+

WO1991012849A1 - Greffe de tissus ou organes allogenes utilisant l'interleukine - Google Patents

Greffe de tissus ou organes allogenes utilisant l'interleukine Download PDF

Info

Publication number
WO1991012849A1
WO1991012849A1 PCT/US1991/001119 US9101119W WO9112849A1 WO 1991012849 A1 WO1991012849 A1 WO 1991012849A1 US 9101119 W US9101119 W US 9101119W WO 9112849 A1 WO9112849 A1 WO 9112849A1
Authority
WO
WIPO (PCT)
Prior art keywords
marrow
gvhd
allogeneic
tcd
bmc
Prior art date
Application number
PCT/US1991/001119
Other languages
English (en)
Inventor
Megan Sykes
David H. Sachs
Original Assignee
The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce
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 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce filed Critical The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce
Publication of WO1991012849A1 publication Critical patent/WO1991012849A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/20—Interleukins [IL]
    • A61K38/2013—IL-2
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin

Definitions

  • the present invention relates to methods for facilitating transplantation of allogeneic tissues or organs [i.e., where the major histocompatibility complex (MHC) and/or the minor histocompatibility determinants of the donor do not match those of the recipient] .
  • this invention relates to a method for facilitating engraftment of an allogeneic tissue or organ using interleukin 2 (IL-2) , either alone or in combination with T cell depleted syngeneic bone marrow, to prevent graft-versus-host disease (GVHD) .
  • IL-2 interleukin 2
  • One approach to achieving specific transplantation tolerance for any tissue or organ from a particular donor is to replace MHC-reactive elements of the immune system in the recipient with those from the donor.
  • elements of the immune system may be transferred permanently from an allogeneic donor by transplanting bone marrow into a recipient after ablation of the endogenous bone marrow and other hemopoietic elements (by lethal irradiation, for example) .
  • Allogeneic bone marrow transplant (BMT) recipients are called chimeras because their hematopoietic cells are derived from the bone marrow of the donor which is antigenically different from the other recipient cells.
  • transplanted immune systems of these chimeras are fully tolerant of tissue or organ transplants from the donor of the allogeneic bone marrow.
  • tissue or organ transplants from the donor of the allogeneic bone marrow.
  • MHC- reactive immune system components of the donor particularly certain T lymphocytes, which leads to GVHD
  • allogeneic bone marrow is often depleted of donor T cells by cell-specific cytocidal methods.
  • bone marrow transplantation in humans generally has been confined to treatment of acute life- threatening conditions, such as certain leukemias or severe bone marrow depletion (e.g., by accidental irradiation) , where replacement of the bone marrow is the only effective therapy.
  • acute life- threatening conditions such as certain leukemias or severe bone marrow depletion (e.g., by accidental irradiation)
  • replacement of the bone marrow is the only effective therapy.
  • the MHC profiles of the recipient patient and the donor must be closely matched to reduce the chances of graft rejection and GVHD. Consequently, in this country alone, thousands of BMT candidates die each year for want of an acceptable MHC-matched donor.
  • the treatment may fail to eliminate the cancer.
  • pretransplantation ablation of the recipient hemopoietic system often leaves some viable leukemic cells in the body which may then be eliminated by the transplanted immune system. Failure of the transplant to regenerate full immune functionality thus may lead to resurgence of the leukemia.
  • the donor bone marrow may not contain sufficient numbers of certain T cells that are critical for providing full anti-leukemic activity in the transplanted immune system.
  • Interleukin-2 (IL-2; originally known as T cell growth factor) is a soluble protein factor having lymphopoietic cell-specific regulatory capacities.
  • IL- 2 is known as a cytokine, and more particularly as a lymphokine, owing to its production by lymphocytes.
  • IL-2 has been found to play a variety of important roles in regulating the responsiveness of the immune system.
  • the availability of.,large amounts of purified IL-2 from specialized human cell cultures or, more recently, from recombinant DNA systems, has facilitated considerable research on the potential therapeutic utility of L-2.
  • L3T4 * cells can either augment or retard GVHD elicited by Lyt-2 + cells in class I- different hosts. J. EXP. Med. 16_7:556-569) .
  • IL-2 in bone marrow transplantation teaches that administration of high doses of this multifunctional cytokine after transplantation of allogeneic bone marrow aggravates GVHD, particularly acute forms of this disease, unless the allogeneic marrow is depleted of T cells prior to transplantation.
  • the present inventors have recently demonstrated a new approach to induction of specific transplantation tolerance across major MHC barriers, as evidenced by specific acceptance of donor-type skin grafts.
  • This method involves the use of syngeneic T cell depleted (TCD) bone marrow to ameliorate GVHD.
  • TCD syngeneic T cell depleted
  • Efficacy has been demonstrated by reconstitution of lethally irradiated mice with syngeneic TCD marrow in conjunction with untreated allogeneic bone marrow (Ilstand, S. T., et al., 1986, J. Immunol. 136:28-33).
  • transplantation of syngeneic TCD marrow with untreated allogeneic marrow could be accomplished without evidence of immediate GVHD or immunoincompetence.
  • This invention is based on the discovery by the present inventors that administration in vivo of IL-2 immediately after transplantation of lymphocyte- enriched (with spleen cells) allogeneic bone marrow in lethally irradiated mice protected against GVHD mortality from allogeneic lymphocytes.
  • Dose rates of IL-2 effective for providing protection against GVHD according to the present method approach the upper limit of tolerance of IL-2 for the transplant recipient and may vary from one individual to another within a given species, depending, inter alia, on previous therapy (e.g., radiation treatment) and basic health condition.
  • effective dose rates ranged from about 4 x 10 5 units to about 2 x 10 6 units per kilogram of body mass per 0.5 day.
  • IL-2 treatments at these rates for durations of from about 2.5 to 5 days were effective for providing protection against GVHD.
  • delaying administration of the same IL-2 treatments until one week after allogeneic bone marrow transplantation resulted in acceleration of GVHD mortality, as taught in the prior art (Sprent, J. , M. Schaefer, E. Gao, and R. Korngold, 1988, supra).
  • IL-2 treatment that protected against GVHD did not inhibit complete allogeneic repopulation of the lymphopoietic systems of most transplant recipients, nor did it diminish the anti-leukemic effects of allogeneic lymphocytes.
  • the anti-GVHD efficacy of this IL-2 treatment in such transplant procedures was increased by combination with the known anti-GVHD effect of co-transplantation of TCD syngeneic bone marrow with the allogeneic marrow.
  • maximal protection from GVHD was achieved when TCD syngeneic marrow was also administered.
  • Survivors protected from GVHD with this combined treatment also demonstrated complete allogeneic lymphopoietic repopulation, as did the lymphopoietic systems of similar but less frequent chimeric survivors of this allograft procedure having no IL-2 treatment.
  • the present invention relates to a method for facilitating engraftment of allogeneic bone marrow in a mammal comprising administration of interleukin 2 to that mammal beginning about the time of marrow transplantation, wherein the dose rate and duration of IL-2 administration is effective to provide protection against graft-versus-host disease.
  • the IL-2 may be derived from any mammalian cell or tissue source or recombinant DNA production system. (See, for examples, U.S.
  • the IL-2 is of the same mammalian species as the transplant recipient to be treated with the IL-2.
  • the IL-2 may be administered by any method which is convenient and effective, for example, by injection or by infusion according to regimens known in the art for administration of IL-2 for other purposes.
  • the IL-2 is administered in the-form of multiple discrete uniform doses at regular intervals.
  • the first of these doses is administered within the period extending inclusively from about three hours before marrow transplantation to about one hour after transplantation, and subsequent doses are administered approximately every twelve hours thereafter.
  • IL-2 treatments effective for providing protection against GVHD were administered according to the above schedule in ive or ten uniform doses over a total period of about 2.5 days or 5 days; a single dose given at the time of transplantation failed to protect against GVHD.
  • Uniform dose rates of IL-2 that were found to be effective for providing protection of a mouse against GVHD ranged from about 10,000 to 50,000 units of recombinant human IL-2 every twelve hours for a body mass on the order of 0.025 kilogram, corresponding to a dose rate range of from about 4 x 10 5 to about 2 x 10 6 units of IL-2 per kilogram of body mass per 0.5 day.
  • the present invention particularly relates to the method of facilitating engraftment of allogeneic bone marrow in a mammal, as described above, in which the total number of uniform doses of interleukin 2 is greater than one but not more than ten doses. Further, each of these uniform doses advantageously consists essentially of from about 4 x 10 s to about 2 x 10 6 units of interleukin 2 per kilogram of body mass. In the present mammalian model, the largest tested amount of IL-2 (10 doses of 50,000 units) was most effective for providing protection against GVHD but was also found to cause symptoms indicative of cumulative toxicity of IL-2.
  • the upper limit of tolerance of IL-2 dose rate is determined based on symptoms of IL-2 toxicity which are characteristic of the selected species (e.g., see Hank, J.A. , Kohler, P.C., Weil-Hillman, G. , et al., 1988, supra).
  • symptoms of GVHD are well known in the art (e.g., see the Detailed Description, below; C. Hershko and R. P. Gale, 1980. GVHD scoring systems for predicting survival of specific mortality in bone marrow transplant recipients. In Gale, R.P., Fox, C.
  • toxicity of IL-2 rather than concern for the possibility of aggravation of GVHD is the limiting factor in determining the maximum duration of IL-2 treatment that is efficacious in the facilitation of transplantation according to the method of the present invention.
  • the amount of IL-2 needed for protection against GVHD can be reduced by co-transplantation of TCD syngeneic bone marrow with the allogeneic marrow.
  • the present invention also relates to the method of facilitating engraftment of allogeneic bone marrow described above, further comprising transplanting T cell depleted syngeneic bone marrow to the mammal prior to or during transplantation of the allogeneic bone marrow.
  • the two marrow specimens are admixed and cotransplanted; in a clinical setting, however, one skilled in the art would recognize that practical considerations might require the two marrow specimens to be administered at different times.
  • the syngeneic marrow is transplanted first and may be transplanted at least eight days prior to transplantation of the allogeneic marrow.
  • IL-2 treatment is initiated at about the time of transplantation of the allogeneic marrow.
  • the present inventors have shown that the protective effect of IL-2 or IL-2 in combination with TCD syngeneic bone marrow is sufficient to prevent GVHD, which is caused by allogeneic T cells, even when allogeneic mouse bone marrow is supplemented with additional T cells from the donor spleen.
  • the level of T cells introduced by way of the allogeneic bone marrow alone is not nearly as high as T cell levels obtained by the usual process of harvesting human bone marrow for transplantation.
  • the human method results in greater enrichment of the marrow with T cells from contaminating blood than in mouse bone marrow that is harvested surgically.
  • addition of spleen cells to mouse marrow is necessary
  • SUBSTITUTE SHEET to achieve levels of allogeneic T cells in the mouse transplant that are comparable to T cell levels in human marrow. Yet the present inventors have shown that, with IL-2 treatment according to the present -method, supplementing the allogeneic mouse marrow with allogeneic lymphopoietic (spleen) cells advantageously enhances the immunocompetence of the repopulated lymphopoietic system, as in treatment of leukemia, for example.
  • the present invention also relates to the method of facilitating engraftment of allogeneic bone marrow in a mammal by administering IL-2, as described above, further comprising the transplantation of allogeneic lymphopoietic cells in addition to the allogeneic bone marrow.
  • allogeneic lymphopoietic cells are ordinarily contained in human marrow and thus marrow and lymphopoietic cells are obtained from a single donor.
  • the present inventors have demonstrated previously that survivors protected from GVHD by TCD syngeneic marrow without IL-2 treatment also show complete lymphopoietic repopulation with allogeneic cells, as do the similar chimeric survivors of the IL-2 treatment, and that such fully allogeneic lymphopoietic systems exhibit full tolerance upon subsequent tissue or organ transplantation from the donor of the allogeneic lymphopoietic cells (Ildstad, S.T., S.M. Wren, J.A. Bluestone, S.A. Barbieri, D. Stephany, and D.H. Sachs. 1986.
  • the present invention also relates to a method for facilitating engraftment of an allogeneic organ or tissue other than bone marrow in a mammal in which allogeneic bone marrow cells and, optionally, additional allogeneic lymphopoietic cells from the proposed organ or tissue donor are first engrafted into that mammal by administering to that mammal, within the period from about three hours before to five days after marrow transplantation, an effective amount of interleukin 2 to provide protection against graft- versus-host disease, whereby said mammal becomes tolerant to organ or tissue transplants from said donor.
  • the methods of this invention are of general utility for the facilitation of transplantation of any allogeneic organ or tissue.
  • FIGURES Figure 1 Effect of IL-2 and TCD syngeneic marrow on GVHD mortality from A/J lymphocytes.
  • Lethally irradiated BIO mice received 8xl0 6 A/J spleen cells plus 15xl0 6 A/J BMC, with or without 5xl0 6 TCD BIO BMC and IL-2, 50,000 U i.p. twice daily for five days.
  • A/J BMC plus spleen cells (n-15) .
  • Figure 2 Effect of IL-2 and TCD syngeneic marrow on rapid, acute GVHD mortality produced by A/J lymphocytes.
  • Lethally irradiated B10 mice received 9xl0 6 A/J spleen cells plus llxlO 6 A/J BMC, with or without 5xl0 6 TCD B10 BMC and IL2, 10,000 U twice daily for 5 days.
  • panels B, C, and D indicates the survival of animals receiving A/J
  • SUBSTITUTE SHEET Figure 3 IL-2 alone prevents acute GVHD mortality, but maximal early survival is achieved in recipients of TCD syngeneic marrow plus IL-2.
  • Top and bottom panels two independent experiments showing survival in lethally irradiated mice reconstituted with similar inocula containing A/J BMC plus A/J spleen cells, along with: no additional treatment ; TCD syngeneic marrow co-administered in the reconstituting inoculum on day 0 ---- ; IL-2, 50,000 U twice daily on day 0-4 (top) or day 0-2 (bottom) plus TCD syngeneic marrow co-administered in the reconstituting inoculum on day 0 . Each group contained 8 to 10 animals.
  • Figure 5 Assessment of the number of IL-2 doses required for protection against GVHD mortality.
  • Lethally irradiated B10 mice received 5 X 10 6 TCD BIO BMC, lOxlO 6 A/J BMC, and 9xl0 6 A/J spleen cells.
  • FIG. 6 Examples of the phenotype of lymphopoietic cells repopulating lethally irradiated B10 mice treated or not treated with IL-2, 50,000 U twice daily for 5 days beginning on the day of BMT.
  • B10 mice were lethally irradiated and reconstituted with either TCD B10 BMC plus B10.D2 BMC, or with TCD B10 BMC, B10.D2 BMC and B10.D2 spleen cells, or with B10.D2 BMC and spleen cells alone, as indicated.
  • PBL were obtained 15 weeks after BMT, stained with mAbs, and analyzed using FCM, as indicated in the Materials and Methods section. Staining with K b -specific mAb 5F1 ; staining with D d -specific mAb 34-2-12 .
  • Figure 7 A. Survival of lethally irradiated B10 mice receiving intravenous inocula containing: TCD
  • BMC plus EL4 leukemia cells and i.p. IL-2 ( ;n 4) ; A/J BMC, A/J spleen cells, EL4 leukemia cells, and i.p.
  • Figure 8 A. Survival of lethally irradiated BIO recipients of: TCD BIO BMC plus * EL4 cells and i.p. IL-2 ( ; n-10) ; TCD BIO BMC, EL4 cells, plus A/J BMC ( ; n-10) ; TCD BIO BMC, EL4 cells, A/J BMC and
  • Figure 9 Survival of lethally irradiated BIO recipients of: TCD BIO BMC, EL4 cells, and i.p. IL-2 ( ; n-10) ; TCD BIO BMC, 30xl0 6 A/J BMC, plus EL4 cells
  • A/J BMC ( — '—; n-4); TCD BIO BMC, 30xl0 6 A/J BMC, and i.p. IL-2 ( — • • — ; n-4).
  • the present invention relates to a method for facilitating engraftment of allogeneic bone marrow in a mammal by administering to that mammal, within the period from about three hours before to five days after marrow transplantation, an effective amount of IL-2 for providing protection against GVHD.
  • SUBSTITUTE SHEET describes the details of the experiments which demonstrate that IL-2 administered in vivo at the time of BMT has a potent effect in preventing mortality due to both acute and chronic GVHD. Furthermore, evidence is presented that the combination of IL-2 and TCD syngeneic marrow provides optimal protection against acute GVHD mortality. Neither IL-2 alone nor IL-2 plus TCD syngeneic marrow prevented complete lymphopoietic reconstruction by co-administered allogeneic BMC plus spleen cells.
  • TCD syngeneic marrow can delay mortality from acute GVHD (Ildstad, S.T., et al., 1986, supra) .
  • a protective effect of TCD syngeneic marrow against acute GVHD has been detected only when the GVHD was mild in severity (e.g., Figure 1).
  • TCD syngeneic marrow alone did not prevent late mortality from chronic GVHD.
  • TCD syngeneic marrow alone has a limited ability to prevent acute GVHD mortality and no detectable effect on chronic GVHD.
  • IL-2 In the absence of TCD syngeneic marrow, IL-2 also has significant protective activity against GVHD mortality, but, in every instance, such protection was increased when TCD syngeneic marrow was co-administered (e.g.. Figures 2, 3).
  • the capability of TCD syngeneic marrow to increase the protective effect in recipients of IL-2 was most apparent in experiments in which IL-2 alone provided sub-optimal protection (e.g.. Figure 2).
  • the degree of protection afforded by IL-2 alone was so potent that there was little room for improvement by the addition of TCD syngeneic marrow (e.g., Figure 3).
  • the reasons for the variability in the degree of protection afforded by similar doses of IL-2 alone are as yet unclear.
  • the present application discloses a new approach to the problem of preventing mortality from acute and chronic GVHD which does not prevent alloengraftment and does not require T cell depletion of allogeneic bone marrow.
  • This depletion is associated with an increased incidence of failure of alloengraftment, increased probability of leukemic relapse (P.J. Martin, J.A. Hansan, B. Torok-Storb, et al., 1988, Bone Marrow Transplant 3:445; N.A. Kernan, N. Flomenberg, B. Dupont, R.J. O'Reilly, 1987, Transplantation 43.:842; K.M. Sullivan, P.L. Weiden, R.
  • IL-2 appears to be the first agent without known immunosuppressive properties with anti-GVHD activity. This observation prompted attempts to apply the present invention in a murine leukemic model.
  • T cell depletion which remains the most effective known method of abrogating GVHD in human BMT recipients, has been associated with an increased probability of leukemic relapse in several hematologic malignancies.
  • evidence has also been obtained that the widely utilized immunosuppressive agent, cyclosporin, may also increase leukemic relapse probability (K. Atkinson, J.C Biggs, A. Concannon, A. Dodds, 1989, Aust. N. Z. J. Med. 4.:587).
  • IL-2 appears to represent the first agent for abrogating GVHD which lacks known non-specific immunosuppressive properties. On the contrary, IL-2 is known to be capable of shrinking solid tumors in humans and animals (S.A. Rosenberg, M.T. Lotze, J.C. Yang, et al, 1989, Ann. Surq. 210:474; S.A. Rosenberg, J.J.
  • mice Male and female C57BL/10SnJ (BIO, H- 2 b , I ⁇ I ⁇ D 13 ) , B10.D2/nSn (B10.D2, H-2 d , I ⁇ I ⁇ V) , and A/J (H-2 a , K k I k S d D d ) mice were obtained from Jackson
  • Bone marrow transplantation was performed as previously described (Ildstad, S.T., S.M. Wren, J.A. Bluestone, S.A. Barbieri, and D.H. Sachs. 1985. Characterization of mixed allogeneic chimeras. Immunocompetence, in vitro reactivity, and genetic specificity of tolerance. J. Ex . Med. 162:231) . Briefly, recipient B10 mice, aged 12 to 16 weeks, were lethally irradiated (1025R, 137 Cs source, HOR/min) and reconstituted within 8 to 12
  • All BMC and spleen cells were co-administered in a single 1 ml intravenous injection. Irradiation controls received no BMC or spleen cells and died 7 to 12 days after irradiation. In order to avoid any cage-related effects on experimental results, animals were randomized both before assigning the experimental groups, and after BMT, so that animals from different experimental groups were randomly mixed in each cage. Survival was checked on a daily basis for 100 days.
  • Leukemia model 5x10 6 syngeneic (BIO) BMC, TCD as previously described (S.T. Ildstad, et al., 1985, supra) , 10 to 15 million untreated A/J BMC (except where indicated) , 6- a subline of the B6 T cell leukemia/lymphoma EL4, were thawed from frozen vials and maintained in culture for 4 to 14 days prior to each experiment.
  • BMC, EL4 cells, and spleen calls were co-administered in a single 1 ml intravenous injection, as described (M. Sykes, Z. Bukhari, D.H. Sachs, 1989, Bone Marrow Transplant 4.:465). Animals were randomized as described (M. Sykes, Z. Bukhari, D.H. Sachs, 1989, Bone Marrow Transplant 4.:465), and survival was checked
  • IL-2 administration The indicated doses of recombinant human IL-2, provided by Cetus corporation (Emeryville, CA) , were injected intraperitoneally in 0.2 ml of Hanks Balanced Salt Solution. Unless otherwise indicated, the first dose of IL-2 was administered one to three hours before BMT, and approximately every 12 hours thereafter for a total of 10 doses. As a control for IL-2 toxicity, additional irradiated animals received IL-2 plus TCD syngeneic marrow with or without allogeneic marrow, and without allogeneic spleen cells.
  • Monoclonal antibodies (Abs) : FITC-conjugated mAb (anti-K b ) (Sherman, L.A. , and CP. Randolph. 1981. Monoclonal anti-H-2Kb antibodies detect serological differences between H-2kb mutants. Immunoqenetics .12.:183) and biotinylated mAb 34-2-12 (anti-D d ) (Ozato, K. , N.M. Mayer, and D.H. Sachs. 1982. Monoclonal antibodies to mouse major histocompatibility complex antigens. Transplantation. 34.:113) were prepared by standard methods using antibodies purified from ascites using Protein A-Sepharose 4B beads (Pharmacia, Uppsala, Sweden) .
  • Phenotyping of chimeras Phenotyping was performed 9 to 15 weeks after BMT. Animals were bled and peripheral blood mononuclear cells (PBMC) were isolated as described (Ildstad, S.T., et al., 1985, supra) . PBMC from each animal were then split into two tubes, and staining with mAbs were performed as described (Sykes, M. , M. Sheard, and D.H. Sachs. 1988. Effects of T cell depletion in radiation bone marrow chimeras. I. Evidence for a donor cell population which increases allogeneic chimerism but which lacks the potential to produce GVHD. . Immunol. 141.:2282).
  • PBMC peripheral blood mononuclear cells
  • FCM flow cytometry
  • the percentage of cells considered positive after staining with a mAb was determined using a cutoff for positivity chosen as the fluorescence level at the beginning of the positive peak of the positive control strain.
  • the relative percent staining of a chimera with a mAb was calculated using the formula:
  • Example l IL-2 +/- TCD syngeneic BMT prevents moderate GVHD
  • FIG. 1 show the effects of IL-2 and TCD syngeneic marrow on mortality from a moderately severe GVHD, which caused early mortality in one third of control animals.
  • the survival of lethally irradiated BIO control mice given 15xl0 6 A/J BMC plus 8xl0 6 A/J spleen cells is shown by the solid line in Figure 1, panels B to D. All animals presumably succumbed to GVHD, since control animals not receiving A/J spleen cells demonstrated excellent survival (Figure 1A) .
  • Figure IB demonstrate that, while TCD syngeneic marrow prevented early GVHD mortality, all animals eventually succumbed to chronic GVHD, and the overall survival curve was not significantly different from that of the controls.
  • Figure ID shows the effect of combined treatment with TCD syngeneic marrow plus IL-2, 50,000 units (U) twice daily from days 0 to 4, on GVHD mortality.
  • This combined regimen significantly reduced both early and late GVHD mortality, so that 63% of animals survived greater than 100 days, compared with only 7% survival among animals receiving neither IL-2 nor TCD syngeneic marrow (P ⁇ 0.0006).
  • Similar protection from late GVHD mortality by this treatment regimen has been reproducibly observed in another strain combination, B10.D2 into B10 (P ⁇ 0.003 for the combined results of three experiments; N-27 in each group) .
  • Example 2 IL-2 +/- TCD syngeneic BMT prevents acute GVHD
  • the indicated dose was administered twice daily for five days beginning immediately prior to bone marrow transplantation.
  • b MST median survival time determined from Kaplan-Meier plots.
  • c P value comparing group above and below the indicated value. For group receiving 50,000 U IL-2, P ⁇ 0.0001 compared with group not receiving IL-2. All P values were determined using the method of Wilcoxon and
  • lethally irradiated B10 recipients of TCD B10 marrow plus A/J BMC and spleen cells were treated with two courses of IL-2 administered twice daily for 5 days, with the first course beginning on the day of irradiation and BMT, and the second 7 days later (data not shown) .
  • the protective effect of the initial IL-2 course against GVHD was not obviated by the later course.
  • Example 5 Effect of IL-2 on engraftment
  • the PBL of long-term BMT survivors were phenotyped using mAbs and FCM. No differences were observed in the level of allogeneic reconstitution between animals receiving or not receiving IL-2 (10,000 to 50,000 U twice daily for 5 days for one or two courses) along with allogeneic (A/J or B10.D2) spleen cells, BMC, and TCD syngeneic marrow. Examples of FCM profiles from such animals are shown in Figure 6. Most animals in all groups, regardless of whether or riot spleen cells were administered, demonstrated complete allogeneic lymphopoietic repopulation, similar to the results shown in Figure 6.
  • Example 6 IL-2 permits an anti-leukemic effect of BMT
  • mice Female C57BL/10nCR (B10) H-2 b mice were lethally irradiated and reconstituted with 5xl0 6 T cell- depleted (TCD) syngeneic bone marrow cells (BMC) plus 5xl0 2 EL4 leukemia cells (M. Sykes, Z. Bukhari, D.H. Sachs, 1989, Bone Marrow Transplant. 4:465). All such recipients died of tumor by day 19 ( Figure 7A) .
  • TCD T cell- depleted
  • BMC syngeneic bone marrow cells
  • Figure 7 required the co-administration of A/J spleen cells, since, as shown in Figure 8A, A/J BMC plus TCD syngeneic marrow administered without A/J spleen cells produced a minimal prolongation (P ⁇ 0.05) of survival (median survival time [MST] 24 days) compared to recipients of TCD syngeneic marrow, EL4 and IL-2 (MST 22 days) .
  • MST median survival time
  • animals receiving 6xl0 6 A/J spleen cells in addition to EL4, A/J BMC, TCD syngeneic marrow and IL-2 demonstrated markedly improved survival (MST 39 days: P ⁇ 0.0001), with 3 of 10 animals surviving longer than 50 days ( Figure 8A) .
  • Figure 8B demonstrates in this experiment that IL-2 was necessary to prevent acute GVHD mortality, since most control recipients of EL4 plus A/J BMC and spleen cells without IL-2 were dead by day 12, before leukemic deaths even began in recipients ⁇ f syngeneic marrow, EL4, and IL-2. Recipients of similar A/J inocula without EL4 cells showed an almost identical mortality pattern, indicating that EL4 cells had no effect on GVHD. " The addition of IL-2 to this regimen was associated with significant protection from GVHD (P ⁇ 0.01), and a highly significant anti-leukemic effect could be detected; recipients of the same treatment plus TCD syngeneic marrow again enjoyed optimal survival (Figure 8B) (P ⁇ 0.002).
  • TCD syngeneic marrow administered with A/J BMC and spleen cells in the absence of IL-2 did not protect against GVHD mortality ( Figure 8B) , confirming the lack of efficacy of TCD syngeneic marrow alone against very severe GVHD noted above.
  • Control animals receiving A/J BMC plus TCD syngeneic marrow with or without IL-2 demonstrated 100% survival (not shown) .
  • Example 7 IL-2 does not reduce anti-leukemic effect of BMT
  • IL-2 can protect " against GVHD mortality without reducing the magnitude of even a weak anti-leukemic effect.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biomedical Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Anesthesiology (AREA)
  • Medical Informatics (AREA)
  • Dermatology (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

Procédé de traitement in vivo d'un mammifère à l'aide d'interleukine 2 (IL-2) qui protège contre la mortalité due à la réaction aiguë du greffon contre l'hôte (GVHD) causée par des cellules lymphoïdes incompatibles MHC. Des doses de 10.000 à 50.000 unités de IL-2 deux fois par jour pendant les cinq premiers jours suivant une greffe de moelle osseuse allogène chez des souris soumises à une dose létale de radiations ont réduit considérablement la mortalité due à la GHVD à la fois aiguë et chronique induite à travers des barrières MHC complètes et ont fréquemment conduit à la survie à long terme. Une reconstitution médullaire allogène complète a été démontrée chez tous les survivants à long terme ayant bénéficié de ce traitement. Si l'administration soit de IL-2, soit de moelle syngénique exempte de cellules T (TCD) a fourni une protection dans certaines expériences, une protection maximale a été observée en administrant à la fois de la IL-2 et de la moelle syngénique TCD, en particulier lorsque les effets de la IL-2 étaient suboptimaux. Le moment de l'administration de IL-2 est critique pour la protection puisqu'un retard de sept jours dans le début du traitement à la IL-2 a été associé à une accélération de la mortalité due à la GVHD. Les effets anti-leucémiques des lymphocytes allogènes ne sont pas diminués par l'administration simultanée de IL-2 et de moelle syngénique TCD.
PCT/US1991/001119 1990-02-27 1991-02-27 Greffe de tissus ou organes allogenes utilisant l'interleukine WO1991012849A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US48587990A 1990-02-27 1990-02-27
US485,879 1990-02-27

Publications (1)

Publication Number Publication Date
WO1991012849A1 true WO1991012849A1 (fr) 1991-09-05

Family

ID=23929794

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1991/001119 WO1991012849A1 (fr) 1990-02-27 1991-02-27 Greffe de tissus ou organes allogenes utilisant l'interleukine

Country Status (2)

Country Link
AU (1) AU7481391A (fr)
WO (1) WO1991012849A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993008843A1 (fr) * 1991-10-30 1993-05-13 Universite Pierre Et Marie Curie (Paris Vi) Cellules piegees et leur utilisation comme medicament
WO1996037208A1 (fr) * 1995-05-25 1996-11-28 Baxter International Inc. Therapie cellulaire allogenique anticancereuse suivant une transplantation de cellules souches allogeniques

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4388309A (en) * 1981-06-10 1983-06-14 Shanksville Corporation, N.V. Process for suppressing graft rejection in organ transplantation
US4489710A (en) * 1981-06-23 1984-12-25 Xoma Corporation Composition and method for transplantation therapy
US4778879A (en) * 1982-04-20 1988-10-18 Sloan-Kettering Institute For Cancer Research Highly purified human interleukin 2 and method
US4808611A (en) * 1986-07-30 1989-02-28 Immunex Corporation Use of interleukin-1 to induce development of multipotent hemopoietic cell populations
US4863727A (en) * 1986-04-09 1989-09-05 Cetus Corporation Combination therapy using interleukin-2 and tumor necrosis factor

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4388309A (en) * 1981-06-10 1983-06-14 Shanksville Corporation, N.V. Process for suppressing graft rejection in organ transplantation
US4489710A (en) * 1981-06-23 1984-12-25 Xoma Corporation Composition and method for transplantation therapy
US4778879A (en) * 1982-04-20 1988-10-18 Sloan-Kettering Institute For Cancer Research Highly purified human interleukin 2 and method
US4863727A (en) * 1986-04-09 1989-09-05 Cetus Corporation Combination therapy using interleukin-2 and tumor necrosis factor
US4808611A (en) * 1986-07-30 1989-02-28 Immunex Corporation Use of interleukin-1 to induce development of multipotent hemopoietic cell populations

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
BLOOD, Volume 64, No. 2, (August), 1984, WELTE et al., "Defective Interleukin 2 Production in Patients After Bone Marrow Transplantation and In Vitro Restoration of Defective T Lymphocyte. Proliferation by Highly Purified Interleukin 2", see pages 380-385. *
BLOOD, Volume 74, No. 4, (September), 1989, HELSOP et al., "In Vivo Induction of Gamma Interferom and Tumor Necrosis Factor by Interleukin-2 Infusion Following Intensive Chemotherapy or Autologous Marrow Transplantation", see pages 1374-1380. *
BONE MARROW TRANSPLANTATION, (1989), 4, SYKES et al., "Graft-Versus-Leukemia, Effect Using Mixed Allogeneic Bone Marrow", see pages 465-474. *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993008843A1 (fr) * 1991-10-30 1993-05-13 Universite Pierre Et Marie Curie (Paris Vi) Cellules piegees et leur utilisation comme medicament
US6048525A (en) * 1991-10-30 2000-04-11 Universite Pierre Marie Curie Cells designed as traps and their use as medicines
EP1005871A3 (fr) * 1991-10-30 2001-04-18 Universite Pierre Et Marie Curie Paris Vi Cellules piegées et leur utilisation comme médicament
WO1996037208A1 (fr) * 1995-05-25 1996-11-28 Baxter International Inc. Therapie cellulaire allogenique anticancereuse suivant une transplantation de cellules souches allogeniques
US6143292A (en) * 1995-05-25 2000-11-07 Baxter International Inc. Allogeneic cell therapy for cancer following allogeneic stem cell transplantation

Also Published As

Publication number Publication date
AU7481391A (en) 1991-09-18

Similar Documents

Publication Publication Date Title
Sykes et al. In vivo administration of interleukin 2 plus T cell-depleted syngeneic marrow prevents graft-versus-host disease mortality and permits alloengraftment.
EP1030675B1 (fr) Traitement de troubles hematologiques
Kupiec-Weglinski et al. Therapy with monoclonal antibody to interleukin 2 receptor spares suppressor T cells and prevents or reverses acute allograft rejection in rats.
Sykes et al. Interleukin-12 inhibits murine graft-versus-host disease
Jagannath et al. Low-risk intensive therapy for multiple myeloma with combined autologous bone marrow and blood stem cell support
EP0750505B1 (fr) Immunotherapie du cancer au moyen de lymphocytes allogeniques
US5573764A (en) Use of interleukin-12 to prevent graft versus host disease
Merluzzi et al. Expansion of cyclophosphamide-resistant cytotoxic precursors in vitro and in vivo by purified human interleukin 2.
Hoelzer Therapy of the newly diagnosed adult with acute lymphoblastic leukemia
Shiohara et al. The lichenoid tissue reaction. A new concept of pathogenesis.
Gurevitch et al. TRANSPLANTATION OF ALLOGENEIC OR XENOGENEIC BONE MARROW WITHIN THE DONOR STROMAL MICROENVIRONMENT1
Takeyama et al. PBSC mobilization
Chao et al. A randomized study of erythropoietin and granulocyte colony-stimulating factor (G-CSF) versus placebo and G-CSF for patients with Hodgkin’s and non-Hodgkin’s lymphoma undergoing autologous bone marrow transplantation
US20020142462A1 (en) Methods for mobilizing hematopoietic facilitating cells and hematopoietic stem cells into the peripheral blood
Kozlowski et al. Effect of pig‐specific cytokines on mobilization of hematopoietic progenitor cells in pigs and on pig bone marrow engraftment in baboons
De Fabritiis et al. CD34+ cell mobilization for allogeneic progenitor cell transplantation: efficacy of a short course of G–CSF
Ji et al. The effect of G‐CSF‐stimulated donor marrow on engraftment and incidence of graft‐versus‐host disease in allogeneic bone marrow transplantation
WO1991012849A1 (fr) Greffe de tissus ou organes allogenes utilisant l'interleukine
MacVittie et al. Myelopoietin, a chimeric agonist of human interleukin 3 and granulocyte colony-stimulating factor receptors, mobilizes CD34+ cells that rapidly engraft lethally x-irradiated nonhuman primates
Merluzzi et al. Production and response to interleukin 2 in vitro and in vivo after bone marrow transplantation in mice.
Varkila Depletion of asialo-GM1+ cells from the F1 recipient mice prior to irradiation and transfusion of parental spleen cells prevents mortality to acute graft-versus-host disease and induction of anti-host specific cytotoxic T cells
Verma et al. Effect of the in vivo priming regimen for peripheral blood stem cells (PBSC) mobilization on in vitro generation of cytotoxic effectors by IL-2 activation of PBSC in a murine model
Perillo et al. Peripheral blood progenitor cell collection after epirubicin, paclitaxel, and cisplatin combination chemotherapy using EPO‐based cytokine regimens: a randomized comparison of G–CSF and sequential GM–/G–CSF
Rosenfeld et al. Ex vivo purging of allogeneic marrow with L-leucyl-L-leucine methyl ester: a phase I study
Indovina et al. Engraftment kinetics and long-term stability of hematopoiesis following autografting of peripheral blood stem cells

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载