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WO1993007268A1 - Formes solubles de la molecule d'adhesion des leucocytes endotheliaux, leurs utilisations et procedes diagnostiques les utilisant - Google Patents

Formes solubles de la molecule d'adhesion des leucocytes endotheliaux, leurs utilisations et procedes diagnostiques les utilisant Download PDF

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Publication number
WO1993007268A1
WO1993007268A1 PCT/GB1992/001790 GB9201790W WO9307268A1 WO 1993007268 A1 WO1993007268 A1 WO 1993007268A1 GB 9201790 W GB9201790 W GB 9201790W WO 9307268 A1 WO9307268 A1 WO 9307268A1
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WIPO (PCT)
Prior art keywords
elam
soluble
domain
preparation
bound
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PCT/GB1992/001790
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English (en)
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Andrew John Hubert Gearing
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British Bio-Technology Limited
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Publication of WO1993007268A1 publication Critical patent/WO1993007268A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/7056—Lectin superfamily, e.g. CD23, CD72
    • C07K14/70564—Selectins, e.g. CD62
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to material useful in certain diagnostic and prognostic methods and to methods of diagnosis and prognosis using the material.
  • Endothelial leukocyte adhesion molecule which is also known as E-selectin, is a membrane-bound protein which is expressed on cytokine- activated vascular endothelial cells (Bevilacqua M.P., Proc. Natl. Acad. Sci. USA. (1987) 84: 9238-9242) .
  • ELAM functions as an adhesive structure for cells bearing appropriate ligands including sialylated Lewis x and leukocyte endothelial cell adhesion molecule 1 (LECAM-1) (Phillips M.L. et al.. Science (1990) 250: 1130-1132) .
  • ELAM Cells which bind to ELAM include neutrophils, monocytes, a sub-population of lymphocytes and some tumour cells.
  • ELAM is a member of the LECAM or selectin family characterised by an N-terminal C-type lectin domain, an EGF-like domain and a variable number of complement regulatory protein (CRP) repeats with a transmembrane region and intracytoplasmic tail (Bevilacqua M.P. et al.. Science (1989) 2.3:1160-1165); ELAM has 6 CRP repeats. Expression of ELAM is restricted to endothelial cells.
  • CRP complement regulatory protein
  • ELAM is not detectable on normal endothelium but is expressed on cytokine- or endotoxin-treated endothelium in vitro, and on endothelium in tissues which are undergoing an inflammatory response in vivo (Ruco L.P. et al. Am. J. Pathol. (1990) 137: 1163-1171 ).
  • ELAM is therefore a specific marker of endothelial activation. As access to endothelium would normally require some form of invasive sampling technique or biopsy, measurement of cell surface ELAM is not a feasible routine assay for endothelial activation. If ELAM expression could be more readily detected, there would be considerable potential for ELAM-based diagnoses and prognoses. Detection of ELAM would be of particular use in a disease such as diabetes, which is characterised by damage to the vascular endothelium, as a measure of disease progression and indicator for therapeutic intervention.
  • soluble ELAM construct designated CH101 which comprises amino acids 1 to 139, inclusive
  • CH102 which comprises amino acids 1 to 177, inclusive
  • WO-A-9005786 discloses the molecular cloning of full length ELAM-1 of 610 amino acid residues; the deduced amino acid sequence is said to show "typical features of a transmembrane protein". Fragments of ELAM-1 are mentioned generically.
  • EP-A-0408859 discloses a fragment of ELAM-1, which is said to comprise "about the N-te ⁇ inal 31% of the ELAM-1 antigen". If this is referring to part of the ELAM-1 full length molecule as disclosed in O-A-9005786, then about 189 amino acid residues will be present in the fragment.
  • a preparation of soluble endothelial leukocyte adhesion molecule (ELAM) - substantially free of proteins with which it is naturally associated, as an assay standard for soluble ELAM.
  • a preparation as described above also enables antibodies, whether poiyclonal or monoclonal, to be prepared against soluble ELAM.
  • a preparation of soluble ELAM may be regarded as substantially free of proteins with which it is naturally associated when at least 70%, 80%, 90%, 95%, 98% or 99% by weight, in increasing order of preference, of the total protein present is soluble ELAM.
  • the preparation of soluble ELAM may be as pure as is obtainable by affinity chromatography, as will be described below.
  • Other components such as water, salts and buffering materials and any ether non-interfering materials, may be present in the preparation.
  • Soluble ELAM as used in the present invention may be in the form in which it is naturally present in human serum and plasma, from which it can be isolated.
  • soluble ELAM may correspond to membrane-bound ELAM but with the functional absence of that portion of the protein which effects binding to the membrane.
  • functional absence is meant that the relevant portion either is actually absent or is so modified or masked as to be ineffective for binding the protein to the membrane.
  • At least the transmembrane domain, and preferably the intracytoplasmic domain, of full length, membrane-bound ELAM is functionally absent in preferred soluble ELAM useful in this invention. In preferred embodiments of the invention, only those regions are absent.
  • Soluble ELAM may be prepared in a variety of ways. First, it may be prepared from membrane-bound ELAM by detergent extraction: membrane-bound ELAM may be prepared either in cells which naturally produce ELAM (such as activated human umbilical vein endothelial cells (HUVEC) ) or in cells which are modified to produce ELAM (such as Chinese hamster ovary (CHO) or other suitable host cells transformed or transfected to express ELAM) . Detergents used may for example be anionic or nonionic or a mixture thereof.
  • ELAM activated human umbilical vein endothelial cells
  • CHO Chinese hamster ovary
  • Detergents used may for example be anionic or nonionic or a mixture thereof.
  • soluble ELAM may be produced by proteolytic cleavage or other protein engineering of membrane-bound ELAM.
  • soluble human ELAM can also be found in human serum and plasma, from which it can be purified, for example by affinity chromatography.
  • soluble ELAM may also be prepared by recombinant DNA technology.
  • a process for the preparation of soluble ELAM comprising expressing DNA coding for an ELAM molecule in which the transmembrane domain is functionally, and preferably actually, absent and in which the lectin domain and a ⁇ _ least part of the EGF-like domain are present.
  • the intracytoplasmic domain may also be functionally absent.
  • At least the lectin domain, and preferably also most if not all of the EGF-like domain, will be present.
  • the N-terminus of ELAM up to but not including Leu 536 is present.
  • Other embodiments include, in increasing order of preference, at least residues 1 to 200, 1 to 300, l to 400, 1 to 450 and 1 to 500, inclusive.
  • DNA useful in such a process also forms part of the invention, according to a further aspect of which there is provided DNA, which will usually be recombinant or isolated, coding for a an ELAM molecule in which the transmembrane domain is functionally, and preferably actually, absent and in which the lectin domain and at least part of the EGF-like domain is present.
  • Recombinant DNA in accordance with the invention may be in the form of a vector.
  • the vector may for example be a plasmid, cosmid or phage.
  • Vectors will frequently include one or more selectable markers to enable the selection of host cells transformed (or transfected: the terms are used interchangeably in this specification) with them and, preferably, to enable selection of cells harbouring vectors incorporating heterologous DNA. Appropriate start and stop signals will generally be present. Additionally, if the vector is intended for expression, sufficient ⁇ regulatory sequences to drive expression will be included. Vectors not including regulatory sequences are useful as cloning vectors.
  • Cloning vectors can be introduced into E. coli or any other suitable hosts which facilitate their manipulation.
  • Expression vectors can be introduced into host cells suitable for the expression of soluble ELAM.
  • prokaryotic cells may be used, eukaryotic and, particularly, mammalian cells such as CHO cells may be preferred for this purpose.
  • a host cell transfected or transformed with DNA described above there is- provided a host cell transfected or transformed with DNA described above.
  • DNA in accordance with the invention can be ⁇ prepared by any convenient method involving coupling together successive nucleotides, and/or ligating oligo- and /or poly-nucleotides, including in vitro processes, but recombinant DNA technology forms the metho -of choice.
  • soluble ELAM Purification of soluble ELAM, however prepared, can be achieved by any suitable method, but a particularly preferred technique is affinity chromatography, which may be embodied in the following method: (A) providing a crude source of soluble ELAM (for example the supernatant of cells expressing a soluble ELAM protein) ; (B) introducing the crude source to an affinity matrix containing immobilised specific binding molecules for ELAM; (C) permitting soluble ELAM to bind to the matrix; (D) washing the matrix to remove unbound contaminants; and (E) recovering soluble ELAM in substantially pure form by elution from the matrix.
  • affinity chromatography which may be embodied in the following method: (A) providing a crude source of soluble ELAM (for example the supernatant of cells expressing a soluble ELAM protein) ; (B) introducing the crude source to an affinity matrix containing immobilised specific binding molecules for ELAM; (C) permitting soluble ELAM to bind to the
  • Affinity columns can be prepared in a number of ways well known to those skilled in the art.
  • Specific binding molecules which have an affinity for soluble ELAM include anti-ELAM antibodies and natural ligands for ELAM such as sialylated Lewis x antigens.
  • Cyanogen bromide-activated SEPHAROSE for example, can readily be used to couple antibodies or other specific binding molecules. (The word SEPHAROSE is a trade mark.)
  • a soluble ELAM preparation in accordance with the invention is useful in immunological and other assays for the detection of naturally released soluble forms of ELAM.
  • a method of assaying a sample of body fluid for the presence, and optionally the amount, of soluble ELAM comprising contacting the sample with a specific binding molecule for ELAM, detecting the amount of ELAM bound to the specific binding molecule and calibrating the assay with a soluble ELAM preparation as described above.
  • the specific binding molecule may be a monoclonal or poiyclonal antibody.
  • an ELAM enzyme-linked im unosorbant assay can be prepared which detects soluble ELAM in plasma and serum samples and other body fluids.
  • antibody-based assay formats including but not restricted to radioimmunoassays, two site radiobinding assays and immunoblotting assays, can equally be applied to the detection of ELAM in samples.
  • assay formats are well known to those skilled in the art and are described by Johnstone and Thorpe in 'Immunochemistry in Practice 7 , Blackwells Scientific, Oxford (1989).
  • the ligands for ELAM could also form the specific binding molecule of an assay system.
  • ELAM is known to bind to the sialylated Lewis x antigen, a complex carbohydrate which is carried on several proteins, and which acts as the physiological ligand for ELAM.
  • Sialylated Lewis x could function in an analogous fashion to an anti-ELAM antibody in any of the assay formats described above, but would have the advantage of only binding to and therefore detecting biologically active ELAM.
  • the levels of soluble ELAM in biological samples such as serum, plasma, bronchiolar lavage fluid, ascites, cerebro-spinal fluid or synovial fluid can be determined quantitatively or semi-quantitatively using any of the assay formats described above.
  • kits for use in an assay for soluble ELAM comprising (a) means for detecting soluble ELAM and (b) an assay standard comprising a preparation of soluble ELAM as described above.
  • the means for detecting soluble ELAM will depend on the particular assay method used, but may include a detecrably labelled specific binding molecule for ELAM.
  • Components (a) and (b) may be provided in separate containers within the kit.
  • Measurement of elevated levels of soluble ELAM in body fluids would provide an indication as to the state of activation or damage to the endothelium and could provide diagnostic or prognostic information.
  • a method of assessing the state of activation of, and/or damage to, the endothelium of an individual comprising assaying body fluid from the individual for the presence of, and optionally the amount of, ELAM.
  • the amount of ELAM present need not be determined absolutely: it will often be sufficient to compare the amount of ELAM detected in the individual with that detected in a control (normally healthy) individual.
  • Such a method would be particularly relevant to diseases and other conditions which affect the vasculature.
  • Vascular damage is a hallmark of diabetes leading to complications such as retinopathy; early indication of vascular damage could lead to a more rapid therapeutic intervention and possibly avert blindness in these patients, other indications could include local and systemic inflammatory conditions including, but not restricted to, transplant rejection, septic shock (in which local and systemic release of cytokines can activate endothelium) and viral infections such as cytomegalovirus , which can infect endothelium.
  • Figure l shows the detection of ELAM, expressed as optical density at 405 nm, in the supernatant of CHO cells transfected with a recombinant ELAM gene truncated at the transmembrane region so as to encode a soluble form of ELAM ;
  • Figure 2 shows the detection of ELAM, expressed as optical density at 405 nm, in human serum and the enrichment of ELAM from the serum by affinity purification on an anti-ELAM monoclonal antibody affinity column.
  • Figure 3 shows the levels of ELAM in septic shock patients and controls and demonstrates that the ELAM levels are much higher in those septic shock patients who go on to develop multiple organ failure (SIRS) .
  • SIRS multiple organ failure
  • Figure 4 shows the levels of serum creatinine and serum ELAM in renal transplant patients. Preparation: Monoclonal Antibodies to ELAM
  • Monoclonal antibodies to ELAM were raised by immunisation of mice with human umbilical vein endothelial cells (HUVEC) which have been activated to express ELAM using a mixture of interleukin-1 (IL-1) , tumour necrosis factor (TNF) and endotoxin. Standard procedures were used to derive hybridoma lines from the spleen cells of immune mice, which were screened for reactivity with the activated HUVEC. ELAM reactivity was confirmed by specific binding to COS cells transfected with the expression vector pCDM8 containing a full length human ELAM gene. The derivation and characterisation of anti-ELAM monoclonal antibodies have been described by Piggott R. et al.. Journal of Immunology, (1991) 147: 130-135.
  • Monoclonal antibodies could also be raised using the ELAM-transfeeted mammalian cells as immunogens, using ELAM which has been detergent extracted from activated HUVEC or ELAM-transfected mammalian cells, and using recombinant genetically engineered soluble forms of ELAM, or soluble ELAM from human serum or plasma.
  • Poiyclonal antibodies can be raised in mammals or birds using standard procedures with purified ELAM as immunogen.
  • Example l Preparation of Purified soluble ELAM by recombinant DNA technology
  • a recombinant soluble ELAM has been constructed by deletion of the transmembrane and intracytoplasmic domains of full length ELAM.
  • This construct can be expressed in mammalian cells using any of several expression vectors known to those skilled in the art.
  • this process is exemplified using CHO cells with the pGWIHG expression vector, disclosed in WO-A-9109118, to give a supernatant containing soluble ELAM.
  • the full length ELAM gene was isolated by an antibody panning technique using anti-ELAM monoclonal antibodies to select COS cells expressing ELAM from a population of cells which had been transfected with a cDNA library from cytokine-activated HUVEC in the expression vector pCDM8 (method described in Pigott R. et al., J. Immunol. (1991) 147: 130-135).
  • the ELAM-1 gene is available commercially, for example from the Laboratory Products Division of British Bio-technology Limited, Abingdon, Oxfordshire, UK.
  • the fragment was cloned into a pUC18 vector (Pharmacia) containing the full length ELAM cDNA sequence on an Xbal fragment from pCDM8 containing the full length ELAM gene, which had also been cut with Drain and NotI, thus replacing the DNA coding for the transmembrane domain and intracytoplasmic tail and also the 3' untranslated sequence.
  • the new fragment encoding the truncated ELAM was excised with the restriction enzyme Xbal and cloned into pG IHG also cut with Xbal.
  • the plasmid vector was purified from a large preparation in E. coli.
  • the vector was linearised with the restriction enzyme Nael and electroporated into CHO cells for expression. Expression resulted in ELAM being released into the supernatant, as indicated in Figure 1.
  • the soluble ELAM produced by these cells is used as a laboratory unitage standard for ELAM.
  • soluble ELAM was prepared from a detergent extract of CHO cells which had been transfected with a full length ELAM gene in the expression vector pGWIHG.
  • Example_3 Purification of Soluble ELAM Preparation
  • Soluble ELAM as prepared in Example 1 or 2 was purified by affinity chromatography, using CNBr-activated SEPHAROSE to couple the antibodies. (The word SEPHAROSE is a trade mark.)
  • CNBr-activated SEPHAROSE (lg, giving about 3.5ml swollen gel) was washed thoroughly with ImM HCI.
  • the lyophilised capture antibody as prepared in in the preparation above, was dissolved in coupling buffer (0.1M NaHC0 3 , pH8.3, containing NaCl), to give a 5mg/ml solution.
  • the antibody was mixed with the gel (1ml antibody per ml swollen gel) in a stoppered vessel and rotated end over end for 2 hours at room temperature. Excess antibody was washed away with coupling buffer and remaining active sites blocked with Tris-HCl buffer (0.1M, pH8.0) for 2 hours at room temperature. The product was washed with 3 cycles of alternating pH.
  • Each cycle consisting of a wash with acetate buffer (0.1M, pH4) containing NaCl (0.5M) followed by a wash with Tris buffer (0.1M, pH8) containing NaCl (0.5M) .
  • the product was packed into glass chromatography columns and stored at 4"C.
  • Example 1 or 2 containing soluble ELAM were centrifuged at high speed to remove all cell debris. Buffering reagents were added to the sample solution to produce a solution containing 0.1M Tris and 0.5M NaCl (pH8.5). This was loaded on to the appropriate column at a rate of 2ml/hr. using a peristaltic pump. The column was washed with 20 column volumes .of wash buffer (0.1M Tris, 0.5M NaCl, pH8.5) or until a baseline reading had been attained on a chart recorder. Bound proteins were eluted using glycine-HCl (0.1M, pH2.5) at lml/min using a peristaltic pump.
  • Soluble ELAM purified by affinity chromatography is indicated in Figure 2.
  • Antibodies can be used to construct assay systems for the detection of soluble ELAM in samples.
  • This example relates to a two site enzyme linked immunoassay (ELISA) for the detection of soluble ELAM.
  • ELISA enzyme linked immunoassay
  • antibody at lO ⁇ g/ml is allowed to bind in 50mM sodium carbonate buffer at pH 9.0 to the surface of wells in plastic culture plates. Unbound antibody is removed by washing with phosphate buffered saline containing 0.1% Tween (PBS-T) (the word TWEEN is a trade mark) , and any residual protein binding capacity of the plastic is blocked by an excess of another protein for example 1% casein in PBS. Samples containing ELAM are added to the wells where the ELAM specifically binds to the capture antibody.
  • PBS-T phosphate buffered saline containing 0.1% Tween
  • Excess ELAM is washed off with PBS-T, and a second antibody to ELAM is added which binds to the captured ELAM.
  • This second antibody is directly chemically conjugated to NHS-LC biotin according to the manufacturers' instructions (Pierce NHS-LC Biotin instructions 21335 ) . After 1 hour the unbound antibody is washed off with PBS-T and the biotinylated antibody is subsequently allowed to bind to a 1:1000 dilution of a streptavidin-horseradish peroxidase conjugate.
  • the enzyme After washing with PBS-T, the enzyme is revealed by conversion of a substrate, 2,2-azino-bis-(3-ethyl- benzthiazoline sulphonic acid, at 1 mg/ml in phosphate buffer pH 9.0, to give a coloured reaction product, which is detected by absorbtion of light at 405nm to give an optical density (O.D.) reading.
  • the reaction is allowed to proceed for 30 minutes before stopping by the addition of 0.2M citric acid.
  • an ELISA is used to measure ELAM levels in a variety of serum samples.
  • the assay was calibrated using a standard preparation of recombinant soluble ELAM which has been aliquoted and stored at -20 ⁇ C. This standard has been given a defined arbitrary unitage. ELAM can be detected using this assay in normal apparently healthy individuals. 1 Serum from two groups of individuals was examined. One
  • NBTS normals were normal blood donors; 24 "CAPD” were patients on chronic ambulatory peritoneal
  • the most significant association of high ELAM levels is 27 with the presence of diabetes.
  • the source of the 8 soluble ELAM is likely to be the vascular endothelium 9 as soluble ELAM has been detected in the supernatant of 0 HUVECs activated with cytokines.
  • 1 2 3 Normal levels of soluble ELAM are detected in the plasma or serum of normal individuals; elevated levels are detected in samples from patients with a variety of diseases which are characterised by activation of the immune system and activation or damage of the vascular endothelium; particularly elevated levels are detected in diabetic patients. Measurement of soluble ELAM in pathological samples is therefore of diagnostic or prognostic value in such diseases.
  • Serum levels of ELAM were measured using an ELISA test as described in Example 5.
  • the levels of soluble ELAM were monitored in the serum of a group of control patients, a group of 36 patients entering intensive care with symptoms of sepsis and a group of other patients entering intensive care with no symptoms of sepsis.
  • the results are shown in Figure 3 in which the upper and lower limits of the range of 95% confidence of the control region are shown by dotted horizontal lines. 22 of the 36 patients with symptoms of sepsis had sepsis syndrome and a poor prognosis and, of those 22 patients, 14 had initial levels of ELAM >50 U/ml or a maintained level of ELAM >30 U/ml for longer than 2 days. Peak levels of ELAM were elevated in 19/22 patients.
  • Serum levels of ELAM were measured using an ELISA test as described in Example 5.
  • the levels of soluble ELAM were measured in the serum of 15 patients following renal allograft. Rejection episodes (defined by a rise in the level of creatinine) are accompanied by a rise in the level of soluble ELAM (see Figure 4) . Confirmation of a rejection episode by measurement of ELAM will enable immunosuppressive treatment to be given with greater confidence.

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Abstract

La présence de molécules d'adhésion des leucocytes endothéliaux (ELAM) dans des liquides organiques est un indicateur de l'état d'activation et/ou de détérioration de l'endothélium. Des préparations homogènes d'ELAM soluble, qui sont utiles comme normes de dosage dans des méthodes de dosage diagnostiques et pronostiques utilisées pour des maladies affectant le système vasculaire (en particulier des complications du diabète produisant la rétinopahtie), sont obtenues selon une variété de procédés, y compris l'extraction détersive de cellules exprimant l'ELAM à fixation membranaire et l'expression d'ADN recombiné codant pour une forme tronquée d'ELAM. L'ELAM soluble et tronquée comprend le domaine de lectine et au moins une partie du domaine analogue du facteur de croissance épidermique (et de préférence la totalité de ce domaine). La chromatographie par affinité constitue une étape de purification finale utile.
PCT/GB1992/001790 1991-09-30 1992-09-30 Formes solubles de la molecule d'adhesion des leucocytes endotheliaux, leurs utilisations et procedes diagnostiques les utilisant WO1993007268A1 (fr)

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GB919120768A GB9120768D0 (en) 1991-09-30 1991-09-30 Diagnostic and prognastic materials and methods
GB9120768.8 1991-09-30

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5643873A (en) * 1992-05-06 1997-07-01 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelial leukocyte adhesion molecule 1
US5648458A (en) * 1992-05-06 1997-07-15 Affymax Technologies N.V. Peptides and compounds that bind to ELAM-1
US5728802A (en) * 1992-05-06 1998-03-17 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelium leukocyte adhesion molecule 1 (ELAM-1)
US5891645A (en) * 1994-06-01 1999-04-06 Alexion Pharmaceuticals, Inc. Porcine E-selectin

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005539A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Anticorps specifiques contre elam-1 et leur utilisation
WO1990005786A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Genes clones codant elam-1 et leurs proteines de fusion, produits proteiques qui en sont exprimes, compositions pharmaceutiques et leur utilisation
WO1990013300A1 (fr) * 1989-04-28 1990-11-15 Biogen, Inc. Molecules d'adhesion leucocytes-cellules endotheliales (elam) et molecules impliquees dans l'adhesion des leucocytes (mila)

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005539A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Anticorps specifiques contre elam-1 et leur utilisation
WO1990005786A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Genes clones codant elam-1 et leurs proteines de fusion, produits proteiques qui en sont exprimes, compositions pharmaceutiques et leur utilisation
WO1990013300A1 (fr) * 1989-04-28 1990-11-15 Biogen, Inc. Molecules d'adhesion leucocytes-cellules endotheliales (elam) et molecules impliquees dans l'adhesion des leucocytes (mila)

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JOURNAL OF IMMUNOLOGY. vol. 147, no. 1, 1 July 1991, BALTIMORE US pages 124 - 129 Lobb RR;Chi-Rosso G;Leone DR;Rosa MD;Bixler S;Newman BM;Luhowskyj S;Benjamin CD;Dougas IG;Goelz SE;et al 'Expression and functional characterization of a soluble form of endothelial-leukocyte adhesion molecule 1.' *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA. vol. 88, no. 14, 15 July 1991, WASHINGTON US pages 6224 - 6228 Polley MJ;Phillips ML;Wayner E;Nudelman E;Singhal AK;Hakomori S;Paulson JC 'CD62 and endothelial cell-leukocyte adhesion molecule 1 (ELAM-1) recognize the same carbohydrate ligand, sialyl-Lewis x' *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5643873A (en) * 1992-05-06 1997-07-01 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelial leukocyte adhesion molecule 1
US5648458A (en) * 1992-05-06 1997-07-15 Affymax Technologies N.V. Peptides and compounds that bind to ELAM-1
US5728802A (en) * 1992-05-06 1998-03-17 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelium leukocyte adhesion molecule 1 (ELAM-1)
US5786322A (en) * 1992-05-06 1998-07-28 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelium leukocyte adhesion molecule 1
US5891645A (en) * 1994-06-01 1999-04-06 Alexion Pharmaceuticals, Inc. Porcine E-selectin

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