US4404280A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for preparing murine interleukin 2 in the presence of interleukin 1 from an interleukin 2 nonproducer malignant cell line and cell line therefor
Assignee: Immunex Corp
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Filing: 1981-07-16
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Granted: 1983-09-13
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Expires: 2001-07-16
Abstract
Abstract A process for producing murine IL-2 from malignant neoplastic cells which are incapable of IL-2 production by mitogen stimulation alone includes culturing murine leukemia or lymphoma cells in vitro in a protein-containing medium supplemented with various additives. A T cell mitogen and IL-1 are added to the culture medium as co-stimulants inducing the production of a supernate which contains IL-2. After a period of time, the supernate is collected and then assayed for IL-2 activity. Also in the present invention, IL-1 is utilized as a co-stimulant together with a suboptimum concentration of a T cell mitogen to induce IL-2 production in cell lines capable of generating IL-2 by mitogen stimulation alone. The use of IL-1 in these instances reduces the quantity of mitogen required to produce maximum levels of IL-2.
Inventors
- 👤 Steven Gillis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 06:49
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
57 words
A process for the production of IL-2, comprising culturing malignant murine T cells from a cell line which is incapable of producing IL-2 by T-cell mitogen stimulation alone, is capable of producing IL-2 and requires IL-1 for IL-2 production, in a culture medium containing a T-cell mitogen and IL-1; and recovering the IL-2 from the culture medium.
Claim 2
Dependent Claim
57 words
A process for the production of IL-2, comprising culturing malignant murine T cells from a cell line which is incapable of producing IL-2 by T-cell mitogen stimulation alone, is capable of producing IL-2 and requires IL-1 for IL-2 production, in a culture medium containing a T-cell mitogen and IL-1; and recovering the IL-2 from the culture medium.
Claim 3
Dependent Claim
43 words
The process of claim 1, wherein said malignant murine cells which are not capable of producing IL-2 by T-cell mitogen stimulation alone, are capable of producing IL-2 and require IL-1 for IL-2 production, are composed of T leukemic cells or T lymphoma cells.
Claim 4
Dependent Claim
44 words
The process of claim 2, wherein said malignant murine cells which are not capable of producing IL-2 by T-cell mitogen stimulation alone, are capable of producing IL-2 and require IL-1 for IL-2 production, are clones of said T leukemic cells or T lymphoma cells.
Claim 5
Dependent Claim
36 words
The process of claim 2, wherein said malignant murine cells which are not capable of producing IL-2 by T-cell mitogen stimulation alone, are capable of producing IL-2 and require IL-1 for IL-2 production, are LBRM-33-1A5 cells.
Claim 6
Dependent Claim
33 words
The process of claim 2, 3, or 4, wherein the initial T leukemic cell or T lymphoma cell concentration is in the range of about 6×10 5 to 3×10 6 cells per milliliter.
Claim 7
Dependent Claim
33 words
The process of claim 1, 2, 3, or 4, wherein the T cell mitogen in the culture medium is a compound selected from the group consisting of phytohemagglutinin, concanavalin A or pokeweed mitogen.
Claim 8
Dependent Claim
18 words
The process of claim 6, wherein the concentration of phytohemagglutinin mitogen is between 0.1 to 2.0% by volume.
Claim 9
Dependent Claim
20 words
The process of claim 6, wherein the concentration of concanavalin A mitogen is between 10 to 100 micrograms per milliliter.
Claim 10
Dependent Claim
23 words
The process of claim 6, wherein the culture medium further includes either Roswell Park Memorial Institute medium 1640, Click's medium or Dulbecco's medium.
Claim 11
Dependent Claim
35 words
The process of claim 9, wherein the culture medium further contains one or more compounds selected from the group consisting of penicillin, streptomycin, fresh glutamine, Hepes buffer, NaHCO 3 , fetal calf serum and 2-mercaptoethanol.
Claim 12
Dependent Claim
21 words
The process of claim 6, comprising recovering the IL-2 from the culture medium after approximately sixteen to forty-eight hours of culturing.
Claim 13
Dependent Claim
21 words
The process of claim 3, wherein said clones of said T lymphoma cells are clones of LBRM-33 lymphoma murine T cells.
Claim 14
Dependent Claim
38 words
A malignant murine cell line which is incapable of producing IL-2 by T-cell mitogen stimulation alone but which produces IL-2 in the presence of both a T-cell mitogen and IL-1, having the identifying characteristics of ATCC CRL 8079.
Claim 15
Dependent Claim
38 words
A malignant murine cell line which is incapable of producing IL-2 by T-cell mitogen stimulation alone but which produces IL-2 in the presence of both a T-cell mitogen and IL-1, having the identifying characteristics of ATCC CRL 8079.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-09-12 | 1987-03-14 | 1987-09-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-09-12 | 1991-03-14 | 1991-09-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-09-12 | 1995-03-14 | 1995-09-13 | $7700.00 | Window Closed / Lapsed |