Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Lymphokines – e.g. – interferons – interlukins – etc.
Patent
1988-03-01
1991-12-10
Moskowitz, Margaret
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Lymphokines, e.g., interferons, interlukins, etc.
530413, 530417, 424 851, C07K 1500, C07K 1502, C07K 320, C07K 328
Patent
active
050719599
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The present invention relates to a novel lymphokine which suppresses platelet activation, to its process of isolation and purification and to medicaments containing it.
It is known that stimulated T cells excrete a certain number of soluble factors among which certain are capable of regulating the functions of effector cells.
It has been demonstrated recently (cf. M. JOSEPH, C. AURIAULT, A. CAPRON, M. VORNG and P. VIENS, NATURE 303:8IO) that blood platelets taken up in rats infected with Schistosoma mansoni, express antiparasite properties destructive in vitro, which increase in the course of the infection. The maximum cytoxicity has been observed when the rats have expressed a high level of immunity with respect to re-infection. This platelet cytoxicity declines rapidly after eight weeks of infection, despite the presence of IgE antibodies in the serum of these infected rats. This is the reason why the inventors have been led to consider that the factors of the T cells, produced after stimulation of suppressor T cells by circulating antigens of S. mansoni, play a role in this diminution, and this mechanism can appear as a regulation in return of immune functions of the platelets in the infection of the rat. In the same way, it has been shown that human platelets can be induced in cytoxic effectors against schistosomules, by incubation with serum rich in IgE of patients afflicted with schistosomiasis or asthmatic allergy patients, the cytoxic process being retarded in the latter case by the addition of anti-IgE or of a specific allergen (cf. M. JOSEPH, J. C. AMEISEN, J. P. KUSNIERZ, V. PANCRE, M. CAPRON and A. CAPRON, 1984, C. R. ACAD. SC. PARIS 298:55).
It is an object of the present invention to isolate a suppressor factor of the activity of the blood platelets, which is capable of playing, among other things, an immunomodulator role of allergic manifestations.
The present invention relates to a factor obtained from T cells stimulated by Concanavaline A or by an antigen, which can inhibit the IgE-dependant platelet cytotoxicity with respect to young larvae of S. mansoni, of greatly diminishing the chemoluminescence of platelets in an IgE-anti/IgE reaction, which is in correlation with the antiparasitic cytotoxicity and of inhibiting platelet activation in non-IgE dependant intolerances, which factor is denoted by the name of lymphokine for the suppression of platelet activation (LSPA) and is produced by T OKT8.sup.+ lymphocytes.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the percentage inhibition of the cytotoxicity as a function of the amount of Concanavaline A used as a stimulant.
FIG. 2 shows the percentage inhibition of the cytotoxicity as a function of the amount of supernatant employed.
FIG. 3 shows the percentage inhibition of the cytotoxicity represented as a function of the addition of supernatant in time, expressed in hours.
FIG. 4 shows the effect of the supernatant of T lymphocytes stimulated by Concanavaline A on the chemoluminescence of platelets.
FIG. 5 illustrates the level of respective suppressor activity of the LSPA obtained from T cells, from OKT8.sup.+ OKT4.sup.- cells and from OKT8.sup.- OKT4.sup.+ cells, stimulated by Concanavaline A.
FIG. 6 illustrates the percentage inhibition of the cytotoxicity of supernatants treated by trypsin, protease K and neuraminidase.
FIG. 7 shows a chromatographic profile of the inhibitor lymphokine of blood platelets according to the present invention.
FIG. 8 shows the percentage inhibition of cytotoxicity as a function of pH.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
In one embodiment of LSPA, the latter is isolated from supernatants T OKT8 lymphocyte cultures stimulated by Concanavaline A or an antigen.
In another embodiment of LSPA, in the case where it is isolated from supernatants of T OKT8.sup.+ lymphocyte cultures stimulated by an antigen, the T lymphocytes have been stimulated by Echinococcus granulosus or Schistosoma mansoni antigens. The present invention also relates to a process of preparation
REFERENCES:
Takakuma et al., J. of Immunol. 120(2), pp. 481-486, (1978).
Aggarwal, "Micropurification of Cytokines" in Protein Purification: Microacro, Burgess, ed., Alan R. Liss, Inc. (New York) 1987.
The Journal of Immunology, vol. 118, No. 6, Jun. 1977, The Williams & Wilkins Co. (Baltimore), U.S.), H.G. Remold et al., "Two Migration Inhibitory Factors With Different Chromatographic Behavior and Isoelectric Points", pp. 2015-2019.
The Journal of Immunology, vol. 131, No. 6, Dec. 1983, The Williams & Wilkins Co. (Baltimore, U.S.) T. M. Aune et al, "Purification and Initial Characterization of the Lymphokine Soluble Immune Response Suppressor", pp. 2848-2852, p. 2848 Abstract, rt-hand col., 4 & 5, p. 2849 left-hand column 1 & 2.
Nature, vol. 303, 30-Jun.-1983, Macmillan Journals Ltd (Chesham, Bucks, GB), M. Joseph et al.: "A New Function for Platelets IgE-Dependent Killing of Schistosomes", pp. 810-812, see the whole document (cited in the application).
Auriault Claude
Capron Andre
Joseph Michel
Pancre Veronique
Guest Shelly J.
Institut National de la Sante et de la Recherche Medicale - INSE
Institut Pasteur de Lille
Moskowitz Margaret
LandOfFree
Novel lymphokine for suppressing platelet activation does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Novel lymphokine for suppressing platelet activation, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Novel lymphokine for suppressing platelet activation will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1041604