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Biomedical subjects

M Vandeputte

Publications and source records attributed to M Vandeputte.

At least 109 records · Page 6Linked to original sources

Inhibition of lymphocyte aggregation by progesterone.

Investigations were carried out on the influence of progesterone and murine trophoblast culture supernatants on the capacity of lymphocytes to form aggregates (clusters), an important feature in immunologic recognition and response. Both progesterone and trophoblast supernatants inhibited this cluster formation in a dose-dependent way. The effect of trophoblast supernatants appeared to be mediated mainly by progesterone since they lost their inhibitory effect on the cluster formation after treatment with anti-progesterone serum (APS). Preparations with I1-2 activity of rat and mouse origin could either prevent or restore the suppressive effect of both progesterone and trophoblast supernatants on lymphocyte aggregation. The interference with lymphocyte interaction by trophoblast may represent one of the mechanisms by which the fetal allograft is protected against maternal recognition.

Animals↗

Histochemical and autoradiographic study of the cultured rat visceral yolk sac.

The proliferation and differentiation potentiality of the rat visceral yolk sac was investigated both in organ culture and after grafting in vivo. Using alkaline phosphatase as a marker for germ cells, it was shown that these cells are absent in the 12-day-old visceral yolk sac examined before and after organ culture. Therefore, the only cells that proliferate and differentiate must be of endodermal and/or mesodermal origin. By labelling the cells with [3H]thymidine both the endodermal and mesodermal cells were found to proliferate. After 10 days in organ culture or implantation in vivo differentiated tissues (e.g. squamous epidermis, endodermal cysts and giant trophoblast cells) were regularly detected. Several of these differentiated cells contained the radiolabel indicating that they derived from the initial proliferating endodermal and/or mesodermal cells.

Alkaline Phosphatase↗

Multipotentiality of yolk-sac carcinoma cell clones.

Yolk-sac carcinoma (YSCa) clones were isolated by manual cloning under visual control. By morphology, immunohistology and histochemistry the clones were shown to differentiate in vitro into visceral and parietal yolk-sac cells and in vivo into visceral and parietal yolk-sac cells, trophoblast and mesenchyme. Trophoblast and mesenchyme were only recorded in rat YSCa derived from displaced yolk sac but not in embryo-derived YSCA. Hence YSCa cells displaying a visceral and parietal yolk-sac pattern as well as the trophoblastic and the mesenchymal structures should have a common cell ancestor.

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Rat yolk-sac antigen-2 defined by monoclonal antibodies.

The distribution of the rat yolk-sac antigen-2 (Rat YSA-2) as defined by a monoclonal antibody raised against rat yolk-sac carcinoma cells is described. The antigen is present on rat yolk-sac carcinoma, on parietal yolk-sac endoderm and on the epithelium of fetal and adult gut and of the adult proximal kidney tubules. It is not present on a variety of rat tumors other than yolk-sac carcinomas and not detectable on pre-implantation and post-implantation embryos. Rat YSA-2 differs from YSA-I, other stage-specific embryonic antigens, basement membrane antigens, intestinal and tubular antigens.

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Polyoma virus-induced hemangiomas in grafts of visceral yolk sac and of embryos.

The development of hemangiomas in grafts of visceral yolk sac and embryos after in vitro infection with polyoma virus is described. These hemangiomas were shown to be positive for the polyoma tumor-specific transplantation antigen(s) and to develop as well in recipient virgin as in pregnant rats. In grafts of fetal skin or gut infected with polyoma virus neoplastic transformation was never observed.

Animals↗

Kidney allograft tolerance in diabetic patients after total lymphoid irradiation (TLI).

The value of total lymphoid irradiation (TLI) combined with low dose prednisone as sole immunosuppressive regimen in renal allograft transplantation in humans has been investigated. Seventeen patients with end-stage diabetic nephropathy received TLI to a cumulative dose of 20-30 Gy in fractions of 1 Gy. Cadaver kidneys were grafted as soon as they were available after completion of TLI. Low dose prednisone was given after transplantation. Profound and long-term immunosuppression has been achieved in all patients. Six patients live already more than one year (greater than 2 years in 3 of them) and 7 for less than one year with a functioning kidney graft. Two out of these 13 patients had repeated rejection episodes necessitating a supplementary immunosuppressive treatment with cyclosporine A. One patient returned to chronic hemodialysis 11 months after transplantation and died of pericardial tamponade one month later. One patient had severe acute rejection for which cyclosporine A was administered; he died of septic shock as a consequence of immune deficiency a month later. The other two patients succumbed to other causes (myocardial infarction and hyperglycemia). The amount of steroids and azathioprine administered to these patients was substantially lower than in the case of conventional immunosuppression. The preliminary results are thus encouraging. However, the treatment schedule as used in the present study can not yet be considered as optimal since the majority of patients still had one or more rejection episodes. Further investigations are warranted. The optimal dose of radiation, the importance of the interval between TLI and organ transplantation, the influence of splenectomy on the immunity, etc., are still to be assessed.

Adult↗

Suppression of cellular immune reactions by rat ovarian cells.

The influence on the immunologic response by rat ovarian cells of follicle and stromal tissue origin was investigated using the mixed lymphocyte reaction (MLR), the phytohemagglutinin (PHA) assay, the cytotoxic T lymphocyte (CTL) assay, and a cytotoxic T-cell-mediated microcytotoxicity test. The MLR between BN/Mai Pfd (RT-In) stimulator splenocytes (mitomycin-C treated) and R/A Pfd (RT-1u) responder lymph node cells was markedly suppressed by ovarian cells of follicle and/or stromal tissue origin. A similar in vitro inhibition was observed with cell-free supernatants from ovarian cells, not only in the rat MLR but also in the mouse MLR between Balb/c stimulator splenocytes and C57B1 responder lymph node cells. Moreover, the reactivity of rat lymphocytes was suppressed when they were cocultured with ovarian cells in the PHA and CTL assays. Preincubation of the supernatants with serial diluted antiprogesterone serum (APS) and antiestradiol serum (AES) revealed that the suppressive effect of these supernatants could be completely abolished with APS--and, to a lesser extent, by AES. As shown by the cell-mediated microcytoxicity assay both the ovarian cells and the steroids secreted by these cells were, however, ineffective in suppressing the effector phase. It is suggested that the steroid secretion by the ovary may play an important role in the prolonged survival of ovarian grafts.

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A rat yolk-sac antigen defined by monoclonal antibodies.

The distribution of a rat yolk-sac antigen (Rat YSA-I) as defined by a monoclonal antibody raised against rat yolk-sac carcinoma cells is described. The antigen is present on rat yolk-sac carcinomas, on visceral yolk-sac endoderm and on embryonal endoderm of 9-day-old embryos. It is not present on a variety of rat tumors other than yolk-sac carcinomas and not detectable on pre-implantation embryos, inner cell mass and fetal endoderm. Rat YSA-I differs from alpha-fetoprotein and Forssman antigen and is species-specific. In adult rats the only cells displaying this antigen are the spermatozoa and certain cells of the spermatogenic lineage.

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Immunosuppressive properties of murine trophoblast.

The modification of the immunological response by murine trophoblast cells of different sources was investigated using the mixed lymphocyte reaction (MLR) and the cell mediated lympholysis (CML) test. MLR between C57BL (H-2b) stimulator splenocytes (mitomycin C treated in the unidirectional MLR) and BALB/c (H-2d) responder lymph node cells were markedly suppressed by trophoblast of ectoplacental cone (EPC) and placental origin. The same in vitro effect was observed with supernatants (SN) of trophoblast cells and with supernatants of blastocysts. Addition of anti-progesterone serum (APS), anti-testosterone serum (RAT), and anti-immunoglobulin serum (RAHIg) in serial dilutions to the trophoblast-MLR system revealed that the immunosuppressive effect of trophoblast giant cells and trophoblast giant cell culture supernatants can be abolished with APS. Identical results were obtained with APS added to immunosuppressive doses of progesterone. CML between C57BL responder lymph node cells and mitomycin C-treated BALB/c stimulator spleen cells was also markedly suppressed when trophoblast of EPC origin was added. A similar suppression of cytotoxic T-cell induction was seen when progesterone was added to the system. The immunosuppressive action of trophoblast as detected in vitro is likely to play an important role in the maintainance of pregnancy by protecting the semiallogeneic conceptus against immune aggression by the maternal immune system.

Animals↗

Increased incidence of murine graft-versus-host disease after allogeneic bone marrow transplantation by previous infusion of syngeneic bone marrow cells.

Different groups of BALB/c mice received supralethal total-body irradiation (TBI; 8.5 Gy, day 0). When 30 x 10(6) allogeneic (C57B1) bone marrow (BM) cells were infused with or without 10 x 10(6) syngeneic (BALB/c) bM cells on day 1, many animals (60%) died from graft-versus-host disease (GVHD). Typing of peripheral blood leukocytes for donor antigens showed that, respectively, 22/22 and 17/21 of the mice in both groups became chimeric. When syngeneic bone marrow was given on day 1 and allogeneic bone marrow on day 2 after TBI, a similar number of animals (21/23) became chimeric, but GVHD occurred more frequently in this group (25/26 mice, P less than 0.01). When the syngeneic bone marrow cells were replaced by spleen cells, or when the transplantation of allogeneic bone marrow was delayed till days 3 or 6 after TBI, almost all mice rejected the allogeneic BM graft and became long-term survivors. BALB/c mice receiving 30 x 10(6) C57B1 BM cells after 17 daily fractions of 0.2 Gy of total lymphoid irradiation (TLI), showed a high incidence of chimerism (15/17) and in none of the latter animals was GVHD observed. Despite the high incidence of GVHD in the mice receiving allogeneic BM after TBI and syngeneic BM transplantation, as compared with mice prepared with TLI which do not develop GVHD, suppressor cells were as easily induced after TBI and syngeneic BM transplantation as after TLI.

Animals↗

Comparison of the immunosuppressive effect of fractionated total lymphoid irradiation (TLI) vs conventional immunosuppression (CI) in renal cadaveric allotransplantation.

Beginning in November 1981, eight patients with end stage diabetic nephropathy underwent renal cadaveric transplantation after TLI. Transplantation was done between 2 to 11 days after the end of a fractionated TLI to a total dose of 20 to 30 Gy. During the same observation period, 60 nondiabetic patients with end stage renal disease of different origin also received a cadaveric kidney graft, with a conventional regimen of immunosuppression that consists of anti-lymphocyte-globulin, tapering high doses of prednisone, and azathioprine. In the TLI-treated group only a low maintenance dose of prednisone (15 mg) was given. Immunologic monitoring was performed after transplantation at regular intervals and was compared in both groups. Phytohemagglutinin (PHA)-, concanavalin A (con A)-, and pokeweed mitogen (PWM)-induced blastogenesis, as well as the mixed lymphocyte reaction (MLR) and the cell-mediated lympholysis (CML) decreased progressively during the first months after conventional immunosuppression to 50% of the pretransplantation level, and remained there for the first year after transplantation. These tests were much more impaired after TLI (less than 15% of pre TLI value) and again no recovery occurred during the first year. Natural killer (NK) cell activity progressively decreased from a mean value of 53% lysis before transplantation to 15% lysis at the end of the first year after transplantation in the conventionally treated patients. In TLI-treated patients, however, the NK activity, which declined during irradiation from 46% specific lysis to 12%, recovered rapidly after TLI to reach levels of 35 to 40% of specific lysis from the second month on after TLI. In both groups of patients the ratio of helper-inducer (TH) to suppressor-cytotoxic (TS) lymphocytes, as determined with monoclonal antibodies, progressively declined during the first 2 mo after transplantation to a low value of about 1.2. In TLI-treated patients however, this fall progressed further, so that very low levels (less than 0.6) were noticed from the third month on after TLI. The decline of the TH:TS ratio after TLI is due to an absolute increase in the number of suppressor cells. This is in contrast with the conventionally treated patients, where the low ratio is mainly provoked by a more important decrease of the helper cell population. These changes in the balance between TH and TS subpopulations are more frequently associated with positive functional suppressor cell assays in the TLI-treated patients.(ABSTRACT TRUNCATED AT 400 WORDS)

Cadaver↗

Influence of radiation field and fractionation schedule of total lymphoid irradiation (TLI) on the induction of suppressor cells and stable chimerism after bone marrow transplantation in mice.

When BALB/c mice received 17 daily fractions of 2 Gy each of total lymphoid irradiation (TLI, total dose 34 Gy) and 30 X 10(6) C57 B1 bone marrow cells (BM) on the day after the last fraction, stable bone marrow chimerism without signs of graft-vs-host disease (GVHD) was obtained in 84% of the animals. On the contrary, in BALB/c mice receiving only seven fractions of TLI (total dose 14 Gy), all bone marrow grafts were rejected. When the last two fractions of a 14-Gy TLI course were given without shielding the extra lymphatic tissues (combined total lymphoid + total body irradiation, TLBI), chimerism could be induced in 53% of the animals. When this 14-Gy TLBI schedule was used, it was even possible to administer four fractions per day (multiple fractions per day schedule, MFD), thus reducing the overall treatment time to 2 consecutive days. After this concentrated form of TLBI, chimerism was detected in 35% of the animals. As in the 34-Gy TLI schedule, graft-vs-host reaction could not be prevented in the 14-Gy TLBI schedule when spleen lymphocytes (10 X 10(6) were added to the BM inoculum. Leucopenia or suppression of the phytohaemagglutinin (PHA)-induced blastogenesis could not predict which schedule would result in a successful allogeneic bone marrow take. Suppressor cells of the mixed lymphocyte reaction, on the other hand, were only found in the spleen of BALB/c mice treated with the TLI or TLBI schedules, which also resulted in stable bone marrow chimerism.

Animals↗

Allogeneic bone marrow transplantation in mice after total lymphoid irradiation: influence of breeding conditions and strain of recipient mice.

Different groups of C57BL/ka or BALB/c mice received a dose of 34 Gy or 42 Gy of fractionated total lymphoid irradiation (TLI) before bone marrow transplantation with 30 X 10(6) BALB/c or C57BL nucleated bone marrow cells, respectively. BALB/c mice that were not bred in specific pathogen-free conditions before TLI showed a high morbidity and mortality rate after 34 Gy of TLI and allogeneic bone marrow transplantation as compared with BALB/c or C57BL that were bred in pathogen-free conditions before irradiation. Many of the conventionally bred BALB/c mice had clinical and histologic signs of graft-vs-host disease after TLI and allogeneic bone marrow infusion. Although leucocytosis and lymphocytosis and the immunologic competence as measured with in vitro tests were equally depressed after 34 Gy TLI in BALB/c and C57BL mice, chimerism was nevertheless significantly easier to obtain in BALB/c mice. The incidence of chimerism after TLI could be enhanced in C57BL mice by increasing the total radiation dose from 34 to 42 Gy. This augmentation of chimerism was paralleled by the induction of more suppressor cells after 42 Gy of TLI in C57BL mice.

Animals↗

Differentiation potentiality of rat visceral yolk sac in organ culture.

Visceral yolk sacs removed at day 12 of pregnancy in the rat were kept in organ culture for as long as 28 days. During this in vitro culture, proliferation of the endoderm and the mesoderm as well as of poorly differentiated cells was observed. The latter displayed neither the characteristics of endodermal nor mesodermal cells and their presence was frequently associated with the development of giant trophoblast cells. The hypothesis is proposed that these trophoblast cells originate from these poorly differentiated cells that acquire in vivo and in vitro the potentiality to differentiate.

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Yolk-sac carcinoma of extra-embryonic origin in the 129 Sv/Sl mouse.

Displacement of the visceral yolk-sac in fetectomized rats and hamsters does not only induce benign teratomas but may also lead to the development of malignant yolk-sac carcinoma. In the mouse, displacement of this extraembryonic membrane readily induces teratoma but until now yolk-sac carcinoma has never been observed in this species. In the present study we report the development of yolk-sac carcinoma in 129 Sv/Sl mice after a long latent period (8 months). The tumor is composed of visceral and parietal patterns in the primary neoplasm and in its metastases. It develops from the extra-embryonal part belonging to normal (+/+), heterozygous (Sl/+) or sterile (Sl/Sl) embryos, which proves that these yolk-sac carcinomas are not of germ-cell origin.

Animals↗

Facilitation of allogeneic bone marrow engraftment in mice by total lymphoid irradiation combined with total-body irradiation.

Different groups of C57BL/Ka mice received daily fractions of 2 Gy total lymphoid irradiation (TLI) in a total dose of 34, 24, or 14 Gy. On the day after the last irradiation, 30 X 10(6) allogeneic (BALB/c) nucleated bone marrow cells were infused into the irradiated animals. When the last one or two fractions of the radiation schedule were given to the whole body (combined total lymphoid-total-body irradiation, TLBI): (1) stable bone marrow chimerism with a higher number of donor-type cells in the peripheral blood was induced in a higher percentage of mice that had received 34 Gy TLBI compared with mice that received 34 Gy TLI. (2) bone marrow chimerism could also be induced after 24 Gy or 14 Gy TLBI, whereas 24 and 14 Gy TLI alone were ineffective. The tolerance to the TLBI schedules was excellent and no clinical signs of graft-versus-host disease were noticed. It is concluded that the addition of TBI can facilitate bone marrow engraftment after TLI and drastically reduce the number of radiation fractions needed to obtain successful chimerism after allogeneic bone marrow transplantation in mice.

Animals↗