Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
  • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
  • A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/14—Blood; Artificial blood
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/37—Digestive system
  • A61K35/39—Pancreas; Islets of Langerhans
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
  • A61K41/0023—Aggression treatment or altering

Definitions

• transplanted organs or tissues in a subject may be enhanced by treating donor specific blood with a suitable dose of ultraviolet radiation for an appropriate period of time, transfusing the irradiated blood into the subject during a suitable period of time prior to transplanting the organs or tissues and then transplanting the organs or tissues into the subject.
• transplanted organs or tissues in a subject may also be enhanced by treating the organs or tissues to be transplanted with a suitable dose of ultraviolet radiation for an appropriate period of time and then transplanting the organs or tissues into the subject.
• irradiated donor specific blood, organs and tissues are preferred for use in surgical transplantation.
• FIG. 1 Percentage of graft survival in ACI diabetic recipients of islet allografts.
• Group I (O) transfused Lew UV irradiated blood and Lew islet allografts.
• Group II ( ⁇ ) transfused non-treated Lew blood and Lew islet allografts.
• Group III ( ⁇ ) control recipients without transfusion and Lew islet allografts, and
• Group IV (•) transfused as in Group I but transplanted third-party W/F (RTl u ) islets.
• FIG. 2 The effect of dose of UV irradiation on the MLC stimulatory activity of Lewis dendritic cells (DC).
• DC Lewis dendritic cells
• TDL ACI thoracic duct lymphocytes
• Lewis rat afferent lymph derived DC as stimulators. Methods used for isolation of dendritic cells have been previously described (20).
• abdominal lymph nodes were removed from rats six weeks prior to thoracic duct drainage. Lymph was collected over a 36 hour period and resultant cells were enriched for DC by a high density BSA centrifugation step (44).
• Resultant light density cells had a population of approximately 70% DC with their distinct morphological appearance (45).
• FIG. 3 Effect of dose of UV irradiation on islet function after syngeneic transplantation.
• Lewis rats were made diabetic with i.v. streptozotocin (60 mg/kg) (courtesy of Dr. Dulin, Upjohn, Kalamazoo, Michigan) and used as recipients if blood glucose was 300 mg/dl on three weekly successive measuremens.
• Lewis islets were isolated using collagenase digestion (46), Ficoll gradient separation (47) and subsequent hand-picking under a stereomicroscope. Isolated islets were suspended in HBSS in petri dishes and irradiated with constant stirring with a magnetic bar. UV source was same as described for DC irradiation.
• islets were placed into culture at 37°C, 5% CO 2 in CMRL 1066 with 10% FCS for 24 hours and transplanted via the portal vein.
• Stippled line represents normal blood glucose range.
• FIG. 4 Survival of Lewis UV irradiated and non-irradiated islets in diabetic ACI recipients.
• ACI rats were made diabetic with i.v. streptozotocin as described before.
• Lewis islets were isolated as described for syngeneic transplants and were UV irradiated with 900 J/m 2 exactly as described previously.
• UV and non-UV irradiated islets were cultured for 24 hours prior to intraportal transplantation. Rejection of islets was considered to have occurred if blood glucose was 200 mg/dl on two consecutive daily measurements, o---o Lewis islets to ACI without UV irradiation; •---• Lewis islets to ACI with prior UV irradiation.
• This invention provides a method for enhancing acceptance of transplanted organs or tissues in a subject which comprises treating donor specific blood with a suitable dose of ultraviolet radiation for an appropriate period of time, transfusing the irradiated blood into the subject during a suitable period of time prior to transplanting the organs or tissues into the subject and then transplanting the organs or tissues into the subject.
• the invention is primarily intended for use with human patients requiring an organ or tissue transplant.
• the method may be used with any organ or tissue.
• organs include the kidney, heart, lung, liver and intestine.
• Tissues may be derived from these organs or may be derived from other body tissues, e.g., bone marrow.
• Varying doses of ultraviolet radiation may be employed to irradiate the organ or tissue to be transplanted.
• the presently preferred dosage is a dose less than about 1000 J/m 2 .
• the organ or tissue may be exposed to the ultraviolet radiation for various periods of time.
• the presently preferred time for exposure to the ultraviolet radiation is a time period greater than about ten minutes, e.g., about twenty minutes.
• the irradiated blood is desirably transfused into the subject into whom the organ or tissue is to be transplanted during a suitable period of time prior to the actual transplantation procedure.
• the transfusion with donor specific irradiated blood is performed more than once and at least one week prior to transplantation of the organ or tissue.
• the subject receive transfusion three times: at three weeks, at two weeks and at one week prior to the transplant.
• the organ or tissues is transplanted into the subject using conventional surgical procedures.
• this invention also provides a method for enhancing acceptance of transplanted organs or tissues in a subject which comprises treating the organ or tissue to be transplanted with a suitable dose of ultraviolet radiation for an appropriate period of time and then transplanting the irradiated organ or tissue into the subject.
• the method is applicable to a wide variety of subjects, the primary use presently contemplated being human patients requiring transplant surgery, e.g., a replacement kidney, heart, lung, liver or intestine or replacement tissue derived from such organs or from other body tissues, e.g., pancreatic islet cells or bone marrow.
• transplant surgery e.g., a replacement kidney, heart, lung, liver or intestine or replacement tissue derived from such organs or from other body tissues, e.g., pancreatic islet cells or bone marrow.
• the presently preferred dose is one whose intensity is less than about 1000 J/m 2 and to which the subject is exposed for at least ten minutes, e.g., twenty minutes.
• the irradiated organ or tissue may then be transplanted into the subject.
• the time within which the implant surgery is performed may vary as may the time during which the irradiated organ or tissue is stored prior to use, the limits depending primarily upon the nature of the organ or tissue.
• the irradiated organ or tissue will be employed within a few days after irradiation, e.g., within 24 hours after irradiation.
• the actual transplantation is performed using conventional procedures.
• UV irradiation of the stimulating cell population in a primary mixed-lymphocyte reaction leads to little or no proliferative response (6)
• labearing cells may not need to be eliminated from blood before its use for immunization, but may need to be inactivated with UV light, leading to abrogation of the stimulating allogeneic signal while leaving major histo compatibility complex antigens intact for the induction of donor-specific immunological unresponsiveness.
• Rats of strain ACI were made diabetic with intravenous streptozotocin (60 mg/kg).
• a rat was used as a recipient of blood and islets only if its blood glucose concentration exceeded 300 mg/dl for more than 3 weeks.
• Islet allografts were considered to have been rejected when plasma glucose was greater than 200 mg/dl on two successive daily measurements.
• RTl 1 normal Lewis rats
• the blood was diluted 1:50 in phosphate-buffered saline (PBS), placed with a magnetic stirring bar into 250-ml petri dishes, and irradiated for 20 minutes with two Sylvania FS-20 lamps located 10 cm from the dishes.
• PBS phosphate-buffered saline
• the blood cells were then centrifuged and the resulting pellet was resuspended in PBS to 50 percent packed cell volume.
• Each diabetic ACI rat received 1 ml of UV-irradiated blood or 1 ml of identically treated non- irradiated blood adjusted to 50 percent packed cell volume through the penile vein 3 weeks, 2 weeks, and 1 week before islet transplantation.
• One group of diabetic ACI rats received islets without previous transfusions.
• Pancreatic islets were harvested from Lewis (RTl 1 ) and Wistar Furth (WF) (RTl u ) rats by the collagenase technique (7) and Ficoll gradient separation (8), with subsequent handpicking under a dissecting .microscope. Some 1200 to 1500 freshly prepared allogeneic islets were transplanted intraportally into four groups of diabetic ACI rats. Two groups of islet recipients (groups 1 and 4) were first transfused with UV-irradiated whole blood. One control group (group 3) was not transfused before receiving islets, while a second control group (group 2) was transfused with nonirradiated blood before allografting.
• Lewis peripheral blood lymphocytes obtained from UV-irradiated blood did not stimulate ACI thoracic duct lymphocytes significantly compared to Lewis lymphocytes obtained from nonirradiated whole blood (Table 1).
• Table 2 Effect of UV light on the serological reactivity of Lewis rat PBL surface antigens. Values are mean counts ( ⁇ standard deviations) of 125 I-labeled staphylococcal protein A bound per assay (background, 200 count/min).
• the rabbit antiserum to rat lymphocytes and the monoclonal antibody to rat la showed similar binding to Lewis peripheral blood lymphocytes, regardless of whether uv irradiation was used. Therefore we did not detect allostimulation in the MLR's by peripheral blood lymphocytes that were irradiated, despite the clear demonstration by radioimmunoassay that major histocompatibility antigens are quantitatively unchanged by previous irradiation of lymphocytes.
• diabetic ACI recipients that were transfused with UV-irradiated Lewis whole blood and subsequently transplanted with fresh Lewis islets (group 1) showed 100 percent conversion to normo glycemia. There was no tissue rejection in any of the ten animals in a period of more than 160 days after allografting.
• UV irradiation offers a promising method for the induction of donor-specific immunological unresponsiveness.
• the use of UV irradiation for immunological inactivation of blood products could be easily applicable to allotransplantation in other species for which specific antibodies to la are not available or required.
• This approach may prove useful in the transplantation of human organs, an area where donor-specific blood transfusions are already in use, and may eliminate the possibility of sensitization to major histocompatibility antigens of the donor.
• Prolonged (or indefinite) islet allograft survival and correction of diabetes may be achieved by this simple maneuver without requiring immunosuppression of the diabetic host.
• MHC major histocompatibility complex
• UVB ultraviolet irradiation
• rat dendritic cells have been demonstrated to be extremely powerful as accessory cells in T cell proliferation and in causing acute rejection of otherwise 'passenger leukocyte' -depleted rat kidneys (22).
• Dendritic cells were completely ineffective as stimulators in the MLC with resulting SI of 3. Although dendritic cells are extremely powerful allogeneic stimulators as demonstrated in the MLC (20, 23) and in causing graft rejection (21), they appear to be inactivated by UV irradiation but not by gamma irradiation.
• UV irradiated islets (same dose range) to reverse the diabetic state in syngeneic streptozotocin (STZ)-induced diabetic rats (FIG. 3).
• STZ syngeneic streptozotocin
• Irradiation (UV) with 600 or 900 J/m 2 resulted in indefinite conversion to normoglycemia in all diabetic syngeneic recipients.
• the UV irradiation dose that can abrogate the proliferative response in the MLC using l ⁇ 5 DC as stimulators, has no deleterious effect on the in vivo endocrine function of syngeneic islet grafts irradiated with 900 J/m 2 .
• UV irradiation has a selective effect on antigen presenting cells (APC) (34-36) and that passive transfer of UV irradiated APC can induce antigen specific T suppressor cells (37).
• APC antigen presenting cells
• 38-40 would therefore suggest that improper antigen presentation, i.e., allograft without stimulatory leukocytes may induce preferentially production of T suppressor cells or effect nonrecognition of foreignness until antigens are represented to host T cells by host APC's whereby production of donor-specific T suppressor cells may also occur.
• UV irradiation of peripheral blood lymphocytes does not quantitatively alter cell surface antigens including Class II MHC antigens (41). It would appear that primary allostimulation not only requires Class I and Class II antigen bearing lympho-reticular cells (31, 42), but that they must be metabolically active and are susceptible to inactivation by UV irradiation (43).
• UV irradiation that is effective in abrogating the MLC response, is selectively effective in attenuating the stimulatory activity of the interstitial dendritic cells or other allostimulatory cells present in the islet preparation. Since islets are not single cell suspensions, some allostimulatory leukocytes may escape complete inactivation which might explain a 27% failure rate in prolongation of islet allograft survival. A more precise quantitative method of delivery of UV irradiation and elucidation of its ultimate effect on islet function and simultaneously on the allostimulatory activity of the dendritic and other lympho-reticular cells contained in the islet preparation, is necessary before this approach is uniformly successful.
• the MLR was conducted in 96-well U-bottom microtiter plates (Falcon Plastics) in RPMI 1640 medium supplemented with penicillin and streptomycin (100 ⁇ g/ml each), L-glutamine, and 10 percent rat serum.
• Thoracic duct lymphocytes from ACI ras were used as responders and peripheral blood lymphocytes (purified by Ficoll- Hypaque sedimentation) from ultraviolet-irradiated or untreated blood were used as stimulators at 5 x 10 5 cells per well. Plates were harvested after 96 hours with a 16-hour exposure to thymidine. 13.

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• 1984
  • 1984-08-22 WO PCT/US1984/001347 patent/WO1985000954A1/fr not_active Ceased
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  • 1984-08-22 JP JP59503313A patent/JPS60502103A/ja active Pending
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