Claims
- 1. A continuous hybridoma cell line which produces antigen-specific human glycosylation inhibiting factor (GIF), wherein the antigen is an allergen.
- 2. The hybridoma of claim 1, wherein the allergen is a venom component.
- 3. The hybridoma of claim 2, wherein the venom component is a honey bee verom component.
- 4. The hybridoma of claim 3, wherein the honey bee venom allergen is PLA.sub.2.
- 5. The hybridoma of claim 4 which is cell line AC5 ahaving ATCC accession number HB 10473.
- 6. The hybridoma of claim 1, wherein the allergen is pollen.
- 7. The hybridoma of claim 6, wherein the pollen is tree pollen.
- 8. The hybridoma of claim 7, wherein the tree pollen is cedar pollen.
- 9. The hybridoma of claim 8, which is cell line 31E9 having ATCC accession number HB 11052.
- 10. A method of producing a continuous hybridoma cell line capable of producing antigen-specific human glycosylation inhibiting factor (GIF) wherein the antigen is an allergen, comprising:
- (a) obtaining antigen-primed human T-cells activated to the allergen; and
- (b) combining the activated T-cells by fusion with an immortalized human lymphoblastiod fusion partner cell line to produce a continuous hybridoma cell line which produces antigen-specific human GIF.
- 11. The method of claim 10, wherein the antigen-primed human T-cells of step (a) which are activated to the allergen are produced by:
- (i) acquiring a sample containing antigen-primed human T-cells;
- (ii) activating the T-cells by culturing the T-cells in the presence of the allergen for which the T-cells are primed; and
- (iii) culturing the activated T-cells in the presence of 1L-2 and phospholipase A.sub.2 inhibitor.
- 12. The method of claim 11, wherein the phospholipase A.sub.2 inhibitor is lipocortin.
- 13. The method of claim 11, wherein the allergen for which the T-cells are primed is chemically modified.
- 14. The method of claim 13, wherein the allergen is chemically modified with cyanogen bromide.
- 15. The method of claim 10, wherein the antigen-primed human T-cells are from the peripheral blood mononuclear cell fraction.
- 16. The method of claim 10, wherein the fusion partner cell line is a human lymphoblastoid T-cell line.
- 17. The method of claim 16, wherein the lymphoblastoid cell line is BUC.
- 18. The process for producing substantially pure antigen-specific human glycosylation inhibiting factor (GIF), wherein the antigen is an allergen, comprising:
- (a) culturing a continuous human hybridoma cell line capable of producing antigen-specific human GIF such that the cell line produces antigen-specific human GIF; and
- (b) isolating substantially pure antigen-specific human GIF from the culture.
- 19. The process of claim 18, wherein the continuous hybridoma cell line is produced by the method of claim 10.
- 20. The process of claim 18, wherein during culturing the hybridoma cell line line is exposed to antigen-pulsed syngeneic marcophoges or antibodies to the CD3 or T-cell receptor.
- 21. The process of claim 18, wherein the antigen-specific human GIF is substantially purified by affinity purification using the antigen to which the antigen-specific GIF binds.
- 22. The process of claim 18, wherein the step of isolating substantially pure antigen-specific human GIF includes:
- (i) reacting the hybridoma cell line culture with a monoclonal antibody specifically reactive with human GIF;
- (ii) eluting the human GIF form the monoclonal antibody;
- (iii) reacting the eluted GIF with the allergen to which the antigen-specific human GIF binds;
- (iv) eluting the antigen-specific human GIF from the allergen; and
- (v) recovering the antigen-specific human GIF.
- 23. The process of claim 18 wherein the step of isolating substantially pure antigen-specific human GIF includes:
- (i) reacting the hybridoma cell line culture with the allergen to which the antigen-specific human GIF binds;
- (ii) eluting the human GIF from the allergen;
- (iii) reacting the eluted GIF with a monoclonal antibody specifically reactive with human GIF;
- (iv) eluting the antigen-specific human GIF from the monoclonal antibody; and
- (v) recovering the antigen-specific human GIF.
- 24. The process of claim 18 wherein the step of isolating substantially pure antigen-specific human GIF includes:
- (i) contacting the hybridoma cell line culture medium with an anionic exchange resin;
- (ii) eluting the human GIF from said resin;
- (iii) reacting the eluted GIF with a monoclonal antibody specifically reactive with human GIF or with the allergen to which the antigen-specific human GIF binds, or both;
- (iv) eluting the antigen-specific human GIF from the monoclonal antibody or the allengen in step (iii); and
- (v) recovering the antigen-specific human GIF.
- 25. The process of claim 24, wherein the anion exchange resin is DEAE.
- 26. The process of claim 24, wherein the antigen-specific human GIF is eluted in step (ii) using a salt concentration from about 10 mM to about 100 mM.
- 27. The process of claim 24, wherein the salt is NaCl.
- 28. The process of claim 24, wherein the hybridoma cell line culture is concentrated from about 25-fold to about 1000-fold then diluted from about 3-fold to about 10-fold before contacting with the anionic exchange resin.
- 29. The process as in either of claims 22 or 23, wherein the monoclonal antibody has the specificity of a monoclonal antibody produced by cell line 388F.sub.1 having ATCC accession number HB 10472.
- 30. The process as in either of claims 22 or 23, wherein the monoclonal antibody is produced by cell line 388F.sub.1 having ATCC accession number HB 10472.
Parent Case Info
This application is the national stage of PCT/US 92/04614, filed Jun. 3, 1992, which is a continuation-in-part of Ser. No. 709,375, filed Jun. 3 1991, now abandoned, which is a continuation-in-part of Ser. No. 533,889, filed Jun. 4, 1990 now abandoned.
Government Interests
This invention was made with Government support under grant numbers AI112202, AI14784, and AI32834 awarded by the National Institutes of Health. The Government has certain rights in the invention.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
102e Date |
371c Date |
PCT/US92/04614 |
6/3/1992 |
|
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1/27/1993 |
1/27/1993 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO92/21240 |
12/10/1992 |
|
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US Referenced Citations (1)
Number |
Name |
Date |
Kind |
4946788 |
Delespesse |
Aug 1990 |
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Foreign Referenced Citations (1)
Number |
Date |
Country |
8807577 |
Oct 1988 |
WOX |
Non-Patent Literature Citations (9)
Entry |
Carini et al, "A method to generate autigen-specific suppressor T cells . . . " J. Immunol. Meth. 127(2): 221-233 (Mar. 1990). |
Thomas et al, "Construction of human T cell hybridomas . . . " FASEB J. 4(7): A1937, Abstract #1418 (Apr. 1990). |
Steele et al, "A monoclonal antibody raised to lipomodulin . . . " J. Immunol 142(7): 2213-2220 (Apr. 1989). |
Jardieu et al, pp. 595-604 in Immune Regulation by Characterized Polypeptides, (Alan R. Liss, Inc. 1987). |
Akasaki et al "Immunosuppressive Effects of Glycosylation Inhibiting Factor on the IgE and IgG Antibody Responsive", J. Immunol. 136(9): 3172-3179 (May 1986). |
Iwata et al "Coustruction of Antigen-Specific Suppressor T Cells Hybridomas . . . " i J. Immunol. 141(10): 3270-3277 (Nov. 1988). |
Jardieu et al "Carrier-specific Suppression of Antibody Responses . . . " J. Immunol. 138(5): 1494-1501 (Mar. 1987). |
Ohno et al. "Effect of PLA.sub.2 inhibitors on mouse T lymphocytes . . . " Int. Immunol. 1(4):425 abstract. (1989). |
Takeuchi et al, "Developement of an Antigen-Specific CD8 Suppressor-Effector Clone in Man", J. Immunol. 141(9):3010-3015. |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
709375 |
Jun 1991 |
|
Parent |
533889 |
Jun 1990 |
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