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WO1995013537A1 - Procede permettant de determiner in vivo l'exposition a la silicone - Google Patents

Procede permettant de determiner in vivo l'exposition a la silicone Download PDF

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Publication number
WO1995013537A1
WO1995013537A1 PCT/US1994/008823 US9408823W WO9513537A1 WO 1995013537 A1 WO1995013537 A1 WO 1995013537A1 US 9408823 W US9408823 W US 9408823W WO 9513537 A1 WO9513537 A1 WO 9513537A1
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WIPO (PCT)
Prior art keywords
lymphocytes
silicone
set forth
method set
patient
Prior art date
Application number
PCT/US1994/008823
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English (en)
Inventor
David L. Smalley
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Baptist Memorial Healthcare Foundation
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 Baptist Memorial Healthcare Foundation filed Critical Baptist Memorial Healthcare Foundation
Priority to AU74517/94A priority Critical patent/AU7451794A/en
Publication of WO1995013537A1 publication Critical patent/WO1995013537A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966—Animal cells
    • G01N33/56972—White blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2430/00—Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
    • G01N2430/60—Synthetic polymers other than synthetic polypeptides as analytes

Definitions

  • the present invention relates generally to the field of diagnostic medicine and, more particularly, to a method for determining if immune system stimulation has occurred as a result of ⁇ n vivo exposure to silicone such as may be released from a silicone implant.
  • Silicones are complex polymers of organo- silicone chlorides made with dimethylsilyl chloride. They have a wide range of physical properties depending on polymer length, number and length of side chains and viscosity. As indicated in the article by Shanklin, D.R. entitled Late Tissue Reactions to Silicone and Silica: The Natural History of Silicone-Associated Diseases with Special Reference to Infections and Immunological Markers In Tissue (USHHS/FDA, Conference Proceedings, pp. 103-125, Baltimore, Feb., 1991), silicones are not biological or natural substances. Thus, in the past silicones have been presumed to be biologically inert.
  • silicones have been utilized as the basis for certain medical replacement implants used as a lens in the eye, for small joint surfaces, to replace deviated nasal septums, to enhance contours of skin and more recently and widely, as mammary replacement or enhancement prosthesis (ie. breast implants) .
  • rheumatoid arthritis One of the more common is in the form of rheumatoid arthritis. This condition is debilitating to an advanced degree. Shoulder involvement is the most prominent followed by hip or knee. Further, patients often develop periarticular inflammation, bone loss and rheumatoid signs in other joints.
  • Lupus syndromes are also common. Lichen planus and scleroderma-like syndromes have been reported. These are more generally referred to in the art as human adjuvant disease. Many cases report findings similar to Reiter's disease. Cases of thyrotoxicosis have been reported and a very large number of women have symptoms suggestive of chronic fatigue syndrome. The spread of silicone to other parts of the body has also clearly been evidenced by migratory nodules under the skin down to the thighs and to the back and by the finding of silicone in lymph nodes beyond the axilla, ovaries, uterine tubes, liver, spleen, brain, and peripheral nerves. The finding of bulk deposits of silicone in the tubal and paratubal tissues of some patients indicates a potential interference with fertility.
  • immune system response has two aspects, one is humoral and the other is cellular.
  • the humoral aspect of the response relates to the production of serum antibodies.
  • an antigen when injected into an animal, a complex series of events occurs leading to the production of antibodies by cells called lymphocytes.
  • Antibodies are protein molecules that are specifically adapted to combine with a particular antigen to which the immune system has been exposed thereby neutralizing toxins, agglutinating bacteria or cells and precipitating soluble antigens from the body.
  • Lymphocytes may be divided into at least two classes or types called B cells and T cells.
  • the B cells are responsible for the synthesis and secretion of antibody molecules.
  • the T cells are responsible for helping the B cells but T cells are also able to carry out a whole series of reactions on their own. Generally these are reactions that involve tissue destruction known as cell-mediated reactions. These cell mediated reactions have ranges of specificity similar to humoral or antibody reactions however, the cells may react directly with an antigen on the foreign tissue and cause destruction of that tissue.
  • autoimmune process derived from silicone is, at least initially, cellular in nature rather than humoral, relatively easy to perform antibody assays are not an effective procedure for the early detection of immune system stimulation resulting from in vivo exposure to silicone. Accordingly, a need is identified for an advanced assay procedure to identify cellular aspects of immune stimulation occurring as a result of in vivo exposure to silicone such as results from implant failure.
  • Yet another object of the present invention is to provide a method for determining silicone implant failure in a patient wherein that method is characterized by high sensitivity so as to allow the early detection of immune system stimulation resulting from in vivo exposure to silicone.
  • Yet another object of the present invention is to provide a method for determining silicone implant failure in a patient wherein the method is particularly adapted to detect the cellular aspect of immune system stimulation and hypersensitivity resulting from in vivo exposure to silicone.
  • the autoimmune process derived from silicone at least initially, is cellular not humoral, early detection is ensured. Since it is important to identify autoimmune diseases at an early stage when implant removal can reduce the risks and severity of long term effects, the present invention represents a significant and important diagnostic tool to aid the doctor in monitoring and providing the best treatment for the patient.
  • an improved method for determining in vivo primary immune stimulation resulting from exposure to silicone includes the step of collecting lymphocytes from the patient. This may be done by drawing a blood specimen from the patient and separating the lymphocytes from that specimen in a manner known in the art.
  • Next is the step of dividing the separated lymphocytes into a control group and a test group. Then follows the exposing of the lymphocytes in the test group to a silicone antigenic determinant that may, for example, be selected from a group including silica and silicon dioxide. Next is quantitatively measuring the DNA synthesis and/or protein synthesis by the lymphocytes in both the control and test groups and the comparing of results. A relative increase in DNA synthesis and/or protein synthesis by the lymphocytes of the test group indicates hypersensitivity resulting from silicone implant failure with n vivo primary immune stimulation.
  • the lymphocytes collected from the patient are diluted with a tissue culture media, such as RPMI-1640 media, to a concentration of substantially 5xl0 5 lymphocytes/ml.
  • a tissue culture media such as RPMI-1640 media
  • the lymphocytes in each reaction well number substantially 1x10 s .
  • Human AB+ serum is then added to each control and test group reaction well.
  • the lymphocytes of the control group in the reaction wells are then exposed to at least one mitogen.
  • a mitogen is an agent that causes a wide range of T or B lymphocytes to undergo cell division.
  • the mitogen is preferably selected from a group including pokeweed mitogen, concanavalin A and phytohemagglutinin P. Such mitogens provide a known level of stimulation that has been documented in experiments over time.
  • control group lymphocytes in at least one reaction well are individually exposed to each of these mitogens.
  • control group lymphocytes in at least one of the reaction wells are exposed only to tissue culture media.
  • the test group lymphocytes in their reaction wells are exposed to a silicone antigenic determinant in the form of silica or silicone dioxide. Any remaining reaction wells in the microtiter plate are then filled with tissue culture media to ensure proper humidity and pH.
  • the plates are then covered and incubated for a set period of time at a set temperature. For example, the plates may be incubated for three to five days at approximately 35- 39°C and more preferably for four days at 37°C in a incubator.
  • tritiated thymidine is then added to each of the reaction wells of the control group and test group.
  • protein synthesis is to be quantitatively measured, tritiated leucine is added to each of the reaction wells in the control group and test group. After adding either tritiated compound the plates are returned to incubation.
  • the lymphocytes are harvested utilizing distilled water in a manner known in the art. More specifically, the lymphocytes are recovered and collected in a scintillation vial. Scintillation fluid is added and mixed with the lymphocytes. After sitting for approximately 10 minutes in the dark, the activity is counted in a beta scintillation counter using tower 2 and recording counts per minute from channel 2 after 1 minute. The counts are then averaged. The result is then expressed as a stimulation index determined by dividing the average counts per minute of each mitogen by the average counts per minute of the unstimulated cell control.
  • the present method may be utilized to provide early and reliable diagnosis of autoimmune disease resulting from in vivo silicone exposure due to, for example, either silicone implant failure and/or a genetic predisposition to autoimmune disease resulting from in vivo contact with silicone even in small quantities.
  • the present method measures the cellular aspect of immune system response and the silicone induced autoimmune process is initially cellular not humoral, early diagnosis is made possible.
  • Figure 1 shows the reaction well arrangement for a microtiter plate utilized in the present method.
  • the present invention relates to a method of accurately and reliably determining if immune system stimulation has occurred as a result of in vivo exposure to silicone such as may be released from a failed silicone gel filled implant.
  • the present method is characterized by high sensitivity so that any developing autoimmune disease resulting from in vivo exposure to silicone may be detected early on before any serious damage results.
  • our studies have demonstrated that the immune system stimulation resulting from in vivo exposure to silicone is primarily of T lymphocytes and not B lymphocytes which produce antibodies. Since the effect on B lymphocytes is only a secondary reaction, silicone antibody assays are only effective in detecting autoimmune disease where the patient has experienced leakage and prolonged immune stimulation.
  • the present method determines immune system stimulation resulting from in vivo exposure to silicone by measuring the cell mediated immune response (eg. the effect on T lymphocytes) .
  • the cell mediated immune response eg. the effect on T lymphocytes
  • the present method includes the initial step of collecting lymphocytes from the patient. This is done by drawing a blood specimen from the patient. The lymphocytes are then separated from that specimen in any effective manner well known in the art for this purpose. After separation, the lymphocytes are diluted with a tissue culture media to a concentration of substantially 5xl0 5 lymphocytes/ml.
  • tissue culture media that may be utilized for this purpose is RPMI- 1640 media.
  • lymphocytes in the control group and test group are then added to reaction wells in a microtiter plate. More specifically, approximately lxlO 5 lymphocytes are added to each reaction well.
  • the microtiter plate 10 such as available from Falcon Plastics, Lincoln Park, NJ including the reaction wells 12 is shown in Figure 1.
  • human AB+ serum is added to each reaction well containing lymphocytes.
  • a selected mitogen is then added to a selected number of the reaction wells of the control group.
  • the mitogen utilized may, for example, be selected from a group including pokeweed mitogen, concanavalin A, and phytohemagglutinin P.
  • the Figure 1 legend indicates which mitogen is added to which reaction well.
  • tissue culture media RPM-1640 media
  • tissue culture media is also added to blank wells surrounding the test wells in order to ensure proper humidity and pH during the subsequent incubation period.
  • a silicone antigenic determinant is added to the lymphocytes in the reaction wells of the test group.
  • That silicone antigenic determinant may, for example, be selected from a group including silica and silicon dioxide. Preferably two different concentrations of silicone antigenic determinant are used.
  • the microtiter plate is covered and incubated for three to five days at 35-39°C and more preferably, for four days at 37°C in an incubator.
  • DNA synthesis may be measured by adding tritiated thymidine to the reaction wells.
  • the lymphocytes are harvested and placed in a glass scintillation vial. Scintillation fluid is added to the harvested lymphocytes in the vial. After mixing, the vial is allowed to sit in the dark for a period of approximately 10 minutes. Activity is then counted in a beta scintillation counter using tower 2 and recording counts per minute from channel 2 after one minute. The counts are then averaged.
  • lymphocyte stimulation assays completed in accordance with this method should always be done in triplicate or quadruplicate.
  • the values utilized for analysis are the average counts for any given mitogen control, silicone antigenic determinant or tissue media control.
  • the result is expressed as a stimulation index (SI) .
  • SI stimulation index
  • the stimulation index is determined by dividing the average counts per minute of each mitogen or stimulant by the average counts per minute of the unstimulated cell (tissue media) control.
  • the patient result is then compared to a known normal cell control.
  • a similar procedure is utilized to measure protein synthesis. However, instead of adding tritiated thymidine to the reaction wells, tritiated leucine is added.
  • the incubating, harvesting and counting by means of beta scintillation counter are performed in an identical manner.
  • a relative increase in DNA synthesis and/or protein synthesis by the lymphocytes of the test group indicates that immune system stimulation has occurred as a result of in vivo exposure to silicone.
  • the present assaying method measures the cellular aspect of the immune system response, high sensitivity and the possibility of early diagnosis of the condition is provided.
  • a patient may chose to have the silicone implants removed before the consequences of the autoimmune disease or associated conditions become too severe.
  • potential long term complications from the autoimmune disease/associated conditions are reduced to the benefit of the patient.
  • RPMI-1640 media - pre-prepared commercially available IX media may be used (Gibco, Grand Island, NY) . It is best to buy 100 ml bottles since pH change or bacterial contamination may occur after opening tissue culture media. Store at 4°C.
  • Tritiated thymidine may be obtained commercially (New England Nuclear, Wilmington, DE) ; the specific activity must be 6.7Ci/mmol in a vial containing 1 mci/ml.
  • Stock solution - add 9.0 ml RMP-1640 media to 1.0 ml of commercially obtained T 3 thymidine; mix thoroughly. Dispense into 1.0 ml aliquots and store at -20°C. Stock concentration equal 100 ⁇ Ci/ml.
  • Working solution - add 9.0 ml RMP- 1640 media to 1.0 ml of stock solution and mix well. Final concentration equal 10 ⁇ Ci/ml or 0.5 ⁇ Ci/50 ⁇ l. 3.
  • Scintillation fluid - commercially available scintillation fluid should be purchased (Beckman Instruments, Fullerton, CA) . Scintisol (name and city and state of manufacturer) gives the best and most reliable results and does not appear to vary appreciably from lot to lot. Store at room temperature.
  • Pokeweed mitogen - may be obtained commercially (Gibco, Grand Island, NY) .
  • Stock solution - reconstitute with 5.0 ml sterile distilled water. Dispense in 1.0 ml aliquots and store at -20°C.
  • Working solution - add 4.0 ml RPMI- 1640 media to 1.0 ml of stock solution for a final dilution of 1:5. Discard any surplus working solution: do not refreeze.
  • Concanavalin A - may be obtained commercially (ICN Biomedicals, Inc., Costa Mesa, CA) .
  • Phytohemagglutinin P - may be obtained commercially (Gibco, Grand Island, NY) .
  • Stock solution - reconstitute with 5.0 ml sterile distilled water. Dispense in 0.5 ml aliquots and freeze at -20°C. Stock solution must be discarded after two weeks.
  • Working solution - add 0.1 ml stock solution to 9.9 ml RPMI-1640 media for a final dilution of 1:1,000 of the stock solution or 1:5,000 dilution of the original material. Discard any surplus working solution after initial use; do not refreeze.
  • Silicon dioxide - may be obtained commercially (Paddock Laboratories, Dayton, OH) Two working solutions are prepared at concentrations of 0.125 mg/ml and 0.335 mg/ml.
  • Penicillin - 1,000 U/ml stock solution store at 4°C. May be obtained commercially (Gibco, Grant Island, NY) .
  • Streptomycin - 200 mg/ml stock solution (store at 4°C. May be obtained commercially (Gibco, Grand Island, NY)
  • Ficoll-Paque - obtained commercially (Pharmacia LKB, Uppsala, Sweden) ; stored at room temperature.
  • Blood was collected by syringe from the cephalic or basilica vein in the ante brachial fossa of the left arm of the patient.
  • the blood was transported in sterile yellow top tubes containing ACD anticoagulant.
  • a minimum volume of 10 ml blood is required to perform this procedure. The actual volume necessary is dependent upon the white blood cell count and differential.
  • a normal control non- silicone exposed must be run each time a patient specimen is being tested. This control should be handled in the same manner as the patient specimen throughout the testing procedure.
  • the lymphocytes were then separated from the blood by thoroughly mixing equal amounts of anticoaggulated blood with RPMI-1640 media.
  • 4.0 ml of Ficoll-Paque was added to a 15 ml sterile disposable centrifuge tube.
  • the blood-RPMI mixture was then carefully overlayed over the Ficoll-Paque.
  • the centrifuge tube was then spun for 30-40 minutes in a centrifuge at 500 RPM.
  • the lymphocyte layer was then immediately removed and washed three times in RPMI-1640 media. The immediate washing of the specimen is 5 necessary to prevent the formation of a fibrin clot since the platelets are in the white cell layer.
  • the lymphocytes are resuspended to a final concentration of 5xl0 5 cells per ml in RPMI-1640 media with the antibiotics pencillin and streptomycin (1:100
  • microtiter plates are prepared. 0.2 ml (200 ⁇ l) of lymphocyte cell suspension is added to each reaction well as shown in Figure 1 so that final cell concentration per well is approximately lxlO 5 cells.
  • a mitogen selected from a group including pokeweed mitogen, concanavalin A and phytohemagglutinin P or a silicone antigenic determinant solution (concentration 0.125 or
  • microtiter plates 25 RPMI-1640 media to ensure proper humidity and pH.
  • the microtiter plates are then covered and incubated for four days at 37°C in an incubator.
  • Non ⁇ specific mitogens transform a larger population of lymphocytes than do specific antigens and do not require prior sensitization.
  • Antigen responsiveness is dependent upon previous sensitization to silicone dioxide and is actually a secondary response in vitro. The in vitro responses correlate with other tests of delayed hypersensitivity and therefore may be used to test response to silicone that cannot be tested in vivo.
  • Standard mitogens routinely tested are pokeweed mitogen, concanavalin A and phytohemagglutinin P.
  • Concanavalin A and phytohemagglutinin P transform T cells.
  • Pokeweed mitogen transforms B cells in the presence of normal T cells. Silicon dioxide stimulates lymphocyte stimulation but T or B cell activity has not been determined.
  • the measurement of DNA synthesis may be determined by adding 50 ⁇ l of tritiated thymidine (working solution, 0.5 ⁇ Ci) to each reaction well. This study is pulsed on day 4. After adding the tritiated thymidine, the lymphocytes are returned to incubation at 37°C in an incubator (NAPCO, Portland, Oregon) . The lymphocytes are then harvested 12-24 and preferably 18 hours after labeling with the thymidine. The harvesting may be completed on a multiple automated cell harvester (WALLAC, Gaithersburg, MD) utilizing distilled water.
  • WALLAC Gaithersburg, MD
  • the electrical supply is switched on.
  • the vacuum is then turned on and the pump is allowed to run for at least five minutes for purposes of vacuum build-up.
  • the distilled water line is also opened.
  • a filter mat is placed in position rough side down and locked in place.
  • the filter is then pre-wet by rinsing for five minutes using a blank plate.
  • the suction head is positioned in the reaction wells of the microtiter plate. As the suction head is pressed down lightly the reaction wells are rinsed for five seconds. The rinse is then repeated for a second five second interval.
  • the suction head is then turned up and the air switch is pressed for ten seconds. The air cycle causes a preliminary drying effect of the filter mat. Then the filter mat is dried overnight at 37°C. The remaining cells in the reaction wells are then loosened with a disposable plastic pipet and the residue that is recovered is harvested on a second mat.
  • the filter disks are then removed from the mat carefully and placed in a 7ml glass scintillation vial.
  • the disk from the mats are both pooled.
  • 2.0 ml of scintillation fluid is then added and mixed with the cells. After allowing this mixture to sit for 10 minutes in the dark, activity is counted (in the dark) in a beta scintillation counter (Beckman Instruments, Fullerton, CA) using tower 2 and recording counts per minute from channel 2 after 1 minute. The counts are averaged and any counts with greater than 20% difference in total count are discarded.
  • the lymphocyte stimulation assays are always done in triplicate or quadruplicate. Values are the average counts for any given mitogen control, silicone antigenic determinant test or tissue media control.
  • the result is expressed as a stimulation index (SI) .
  • SI stimulation index
  • the stimulation index is determined by dividing the average counts per minute of each stimulant (eg. mitogen, silicone antigenic determinant) by the average counts per minute of the unstimulated cell (tissue media) control.
  • the patient result is then compared to the normal cell control.
  • tritiated leucine is substituted for the tritiated thymidine. Otherwise, the same procedure described above is followed. Relative increases in either DNA synthesis or protein synthesis in the lymphocytes of the test group subjected to the silicone antigenic determinant indicates that the immune system of the patient has been stimulated as a result of in vivo exposure to silicone.
  • Example 1 The diagnostic method or assay set forth in Example 1 was followed for two patients.
  • the first patient was a 33-year old female who had silicone breast implants for six years. Approximately 2 years prior to undergoing this procedure, the patient experienced generalized aching and fatigue. She was subsequently diagnosed as having scleroderma and was treated for a two year period. Just prior to this first patient undergoing the present diagnostic method she was demonstrated to have leakage of the implants by ultrasonography and saline replacement surgery was done within days of examination.
  • the second patient was a 44 year old female who had silicone breast implants for nearly 18 years. She also had experienced severe fatigue, arthralgia and had been seen in several medical facilities for these symptoms. She had subsequently been diagnosed for scleroderma, lupus-like disease and chronic fatigue syndrome. Upon recognition of implant leakage by ultrasonography, she had saline implant replacement. During surgery, extensive rupture and silicone leakage was verified. She continued demonstrating the prior symptoms for nearly two years and then had blood samples tested utilizing the present diagnostic method. As a comparison of reliability of the in vitro test, blood from a non-implant scleroderma patient and two normal females of similar age and no history of autoimmune disease or implants were tested.
  • the first patient demonstrated normal lymphocyte stimulation with pokeweed mitogen, phytohemagglutinin P and concanavalin A.
  • the stimulation index of the patient's lymphocytes with low-level silicone dioxide (0.125 mg/ml concentration) was over 100 times that of normal controls and in saturated silicone levels (0.335 mg/ml concentration) over 200 times that of normal controls.
  • the second patient demonstrated normal lymphocyte stimulation with pokeweed mitogen, phytohemagglutinin P and concanavalin A. Further, the second patient had stimulation of 63 times the normal control in low-level silicone and 140 times the normal control in saturated silicone levels.
  • the normal control patients and the patient with scleroderma but no implants showed no silicone stimulation by either level of silicone dioxide antigenic determinant. Since the normal adults and known autoimmune patients failed to respond to the silicone stimulation, specificity of the immune response is clearly well defined.
  • a reliable diagnostic tool or assaying method for efficiently, accurately and reliably determining when a patient is suffering from autoimmune diseases and/or conditions associated with in vivo exposure to silicone.
  • the method measures the cellular aspect of the immune system response. As the autoimmune process derived from silicone is primarily cellular rather than humoral, this insures better assay sensitivity so as to allow reliable, early diagnosis of the medical condition.

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Abstract

L'invention se rapporte à un procédé permettant de déterminer la défectuosité d'un implant de silicone, ce procédé comprenant les étapes consistant à prélever des lymphocytes chez un patient puis à diviser ces lymphocytes en un groupe témoin et un groupe test. On expose les lymphocytes du groupe test à un déterminant antigénique de la silicone. Après incubation, on mesure quantitativement l'ADN et/ou la synthèse de protéine, à la fois dans le groupe test est dans le groupe témoin, puis on compare les résultats. Une augmentation relative de l'ADN et/ou de la synthèse de protéine provoquée(s) par les lymphocytes du groupe test indiquent une hypersensibilité résultant de la défectuosité de l'implant de silicone.
PCT/US1994/008823 1993-11-12 1994-08-04 Procede permettant de determiner in vivo l'exposition a la silicone WO1995013537A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU74517/94A AU7451794A (en) 1993-11-12 1994-08-04 Method for determining (in vivo) silicone exposure

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US15148593A 1993-11-12 1993-11-12
US08/151,485 1993-11-12

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WO1995013537A1 true WO1995013537A1 (fr) 1995-05-18

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PCT/US1994/008823 WO1995013537A1 (fr) 1993-11-12 1994-08-04 Procede permettant de determiner in vivo l'exposition a la silicone

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0979306A4 (fr) * 1997-02-14 2004-09-01 Univ George Washington Dosage destine a la mesure des taux de synthese d'adn

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
A. VOJDANI ET AL.: "Immune functional impairment in patients with clinical abnormalities and silicone breast implants.", TOXICOLOGY AND INDUSTRIAL HEALTH, vol. 8, no. 6, 1992, PRINCETON NJ USA, pages 415 - 429 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0979306A4 (fr) * 1997-02-14 2004-09-01 Univ George Washington Dosage destine a la mesure des taux de synthese d'adn

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AU7451794A (en) 1995-05-29

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