WO2002036833A1 - Methode et kits de detection de cellules infectees par l'hepatite - Google Patents

Methode et kits de detection de cellules infectees par l'hepatite Download PDF

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Publication number
WO2002036833A1
WO2002036833A1 PCT/US2001/042919 US0142919W WO0236833A1 WO 2002036833 A1 WO2002036833 A1 WO 2002036833A1 US 0142919 W US0142919 W US 0142919W WO 0236833 A1 WO0236833 A1 WO 0236833A1
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Prior art keywords
hcv
antigen
cells
binding partner
antibody
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PCT/US2001/042919
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English (en)
Inventor
Robert A. Hallowitz
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Bio-Tech Imaging, Inc.
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Priority to AU2002214689A priority Critical patent/AU2002214689A1/en
Publication of WO2002036833A1 publication Critical patent/WO2002036833A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
    • G01N33/5767—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis non-A, non-B hepatitis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value

Definitions

  • the present invention relates to all aspects of detecting, identifying, isolating, etc., cells infected with a hepatitis type virus, such as hepatitis C virus ("HCN").
  • a hepatitis type virus such as hepatitis C virus ("HCN").
  • the invention relates to methods, reagents, kits, etc., for detecting the presence of antigens associated with virally-infected cells, especially antigens which are displayed or expressed on the surface and/or interior compartments of productively-infected cells.
  • Cells displaying such antigens can be detected, isolated, separated, cultured, subjected to treatment, etc., depending upon the desired purpose.
  • HCN liver disease in the world.
  • HCN is a rapidly worsening problem, epidemiologically much larger than the spread of HIV.
  • plasma viral load not proving adequate.
  • disease progression of HCN can be monitored by detecting the appearance of productively-infected cells in the liver, blood, lymph nodes, and other body compartments.
  • Productively-infected cells contain active virus capable of producing HCN antigens, as well ' as complete infectious virions. The appearance of extrahepatic productively-infected cells provides evidence that the disease has progressed from the primary site of infection.
  • PBMCs peripheral blood mononuclear cells
  • CAH active liver disease
  • the PBMCs were positive for both strands of HCN.
  • the patients with active liver disease had higher positive rates of negative strands of HCN-R ⁇ A in PBMCs.
  • HCN may not only infect PBMCs, but may also replicate in PBMC, and that the occurrence of the negative strand of HCN-R ⁇ A (negative-strand R ⁇ A is a replicative intermediate of HCN) is associated with activity of liver disease.
  • the HCN-infected cell isolation system of the present invention provides a number of independent and tangible benefits to the clinician in monitoring the clinical progression of the disease. For instance, since blood cells can be assayed for HCN, providing direct evidence of actively progressing disease, the performance of invasive liver biopsies on patients can be reduced an/or eliminated.
  • HCV-containing cells at sites distant from the liver suggests the immediate need to use more aggressive therapies, such as intensive interferon, ribavirin, and another antiviral strategies, thus providing a more detailed clinical description of the disease, its prognosis, and intervention strategies.
  • therapies such as intensive interferon, ribavirin, and another antiviral strategies
  • the methods also identify very early the patients who, notwithstanding intensive therapy, will progress to the requirement for liver transplant, and hence the search for donors could begin much earlier than is currently possible.
  • the methods would help monitor the efficacy of intensive treatment, e.g., a drop in the number of infected PBMCs correlates with benefit.
  • the methods are useful for describing the disease state, for providing valuable information to guide the . selection of therapeutic regimes, and in assessing the effectiveness of such treatment strategies.
  • the HCN viral genome is a positive-sense, single-stranded, 9.6 kb long R ⁇ A molecule which is translated into a single polyprotein of about 3000 amino acids.
  • the viral polyprotein is proteolytically processed by a combination of cellular and viral proteinases producing the mature viral gene products.
  • the genomic order of HCN has been shown to be C-El-E2-p7- ⁇ S2- ⁇ S3- ⁇ S4A- ⁇ S4B- ⁇ S5A- ⁇ S5B.
  • C, El and E2 are structural proteins which encapsidate and form part of the hepatitis C virion.
  • El can be detected as a 32-35 Da species and is converted into a single endo H-sensitive band of > approximately 18 kDa.
  • E2 displays a complex pattern upon immunoprecipitation consistent with the generation of multiple species. See, e.g., Grakoui et al., J. Nirol., 67:1385-1395, 1993; Tomei et al, J. Nirol., 67:4017-4026, 1993.
  • the HCN envelope glycoproteins El and E2 form a. stable complex that is co-immunoprecipitable. See, e.g., Grakoui et al, J.
  • NS3 also contains an RNA helicase domain at its C-terminus.
  • NS3 forms a heterodimeric complex with NS4A.
  • the latter is a membrane protein that has been shown to act as a cofactor of the proteinase.
  • NS5A appears to be involved in mediating the resistance of the hepatitis C virus to the action of interferon.
  • the NS5B protein has been shown to be the viral RNA-dependent RNA polymerase.
  • the present invention relates to a method of separating cells expressing a hepatitis C viral (HCV) antigen, comprising one or more of the following steps, in any effective order, e.g., a) combining (i) an effective amount of anti-HCV antigen first specific- binding partner, e.g., an antibody, optionally conjugated to a detectable label, (ii) an effective amount of a second specific-binding partner specific for said first binding partner, and (iii) an aqueous sample containing HCV-infected cells displaying said HCV antigen, to form a mixture; b) incubating said mixture under conditions effective for binding of said anti-HCV first specific -binding partner to said HCV antigen displayed by said infected cell, and for binding of said second specific binding partner to said first binding partner, form a complex; and c) separating said complex.
  • HCV hepatitis C viral
  • the present invention also relates to methods of separating out cells infected with HCV, e.g., from the blood, without using an enrichment step.
  • cells can be directly or indirectly labeled with a detectable label, and then directly counted, e.g., using FACS as described below.
  • a hepatitis C antigen can be any entity that is displayed by cells when infected by hepatitis C virus, including nucleic acids, polypeptides, and complexes thereof.
  • the hepatitis C genome codes for a number of different core, envelope, and nonstructural proteins, including, but not limited to, C, El, E2, NS1, NS2, NS3, NS4, NS5, etc.
  • a hepatitis C antigen can be one or more such proteins, fragments thereof, and nucleic acids which code therefor.
  • the detection can be achieved with an "anti-HCV antigen specific-binding partner.”
  • a specific-binding partner is a molecule, which through chemical or physical forces, selectively binds or attaches to a polynucleotide or polypeptide.
  • Specific binding partners generally are referred to in pairs, e.g., antigen and antibody, ligand and receptor.
  • Specific- binding partners include, e.g., polypeptides, such as antibodies and receptors, and polynucleotides, such as aptamers (Pitrung et al., U.S. Pat. No. 5,143,854; Geysen et al., J. Immunol. Methods, 102:259-274, 1987; Scott et al., Science, 249:386, 1990; Blackwell et al., Science, 250:1104, 1990; Tuerk et al, Science, 249: 505, 1990).
  • polypeptides such as antibodies and receptors
  • polynucleotides such as aptamers
  • an antibody specific for a polypeptide means that the antibody (e.g., the phrase "anti-HCV antigen antibody” is a shortened form, but has the same meaning) recognizes a defined sequence of amino acids within or including the polypeptide, e.g., a sequence of El, E2, etc.
  • a specific antibody will generally bind with higher affinity to the defined amino acid sequence to which it was prepared, e.g., an epitope found in El as compared to an epitope found in a different polypeptide, e.g., as detected and/or measured by an immunoblot assay or other conventional immunoassays.
  • an antibody which is specific for an epitope of a polypeptide is useful to detect the presence of the epitope in a sample, e.g., a sample of tissue containing human polypeptide product, distinguishing it from samples in which the epitope is absent.
  • Epitopes can be linear or confo.rmational.
  • a comfortnational epitope is an epitope in which the binding partner attaches to non-linear segments along the primary structure of a polypeptide, based upon its conformation in three-dimensions. Unlike a linear epitope, which is solely dependent upon the number and order of amino acids in a particular sequence, a comfortnational epitope can possess primary, secondary and/or tertiary structural requirements.
  • Antibodies e.g., polyclonal, monoclonal, recombinant, chimeric, humanized, single-chain, Fab fragment, etc.
  • a HCV envelope protein such as El or E2
  • a HCV envelope protein can be administered to mice, goats, or rabbits subcutaneously and/or intraperitoneally, with or without adjuvant, in an amount effective to elicit an immune response.
  • the antibodies can be IgM, IgG, subtypes, IgG2a, IgGl, etc.
  • Antibodies, and immune responses can also be generated by administering naked DNA See, e.g., U.S. Pat. Nos. 5,703,055; 5,589,466; 5,580,859.
  • Antibodies to HCV antigens can be prepared conventionally, e.g., as described above, and below, using whole or parts of antigen, naked DNA, phage-selection methods, etc.
  • An antibody which is specific for an HCV antigen can be conjugated to a detectable label or a capture moiety. Conjugation includes any physical association, including chemical bonding and in-frame polypeptide fusions, e.g., such as a fusion between an antibody and GFP.
  • detectable label an entity whose existence can be determined.
  • detectable labels include, but are not limited to, avidin, biotin, radioactive elements, fluorescent tags and dyes, energy transfer labels, energy-emitting entities, such as a fluorochrome, e.g., FITC, TRITC, R-phycoerythrin, Quantum Red, or Cy3, gold, ferritin, biotin, avidin, streptavidin, green fluorescent protein GFP (Chalfie et al., 1994, Science, 263:802; Cheng et al., 1996, Nature Biotechnology, 14:606; Levy et al., 1996, Nature Biotechnology, 14:610), alkaline phosphatase, peroxidase, HRP, ⁇ rease, etc.
  • a “capture moiety” is any entity that can be used to isolate, or grab on to, the moiety to which it is attached.
  • Capture moieties can be any of the above mentioned labels, but also moieties, such as nitrocellulose, magnetic or paramagnetic microspheres (e.g., as described in U.S. Pat. No. 5,411,863; U.S. Pat. No. 5,543,289; for instance, comprising ferromagnetic, supermagnetic, paramagnetic, superparamagnetic, iron oxide and polysaccharide), nylon, agarose, diazotized cellulose, latex solid microspheres, polyacrylamides, etc. See also, e.g., U.S. Pat. Nos.
  • a preferred magnetic particle is comprised of iron oxide and polysaccharide.
  • a preferred magnetic particle has a diameter which is less than the diameter of the cell which is to be . captured, e.g., about 1-300 nm, about 5-200 nm, about 10-150 nm, preferably, about 20- 150 nm, more preferably, about 50-120 nm.
  • the magnetic particles are of a sufficient size that they can form a coating around the cell, e.g., having more than one bead attached to the cell, such as about 10 beads, about 100 beads, about 1000, or about 100-1000 etc. These beads can be manufactured or commercially obtained e.g., Miltenyi Biotech, Germany.
  • the specific-binding partners can be conjugated to capture moieties.
  • a double-labeling method is used, e.g., where a first specific binding partner, specific for a viral associated antigen, is conjugated to a detectable label, and then a second specific binding partner, specific for the detectable label, is conjugated to a capture moiety.
  • the first binding partner is specific for an antigen associated with a cell and the second binding partner is specific for the detectable label.
  • other embodiments can also be used, e.g., direct labeling, where the capture moiety is conjugated to an antibody which is specific for a viral associated antigen.
  • the specific binding partner recognizes the detectable label selectively in analogy to the specificity observed with .
  • antibodies "specific for" polypeptides In this embodiment, the first and second binding partner are combined, in any effective order, with an aqueous sample containing HCN-infected cells displaying said HCV antigen, to form a mixture.
  • the binding partners can be added sequentially,. at the same time, sequentially after appropriate incubation times allowing binding to take place, sequentially with wash steps, etc.
  • one or more antibodies which are specific for a viral- associated antigen can be used.
  • the antibodies can recognize conformational or linear epitopes.
  • antibodies which recognize conformational epitopes since these are more likely to recognize antigens which are being produced by active virus, rather than adventitious virus (i.e., virus which is present in the cell, but which is not replicative).
  • Such antibodies can distinguish from native-folded or mature polypeptide, as opposed to polypeptide fragments which have been endocytosed and in the process of being degraded.
  • One or more antibodies can be used. For instance, it may be desirable to use a . mixture or cocktail of antibodies.
  • a cocktail can comprise antibodies that recognize different epitopes on the same antigen, different antigens, or combinations thereof.
  • a particular antigen can be targeted with a several different antibodies, e.g., antibodies which label: different epitopes on the same antigen, the same generic epitope, but variations, polymorphisms, hypervariable regions, etc., within it, linear epitopes, conformational epitopes, etc.
  • Various antibodies can be used, e.g., a conformation- sensitive E2 -reactive monoclonal antibody, such as H2 as described in Deleersnyder et al., J Virol, 71(l):697-704, 1997, or, Hadlock et al., J Virol, 74(22):10407-10416, 2000; antibodies to the hypervariable region 1 (HVR1) in E2, such as described in Zhou et al., Virology, 269(2):276-83, 2000;. core-protein specific antibodies, e.g., as in Heintges et al, Biochem. Biophys. Res. Commun. 263 (2) :410-8, 1999.
  • H2 conformation- sensitive E2 -reactive monoclonal antibody
  • H2 as described in Deleersnyder et al., J Virol, 71(l):697-704, 1997, or, Hadlock et al., J Virol,
  • Antibodies can also be prepared and of the.type described in, e.g., U.S. Pat. Nos. 6,136, 527, 6,121,020, 6,110,465, 6,103,485, 6,096,541, 6,074,852, 6,063,904, 6,051,696, 5,747,241, 5,698,390, 5,443,965, 5,436,318, etc., using whole antigen, phage display (e.g., Tafi et al., J. Biol Chem., 378(6):495-502, 1997), etc.
  • phage display e.g., Tafi et al., J. Biol Chem., 378(6):495-502, 1997), etc.
  • the detection and/or separation method can be accomplished on any sample comprising cells, e.g., whole blood, fractionated blood, biopsied specimens (e.g., liver), tissue sections, serum, urine, body fluids, cultured cell samples, matrices comprising cells, multi-well plates, slides, etc.
  • Cells which can be assayed in accordance with the present invention included, e.g., peripheral blood cells, lymphocytes, T-cell, B-cells, peripheral mononuclear blood cells, macrophages, liver cells, tissue culture cells, etc.
  • Control cell lines can be engineered by an effective method to express specific HCV antigens.
  • cell lines expressing E2, etc. can be transfected with coding sequences for E2, etc., where the coding sequence is operably linked to an expression control sequence (e.g., a promoter and other transcription regulatory sequences).
  • an expression control sequence e.g., a promoter and other transcription regulatory sequences.
  • the binding reaction between antibody and antigen generally occurs in aqueous media; however, any effective solvent can be used.
  • the mixture is incubated under conditions effective for binding of the at least one anti-HCV antibody to the HCV antigen displayed by said infected cell, and for binding of said antibody specific for said detectable label to said detectable label, to form a complex.
  • condition effective for means, e.g., a milieu in which the desired effect is.achieved. Such a milieu includes, e.g., appropriate buffers, oxidizing agents, reducing agents, pH, co-factors, temperature, ion concentrations, appropriate incubation times, etc.
  • the conditions are selected such that the recited outcome is accomplished, e.g., as here, binding between antigen and antibody.
  • the binding reaction is "specific" in that the antibody selectively attaches to the target antigen, e.g., an anti-E2 antibody selectively attaches ("binds") to E2 antigen, or anti-detectable label antibody selectively attaches ' ("binds") to the detectable label.
  • the viral-infected cell displays the viral-associated antigen.
  • display it is meant that the cell expresses or produces the viral antigen in detectable amounts. For instance, if the viral antigen is encoded by the viral genome, then the viral antigen is displayed by the cell when the gene is transcribed and translated into a polypeptide.
  • HCV include the viral envelope proteins, such as El and E2, and the nonstructural proteins, such as NS 1 , NS2, etc.
  • tissue culture cells transfected with HCV genes can show core protein in . a globular pattern near the nucleus, ⁇ S2 concentrated in the perinuclear space, and NS4A accumulated in the ER and the Golgi regions.
  • NS3 was displayed in the nucleus, as well as the cytoplasm, when it -was expressed by itself.
  • NS3 became restricted to the cytoplasm, when it was produced together with NS4A.
  • NS4B showed a spot-like pattern throughout the cytoplasm.
  • NS5 A and NS5B were distributed throughout the cytoplasm in a mesh-like pattern.
  • El and E2 are envelope proteins can be complexed with the endoplasmic reticulum. See, e.g., Kila et al., JBiol Chem., 275(31):24070-9, 2000.
  • Separating viral-infected cells can be accomplished by any suitable means.
  • a capture moiety conjugated to a specific-binding partner such as an antibody
  • a specific-binding partner such as an antibody
  • the principle is that infected cells will be decorated more intensely with capture moiety as a result of the display and recognition of viral-associated antigen by the specific-binding partner to which the capture moiety is attached.
  • direct or indirect labeling can be employed.
  • the nature of the capture moiety will depend upon the separation means. If a magnetic field is utilized, then the capture moiety can be ⁇ a entity which is acted upon by a magnetic field, e.g., paramagnetic or magnetic particles, such as those described above.
  • the capture moiety is a fluorescent tag, such as rhodamine or fluorosceine, which can be detected and sorted.
  • labeled cells can be detected and identified in any effective method, including by immunofluorescence, ELISA formats, by an automatic plate reader that detects label, etc.
  • the sample comprising the target cells infected with hepatitis C, or suspected of infection can be live, untreated cells, or can be cells which have been treated in any desired fashion. For instance, if the hepatitis C antigen is present in an interior compartment of the cells, it may be desired to permeabilize the cell in order to enhance access by the specific-binding partner to its target.
  • permeabilize it is meant any treatment which enhances its penetrability to a specific-binding partner, e.g., an antibody.
  • the permeabilization treatment and extent will depend upon the binding partner which is utilized, e.g., a whole antibody, an Fab fragment, a single-chain antibody, etc.
  • Permeabilization can be accompanied by fixation to preserve the structure and integrity of the cell. Methods of fixation and permeabilization are known in the art. Fixatives, include, but are not limited to, formaldehyde, paraformaldehyde, alcohol, ethanol, Boulin reagent, picric acid, methanol, acetic acid, osmium tetroxide, etc.
  • Permeabilization agents include, but are not limited to, detergents, such as non-ionic detergents, e.g., Triton X-10 . 0, CHAPS, octylglucoside, or Igepal CA-630.
  • the present invention also relates to methods of identifying an agent which interferes with hepatitis C virus (HCV) infection of a cell, such as antibodies, antiviral agents, agents which inhibit viral entry, replication, processing, etc.
  • HCV hepatitis C virus
  • Such a method can comprise one or more of the following steps in any effective order, e.g., contacting a test cell with HCV, under conditions effective for achieving infection of the cell with the virus,to form a mixture; b) adding to the resultant mixture formed in a), a test sample containing an agent suspected with interfering with said HCV infection of the test cell; c) adding to the mixture of b), a first binding partner specific for an antigen coded for by HCV which is displayed by the test cell upon viral infection, under conditions effective for the binding partner to bind to the HCV antigen; d) adding to the resultant mixture formed in c), a second binding partner specific for the first binding partner and to a magnetic bead, under 5 conditions effective for the second binding partner to bind to the first binding partner, .
  • test cell when the latter is bound to the HCV antigen displayed by the test cell, to form a complex; e) separating test cells containing said complex, whereby said separation is achieved by a magnetic field; and f) determining the number of cells infected with said virus in the presence and the absence of said test agent.
  • Viral infection of cells can be accomplished by any effective method. For instance,
  • HCV can be purified as a virion from sera of infected patients and concentrated.
  • the viral concentrate can then be combined with cells receptive for infection, under conditions in which the virus can gain entry into the cell in an infectious form.
  • cells Prior to infection, cells can be treated in such a manner as to facilitate their receptivity to virus.
  • PMBCs isolated from blood or cultures
  • an agent which promotes cell replication and/or differentiation such as PHA, CD3/CD28 ' ligand, to activate the cells, producing HCV receptivity.

Abstract

L'invention concerne des méthodes d'identification et/ou de séparation de cellules exprimant un antigène viral de l'hépatite C (HCV), par exemple des cellules sanguines et des cellules des ganglions lymphatiques. La méthode de l'invention consiste à marquer des cellules exprimant l'antigène HCV et à détecter des cellules marquées, avec ou sans étape d'enrichissement.
PCT/US2001/042919 2000-11-06 2001-11-06 Methode et kits de detection de cellules infectees par l'hepatite WO2002036833A1 (fr)

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US60/245,666 2000-11-06

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015112382A1 (fr) 2014-01-21 2015-07-30 Morehouse School Of Medicine Dépistage d'infections et de maladies médié par des exosomes

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5308750A (en) * 1989-12-22 1994-05-03 Abbott Laboratories Monoclonal antibodies to putative HCV E2/NS1 proteins and methods for using same
WO1997036004A1 (fr) * 1996-03-26 1997-10-02 Fodstad Oeystein Procede immuno-magnetique de separation cellulaire utilise pour l'identification de genes associes a la formation de metastases cancereuses a preference de site
WO1998028622A1 (fr) * 1996-12-20 1998-07-02 Fodstad Oeystein Methode de caracterisation de cellules anormales
US5817458A (en) * 1996-10-15 1998-10-06 The Avriel Group, Amcas Division Inc. Reagent system for detecting HIV-infected peripheral blood lymphocytes in whole blood

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5308750A (en) * 1989-12-22 1994-05-03 Abbott Laboratories Monoclonal antibodies to putative HCV E2/NS1 proteins and methods for using same
WO1997036004A1 (fr) * 1996-03-26 1997-10-02 Fodstad Oeystein Procede immuno-magnetique de separation cellulaire utilise pour l'identification de genes associes a la formation de metastases cancereuses a preference de site
US5817458A (en) * 1996-10-15 1998-10-06 The Avriel Group, Amcas Division Inc. Reagent system for detecting HIV-infected peripheral blood lymphocytes in whole blood
WO1998028622A1 (fr) * 1996-12-20 1998-07-02 Fodstad Oeystein Methode de caracterisation de cellules anormales

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
USHIJIMA H. ET AL.: "Removal of HIV antigens and HIV-infected cells in vitro using immunomagnetic beads", JOURNAL OF VIROLOGICAL METHODS, vol. 29, 1990, pages 23 - 32, XP002908457 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015112382A1 (fr) 2014-01-21 2015-07-30 Morehouse School Of Medicine Dépistage d'infections et de maladies médié par des exosomes
EP3097202A4 (fr) * 2014-01-21 2017-06-21 Morehouse School of Medicine Dépistage d'infections et de maladies médié par des exosomes
EP3517959A1 (fr) * 2014-01-21 2019-07-31 Morehouse School of Medicine Dépistage d'infections et de maladies médié par des exosomes

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