WO1983001837A1 - Anticorps monoclonaux contre le schistosome - Google Patents

Anticorps monoclonaux contre le schistosome Download PDF

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Publication number
WO1983001837A1
WO1983001837A1 PCT/US1982/001620 US8201620W WO8301837A1 WO 1983001837 A1 WO1983001837 A1 WO 1983001837A1 US 8201620 W US8201620 W US 8201620W WO 8301837 A1 WO8301837 A1 WO 8301837A1
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antibody
produced
cell line
cells
clone
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PCT/US1982/001620
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English (en)
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And Women's Hospital Brigham
John R. David
Donald A. Harn
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Brigham & Womens Hospital
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Priority to AU1044982A priority Critical patent/AU1044982A/xx
Publication of WO1983001837A1 publication Critical patent/WO1983001837A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/20—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans from protozoa
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to hybridoma cell lines and to monoclonal antibodies produced thereby that are species and stage specific for members of the trematode parasite Schistosoma.
  • Helminth (worm) infections are the most prevalent diseases of humans and animals when considered on a world-wide scale.
  • Tre atodes (flukes) , cestodes (tapeworms), and trematodes (roundworms) are the three major parasitic classes of helminths.
  • the major human trematode disease is schistosomiasis, which infects about 250 million persons in the tropics and subtropics.
  • Adult schistosomes live as pairs in the visceral veins, Schistosoma mansoni mainly in the inferior mesenteric vein and its tributaries.
  • the female swims against the blood flow to capillaries and venules in the intestine or rectum where eggs are deposited.
  • Koprowski et al U.S. Patent 4,196,265, describes continuous cell lines of genetically stable fused cell hybrids capable of producing large amounts of IgG antibodies against specific viruses, such as influenza virus, rabies, mumps, SV40, and the like. Zurawski et al, Federation Proceedings 39:4922 (1980), discloses hybrodomas producing monoclonal IgG antibodies against tetanus toxin. Monoclonal antibodies have also been described against human tumor cells, Yeh et al, Proc. Nat. Acad. Sci 76.2927 (1979); human T lymphocyte subsets, Reinberz et al, Proc. Nat. Acad. Sci 76.4062 (1979); and malaria parasites, Nussenzweig et al, Science 207; 71 (1980).
  • an object of this invention to provide a rapid, immunological method of assaying for the presence of Schistosoma mansomi in biological fluids. It is a further object to provide a method of assaying for the presence of Schistosoma mansoni that does not cross-react to a significant extent with other genera of trematodes. It is a yet further object of this invention to provide a method of assaying for any of the stages of the life cycle of S. mansoni using one monoclonal antibody.
  • Monoclonal antibodies of the invention are reactive with all tested stages of the life cycle of Schistosoma.
  • specific is meant that antibodies bind to antigens derived from an organism to which the antibody is said to be specific so that at least three times as many antibodies bind to the specific organism as to any other trematode for any given concentrations of antibody and trematode antigen in which the concentration of antibody is the limiting factor.
  • SWAP soluble adult worm extract
  • the process comprises sensitizing an animal with an antigen to induce an immune response, obtaining immune cells from the animal, and fusing the immune cell with a malignant cell line using one of a variety of fusing agents. Resulting hybrids are then grown in a medium which precludes the expansion of the original malignant cell line. After an initial period during which non-hybrid cells die, growing hybrids may be observed microscopically. Each of these colonies is assayed for the immune function sought. Colonies which demonstrate antibody secretion against one or more stage of S. mansoni are cloned.
  • the first step of raising monoclonal hybrids is generally immunization of an animal. Because fusion occurs preferentially with proliferating cells, it is preferred to schedule immunization to obtain as many immunoblasts as possible, with harvesting occurring about 3-4 days after the last inoculation being most preferred.
  • suitable antigens for use in immunization include proteins, glycoproteins, lipoproteins, and other macromolecules present on the surface of or excreted by any stage of the life cycle of S. mansoni. Antigenic macromolecules may be used in purified form, but suitable results are also obtained using either whole, killed trematodes or membrane prepartions derived from all or part of the trematode.
  • Whole worms are preferrably rendered non- viable by chemical treatment, e.g. formaldehyde or glutaraldehyde fixation, heat or irradiation, in order minimize biohazard problems.
  • Membrane preparations can be obtained by any methods that disrupt the nematode and allow purification of the membrane faction. Suitable methods include grinding or freeze-thawing followed by differential centrification or density gradient purification.
  • a particularly preferred antigenic preparation consists of mechanically sheared tails from the bodies of circavia, suitably by vortexing, and using the tails, which can be separated from the bodies by gradient centrifugation, to immunize an animal.
  • the circarian tails are non-infective.
  • particulate antigens such as membrane preparations and circarian tails, which are often strong immunogens
  • intraperitoneal, intravenous, subcutaneous, or intra-foot pad injection can be used with success.
  • the immunization schedule may entail two or more injections, at interavls of up to a few weeks, with the last injection being three to four days before the fusion.
  • Adjuvants may be included if desired and are preferred when soluble antigens are used.
  • Circarian tails may be used without adjuvants. It is also possible to employ immune cells which have been sensitized naturally, to use cells stimulated by polyclonal activators such as lipopolysaccharides, or to carry out in vitro sensitization of either B or T lymphocytes.
  • the cell line chosen for hybridization should be capable of rapid growth, be deficient in its metabolism for a component of the growth medium, and have potential for good fusion frequency.
  • the species from which the immortalizing line is derived should be closely related to the species from which the antibody- producing cell is obtained. Intraspecies fusions, particularly between like strains, work better than
  • OMPI interspecies fusions are especially preferred. Especially preferred are plasmocytoma-derived cell lines obtained from the same species and strain as the immune cells.
  • P3-NSl-l-Ag4-l, or NSl a variant of the P3 (M0PC 2 ⁇ _) mouse myeloma line.
  • MPC-, T-X45-6TG, or X45 a Balb/c mouse plasmacytoma.
  • P3-X63-Ag8, or X63 the mouse myeloma cell line originally used by Kohler and Milstein. Recent mutants have been developed (e.g. X63-Ag8.653) which no longer secrete immunoglobulins.
  • sp2/0-Agl4 another BALB/c mouse myeloma line.
  • GD-36-A.Agl lymphoblastoid cell line obtained by injection of SV40 virus into Syrian hamsters.
  • the cells are fused to form the hybrid cell line.
  • Various techniques are available for inducing fusion and include virus-induced fusion and polyethyleneglycol-induced fusion. Inactivated Sendai virus is preferred for virus-induced fusion. Other viruses can induce fusion of somatic cells if the two parental cells are both susceptible to the virus.
  • suitable viruses are HVJ and Epstein-Barr virus.
  • Polyethyleneglycol (PEG) can also be used as a
  • O PI fusing agent PEG itself is toxic for cells at high concentrations and various concentrations should be tested for effects on viability before attempting fusion.
  • PEG having molecular weights varying from 1000 to 6000 may be used.
  • PEG should be diluted with 30-50% saline or serum-free medium. Since PEG is toxic for the cells, the time exposure to PEG should be limited. Exposure to PEG for 1-10 minutes is best for many cell lines. Other conditions that should be controlled for increased fusion efficiency are temperature and cell ratios. Extremes of temperature should be avoided during fusion. Preincubation of each component of the fusion system at about 37° prior to fusion is preferred. The ratio between spleen cells and malignant cells should be optimized to avoid self- fusion among spleen cells.
  • Myeloma/spleen-cell ratios ranging from 2:1 to 1:20 are suitable, with 1:8 to 1:10 being preferred.
  • the mixture of cells obtained after fusion contains hybrids, fused and unfused parental spleen cells, and malignant cells. Spleen cells cannot maintain growth in routine culture medium and will eventually die out. Malignant cells would keep on dividing and soon overgrow the hybrids unless a selective medium is used that will allow only the growth of hybrids. The malignant cell lines must therefore be selected so that they are unable to grow on the chosen culture medium.
  • hypoxanthine guanine phosphoribosyl transferase HGPRT
  • HGPRT + revertants may occur among the malignant cells and these should be periodically purged with 8-azaguanine.
  • the selection medium used to allow only growth of hybrids is composed of hypoxanthine, 1 x 10 ⁇ 4 M; aminopterine, 4 x 10 M; and thymidine,
  • the fusion mixture can be grown in HAT medium immeditely after fusion or at a later time.
  • the feeding schedules for the fused cells may vary, but obligatory feeding of HAT medium (or another deficient medium) at intervals, for example on days 1, 6, and 11, is required, followed by growth in either regular culture medium or a medium containing hypoxathine and thymidine ⁇
  • Standard tissue culture medium may be used to support the growth of hybrids. Good results may be obtained with Iscove's medium, Dulbecco's modified Eagle's medium (DMEM), or HY medium: DMEM enriched with 4.5g glucose/liter, 10% NCTC 109, 20% serum, and 0.15% glutamine. HY medium is preferred.
  • Hybrids Serum used in media should be tested for its ability to support the growth of the malignant cell line prior to use.
  • Hybrids may grow in, for example, horse or calf serum, but fetal bovine serum has no immunoglobulin, an important consideration that makes screening for antibody-producing cells much easier, and is therefore preferred.
  • Hybrids may also be grown in serum-free media supplemented with 10-20% of a serum albumin, e.g., bovine serum albumin, and trace elements.
  • Feeder cells may be used in the initial stages of cell growth to enhance the survivability of the isolated cells.
  • Irradiated thymocytes, spleen cells, myeloma cell lines, and mouse peritoneal macrophages may be used for feeder layers.
  • Preferred feeder cells for use with mouse hybrids are mouse pertinoneal macrophages from the same species as the spleen donor, obtained by washing the peritoneal cavity of a mouse with aqueous sucrose. Macrophages harvested and plated the day before fusion are preferred (about 5 x 10 /ml x l0 4 / ml ) •
  • Rapid identification of antibody-producing hybrids is important in order to avoid expenditure of time and resources on cultivation of extraneous cells.
  • Early detection of hybridoma antibodies may be performed using standard immunological assays, for example, where the antigen is bound to a solid support and allowed to react with hybridoma supernatants.
  • the presence of antibodies may be detected by sandwich techniques using a variety of indicators, such as rabbit anti-mouse antibodies that are labelled with radioactive isotopes or enzymes. Most of the common methods for detecting immunological activity are sufficiently sensitive for use in detecting antibody-producing cells.
  • assay systems are discussed in more detail in a later section.
  • a solid phase assay system is used, several methods are available to bind the immunogen to a solid phase. For example, many soluble antigens bind by adsorbtion to plastic surf ces. Such plates or wells may be rigorusly washed without affecting antigen binding. When whole cells or particulate fractions comprise the antigen, glutaraldehyde or an underlayer of antibody can be used to fix cells or membranes to a plastic surface. Dessication of cells under vacuum can also be used. This technique also perserves cells for prolonged periods of time if they are used as an antigen.
  • the reagents used to detect the presence of the antibody/immunogen complex is chosen according to the species involved in the fusion. For example, when mouse cells are used, anti-mouse immunoglobulins may be used. Protein A can also be used because of its ability to bind to the Fc portion of IgG. These reagents may be labeled with radioactive isotopes
  • RIA radio-immunoassay
  • ELISA enzyme-linked immunoassay
  • EIA enzyme-linked immunoassay
  • Hybrids obtained by fusion are initially heterogenous colonies. In order to obtain a homogeneous cell line these colonies must be cloned. By this is meant the process of achieving growth of a cell line from a single parental cell. Cloning of hybrids is preferably performed after 5-16 days of cell growth in selective medium. Later cloning of hybrids usually results in colonies which are slow growers and low yielders of antibody.
  • Cloning is performed by the limiting dilution method in fluid phase or by directly selecting single cells growing in semi-solid agarose.
  • cell suspensions are diluted serially to yield samples which have a statistical probability of having only one cell per well.
  • the agarose technique begins with seeding of cells in a semi-solid upper layer of agarose over a lower layer containing feeder cells. The colonies from the upper layer are picked up and transferred to individual wells. Feeder cells, such as peritoneal macrophages, can be used to improve the cloning efficiency.
  • OMFI Antibody secreting hybrids grown in tissue culture flasks generally yield a supernatant with an antibody concentration in the range of 10-100 ⁇ g/ml. In order to obtain higher concentrations, hybrids may be transferred into animals with inflammatory ascites.
  • Ascites may be induced, for example, by intraperitoneal injection of mineral oil or 2,6,10,14- tetamethylpentadecane (pristane) 10-30 days in advance of inoculation with hybridoma cells. Under these conditions, antibody-containing ascites can be harvested 8—12 days after intraperitoneal injection of about 10 to 10 cells.
  • the ascites contains a higher concentration of antibodies (1-15 mg/ml) , but includes both monoclonal antibodies and immunoglobulins from the inflammtory ascites.
  • Antibodies may be purified by any of the standard techniques of protein separation such as differential precipitation using, for example, ammonium sulfate; electrophoresis; chromatographic separation based on molecular size, such as Sephadex chromatography? or various techniques based on binding properties of or to the antibodies, much as affinity chromatography. Complete purification is not required since only the desired immunoglobulin is present and other components do not generally interfere with its immunological
  • BALB/c or CBA/J mice are immunized with the previously described circarian tail membrane preparation. The mice are allowed to rest weeks and then immunized again. Two or more inoculations are used. Mouse spleens are removed 3-4 days after the first inoculation in order to obtain dividing B cells for fusion.
  • a preferred myeloma cell line is P3-NSI/-AG 4-1 (NS1), which is an azaguanine resistant, non-secretor myeloma line previously described by Kohler and Milstein, Eur. J. Immunol. _6_:511 (1976). When fused it produces K chains.
  • Sp210-Ag is also a preferred cell line. Thirty percent polyethylene glycol (PEG-1000, Gallard-Schlesinger) is preferred as a fusion promotor. Spleen cells are fused with the NSI cell line at a ratio of 8-10 spleen cells to 1 NSI cell.
  • the NSl/spleen-cell mixture is washed in serum-free MEM medium and suspended in 30% PEG in MEM buffered with 0.02M Hepes, pH 7.2, and centrifuged at 800 rpm for 6 min after a total of 8 min in the PEG-containing medium.
  • the PEG medium is removed, and the cells are plated onto microtiter plates in Dulbecco's MEM with high glucose (4.5 g/l) supplemented with 20% fetal calf serum, 10% NCTC 109 medium, 0.150 mg/ml oxalacetate, 0.050 mg/ml pyruvate, 0.200 units/ml bovine insulin, and 20 mM glutamine and containing 1.6 x 10 M thymidine and 10 "*4 M hypoxanthine.
  • An equal volume of the above medium containing 8 x 10 M aminopterin is added 24-48 hr later to make hypoxanthine-aminopterin- thymidine (HAT) selective medium.
  • a 125I- rabbit anti-mouse F(ab' ) 2 with at least 2 x 10' cpm/ ug is used to detect the antibodies produced by the hybridoma that bind to the target parasite cell.
  • the adult or larval parasite forms or eggs are isolated and disrupted by freeze-thawing and homogenization.
  • the membranous and soluble fraction (100,000 x g supernatant) is hydrophobically attached to polyvinylchloride "U" bottom microliter plates or attached to polylysine-coated (M.W. 150,000) "U” bottom microliter plates and used for assay after blocking remaining sites with fetal calf sera. Culture supernatants (25 ⁇ l)are incubated overnight at 5°C.
  • the plates are then washed five times with phosphate- buffered saline containing 5% fetal calf serum. Then
  • mice Positive wells are cloned by limiting dilution technique. After 10-14 days, the surviving clones are selected and transferred to microliter wells. As the clones grow, they are transferred to larger wells and finally to Falcon flasks (25 cm ) .
  • the hybrids can be grown in tissue culture or as ascites cells in mice. For the latter, mice are prepared by injecting 0.5 cm Pristane (2, 6,10,14-tetramethylpentadecane) intraperitoneally 3 to 4 weeks before injecting hybrid cells intraperitoneally. Karryotype analysis is carried out on the clones, and their products are subjected to isoelectric focussing to verify the presence of monoclonal antibodies.
  • OMPI antibodies of the present invention can be used in any of the array of available immunoassay techniques which utilize the binding interaction between the antibody and an antigen.
  • the present invention is not limited to any of these techniques in particular.
  • the most common of these is radioimmunoassay (RIA) .
  • RIA is a well- known technique and will not be described in detail here.
  • Chard "An Introduction to Radioimmunoassy and Related Techniques", North-Holland Publishing Company, 1978, which is herein incorporated by reference.
  • Any of the many variations of RIA can be used, such as homogenous phase RIA, heterogeneous or solid phase RIA, single antibody methods or double antibody methods, and direct (forward) or reverse sandwich assays.
  • solid phase systems wherein the antibody (IgG or IgM) is covalently coupled to an insoluble support so that both the antibody and the bound complex after incubation can be readily separated from the soluble free fraction.
  • solid phase supports include particles of dextran, cellulose, continuous surfaces such as polystyrene or polypropylene discs, walls of plastic tubes, glass discs, glass particles, and the like. Particulate solid phases are widely used for a variety of different assays and are included in the present invention.
  • Antibodies are attached to the particles by any of a number of techniques designed to yield a non- reversible covalent or non-covalent link between protein and particle, for example, directly or by cyanogen bromide activation.
  • an assay tube is set up containing either sample or standard, along with the tracer and an appropriate amount of solid phase bound antibody, plus a detergent to prevent aggregtion of the particles and non-specific absorption of the tracer. After an incubation period during which the tubes are continuously mixed, the solid phase is sedimented by centrifugation; the supernatant is removed and the solid phase subject to two or more washes with buffer in order to remove free tracer trapped within and between the particles. The counts on the solid phase (bound fraction) are then measured. Immunoradiometric assays, as described in Chards at page 423, can also be used. When a second antibody radioimmunoassay system is used, the second antibody may be IgM or may be IgG.
  • Another immunoassay technique useful with the antibodies of the present invention is enzyme immunoassay. This technique is also well known to the art and reference is made to Schuurs and VanWeemen, Clinica Chimica Acta, 81:1-40 (1977), which is herein incoporated by reference. In this technique, enzymes are applied as labels on antigen or antibodies for identification and localization of the immunoreactants. Any method in which the extent of binding of enzyme—labeled antigen or enzyme-labeled antibody to its immunoreactant is measured is included in this invention.
  • Enzyme immunoassays can be classified as homogenous or heterogeneous, depending on whether the labeled reagent behaves differently or identically whether or not it is bound to specific counterparts in the immunoreaction, and which therefore may or may not require physical separation of the reactants into two fractions.
  • the variety of enzymes used, methods of linking enzymes to the immunological components, purification of the conjugates, as well as various assay principles and methods are well described
  • Another immunoassay method useful in the present invention is the latex agglutination method.
  • latex particles are coated with antigen derived from Schistosoma and incubated with hybridomally produced IgM antibodies. Inhibition of agglutination will occur when a sample of physiological fluid containing the same Schistosoma antigen is incubted with this mixture. The inhibition of agglutination can either be followed with a counter or by recently developed infrared absorption techniques.
  • An alternative is to coat the latex particles with the hybridomally-produced anti-Schistosoma antibodies . Incubation of these coated particles with physiological fluid comprising Schistosoma antigen will cause aggutination.
  • animal cells such as red blood cells can of course be used.
  • the technique becomes a variation of the well known hemagglutination technique used with IgG antibodies and red blood cells.
  • Other useful immunoassay techniques are those using other labeling techniques that result in other • types of detectably-tagged antigens such as : fluorescent dyes, Aalbeses, R.C., Clin. Chim. Acta 48_:109-111 (1973); electron-dense compounds (such as ferritin) ,
  • the detectably-tagged antibodies of the invention can be prepared in kit form ready for use in an assay procedure.
  • the kit will contain the antibody in any stable form, for example, lypholized, frozen, or in solution, along with any other necessary reagents and accessories, such as Schistosoma antigen standards, reaction vessels, such as plastic tubes with or without antigen or antibody coatings, and sample transfering devices, such as pipets.
  • Many such kits are now available for other immunoassays, and the manufacturing of such kits is now standard practice.
  • the monoclonal antibodies of the invention may also be used in the chemotherapy of schistosomiasis as carriers of drugs having anti-Schistosoma activity.
  • the antibody is attached chemically, usually by a covalent bond, to a drug which is administered at the normal rate for that drug, usually intravenously.
  • the antibody acts to concentrate the drug at the location of the trematode infection and thereby to increase its effectiveness. Accordingly, in many cases it is possible to reduce the amount of drug administered in this manner.
  • human hybridoma monoclonal antibodies become available, they will be preferred
  • any method of attachment to the drug that does not destroy the medicinal properties of the drug or the antigen- antibody binding properties of the antibody is suitable. Suitable methods depend on the structure of the drug and can normally be accomplished by a reaction joining a non-essential functional group of the drug with a non-essential amino acid of the antibody. Since only a fraction of the surface of the antibody is involved in binding with the Schistosoma antigen, random attachment will generally preserve binding affinity.
  • Suitable drugs include stibocaptate, antimony tartrate, stibophen, Miracil A-D, lucanthone, hycanthone, miridazole, Furapromidium, and amoscanate.
  • the antibody with the drug attached may be made up into a pharmaceutical composition ready for use, comprising the drug-antibody combination either alone or in the presence of a pharmaceuticaly acceptble carrier.
  • the pharmaceutical carrier in which the composition is suspended or dissolved may be any solvent or solid that is non-toxic to the inoculated animal and chemcially compatable with the drug-antibody complex.
  • Suitable pharmaceutical carriers include liquid carriers, such as normal saline and other non- toxic salts at or near physiological concentrations, and solid carriers, such as talc or sucrose.
  • Hybridomas secreting monoclonal antibodies to S. mansoni were produced using three immunization protocols. The three methods were: 1) low level natural infections followed by larval boosts; 2) repeated intravenous injections with either mechanically transformed or skin-penetration prepared schistosomula; and 3) repeated intravenous injections with cercarial tails.
  • the method of fusion was that of Kennett et al, in Current Topics in Microbiology and Immunology, Vol. 81, 77, Melchers et al, eds.,
  • Antibody production was evaluated between days 14 to 17 after fusion by an indirect radioactive binding assay utilizing schistosoma antigen preparations.
  • the adult or larval parasite forms or eggs were isolated and disrupted by freeze-thawing and homogenization.
  • the membranous and soluble fraction (100,000 x g supernatant) were hydrophobically attached to polyvinylchloride " ⁇ " bottom microliter plates or attached to polylysine- coated (M.W. 150,000) "U” bottom microliter plates and used for assay after blocking remaining sites with fetal calf sera.
  • Culture supernatants 25 ⁇ l were inocubated overnight at 5 C C.
  • mice immunized with the three protocols previously mentioned served as the spleen cell donors can be summarized as follows.
  • clones beginning with T refer to cerearial tail protocol, those with WW to low dose natural infection, and those with XX or E to membrane extract. Plus (+) indicates that these clones are at least two times negative control values.

Abstract

Des clones immortels produits de manière hybride produisent des anticorps qui réagissent de manière spécifique avec des antigènes produits par le parasites trématode Schistosoma mansoni sans réaction croisée avec d'autres parasites de proche parenté. Des procédés de production de clones et d'anticorps et des procédés d'utilisation des anticorps dans des tests diagnostics et des préparations pharmaceutiques sont également décrits.
PCT/US1982/001620 1981-11-17 1982-11-17 Anticorps monoclonaux contre le schistosome WO1983001837A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0288388A1 (fr) * 1987-04-24 1988-10-26 Sanofi S.A. Immunodiagnostic des parasitoses dues à un protozoaire chez les mollusques
US5338660A (en) * 1991-07-12 1994-08-16 Board Of Supervisors Of Louisiana State University And Agricutural And Mechanical College Diagnosis of Fasciola infections by detection of antigens in feces or intestinal content
US8734807B1 (en) 2013-04-06 2014-05-27 Gabriel Langlois-Rahme Preventing and curing Schistosomiasis mansoni by inhibiting Trk receptors on female Schistosoma

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4008002C2 (de) * 1990-03-13 1994-04-28 Siemens Ag Verfahren und Vorrichtung zur Steuerung eines motorbetriebenen Stellantriebes

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4150107A (en) * 1974-04-29 1979-04-17 Behringwerke Aktiengesellschaft Antigen from schistosomes and process for obtaining same
US4178365A (en) * 1977-07-11 1979-12-11 Institut National de la Sante et de la Recherche Medicale - Centre de Technologie Biomedicale I.N.S.E.R.M. Method of preparing an antiparasitic immunological agent, the product obtained, and its application

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4150107A (en) * 1974-04-29 1979-04-17 Behringwerke Aktiengesellschaft Antigen from schistosomes and process for obtaining same
US4178365A (en) * 1977-07-11 1979-12-11 Institut National de la Sante et de la Recherche Medicale - Centre de Technologie Biomedicale I.N.S.E.R.M. Method of preparing an antiparasitic immunological agent, the product obtained, and its application

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
Acta Tropica, 38, 1981, CRUISE et al, "Hybridoma Antibody Immunoassays for the Detection of Parasitic Infection: Further Studies on a Monoclonal Antibody witj Immunodiagnostic Potential" p. 437-47 *
C R Acad. Sci. Paris, 289, 1979, VERWAERDE et al, "Production D"Anticorps Monoclonauxx Anti Schistosoma Mansoni", p. 725-727 *
Current Topics in Microbiology and Immunology, 81, 1979, MELCHERS et al, "Lymphocyte Hybridomas" p. IX-XVII *
Federation Proceedings, 40(3), 3-1981, ZODDA et al, "Characterization of Monoclonal Antibodies Against Schistosoma Mansoni" Abstract # 4433 *
Journal of Immunological Methods, 46, 1981 VOLLER et al. "Diagnostic Serology of Tropical Diseases" p. 1-29 *
Journal of Immunology 117 (5), PELLEY et al, "Schistosoma Mansoni Soluble Egg Antigens" p. 1553-60 *
Proceedings of the National Academy of Sciences, 77(9), 1980, YOULE et al, "Antithy 1.2 Monoclonal Antibody Linked to Ricin is a Potent Cell-Type-Specific Toxin" p. 5483-6 *
Proceedings of the National Academy of Sciences, 78(5) MITCHELL et al, 5-1981, "Hybridoma-Derived Antibody with Immunodiagnostic Potential for Schistosomiasis Japonica" p. 3165-9 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0288388A1 (fr) * 1987-04-24 1988-10-26 Sanofi S.A. Immunodiagnostic des parasitoses dues à un protozoaire chez les mollusques
US5338660A (en) * 1991-07-12 1994-08-16 Board Of Supervisors Of Louisiana State University And Agricutural And Mechanical College Diagnosis of Fasciola infections by detection of antigens in feces or intestinal content
US8734807B1 (en) 2013-04-06 2014-05-27 Gabriel Langlois-Rahme Preventing and curing Schistosomiasis mansoni by inhibiting Trk receptors on female Schistosoma

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