in EAE score; FIGS. 5a-c are bar graphs depicting the effect of E l 2 (pink) or control antibodies (grey) on cytokine secretion from spleen cells of 1 day EA E-indticed mice. Figure 5a - I L-4. Figure 5b - I L- 1 2. Figure 5c - 1 L- 10; FIGS. 6a- f are photographs showing I L- 10 immunostaining of Lumbar spinal cord sections from ΕΛΕ induced mice ( 1 9 days of disease onset) subjected to no treatment (Figure 6a), or treated with i 2 (Figure 6b), or isolype matching control antibody (Figure 6c). Figures 6a-c shows staining with biolinylated E 1 2 for presence of scavenger receptor expressing cells. Figures 6d-f shows staining with anti I L- 10 antibody. Anti-SR-BI therapy reduces the histological score of EA E; FI GS. 7a-e are photographs showing representative histological colon sections obtained at day 12 of disease onset from naive rats (Figure 7a), posi tive control rats suffering form TN BS induced I BD (Figure 7b), rats suffering from TNBS induced I BD that were subjected to repeated administration of isolype matched control IgG (Figure 7c) in comparison to those treated - with mAb E l 2 (Figures 7d-e); and FIGS. 8a-i show representative immuno-histological sections obtained al day 12 of disease onset from control rats suffering from TNBS induced I D (Figures 8a-c), rats suffering form TNBS induced IBD that were subjected to repeated administration of isotype matched control IgG (Figures 8d-l) in comparison to diseased rats treated with inAb E l 2 (Figures 8g-i). Figures 8a, d and g are stained with mAb ED I (macrophages bio-marker); Figures 8b, e and h are stained with anti CD3 (T cell bio-marker) and Figures 8 c, f and I are stained with an anti I L- 10 mAb.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention is of compositions and methods which can be used for the treatment of inflammation. Specifically, the present invention relates to the use of anti scavenger receptor antibodies in treating inflammatory response.

The principles and operation of the present invention may be belter understood with reference lo the drawings and accompanying descriptions.

Before explaining at least one embodiment of the invention in detai l, it is lo be understood that llie invention is nol limited in its application lo the details set forth in the following description or exempli fied by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways. Also, it is to be understood that the phraseology and termi nology employed herei n is for the purpose of description and should not be regarded as l imiting.

Diseases and disorders which have significant inflammatory components are ubiquitous. Skin disorders, bowel disorders, certain degenerative neurological disorders, arthritis, autoimmu e diseases and other illnesses afflict many patients. The factors underlying these disorders are varied and include infectious agents, autoimmune factors, dietary or environmental factors and genetic factors. In the majority of cases, the causative elements have not been defined and many of the key pathophysiological components have not been elucidated. Accordingly, treatment options for the majority of these diseases is suboptimal.

The present inventor has previously shown that the immune system can selectively generate autoimmunity to chemokines and other proinflammatory mediators when such a response is beneficial for the host [9, 10, I I , 1-2, 14, 15] . For example, patients suffering from rheumatoid arthritis (I A) but not osteoarthritis (OA) have significant levels of autoantibodies directed to TNF-a, and therapies that neutralize the function of TNF-a suppress RA but not OA. Studies conducted by the present inventor have shown that selective amplification of these beneficial antibodies by targeted DNA vaccines provided protective immunity in experimental models (9, 10, 1 1 , 12, 14, 15). The present inventor have further shown that subjects suffering from inflammatory disease exhibit elevated levels of autoantibodies to scavenger receptor (SR) and showed that DNA vaccination against SR-131 can prevent such diseases by altering the cytokine profile produced by macrophages from proinflammatory cytokines to anti-inflammatory cytokines (see WO2004/080385).

It is now reported that the present inventor also developed, through laborious experimentations and screening a novel therapeutic anti-SR-B l monoclonal antibody, E 12, which is capable of altering the cytokine profi le and inflammatory activities of macrophages. This antibody which was sequenced is directed against a surface exposed epitope of the scavenger receptor (Figure I ) and is cross-reactive to human CLA-I (human SR-B I ) and also affects the cytokine profi le and in vitro activities of human macrophages (a cell li ne) and as such can be used as a valuable therapeutic and diagnostic tool (see Example I and Figures 1 -3). This anti body was also shown effective in suppressing ongoing ΕΛΕ and TNBS induced IBD (see Figures 4-8). Immunohistological analysis clearly showed that in both diseases and SR-BI tiicrapy altered the cytokine production of invading leukocytes, at the autoimmune site, into high I L- I O producing cel ls. This may explain signi ficant therapeutic effect of this anti body in these diseases. I mmunohistological analysis of CNS sectio s using anti SR-B I uiAb also showed that SR-BI positive leukocytes enter the site o f inflammation (so far detected only for EAE). Thus, it is suggested that anti SR-BI antibodies desrertbed herein affect the cytokine profile and inflammatory functions of inflammatory leukocytes entering the autoimmune site, and thereby the function and polarization of autoimmune T cel ls there.

These findings suggest that the present antibody can be used for targeting scavenger receptor and for treatment of inflammatory diseases, especially I BD and multiple sclerosis.

Thus according lo one aspect of the present invention there is provided an isolated polypeptide comprising an antigen recognition domain capable of specifically* binding a human scavenger receptor, wherein said antigen recognition domain comprises at least three CDR amino acid sequences selected at least 90 % homologous to the group consisting of SEQ I D NOs. : I I . 15. 1 9, 23, 27 and 3 1 .

According to one embodiment of this aspect of the present invention the polypeptide comprises the CDR amino acid sequences are as set forth in SEQ ID NOs. : I I , 1 5, 1 9, 23, 27 and 3 1 .

Preferably, the polypeptide is an antibody. More preferably the antibody is capable of eliciting an anti-inflammatory activity. As used herein "an antiinflammatory activity" refers to any reduction in immune cell inllammaoty activity, such as reduction in pro-inllammalory cytokine (such as TN F-a, I L- 1 and I L- 12) secretion preferably accompanied by induction of anti-inflammatory cytokine (e.g., I L- I O, I L-4 and TCF-b) secretion.

The term "antibody" refers to whole antibody molecules as well as functional fragments thereof, such as Fab, F(ab')2, and Fv that arc capable of binding with antigenic portions of the target polypeptide. These funct ional antibody fragments constitute preferred embodiments of the present invention, and are defined as follows: (1 ) Fab, the fragment which contains a monovalent antigen-binding fragment of an antibody molecule, can be produced by digestion of whole antibody with the enzyme papain to yield an intact l ight chain and a portion of one heavy chain; (2) Fab1, the fragment o an antibody molecule that can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain; two Fab' fragments are obtained pel" antibody molecule; (3) (Fab')2, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab')2 is a dimer of two Fab1 fragments held together by two disulfide bonds; (4) Fv, defined as a genetically engineered fragment containing the variable region of the l ight chain and the variable region of the heavy chain expressed as two chains; and (5) Single chain antibody ("SCA"), a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule as described in, for example, U.S. Patent 4,946,778.

Methods of generating antibodies are well known in the art. Puri fication of serum immunoglobulin antibodies (polyclonal anlisera) or reactive portions thereof can be accomplished by a variety of methods known to those of ski ll including, precipitation by ammonium sulfate or sodium sulfate followed by dialysis against saline, ion exchange chromatography, affinity or immunoaffinity chromatography as well as gel nitration, zone electrophoresis, etc. (see Got!ing in, Monoclonal Antibodies: Principles and Practice, 2nd ed., pp. 04- 126, 1 986, Orlando, Fla., Academic Press). Under normal physiological conditions antibodies are found in plasma and other body lluids and in the membrane of certain cells and are produced by lymphocytes of the type denoted 13 cells or their functional equivalent. Antibodies of the JgG class are made up of four polypeptide chains linked together by disulfide bonds. The four chains of intact IgG molecules are two identical heavy chains referred to as 1-1-chaiiis and two identical light chains referred to as L-ehaiiis. Additional classes include igD, IgE, IgA, IgM and related proteins.

Methods of generating and isolating monoclonal antibodies are wel l known in the art, as summarized for example in reviews such as Tramonlano and Schloeder, Methods in Enzymology 1 78, 55 1 -568, 1989. A recombinant scavenger receptor polypeptide may be used to generate antibodies in vitro (see Example 6 of the Examples section which fol lows). In general, a suitable host animal is immunized with the recombinant polypeptide. Advantageously, the animal host used is a mouse of an inbred strain. Animals are typical ly immunized with a mixture comprising a solution of the recombinant polypeptide in a physiologically acceptable vehicle, and any suitable adjuvant, which achieves an enhanced immune response to the immunogen. By way of example, the primary immunization conveniently may be accomplished with a mixture of a solution of the recombinant polypeptide and Ereund's complete adjuvant, said mixture being prepared in the form of a water in oil emulsion. Typically the immunization will be administered to the animals intramuscularly, intradermal ly, subcutatieously, inlraperitoneally, into the footpads, or by any appropriate route of administration. The immunization schedule of the immunogen may be adapted as required, but customarily involves several subsequent or secondary immunizations using a mi lder adjuvant such as Freund's incomplete adjuvant. Antibody titers and speci ficity of binding to the polypeptide can be determined during the immunization schedule by any convenient method including by way of example radioimmunoassay, or enzyme l inked immunosorbant assay, which is known as the EL1SA assay. When suitable antibody titers arc achieved, antibody-producing lymphocyles from the immunized animals are obtained, and these are cultured, selected and cloned, as is known in the art. Typically, lymphocytes may be obtained in large numbers from the spleens of immunized animals, but they may also be retrieved from the circulation, the lymph nodes or other lymphoid organs. Lymphocytes arc then fused with any suitable myeloma cell line, to yield liybridomas, as is well known in the art. Alternatively, lymphocytes may also be stimulated to grow in culture, and may be immortal ized by methods known in tiic art including the exposure of these lymphocytes to a virus, a chemical or a nucleic acid such as an oncogene, according to established protocols. After fusion, the liybridomas are cultured under suitable culture conditions, for example in multi-well plates, and the culture supernatanls are screened to identi fy cultures containing antibodies that recognize the hapten of choice, l iybridomas that secrete antibodies that recognize the recombinant polypeptide are cloned by limiting dilution and expanded, under appropriate culture condi tions. Monoclonal antibodies are puri fied and characterized in terms of immunoglobulin type and binding affinity.

Anti body fragments according to the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. coli or mammalian cells (e.g. Chi nese hamster ovary cell culture or other protein expression systems) of DNA encoding the fragment.

Antibody fragments can be obtained by pepsin or papain digestion of whole antibodies by conventional methods. For example, antibody fragments can be produced by enzymatic cleavage of antibodies with pepsin to provide a 5S fragment denoted F(ab')2. This fragment can be further cleaved using a thiol reducing agent, and optionally a blocking group for the sul l ydryl groups resulting from cleavage of disul fide l inkages, to produce 3.5S Fab1 monovalent fragments. Alternatively, an enzymatic cleavage using pepsin produces two monovalent Fab' fragments and an Fc fragment directly. These methods are described, for example, by Goldenberg, in U.S. Fat. Nos. 4,036,945 and 4,33 1 ,647, and references contained therein, which patents are hereby incorporated by reference in their entirety (see also Porter, II. R., Biochem. J ., 73 : 1 1 9- 1 26, 1 959). Other methods o cleaving antibodies, such as separation of heavy chains to form monovalent light-heavy chain fragments, further cleavage of fragments, or other enzymatic, chemical, or genetic techniques may also be used, so long as the fragmenls bind to the antigen that is recognized by the intact antibody.

Fv fragments comprise an association of Vn and V| chains. This association may be noncovalenl, as described in Inbar et al. (Proc. Nat'l Acad. Sci. USA 69:2659-62, 1972). Alternatively, the variable chains can be linked by an tnlermolecular disulfide bond or cross-linked by chemicals such as glularaldehyde. Preferably, the Fv fragments comprise Vn and chains connected by a peptide l inker. These single-chain antigen binding proteins (sFv) are prepared by constructing a structural gene comprising DNA sequences encoding the Vt ) and VL domains connected by an oligonucleotide. The structural gene is inserted into an expression vector, which is subsequently introduced into a host cell such as \l. coli. The recombinant host cel ls synthesize a single polypeptide chain with a l inker peptide bridging the two V domains. Methods for producing sFvs are described, for example, by Whitlow and Filpula, Methods, 2: 97- 105, 1991 ; Bird et al., Science 242:423-426, 1988; Pack et al., Bio/Technology I 1 : 1 271 -77, 1 993; unci Ludncr ct ill., U .S. Pal. No. 4,946,778, al l of which are hereby incorporated by reference in its entirety.

Another form of an antibody fragment is a peptide coding for a single complementarity-determining region (CDR). CDI peptides ("minimal recognition 5 units") can be obtained by constructing genes encoding the CDR of an antibody of interest. Such genes are prepared, for example, by using the polymerase chain reaction to synthesize the variable region I om UNA of antibody-producing cells (see, for example, Larrick and Fry Methods, 2: 1 06- 10, 1 1 ).

Humanized forms of non-human (e.g., murine) antibodies are chimeric I t) molecules of immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Huma ized antibodies include human immunoglobulins (recipient antibody) in which residues form a complementary determining region (CDR) of the recipient are replaced by 15 residues from a CDR. of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. I n some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non- human residues. Humanized antibodies may also comprise residues, which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In 0 general, the humanized anti body wi ll comprise substantially al l of at least one, and typically two, variable domains, in which al l or substantially all of the CDR regions correspond to those o f a non-human immunoglobulin and all or substantial ly all of the FR l egions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also wilt comprise at least a portion of an immunoglobulin constant 5 region (Fe), typically that of a human immunoglobulin [Jones et al., Nature, 32 1 :522- 525 ( 1 986); Riechmaiin et al ., Nature, 332:323-329 ( 1 988); and Presta, Curr. Op. Slrucl. Biol., 2:593-596 ( 1 992)].

Methods for humanizing non-human antibodies are well known in the art (see also Example 6 of the xamples section). Generally, a humanized antibody has one or 0 more amino acid residues introduced into it from a source, which is non-human. These non-human amino acid residues are often referred to as import residues, which are typical ly taken from an import variable domain. Humanizalion can be essentially performed following the method of Winter and co-workers [Jones et al., Nature, 321:522-525 (1986); Riechmann ct a!., Nature 332:323-327 (1988); Verhocyen et al., Science, 239:1534-1536 (I988)j. by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S. Pat. No. 4.816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some PR residues are substituted by residues from analogous sites in rodent antibodies.

Human antibodies can also be produced using various techniques known in the art, including phage display libraries Hoogenboom and Winter, .1, ol. Biol., 227:381 (19 1); Marks et al., J. Mol. Biol., 222:5 1 (1991)]. The techniques of Cole et al. and Boerner et al. are also available for the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985) and Boerner et al., J. Immunol., 147(1 ):86-95 (1991)]. Similarly, human monoclonal antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which th endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pal. Nos.5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the following scientific publications: Marks el al., Bio/Technology 10, 779-783 (1 92); Lonberg et al., Nature 368856-859 (1 94); Morrison, Nature 368 812-13 (1994); Fishwild et al., Nature Biotechnology 14, 845-51 (1996); Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol. 1365-93 ( 1 95).

Antibodies of the present invention can be encoded from isolated polynucleotides which comprise nucleic acid sequences such as set forth in SEQ ID NOs.12, 16, 20, 24, 28 and 32.

Polypeptides of the present invention can be synthesized using solid phase peptide synthesis procedures which are well known in the art and further described by John Morrow Stewart and Janis Dillaha Young, Solid Phase Peptide Syntheses (2nd Ed., Pierce Chemical Company. 1984). Synthetic peptides can be purified by preparative high performance liquid chromatography Creighlon T. (1983) Proteins, structures and molecular principles. Wl l Freeman and Co. N.Y.] and the composition of which can be confirmed via amino acid sequencing.

I n cases where large amounts of the polypeptides are desired, they can be generated using recombinant techniques such as described by Bitter et al., ( 1987) Methods in Enzymol. 1 53 :5 1 6-5-44, Sludier el al. ( 1990) Methods in Enzymol. 1 5 :60-89, Brisson et al. ( 1 984) Nature 3 1 0:5 1 1 -5 14, Takumatsu et al. ( 1987) EM. BO J . 6:307-3 1 1 , Coruzzi et al. ( 1 984) EM BO J. 3 : 1 67 1 - 1 680 and Brogli et al., ( 1 984) Science 224:838-843, Gurlcy ct al. ( 1986) Mol. Cell. Biol. 6:559-565 and Wcissbacli & Wcissbacli, 1 988, Methods for Plant Molecular Biology, Academic Press, NY, Section VI I I, pp 421 -463.

As mentioned, the polypeptides (also referred to herein as agents) of the present invention may be used for reducing or treating an inflammatory response (i.e., inflammation) in a subject.

As used herein the term "treating" refers to preventing, curing, reversing, attenuating, alleviating, minimizing, suppressing or halting the deleterious effects of tin inllammatory response.

As used herein the phrase "inllammatory response" refers to an immune response which results in inflammation, typical ly occurring as a result of injurious stimul i including infection, burns, trauma, neoplasia, autoimmune signals and exposure to chemicals, heal or cold or any other harm ful stimulus. An inllammatory response according to the present invention refers to an acute phase response and a chronic inflammation.

As used herein the term "subject" refers to subject who may benefit from the present invention such as a mammal (e.g., canine, fel ine, ovine, porcine, equine, bovine, human), preferably a human subject.

The method of this aspect of the present invention is effected by providing to a subject in need thereof a therapeutically effective amount of the polypeptide of the present invention, thereby reducing the inflammatory response in the subject.

As used herein a "scavenger receptor" refers to a gene product (i.e., RNA or protein) of a scavenger receptor, which is known in the Art. Examples of scavenger receptors include but are not l imited to class A scavenger receptors, class 13 scavenger receptors and class F scavenger receptors. The scavenger receptor is preferably one which is expressed and displayed by macrophages. Preferably, the scavenger receptor of lhc present invenlioii is SR-I3I, a member of the CD36 family, GenBank Accession No. N1 W5496, also known as CLA-I or SR- 1 .

Scavenger receptor aclivily refers lo cell adhesion activity, transporter activity, apoplolic activity, lipid metabol ism activity, signal transduction activity and/or preferably cytokine secretion activity.

An effector of a scavenger receptor refers lo an endogenous molecule which up-regu ales or activates scavenger receptor activity. For example, an effector can be a modi l ed lipid (e.g., oxidized lipid, glycated lipid, alkylated lipid, nitrated lipid, acelylaled l ipid), which binds to the scavenger receptor and activates signaling therefrom.

The above-described agents can be provided to the subject per se, or as part of a pharmaceutical composition where they are mixed with a pharmaceutically acceptable carrier.

As used herein a "pharmaceutical composition" refers lo a preparation of one or more of the active ingredients described herein vvilh other chemical components such as physiologically suitable carriers and excipicnts. The purpose of a pharmaceutical composition is lo faci litate administration of a compound to an organism.

Herein the term "active ingredient" refers lo the preparation accountable for the biological effect.

Hereinafter, the phrases "physiological ly acceptable carrier" and "pharmaceutically acceptable carrier" which may be interchangeably used refer to a carrier or a diluent that does not cause signi ficant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. An adjuvant is included under these phrases. One of the ingredients included in the pharmaceutically acceptable carrier can be for example polyethylene glycol (PEG), a biocompatible polymer with a wide range of solubility in both organic and aqueous media (Mutter el al. ( 1979).

Herein the term "excipient" refers to an inert substance added to a pharmaceutical composition to further Facilitate administration of an active ingredient. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelalin, vegetable oils and polyethylene glycols.

Techniques lor formulation and administration οΓ drugs may be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Gaston, PA, latest edition, vvhich is incorporated herein by reference.

Suitable routes of administration may. for example, include oral, rectal, 5 iransmueosal, especially transnasal, intestinal or parenteral delivery, incl uding intramuscular, subcutaneous and intramedullary injections as well as intrathecal, direct intraventricular, intravenous, inrlaperitoneal, intranasal, or inlraocular injections.

Alternately, one may administer a preparation in a local rather lhan systemic manner, lor example, via injection of the preparation directly into a speci fic region of a t o patient's body.

Pharmaceutical compositions of the present invention may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsi fying, encapsulating, entrapping or lyophi lizing processes. 1 5 Pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipicnls and auxi liaries, vvhich facilitate processing of the active ingredients into preparations which, can be used pharmaceutical ly. Proper formulation is dependent upon the route of administration chosen. 0 For injection, the active ingredients of the i nvention may be formulated in aqueous solutions, preferably in physiological ly compatible buffers such as flank's solution, Ringer's solution, or physiological salt buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in Ihe formulation. Such penetrants are generally known in the art. 5 For oral administration, the compounds can be formulated readi ly by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a patient. Pharmacological 0 preparations for oral use can be made using a solid excipient, optional ly grinding the resulting mixture, and processing the mixture of granules, alter adding suitable auxiliaries if desired, to obtain tablets or dragee cores. Suitable excipicnls arc, in particular, fillers such as sugars, including lactose, sucrose, niannitol, or sorbitol; cellulose preparations such as, ibr example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanlh, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carbomelhylcelhilose; and/or physiological ly acceptable polymers such as polyvinylpyrrolidone (PVP). I f desi red, disintegrating agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, or algi nic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes tuffs or pigments may be added to the tablets or dragee coatings for identi fication or to characterize different combinations of active compound doses.

Pharmaceutical compositions, which can be used orally, include push-lit capsules made of gelatin as wel l as soft, sealed capsules made of gelatin and a plaslicizer, such as glycerol or sorbitol. The push-lit capsules may contain the active ingredients in admixture with filler such as lactose, binders such as starches, lubricants such as talc or magnesi um stearate and, optionally, stabilizers. In soil capsules, the active ingredients may be dissolved or suspended in suitable l iquids, such as fatty oils, liquid paraffin, or ! k|iiid polyethylene glycols. In addition, stabi lizers may be added. All formulations for oral administration should be tn dosages suitable for the chosen ro u te o f ad tn i n ί si rati on .

For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by nasal inhalation, the active ingredients for use according to the present invention arc conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer with the use of a suitable propeliant, e.g., dichlorodilluoromethane, IrichloiOlluoromelhane, dichloro-IctrafiuoiOcthane or carbon dioxide. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin for use in a dispenser may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. The preparations described herein may be formulated for parenteral administration, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multidose containers with optionally, an added preservative. The compositions may be suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulalory agents such as suspending, stabi l izing and/or dispersing agents.

Pharmaceutical compositions for parenteral administration include aqueous solutions of the active preparation in waler-soluble form. Additional ly, suspensions of the active ingredients may be prepared as appropriate oily or water based injection suspensions. Suitable l ipophi lic solvents or vehicles include fatty oi ls such as sesame oi l, or synthetic fatty acids esters such as ethyl oleate, triglycerides or liposomes. Aqueous injection suspensions may contain substances, which increase the viscosity of the suspension, such as sodium carboxymcthyl cellulose, sorbitol or dextran. Optional ly, the suspension may also contain suitable stabilizers or agents which increase the solubility of the active ingredients to allow for the preparation of highly concentrated solutions.

Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water based solution, before use. The preparation of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butler or other glycerides.

Pharmaceutical compositions suitable for use in context of the present invention include compositions wherein the active ingredients are contained in an amount effective to achieve the intended purpose. More specifically, a therapeutically effective amount means an amount of active ingredients effective to prevent, al leviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is wel l within the capability of those skilled in the art.

For any preparation used in the methods of the invention, the therapeutical ly effective amount or dose can be estimated initially from in vitro assays. For example, a dose can be formulated in animal models and such information can be used to more accurately determine useful doses in humans.

Toxicity and therapeutic efficacy of the active ingredients described herein can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals. The data obtained from these in vitro and cell culture assays and animal studies can be used in formulating a range of dosage for use in human. The dosage may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See e.g., Fingl, cl al., 1 ^75, in "The Pharmacological Basis of Therapeutics", Ch. 1 p. I ).

Depending on the seventy and responsiveness of the condition to be treated, dosing can be of a single or a plurality of admi nistrations, with course of treatment lasting from several days to several weeks or until cure is effected or diminution of the disease state is achieved.

The amount of a composilion to be administered wil l, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

Compositions including the preparation of the present invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

Pharmaceutical compositions of the present invention may, i f desired, be prese ted in a pack or dispenser device, such as an FDA approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may, for example, comprise metal or plastic foil, such as a- blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accommodated by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions or human or veterinary administration. Such notice, for example, may be of labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert.

A number of diseases and conditions, which typically cause inflammatory response in individuals can be treated using the methodology described hereinabove. Examples of such diseases and conditions are summarized infra.

Inflammatory diseases - Include, but are not limited to, chronic in flammatory diseases and acute inflammatory diseases. inflammatory diseases associated with hypersensitivity Examples of hypersensitivity include, but arc not limited to, Type I hypersensitivity, Type II hypersensitivity, Type III hypersensitivity, Type IV hypersensitivity, immediate hypersensitivity, antibody mediated hypersensitivity, immune complex mediated hypersensitivity, T lymphocyte mediated hypersensitivity and D I M.

Type I or immediate hypersensitivity, such as asthma.

Type II hypersensitivity include, but are not limited to, rheumatoid diseases, rheumatoid autoimmune diseases, rheumatoid arthritis ( renn V. et ai, Hislol Hislopathol 2000 Jul; 15 (3):791), spondylitis, ankylosing spondylitis (Jan Voswinkel et ai, Arthritis Res 2001; 3 (3): 189), systemic diseases, systemic autoimmune diseases, systemic lupus erythematosus (Erikson J. et ai, Immunol Res 1998; 17 (l-2):49), sclerosis, systemic sclerosis (Renaudineau Y. et ai, Cliti Diagn Lab Immunol. 1999 Mar;6 (2): 156); Chan OT. et ai, Immunol Rev 1999 Jim; 169: 107), glandular diseases, glandular autoimmune diseases, pancreatic autoimmune diseases, diabetes, Type I diabetes (Zimmet P. Diabetes Res Clin Pract 1996 Oct;34 Suppl:S125), thyroid diseases, autoimmune thyroid diseases, Graves' disease (Orgiazzi J. Endocrinol Metab Clin North Am 2000 Jun;29 (2):339), thyroiditis, spontaneous autoimmune thyroiditis (Braley-Mullen H. and Yu S, J Immunol 2000 Dec 15; 165 (12):7262), Hashimoto's thyroiditis (Toyoda N. et ai, Nippon kinsho 1 99 Aug;57 (8): 1810); myxedema, idiopathic myxedema (Milsuma T. Nippon Riusho. 1999 Aug;57 (8): 1759); autoimmune reproductive diseases, ovarian diseases, ovarian autoimmunity (Garza M. et ai, J Reprocl Immunol 1998 l7eb;37 (2):87), autoimmune anti-sperm infertility (Diekman AI3. et ai, Am J cprod Immunol.2000 ar;43 (3): 134), repeated felal loss (Tincani A. et ai, Lupus I 98;7 Suppl 2:S 107-9), neurodegenerative diseases, neurological diseases, neurological autoimmune diseases, multiple sclerosis (Cross AH. et ai, J Ncuoiminunol 2001 Jan l;l 12 (i-2):l), Alzheimer's disease (Oron L. et ai, J Neural Transm Suppl. I997;49:77), myasthenia gravis (Infante A.I. And Kaig E, Int Rev Immunol 1 99; 18 (l-2):83), motor neuropathies ( ornbcrg AJ. J Clin Neurosci. 2000 May;7 (3): 1 1), Guillain-Barre syndrome, neuropathies and autoimmune neuropathies (Kusunoki S. Am J Med Sei.2000 Apr;31 (4):234), myasthenic diseases, Lambert-Eaton myasthenic syndrome (Takamori M. Am J Med Sei. 2000 Apr;3l (4):204), paraneoplastic neurological diseases, cerebellar atrophy, paraneoplastic cerebellar atrophy, non-paraneoplastic stiff man syndrome, cerebellar atrophies, progressive cerebellar atrophies, encephalitis, Rasnuissen's encephalitis, amyotrophic lateral sclerosis, Sycleham chorea, Gi llcs cle la To rettc syndrome, polycndocrinopathies, autoimmune polyendocrinopalhies (Anionic JC. and Honnorat J. Rev Neurol (Paris) 2000 Jan; 156 ( 1 ):23); neuropathies, dysimmune neuropathies (Nobile-Orazio E. et ai, Electroencephalogr Clin Neurophysiol SuppI I 999;50:4 I ); neuromyotonia, acquired neuromyotonia, arthrogtyposis multiplex congenita (Vincent A. et ai, Ann N Y Acad Sci. 1998 May I 3;84 1 :482), cardiovascular diseases, cardiovascular autoimmune diseases, atherosclerosis (Matsuura E. el ai, Lupus. 1998;7 SuppI 2:S 1 35), myocardial infarction (Vaarala O. Lupus. I 998;7 SuppI 2:S 132), thrombosis (Tincani A. et ai, Lupus 1998;7 SuppI 2:S 107-9), granulomatosis, Wegener's granulomatosis, arteritis, Takayasu's arteritis and Kawasaki syndrome (Prapro!nik S. et ai, Wien Klin Wochenschr 2000 Aug 25; 1 12 ( 1 5- 16):660); anti-factor VI I I autoimmune disease (Lacroix-Desmazes S. et ai, Semin Thromb Hcniosl.2000;26 (2): 1 57); vascul itises, necrotizing small vessel vasculitises, microscopic polyatigiilis, Cluirg and Strauss syndrome, glomerulonephritis, pauci-itnmune ocal necrotizing glomerulonephritis, crescentic glomerulonephritis (Noel LH. Ann Med I nterne (Paris). 2000 May; l 5 1 (3): 1 78); anliphospholipid syndrome (Flamholz R. et ai, J Clin Apheresis 1 99; 14 (4): 171 ); heart failure, agonist-like β-adrenoceptor antibodies in heart failure (Wallukal G. et ai, Am J Cardiol. 1 999 J un I 7;83 ( I 2A):75H), thrombocytopenic purpura (Moccia F. Ann ltal Med Int. 1 999 Apr-J un; l 4 (2): 1 14); hemolytic anemia, autoimmune hemolytic anemia (B remov DG. et ai, Leuk Lymphoma 1998 Jan;28 (3-4):285), gastrointestinal diseases, autoimmune diseases of the gastrointestinal tract, intestinal diseases, chronic inflammatory intestinal disease (Garcia l lerola A. et ai, Gastroenterol Hepatol. 2000 Jan;23 ( 1 ): I 6), celiac disease (Landau YE. and Shoenfeld Y. Harefuah 2000 Jan 16; 138 (2): 122), autoimmune diseases of the musculature, myositis, autoimmune myositis, Sjogren's syndrome (Feist E. et ai, hit Arch Allergy Immunol 2000 Sep; 123 ( i ):92); smooth muscle autoimmune disease (Zaul i D. et ai, Biomed Pharmacother 1 99 Jun;53 (5-6):234), hepatic diseases, hepatic autoimmune diseases, autoimmune hepatitis (Manns MP. J Hepatol 2000 Aug;33 (2);326) and primary biliary cirrhosis (Slrassburg CP. et ai, Eur J Gastroenterol Hepatol. 1999 Jun; ] I (6):595).

Type IV or T cell mediated hypersensitivity, include, but are not limited to, rheumatoid diseases, rheumatoid arthritis (Tisch R, McDevitt HO. Proc Natl Acad Sci U S A 1994 Jan I ;9 I (2):437), systemic diseases, systemic autoimmune diseases, systemic lupus erythematosus (Datla SK., Lupus t 98;7 (9):591 ), glandular diseases, glandular autoimmune diseases, pancreatic diseases, pancreatic autoimmune diseases, Type I diabetes (Castano L and Eisenbarth OS. Ann. Rev. Immunol. 8:647); thyroid diseases, autoimmune thyroid diseases, Graves' disease (Sakata S. et al., ol Cell Endocrinol 1 93 Mar;92 ( 1 ):77); ovarian diseases (Garza KM. et al., J Rcprod Immunol 1998 Feb;37 (2):87), prostatitis, autoimmune prostatitis (Alexander RB. et al.. Urology 1 997 Dec;50 (6):893), polyglandular syndrome, autoimmune polyglandular syndrome, Type I autoimmune polyglandular syndrome (l lara T. et l., Blood. 1991 Mar l ;77 (5): 1 127), neurological diseases, autoimmune neurological diseases, multiple sclerosis, neuritis, optic neuritis (Soderstrom M. et al., J Neurol Neurosurg Psychiatry 1 994 May;57 (5):544), myasthenia gravis (Osliima M. et al., Eur J Immunol 1 990 Dee;20 (12):2563), sli ff-man syndrome (Hiemslra MS. et al., Proc Natl Acad Sci U S A 200 1 Mar 27;98 (7):3988), cardiovascular diseases, cardiac autoimmunity in Chagas' disease (Cun!ia-Nelo E. et al., J Clin I nvest 1996 Oct 15;98 (8); 1 709), autoimmune thrombocytopenic purpura (Scmple J W. et al., Blood 1 96 May 1 5;87 ( 1 0):4245), anti-helper T lymphocyte autoimmunity (Caporossi A P. et al., Viral Immunol 1 98; I I ( 1):9). hemolytic anemia (Sal lah S. et al., Ann l-lemalol 1 997 Mar;74 (3): 1 39), hepatic diseases, hepatic autoimmune diseases, hepatitis, chronic active hepatitis (Franco A. et al. , Clin Immunol i mmunopathol 1 990 Mar;54 (3):382), biliary cirrhosis, primary biliary cirrhosis (Jones DE. Clin Sci (Colc ) 1 96 Nov;9 l (5):55 1 ), nephrie diseases, nephric autoimmune diseases, nephritis, interstitial nephritis (Kelly CJ. J Am Soc Nephrol 1 990 Aug; l (2): 140), connective tissue diseases, ear diseases, autoimmune connective tissue diseases, autoimmune ear disease (Yoo TJ. et al.. Cell Immunol 1994 Aug; 1 57 ( l ):249), disease of the inner car (Gloddek 13. et al. , Ann N Y Acad Sci 1 997 Dec 29;830:266), skin diseases, cutaneous diseases, dermal diseases, bul lous ski n diseases, pemphigus vulgaris, bul lous pemphigoid and pemphigus foliaceus.

Examples of delayed type hypersensitivity include, but are not l imited to, contact dermatitis and drug eruption.

Examples of types of T lymphocyte mediating hypersensitivity include, but are not limited to, helper T lymphocytes and cytotoxic T lymphocytes.

Examples of hel per T lymphocyte-mediated hypersensitivity include, but are not limited lo, T|, l lymphocyte mediated hypersensitivity and T|,2 lymphocyte mediated hypersensitivity.

A u toi' ft m ne diseases Include, but are not limited to, cardiovascular diseases, rheumatoid diseases, glandular diseases, gastrointestinal diseases, cutaneous diseases, hepatic diseases, neurological diseases, muscular diseases, ncphric diseases, diseases related to reproduction, connective tissue diseases and systemic diseases.

Examples of autoimmune cardiovascular diseases include, but are not limited lo atherosclerosis (Matsuura E. et aL, Lupus. 1 98;7 Suppl 2:S 135), myocardial infarction (Vaarala O. Lupus. 1 98;7 Suppl 2:8132), thrombosis (Tincani A. et aL, Lupus 1998,7 Suppl 2:S 1 07-9), Wegener's granulomatosis, Takayasu's arteritis, Kawasaki syndrome (Praprotnik S. et aL, Wien Klin Woclienschr 2000 Aug 25; ] 12 ( I 5- I ):660), anti-factor VI I I autoimmune disease (Laeroix-Dcsmazes S. et a , Semin Thromb l lemost.2000;26 (2): 1 57), necrotizing small vessel vascul itis, microscopic polyangiitis, C urg and Strauss syndrome, pauci-immune focal necrotizing and crescenlic glomerulonephritis (Noel LH. Ann Med Interne (Paris). 2000 ay; 1 5 l (3): 1 78), anliphospholipid syndrome (Flambolz R. et aL, J Clin Apheresis 1 99; 14 (4): 1 71 ), antibody-induced heart failure (Wallukat G. et aL, Am J Cardiol . 1999 Jun 17;83 ( 12A):75H), thrombocytopenic purpura (Moccia F. Ann Hal Med Int. 1999 Apr-Jun; 14 (2): I 14; Seniplc J W. et aL, Blood 1 996 May 15;87 ( I 0):4245), autoimmune hemolytic anemia (Ef emov DO. et aL, Leuk Lymphoma 1998 Jan;28 (3-4):285; Sallah S. et aL, Ann l lemalol 1997 Mar; 74 (3): 1 39), cardiac autoimmunity in Chagas' disease (Cunha-Nelo E. et aL, J Clin Invest 1996 Oct 15;98 (8): 1 709) and anti-helper T lymphocyte autoimmunity (Caporossi AP. et aL, Viral Immunol 1 998; l I ( l ):9).

Examples of autoimmune rheumatoid diseases include, but are not limited to rheumatoid arthritis (Krenii V. et aL, Hislol H islopathol 2000 Jul; 1 5 (3):791 ; Tisch R, McDcvill 110. Proc Natl Acad Sci units S A 1 994 Jan 1 8;91 (2):437) and ankylosing spondylitis (Jan Voswinkel et aL, Arthritis Res 2001 ; 3 (3): 1 89).

Examples of autoimmune glandular diseases include, but are not limited to, pancreatic disease, Type 1 diabetes, thyroid disease, Graves' disease, thyroiditis, spontaneous autoimmune thyroiditis, Hashimoto's thyroiditis, idiopathic myxedema, ovarian autoimmunity, autoimmune anti-sperm infertility, autoimmune prostatitis and Type 1 autoimmune polyglandular syndrome, diseases include, but are not limited to autoimmune diseases of the pancreas, Type 1 diabetes (Caslano L. and Eisetibarth GS. Ann. Rev. Immunol. 8:647; Zimmel P. Diabetes Res Clin Pracl 1996 Ocl;34 SupphS 125), autoimmune thyroid diseases, Graves' disease (Orgtazzi J. Endocrinol Metab Clin North Am 2000 Jun;29 (2):339; Sakala S. et al, Mol Cell Endocrinol 1993 Mar;92 ( l ):77), spontaneous autoimmune thyroiditis (Braley-Mullen H. and Y S, J Immunol 2000 Dec 15; 165 ( 12):7262), Hashimoto's thyroiditis (Toyoda N. et al, Nippon Rinsho 1999 Aug;57 (8): 1 10), idiopathic myxedema (Milsuma T. Nippon Rinsho. 1999 Aug;57 (8): 1759), ovarian autoimmunity (Garza KM. el til., J Reprod Immunol 1998 Feb;37 (2):87), autoimmune anti-sperm infertility (Diekman AB. et al., Am J Reprod Immunol. 2000 Mar;43 (3): 134), autoimmune prostatitis (Alexander RB. et al, Urology 1 97 Dee;50 (6):893) and Type 1 autoimmune polyglandular syndrome (Mara T. et al, Blood. 1 1 Mar 1 ;77 (5): 1 127).

Examples of autoimmune gastrointestinal diseases include, but are not limited to, chronic inllammatory intestinal diseases (Garcia l lerola A. et al, Gastroenterol Hepatol. 2000 Jan;23 ( 1 ): 16), celiac disease (Landau YE. and Slioenleld Y. Harefuah 2000 Jan 16; 138 (2): 122), colitis, ileitis and Crohn's disease.

Examples of autoimmune cutaneous diseases include, but are not limited to, autoimmune bullous skin diseases, such as, but are not limited to, pemphigus vulgaris, bullous pemphigoid and pemphigus foliaeeus.

Examples of autoimmune hepatic diseases include, but are not limited to, hepatitis, autoimmune chronic active hepatitis (Franco A. et al , Clin Immunol lmmunoputhol 1990 Mar;54 (3):382), primary biliary cirrhosis (Jones DE. Clin Sci (Colc ) 1996 Nov;9 l (5):551 ; Strassburg CP. et al, Eur J Gastroenterol Hepalol. 1999 Jun; 1 1 (6):595) and autoimmune hepatitis (Manns MP. J Hepalol 2000 Aug;33 (2):326).

Examples of autoimmune neurological diseases include, but are not limited to, multiple sclerosis (Cross AH. et al, J Neuroimmunol 2001 Jan 1 ; 1 12 ( 1 -2): 1 ), Alzheimer's disease (Oron L. et al, J Neural Transm Suppl. 1997;49:77), myasthenia gravis (Infante AJ. And Kraig E, hit Rev Immunol 1 99; 18 (l -2):83; Oshima M. et al, Eur J Immunol 1990 Dec;20 (12):2563), neuropathies, motor neuropathies (Kornberg AJ. J Clin Neurosci. 2000 May;7 (3): 191); Guillain-Barre syndrome and autoimmune neuropathies (Kusunoki S. Am J Med Sci. 2000 Apr; 1 (4):234), myasthenia, Lambert-Eaton myasthenic syndrome (Takamori M. Am J Med Sci. 2000 Apr;3 19 (4):204); paraneoplastic neurological diseas s, cerebellar atrophy, paraneoplastic cerebellar atrophy and stiff-man syndrome (M icmslra I IS. et ai, Proe Natl Aead Sci units S Λ 2001 Mar 27;98 (7):3988); non-paraneoplaslie sti ff man syndrome, progressive cerebellar atrophies, encephalitis, Rasmussen's encephal itis, amyotrophic lateral sclerosis, Sydeham chorea, Gilles de la Tourelte syndrome and autoimmune polyendocrinopathies (Antoine JC. and Honnorat J. Rev Neurol (Paris) 2000 Jan; 156 ( 1 ) :23); dysimmunc neuropathies (Nobi le-Orazio E. et ai, Electrocncephalogr Clin Neurophysiol Suppl 1 99;50:4 ! 9); acquired neuromyotonia, arthrogryposis multiplex congenita (Vincent A. et ai, Ann N Y Acad Sci. 1998 May 1 3;841 :482), neuritis, optic euritis (Sodcrstrom M . et ai, J Neurol Neurosurg Psychiatry 1994 May;57 (5):544) and neurodegenerative diseases.

Examples of autoimmune muscular diseases include, but are not limited to, myositis, autoimmune myositis and primary Sjogren's syndrome (Feist E. ei ai, Inl Arch Al lergy I mmunol 2000 Sep; 123 ( 1 ) 92) and smooth muscle autoimmune disease (Zauli D. et ai, Biomed Pharmaeother 1 99 Jun;53 (5-6):234).

Examples of autoimmune nephric diseases include, but are not limited to, nephritis and autoimmune interstitial nephritis (Kelly CJ. J Am Soc Nephrol 1990 Aug; l (2): 140).

Examples of autoimmune diseases related to reproduction incl ude, but are not limited to, repeated fetal loss (Tincani A. et ai, Lupus 1 998; 7 Suppl 2:S 1 07-9).

Examples of autoimmune connective tissue diseases include, but are not limited to, ear diseases, autoimmune ear diseases (Yoo J. et ai , Cel l Immunol 1994 Aug; 1 57 ( J ):249) and autoimmune diseases o the inner ear (Gloddek 13. et ai , Ann N Y Acad Sci 1 97 Dec 29; 830:266).

Examples of autoimmune systemic diseases include, but are not limited to, systemic lupus erythematosus (Erikson J. et i, Immunol Res 1 998; 1 7 ( l -2):49) and systemic sclerosis (Renaudincau Y. et ai, Clin Diagn Lab Immunol. 1999 Mar;6 (2) : 1 56); Chan OT. et ai, Immunol Rev 1 999 Jun; 1 9: 107).

Infectious diseases Examples of infectious diseases include, but are not limited to, chronic infectious diseases, subacute infectious diseases, acute infectious diseases, viral diseases, bacterial diseases, protozoan diseases, parasitic diseases, fungal diseases, mycoplasma diseases and prion diseases.

Graft rejection diseases Examples of diseases associated wi th transplantation of a graft include, but are not l imited to, graft rejection, chronic grail rejection, subacute graft rejection, hyperacute graft rejection, acute graft rejection and graft versus host disease.

Allergic diseases Examples of allergic diseases include, but are not limited to, asthma, hives, urticaria, pollen allergy, dust mite allergy, venom al lergy, cosmetics allergy, latex allergy, chemical allergy, drug al lergy, insect bile allergy, animal dander allergy, stinging plant allergy, poison ivy allergy and food allergy.

Cancerous diseases Examples of cancer include but are not limited to carcinoma, lymphoma, blasloma, sarcoma, and leukemia. Particular examples of cancerous diseases but are not l imited to: Myeloid leukemia such as Chronic myelogenous leukemia. Acute myelogenous leukemia with maturation. Acute promyelocyte leukemia, Acute nonlyniphocylic leukemia with increased basophils, Acule monocytic leukemia. Acute myelomonocytic leukemia with eosinophi lic!; Malignant lymphoma, such as Birkilt's Non-Hodgkin's; Lymphoctyic leukemia, such as Acule lumphoblaslic leukemia. Chronic lymphocytic leukemia; Myeloproli ferative diseases, such as Solid tumors Benign Meningioma, M ixed tumors of salivary gland, Colonic adenomas; Adenocarcinomas, such as Small cel l lung cancer, Kidney, Uterus, Prostate, Bladder, Ovary, Colon. Sarcomas, Li osarcoma, myxoid, Synovial sarcoma, Rhabdomyosarcoma (al veolar), Extraskeletel myxoid clionodrosarcoma, Ewing's tumor; other include Testicular and ovarian dysgenninoma, Retinoblastoma, Wi lms' tumor, Neuroblastoma, Malignant melanoma, Mesothelioma, breast, skin, prostate, and ovarian.

Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention us delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

EXAMPLES Reference is now made to the following examples, which together with the above descriptions, illustrate the invention in a uon limiting fashion.

Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambiook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-lli Ausubel, R. M.} ed. (1 94); Ausubel el al., "Currenl Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson el at., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); melhodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I-lll Cellis, J. IE., ed. (1994); "Current Protocols in Immunology" Volumes l-Ill Coligan J. L\, ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton & Lange, Norwalk, CT (1994); Mishell and Shiigi (eds), "Selected Methods in Cellular Immunology", W. H. Freeman and Co., New York (1 80); available immunoassays are extensively described in the patent and scientific literature, see, for example, U.S. Pal. Nos. 3,791,932; 3,839,153; 3,850,752; 3,850,578; 3,853,987; 3,867,517; 3,879,262; 3,901,654; 3,935,074; 3,984,533; 3,996,345; 4,034,074; 4,098,876; 4,879,219; 5,011,771 and 5,281,521; "Oligonucleotide Synthesis" Gait, M. J., ed. (1 84); "Nucleic Acid Hybridization" Hames, B. D., and Higgins S. J., eds. (1 85); "Transcription and Translation" Hames, B. D., and Higgins S. J., Eds. (1984); "Animal Cell Culture" Freshney, R. I., ed. (1986); "Immobilized Cells and Enzymes" IRL Press, (1986); "A Practical Guide to Molecular Cloning" Perbal, B., (1984) and "Methods in linzymology" Vol. 1-317, Academic Press; "PCR Protocols: A Guide To Methods And Applications", Academic Press, San Diego, CA (1990); Marshak el al., "Strategies for Protein Purification and Characterization - A Laboratory Course Manual" CSl-IL Press (1 96); all of which are incorporated by reference as if fully set forth herein. Other general references are provided throughout this document. The procedures therein are believed to be well known in the art and are provided for the convenience of reader. All the information contained therein is incorporated herein by reference.

EXAMPLE ! Therapeutic monoclonal human and SR-BI antibody A monoclonal human atiti SR- ! antibody was produced for therapeutic use. MA TERIA LS AND METHODS SR- I encoding piasmt'ds: DNA encoding human SR-B I (CLA-I) was ampli fied using sense primer: 5' CCATGGGCTGCTCCGCCAAA 3' (SBQ ID NO: C), and anti-sense primer: 5' CTACAGTITTGCTTCCTGCAG 3' (SBQ ID NO: 7) The above described reaction mixture was subjected to an ampli fication program of I min at 95 °C, I min at 55 °C and 1 min at 72 °C for 25 cycles, generating 1 .53 kb DNA fragment of SBQ ID NO:8 (Homo sapiens encoding SR-B I niRNA, nucleotides 70- 1 599 from accession number :Z22555). After PCR reaction, the mixture was loaded onto a 5 % polyacrylamide gel in TAB buffer. PCR product was gel-puri fied, cloned into a pUC57/T vector (T-cloning kit 1212; BI Fermenlas, Vilni us, Lithuania) and then used to transform E. coli cel ls. Clones were then set|uenced (Sequenase version 2; Upstate Biotechnology, Cleveland, OH) and transferred into a pcDNA3 vector (Invitrogen. San Diego, CA). Large-scale preparation of plasniid DNA was conducted using Mega prep (Qiagen, Chatsworth, CA).

Cells: Ι ΒΚ.293 (ATCC) were transfected with human SR-B I as described before [Scarselli B, el al .} BM BO J. 2 I ( I ):501 7-25, 2002]. Expression was verif ied by FACS analysis as described before [Scarselli B, ct ul., BM BO J. 2 I ( I 9):501 7-25, 2002].

Production of monoclonal human anti SR-BI antibody: Human anti SR-B I monoclonal antibodies were produced according to one of the two following protocols: Protocol I C57/B6 mice were subsequently immunized (3 weekly immunizations) with the human SR- I (SEQ ID NO:8) encoding DNA plasmid. Two weeks after the last administration, these miee were subjected to active induction of EA . Spleen cells were obtained for production of monoclonal antibodies two weeks later with SP2 cells (ATCC) as a fusion partner as described before (E. Harlow & D. Lane, Antibodies, Cold Spring Harbor Laboratory Press, 1998). Screening of positive 5 hybridoma was done in two steps of selection. The first one selected positi ve antibodies producing cells according to the ability to bind the recombinant SR-B I over expressed by H 293. Supernatant isolated from hybridoma clones ( 1000 wells) was then subjected to FACS analysis for their abi lity to bind SR-B I Protocol II l u The cloned human SR-B I (SEQ I D NO:8), obtained as described above, was re-cloned into a pQE expression vector, expressed in E. colt (Qiagen) and then purified by an Nl-NTA-siippcr flow affinity puri fication of 6xl lis proteins (Qiagen). After puri fication, the purity o the recombinant human SR-B i was veri fied by gel electrophoresis followed by set|ucncing (N terminus) by the Teehnion's sequencing 1 5 services unit (Technion, Hai fa, Israel). This recombinant human SR- I was then injected into 1 0-weeks old BALB/C mice. First immunization was of 50 μ¾ peptide emulsi fied in CFA | incomplete Freund's adjuvant (I FA) supplemented with 1 mg/ml heat-ki lled Mycobacterium tuberculosis H37Ra in oil; Diico Laboratories, Detroit, M I ] at a total volume of 400 μΙ into the peritoneal cavity. Later on, in a 3 weeks 0 interval these mice were administrated with 50 μ^400μ! or recombinant human SR- B I emulsi fied in IFA (Di fco Laboratories, Detroit, Ml). Three weeks after the third interval mice were injected (intravenous) with 50 μ of recombinant human SR-B I in 100 μΙ PBS. Three days later spleen cells were obtained and preparation of monoclonal antibodies was conducted as described above. 5 ELISA - The indirect ELISA was used to screen hybridoinas for antibodies against SR-BI, as follows. Ninety six-welt microliter plates (NUNC) were coated with 50 ng/inl of immuno (recombinant) SR-BI (SEQ I D NO: 8) in phosphate buffered saline (PBS) overnight at 4 °C, followed by blocking with 200 μΐ of 5 % BSA in PBS. Then 100 μΙ of hybridoma supernatants were added and incubated for 0 1 hr at room temperature (RT). The plates were washed 4 ti mes with PBS containing 0.05 % Tween 20 (PBS-T), and then supplemented with peroxidasc-conjugaled goat anti-mouse IgG antibody for I hr at RT, and washed 5 times with PBS-T. Then 1 00 μί of substrate solution 3,3' ,5,5'-tetramethyl Benzedrine liquid (ICN biomedical 2y INC, Germany, TMB) were added. The reaction was stopped using 2.5M M2SO.1 and the absorbance was read by an ELISA reader at a wavelength of 450nm and background of' 63Unm.

Cell binding assay - H EK. 293 cell line was stably transfected with peDNA encoding SR-B1 (peSR-Bi). Positive clones were selected using eomycin (G4 I 8). The positively isolated ELISA hybridoma clones (isolated as described above) were taken into the second screen. N inety six wel l disposable flexible polyvinyl chloride niiciotitration plates (Dynatech laboratories, V irginia) were seeded with l * 1 06 pcSR-Bl-expressing H EK 293 cells. The ceils were washed twice with PBS before 100 μΙ of hybridoinas supernatants were added for 30 minutes on ice. Following 3 washes with PBS, pero idase-conjtigated goat anti-mouse lgG antibody was added for additional 20 min on ice. Following 3 washes with PBS 100 μΐ of substrate solution (TMB) was added. The reaction was slopped using 2.5M l-l2SO.i . After a short centri fugal ion, the reaction was transferred into a clean well and the absorbance was read by an BL1SA reader at a wavelength of 450nm and background of 630nm.

EXAMPLE 2 In vitro characterization of anti Stt-Bl therapeutic antibodies Human SR-B 1 cross-reactive antibodies (with CLA-I) generated as described in Example I according to protocol 2 were in-vitro screened and characterized. The most successful antibody obtained was E l 2, which was further in-vitro cliaraclerized as further described hereinbelow.

Materials and Experimental Procedures Inununoblot analysis - For single-label hninunohistochemistry, standard methodology was used whereby sections were incubated with primary antibodies ( 1 : 100), fol lowed by incubation with secondary antibodies ( 1 : 1 00). Mouse IgG and rabbit polyclonal IgG were used as control antibodies Isotype analysis - I olype analysis was done using Serolec kit www.serotee.com.

Culture 0/ peritoneal macrophages - Resident macrophages were obtained from a peritoneal lavage with PBS. Elicited macrophages were harvested 5 days following i.p. injection of 3 ml of 3 % Thioglycollale (TG, Di lco, Livonia, M I). Peritoneal exudate cel ls were washed, re-suspended in RPM I 1640 medium supplemented with 10 % PCS, I % penicillin, 1 % streptomycin, and incubated in 24 ilal-botlom plates (106 cells per well in 1 ml) for overnight at 37 °C. Nonadherent cells were then removed by vigorous washing (three limes), and macrophages monolayers were incubated for 1-10 days in antibiotic-free IIP Ml containing 10 % PCS. Fresh medium was provided every 3 days.

IL-10 production by macrophage culture The peritoneal macrophages generated as described above were treated with mAb ΙΞ 12 with or without 0.5 ^ig/ml LPS (Sigma) for 24 hr at 37 "C. Supernatants from either treated or untreated macrophages were assayed for the presence of IL-IO or using immunoenzimalic LISA kits (Biolegend).

Nitrite production by macrophage culture - Nitrite formation measurement was done according as described by Katakura, T., M. Miyazaki, M. obayashi, D. N. Herndon, and F. Suzuki. (2004). CCLI7 and IL-10 as effectors that enable alternatively activated macrophages to inhibit the generation of classically activated macrophages. J Immunol 172:1407], Peritoneal macrophages (I*l06/ml) were seeded in 24-well plates as describe above. Following trealment with LPS and/or mAb 12 the supernatant was taken and NO production was assayed by measuring the accumulation of nitrite in the culture medium by Griess reaction using Griess reagent system kit (Promega). Briefly: an equal volume of Griess reagent (Sulfanilamide Solution) and macrophage supernatants was incubated for 10 min at T in a dark room. An equal volume of N-l-naplhylethylenediamine dihydrochloride (NED) was then added for 10 min. An BLISA reader measured ihe absorbance at 550nm. Nitrite concentration was determined using aNC s a standard.

Results Isotype analysis of B12 revealed it to be IgGI. The pu ified E12 was reacted with a nitrocellulose membrane containing various recombinant proteins. As shown in Figure 1, mAb BI2 cross reacted with SR-BI and CLA-1 but not with MI , CXCL6 or IL-27. These resulls indicate that the antibody specifically recognizes scavenger Bl receptor in a cross-species dependent manner and is capable of recognizing the denatured form of the protein indicating that it is directed against an exposed epitope of the native protein, as further demonstrated by its ability to neautralize SR-BI activity.

The iibilily of EI2 to elicit anti-inflammatory activity, was in vitro assayed on cultured peritoneal macrophages. As shown in Figure 2 cultured peritoneal macrophages treated for 24 hours with 0.5 ^ig/ml LPS and with mAb El 2, or with isotype matched control IgO, produced significantly higher IL-10 in the presence of 5 increasing amounts of E12 than compared to control treated cells.

These results were substantiated when following NO levels in the presence of E12 antibody and LPS (0.5 ng/ml). As shown in Figure 3, mAb El 2 suppressed NO synthesis (as determined by nitrite levels) by peritoneal macrophages in a close dependent manner. Control matched isotypes had no effect of NO levels.

It) The variable regions of E12 heavy chain (VII) and light chain (VK) were seciueneed and their CDR composition determined. SEQ ID NO: 9 and 10 show the amino acid and nucleic acid sequences of framework I (FWRl) of E12 light chain, respectively. SEQ ID NO: 11 and 12 show the amino acid and nucleic acid sequences of CD 1 of E12 light chain, respectively. SEQ ID NO: 13 and 14 show the amino 15 acid and nucleic acid sequences of framework 2 (FWR2) of EI2 light chain, respectively. SEQ ID NO: 15 and 16 show the amino acid and nucleic acid sequences of CDR2 of E)2 light chain, respectively. SEQ ID NO: 17 and 18 show the amino acid and nucleic acid sequences of framework 3 (FWR3) of E12 light chain, respectively. SEQ ID NO: 19 and 20 show the amino acid and nucleic acid sequences 0 of CDR3 of E12 light chain, respectively.

SEQ ID NO: 21 and 22 show the amino acid and nucleic acid sequences of framework 1 (FW l) of El 2 heavy chain, respectively. SEQ ID NO: 23 and 24 show the amino acid and nucleic acid sequences of CDR 1 of EI2 heavy chain, respectively. SEQ ID NO: 25 and 26 show the amino acid and nucleic acid sequences of 5 framework 2 (FWR2) ο!Έ12 heavy chain, respectively. SEQ ID NO: 27 and 28 show the amino acid and nucleic acid sequences of CDR2 of EI2 heavy chain, respectively. SEQ ID NO: 29 and 30 show the amino acid and nucleic acid sequences of framework 3 (FWR3) of EI2 heavy chain, respectively. SEQ ID NO: 31 and 32 show the amino acid and nucleic acid sequences of CDR3 of E12 heavy chain, respectively. 0 EXAMPLE 3 A monoclonal antibody to SR-Bl is capable of suppressing ongoing EAE and 1BD The monoclonal antibody generated as taught in Example 1 above was shown highly effective in suppressing ongoing BAB and TNI3S induced 1 I3D, as further described hereinbelow.

MATERIALS AND METHODS Induction of EAE in mice and suppression of (lie ongoing disease with niAh to SR-Bl - A group ο1Ί 8 C57BL/6 mice was subjected to MOGp35-55 induced BAB. At the onset of disease (day 1 3) these mice were separated into three equally sick groups, On this day and on days 1 5 and 17 these groups were intraveneously administered with 500 μ% B 1 2 mAb, isolype matched human IgG (IgG I ), or PBS and followed for clinical manifestation of disease (Figure 4) by an observer blind to the experimental protocol.

Spinal cord histopathology - Histological examination of l l&E-stained sections ol* formal in-fixed, paraffin-embedded sections of the lower thoracic and lumbar regions of the spinal cord was performed. Bach section was eval uated without knowledge of the treatment status of the animal. The fol lowing scale was used: 0, no mononuclear cel l infiltration; 1 , one to five perivascular lesions per section with minimal parenchymal infiltration; 2, live to 1 0 perivascular lesions per section with parenchymal inli llralion; and 3, > I 0 perivascular lesions per section with extensive parenchymal infiltration. The mean histological score ± S B was calculated for each treatment group I munohistochemistry - For single-label immunohistochemislry, standard methodology was used whereby sections were incubated with primary antibodies ( 1 : 100), followed by incubation wilh secondary antibodies ( 1 : 1 00). Mouse IgG and rabbit polyclonal IgG were used as control antibodies Induction of experimental Colitis in Lewis rats - Experimental colitis was induced by intrarectal insti llation of 250 μΙ of 125 mg/ml 2,4,6-lrinilrobenzene sul fonic acid (TNBS) solution (Fluka, cal# 92822) dissolved in 50 % elhanol, using 8 cm neonate feeding tube as described before ['Fiorucci, S. el al., Immunity, 17:769., 2002]. 24 hours post injection al l ruts developed bloody diarrhea and severe diarrhea in the next day, accompanied wilh continuous loss of weight.

Treatment protocol for antibody transfer - On days 6, 8 and 1 0 post induction ol* experimental col itis, 500 ¾ of mAb 121 2 was injected intravenously via a tail vein. Human IgG l (Sigma) was used as a control antibody.

Sample collection - On day 1 2, Ihe rats were ki lled under kelamine-xylashie anesthesia. The terminal colon was then stripped, gently washed with PBS, opened longitudinally and macroscopically evaluated according to a modi fication of the criteria described by Morris Gut (2004);53;99- l 7. Colonic injury was scored on a 0 (normal colon) to 5 (severe damage) scale, (see Table 2, below).

Colon Histopatltology - Tissues (terminal colon, mesentery lymph nodes and spleens) were fixed in 10% neutral buffered formalin and embedded in paraffin. Hematoxyl in and cosin stained sections of the colon were evaluated histologically for four parameters: extent of ulceration, submucosal inllitration, crypt abscesses and wall thickening (see Table 3). The sum of all scores determ ined a rating of slight to severe colonic inflammation.

Immunohistoc!ieinistry - Serial sections from formalin-fixed, paraffin-embedded specimens were de ara ITini zed and rehydrated in decreasing concentrations of ethyl alcohol. Tissue sections were incubated with fresh 3 % H2O2 in methanol for 1 0 min and then washed with PBS. Sections were then treated by microwave for 15 min in 90 "C in citrate buffer and blocked with 10 % donkey serum for 30 min. I miminoislochemical analysis was carried out using primary antibodies against rat 1L-10 (polyclonal goat anli rat I L- 1 0, R&D), CD3 (mAb mouse anti rat, Pharmingen) and ED I (mAb mouse anli rat, Serolec) over night at 4 °C in a humidified chamber. Biotinylatcd donkey anti goat or anli mouse IgG were used as secondary antibodies, followed by a streptavidin-horseradish peroxidase (Zymed). The reaction was developed using aminoethylcarbazole substrate kit (Zymed).

- Macroscopic assessment of colonic damage Results Anti SR-Bl mAb suppress long-term ongoing EAE Three groups ol' mice models of EAE displaying similar cl inical manifestations were subjected to monoclonal antibody therapy and control treatments. As shown in Figure 4, mice treated with PBS or control IgG continued to develop severe EAE, while those treated wilh the and SR-BI mAb E l 2 went into fast remission without residual sign of disease (Figure 4).

On day 1 9, spleen cells were isolated from representative mice of each group and cultured for 72 h with the target antigen wilh which disease was induced. Levels of I L- 10, I L- 12 (p40 subunit) and I L-4 were then recorded using commercially avai lable EL1SA kits. Figures 5a-c summarize the results of this experiment showing a marked elevation in I L- 1 0 production (p<0.001 ), a signi ficant elevation in I L-4 production (p<0.01 ) accompanied by a signi ficant reduction in IL- 12 production (p< 0.01 ). These results are consistent with the in vitro properties of this antibody (see Figures 5a-c and may explain, at least in part the beneficial effect of this therapy (Figure 4).

Spinal cord (lumbar spi nal cord) sections obtained on day 1 9 from control EAE mice and from those subjected to lgG I or 12 therapy (see Figure 4) were subjected to an inuminohistologieal analysis of the expression of SR-BI on leukocytes around high endothelial venules (H EV). Figures 6u-c show representative sections of untreated control EAE mice, EAE mice treated with E 12 and EAE mice treated with control lgG I , respectively. In all sections of sick mice leukocytes entering the CNS highly expressed SR-BI. The reduction in the density of these cells in and SR-BI treated mice could be explained, in part, by the reduced number of invading leukocytes resulting from the decrease in the in flammatory process (i.e. lower histological score).

Finally representative sections from these groups were subjected to immunohistological analysis of I L- 10, using a commercially avai lable and I L- 10 mAb. Figures 6d-f show representative sections of untreated control EAE mice, EAE mice treated with E 12 and EAE mice treated with control lgG I , respectively. The elevation in IL- 10 production in sections of mice treated with E l 2 is apparent compared to each of the control groups. These results support the above in-vitro results, substantiating the anti-inllammalory role of and SR-B I therapy.

And SR-BI m ib suppresses experimental Colitis Similar analysis of the effect of anti-SR-B l monoclonal antibodies on IBD was effected on a rat model of colotis. The fol lowing summarizes macroscopic and microscopic analyses on col itis induced rats (6 rats per group), as well as representative samples of histopalhological analysis, fol lowed by immunohistochemistry detection of ED I positive cells (macrophages), CD3'1' T cells and l L- 10 staining in al l groups.

Table 4 below clearly shows that a significant reduction in macroscopic and microscopic scores ol' disease which is accompanied by a marked reduction in histopalhological changes in the colon.

Table 4 Figures 7a-e show representative histological colon sections obtained at day 12 of IBD onset from naive rats (Figure 7a), positive control rats suffering form TNBS induced I BD (Figure 7b), rats suffering from TNBS induced IBD that were subjected to repeated administration of isotype matched control igG (Figure 7c) in comparison to those treated with mAb 1212 (Figures 7d-e). As shown structural changes between C I 2 treated colon and control are evident. This may be explained by the shi ft in cytokine pro H ie from pro-inflammatory (in control treated animals) to anti-i flammatory cytokines (in E l 2 treated animals) as shown in Figures 8a-L Figures 8a-c show sections of untreated I BD induced rats. Massive submucosal infiltration of macrophages (ED 1 +) and both mucosal and submucosal infiltration of T cel ls (CD3-I-) are shown. I L- 1 0 production was barely detected, mainly in the mucosa.

Figures 8d-f show sections of isotype matching control treated animals. Submucosal infiltration of macrophages (E 1 +), mucosal inllllration of T cells (CD3+) and minor I L- 10 production in the mucosa are detected.

Figures 8g-i show sections of E 12 treated rats. Submucosal infiltration of macrophages (ED H-) in damaged areas is shown, and presence of macrophages in the lamina propria of unaffected areas is detected. CD3+ T ceil infiltrate healthy mucosa, with marked I L- 10 production at the mucosa.

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Claims (11)

1. An isolated polypeptide comprising an antigen recognition domain capable of specifically , binding a human scavenger receptor, wherein said antigen recognition domain comprises CDR amino acid sequences as set forth in SEQ ID NO: 11 , 15, 19, 23, 27 and 31.

2. An isolated polynucleotide comprising a nucleic odd sequence encoding the polypeptide of claim 1.

3. A pharmaceutical composition comprising as an active ingredient the polypeptide of claim 1.

4. A therapeutically effective amount of the polypeptide of claim 1 for use In reducing Inflammation In a subject in need thereof.

5. Use of the polypeptide of claim 1 for the manufacture of a medicament for treating IBD. Θ.

6. Use of the .polypeptide of claim 1 far the manufacture of a medicament for treating multiple sclerosis.

7. Use of the polypeptide of claim 1 for the manufacture of a medicament for treating an autoimmune disease. ·

8. The polypeptide of claim 1 for use in treating IBD,

9. The polypeptide of claim 1 for use In treating multiple sclerosis.

10. Tho polypeptide of claim 1 for use In treating an autoimmune dleea3G. _Dr. Hadassa Waterman Patent Attorney G.E. Ehrlich (1995) Ltd.

11. Munachem Begin Street 52 521 Kainut C n