WO1983002058A1 - Anticorps humains monocloniques ou lymphocines dans la separation de cellules ou dans le diagnostic du cancer du sein - Google Patents

Anticorps humains monocloniques ou lymphocines dans la separation de cellules ou dans le diagnostic du cancer du sein Download PDF

Info

Publication number
WO1983002058A1
WO1983002058A1 PCT/US1982/001712 US8201712W WO8302058A1 WO 1983002058 A1 WO1983002058 A1 WO 1983002058A1 US 8201712 W US8201712 W US 8201712W WO 8302058 A1 WO8302058 A1 WO 8302058A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
cell
human
partner
antibody
Prior art date
Application number
PCT/US1982/001712
Other languages
English (en)
Inventor
University Of South Carolina Medical
Anthony J. Strelkauskas
Original Assignee
Univ South Carolina
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Univ South Carolina filed Critical Univ South Carolina
Publication of WO1983002058A1 publication Critical patent/WO1983002058A1/fr
Priority to DK3251/83A priority Critical patent/DK325183D0/da

Links

Classifications

    • 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
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • 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
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • 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
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3015—Breast
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material
    • C12N5/12—Fused cells, e.g. hybridomas
    • C12N5/16—Animal cells
    • C12N5/163—Animal cells one of the fusion partners being a B or a T lymphocyte

Definitions

  • the non-rosetted, non-fused malignant partner cells are located at the interface and are removed.
  • OMPI these supernatant by conventional immunochemical techniques.
  • a typical subculture producing antibody to one form of mammary carcinoma as evidenced by reactivities to the mammary carcinoma cell line S.W. 527 is A.T.C.C. HB 8143.
  • leucocyte inhibiting factor LIF
  • LIF leucocyte inhibiting factor
  • the PMN were suspended in an agarose medium containing 10% horse serum and 0.1% agarose.
  • Droplets (0.002 ml) containing cells at 10 8 /ml were dispensed with a Hamilton syringe into flat-bottomed icrotitre plate wells, and 0.1 ml of hybridoma supernatant or compared supernatant was added to each of three wells. After incubation for 4-6 hours at 37°C, the areas of migration outside the droplets were calculated using an inverted microscope with a
  • OMPI calibrated lOx ocular The zone of migration from the edge of the droplet to the border of the migrating cells was measured in four perpendicular directions; the radius of the droplet was subtracted from the area of the migration zone. Results were expressed as a migration index calculated as area of migration in presence of mitogen divided by area of migration in absence of mitogen.
  • Example I An antibody prepared in Example I is mixed with serum or ductile secretion from a woman suspected of having mammary carcinoma. There is added a precipitating agent such as goat anti-human antibody, which has been radio-labeled. The mixture is centrifuged at high speed to bring down the precipitate. The precipitate is washed to remove excess radioactivity and the resulting precipitates are counted in a gamma counter.
  • a precipitating agent such as goat anti-human antibody, which has been radio-labeled.
  • the mixture is centrifuged at high speed to bring down the precipitate.
  • the precipitate is washed to remove excess radioactivity and the resulting precipitates are counted in a gamma counter.
  • Lymphocytes are obtained from the blood of patients in an active stage of juvenile rheumatoid arthritis (JRA). The sear of these patients are pre- screened by an assay for binding to T cells from normal donors and their lymphocytes are HLA typed. The lymphocytes are then separated and subjected to the fusion technique as in Example 1 using as the malignant fusion partner lymphoblastoid T cells, e.g., the cell line from J.M. RPMI (other T or B cell lines may also be used).
  • JRA juvenile rheumatoid arthritis
  • OMPI of the clone is conducted as in Example I.
  • a desirable culture medium consists of 90% commercial RPMI medium plus 10% fetal bovine serum.
  • Assays of the supernatants obtained from the subcultures and sera of the donor patients comparing reactivity to isolated T cells from normal donors prove that the clones make the same type of antibody to JRA as the patient's serum.
  • supernatant from a human clone producing leukocyte inhibiting factor was also tested on T cells from these normal donors; a negative result was obtained.
  • mice less than 24 hours old are injected interperitoneally with 0.03 ml of a mixture of 90% commercial RPMI culture medium and 10% fetal bovine serum, the culture medium used in Example V for subculturing. Thirty days later the mice are tested for reaction to this mixture and only the tolerized mice, which do not react, are used. These tolerized mice are injected with 0.5 ml of the supernatant mixture from the JRA clone subculture of Example V. Fourteen days later, the mice are given a booster shoot of 0,5 ml of the same supernatant.
  • Plasmacytoma (e.g. , NS-1 from the Salk Institute) is maintained in continuous culture at 37°C in CO2 and used for the hybridizations.
  • the growth medium consists of a high-glucose modified Eagle's medium (DMEM) (Gibco - Grand Island Biological, Inc. , NY) with 10% fetal calf serum (FCS) and 2% antibiotic mixture containing penicillin, streptomycin, and amphotericin B.
  • DMEM high-glucose modified Eagle's medium
  • FCS fetal calf serum
  • FCS fetal calf serum
  • Cells are cultured in flasks or multi- well culture plates and split, with new medium added every other day. Immunoglobulin is not secreted by this line, thereby alleviating the problem of nonspecific secretion of immunoglobulin.
  • Feeder layers of macrophage are obtained by flushing the peritoneal cavity with 5 ml 0.34M sucrose. Cells are washed in medium with 10% FCS, resuspended to 2-3xl0 4 ml in HAT medium and then 1 ml is added to each well of a 24-well culture plate. Incubation at 37°C in 10% CO2 is carried out for 1 hour to allow feeder cells to adhere.
  • HBSS Hank's balanced salt solution
  • the tubes are gently resuspended and the cells brought up to 48 ml in HAT medium and distributed at 1 ml in each well of 24-well cluster plates containing macrophage feeder layers. Cells are incubated at 37°C in 10% C0 2 « On days 1, 7, 10 and every second day up to 3 weeks, one ml of medium is removed from the wells and replaced by fresh HAT medium up to day 14 and by hybridoma medium without HAT after that.
  • Hybridoma Medium - used for fusions contains:
  • NCTC Nite Collection of Type cultures

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cell Biology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Microbiology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Procédé de séparation de cellules fusionnées résultant de la fusion de cellules humaines dont on sait qu'elle produit un anticorps spécifique ou un lymphocine spécifique associé à des cellules humaines malignes, desdites cellules malignes associées. Le procédé consiste en l'addition d'un antisérum spécifique capable d'identifier des spécificités antigènes propres au clone et ne réagissant pas aux cellules malignes associées non fusionnées. Après réaction de la cellule fusionnée avec l'antisérum, le produit de la réaction est séparé dans les 24 heures qui suivent par un procédé indirect de rosette. Les anticorps monocloniques peuvent être utilisés pour déceler in vitro des malignités.
PCT/US1982/001712 1981-12-08 1982-12-08 Anticorps humains monocloniques ou lymphocines dans la separation de cellules ou dans le diagnostic du cancer du sein WO1983002058A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DK3251/83A DK325183D0 (da) 1981-12-08 1983-07-14 Mumane monoclonale antistoffer og lymfokiner

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US32873881A 1981-12-08 1981-12-08
US328,738 1981-12-08
US39883982A 1982-07-16 1982-07-16
US398,839820716 1982-07-16

Publications (1)

Publication Number Publication Date
WO1983002058A1 true WO1983002058A1 (fr) 1983-06-23

Family

ID=26986489

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1982/001712 WO1983002058A1 (fr) 1981-12-08 1982-12-08 Anticorps humains monocloniques ou lymphocines dans la separation de cellules ou dans le diagnostic du cancer du sein

Country Status (5)

Country Link
EP (1) EP0095502A4 (fr)
JP (1) JPS58502132A (fr)
CA (1) CA1200484A (fr)
DK (1) DK325183D0 (fr)
WO (1) WO1983002058A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0174534A1 (fr) * 1984-08-22 1986-03-19 Tel Aviv University Essai immunologique de cancer de la glande mammaire utilisant les anticorps monoclonaux

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK172110B1 (da) * 1988-04-15 1997-10-27 Gen Hospital Corp Fremgangsmåde til isolation af mutanter af DNA-sekvenser og anvendelsen heraf til identifikation af celleoverfladeproteiner

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4172124A (en) * 1978-04-28 1979-10-23 The Wistar Institute Method of producing tumor antibodies
EP0044722A1 (fr) * 1980-07-18 1982-01-27 The Board Of Trustees Of The Leland Stanford Junior University Hybridomes humains, précurseurs et produits
WO1982001192A1 (fr) * 1980-09-25 1982-04-15 Inst Biolg Studies Salk Anticorps monocloniques specifiques des glycoproteines de surface des cellules hemopoietiques humaines
GB2086937A (en) * 1980-11-07 1982-05-19 Wilstar Inst Production of Human Monoclonal Antibodies by Human Hybridomas

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4172124A (en) * 1978-04-28 1979-10-23 The Wistar Institute Method of producing tumor antibodies
EP0044722A1 (fr) * 1980-07-18 1982-01-27 The Board Of Trustees Of The Leland Stanford Junior University Hybridomes humains, précurseurs et produits
WO1982001192A1 (fr) * 1980-09-25 1982-04-15 Inst Biolg Studies Salk Anticorps monocloniques specifiques des glycoproteines de surface des cellules hemopoietiques humaines
GB2086937A (en) * 1980-11-07 1982-05-19 Wilstar Inst Production of Human Monoclonal Antibodies by Human Hybridomas

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
Cell, Vol 11, issued May 1977 (Cambridge Mass USA) A ROSEN, Double Immunoglobulin Production in Cloned Somatic Cell Hybrids between Two Human Lymphoid Cell Lines, pp 139-147 *
National Academy of Sciences Proceedings, Vol 78, No. 5, issued May, 1981 (Washington D. C, USA) D. COLCHER, A Spectrum of Monoclonal Antibodies Reactive with Human Mammary Tumor Cells, pp 3199-3203 *
National Academy of Sciences, Proceedings Vol 77, No. 11, issued November 1980 (Washington, D.C. USA) J. SCHLOM, Generation of Human Monoclonal Antibodies Reactive with Human Mammary Carcinoma Cells pp 6841-6845 *
National Academy of Sciences, Proceedings Vol 77, No. 9, issued September 1980 (Washington, D.C. USA) L. OLSSON, Human-Human Hybridomas Producing Monoclonal Antibodies of Predefined Antigenic Specificity, pp 5429/5431 *
Nature, Vol 288, issued 04 December 1980 (Londen) CM CROCE, Production of Human Hybridomas Secreting Antibodies to Measles Virus. pp 488-489 *
Tissue Antigens, Vol. 13, issued 1978 (Munksgaard, Copenhagen, Denmark) J.W. STOCKER, Separation of Human Cells Bearing HLA-DR Antigens using Monoclonal Antibody Rosetting Method, pp 212-221 *
Tissue Antigens, Vol. 16, issued 1980, (Munksgaard, Copenhagen, Denmark, TA de KRETSER, The Separation of Cell Populations using Monoclonal Antibodies attached to Sepharose, pp 317-325 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0174534A1 (fr) * 1984-08-22 1986-03-19 Tel Aviv University Essai immunologique de cancer de la glande mammaire utilisant les anticorps monoclonaux

Also Published As

Publication number Publication date
EP0095502A1 (fr) 1983-12-07
DK325183A (da) 1983-07-14
CA1200484A (fr) 1986-02-11
DK325183D0 (da) 1983-07-14
JPS58502132A (ja) 1983-12-15
EP0095502A4 (fr) 1987-01-20

Similar Documents

Publication Publication Date Title
Raidt et al. Cellular events in the immune response: analysis and in vitro response of mouse spleen cell populations separated by differential flotation in albumin gradients
Ortaldo et al. Determination of surface antigens on highly purified human NK cells by flow cytometry with monoclonal antibodies.
Crispe et al. Expression and functional significance of the J11d marker on mouse thymocytes.
Reinherz et al. A monoclonal antibody reactive with the human cytotoxic/suppressor T cell subset previously defined by a heteroantiserum termed TH2.
Nussenzweig et al. Contribution of dendritic cells to stimulation of the murine syngeneic mixed leukocyte reaction.
Minowada et al. Expression of an antigen associated with acute lymphoblastic leukemia in human leukemia-lymphoma cell lines
Morimoto et al. The isolation and characterization of the human suppressor inducer T cell subset.
Oldham et al. Cellular microcytotoxicity in human tumor systems: Analysis of results
EP0018794B1 (fr) Anticorps monoclonal contre les cellules-T-promoteur humaines, procédé pour sa préparation, lignée cellulaire hybride le produisant, ses applications diagnostiques et thérapeutiques et compositions diagnostiques et thérapeutiques le contenant
Kotzin et al. Induction of immunoglobulin-secreting cells in the allogeneic mixed leukocyte reaction: regulation by helper and suppressor lymphocyte subsets in man.
US4363799A (en) Monoclonal antibody to human T cells, and methods for preparing same
EP0025722B1 (fr) Anticorps monoclonal contre les cellules T humaines cytotoxiques et suppressives et son procédé de préparation
Morishima et al. Functionally different T lymphocyte subpopulations determined by their sensitivity to complement-dependent cell lysis with the monoclonal antibody 4A.
JP3593032B2 (ja) ヒト単核球に対するモノクローナル抗体
US4818689A (en) Late differentiation antigen associated with helper T lymphocyte function
Ballas et al. Lymphokine-activated killer (LAK) cells. III. Characterization of LAK precursors and susceptible target cells within the murine thymus.
US4816404A (en) Late differentiation antigens associated with helper T lymphocyte function
Roberts et al. A clonal analysis of human peripheral blood lymphocytes displaying natural killer‐like activity
CA1200484A (fr) Anticorps monoclonaux et lymphokines des humains
US4618585A (en) Hybridoma cell lines producing monoclonal antibodies directed against cervical cancer cells
US4724211A (en) Human monoclonal antibodies and lymphokines and cell lines producing same
US5085985A (en) Monoclonal antibodies and their use in a method for monitoring subsets of activated T cells
US5112735A (en) Detection of lymphocyte amplification
Raziuddin et al. Phenotype, activation and lymphokine secretion by γ/δ T lymphocytes from schistosomiasis and carcinoma of the urinary bladder
EP0060871B1 (fr) Anticorps monocloniques specifiques des glycoproteines de surface des cellules hemopoietiques humaines

Legal Events

Date Code Title Description
AK Designated states

Designated state(s): DK JP KP

AL Designated countries for regional patents

Designated state(s): BE CH DE FR GB NL SE

WWE Wipo information: entry into national phase

Ref document number: 1983900257

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1983900257

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1983900257

Country of ref document: EP