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Biomedical subjects

M Vandeputte

Publications and source records attributed to M Vandeputte.

At least 55 records · Page 3Linked to original sources

Efficacy of (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)-cytosine and 9-(1,3-dihydroxy-2-propoxymethyl)-guanine in the treatment of intracerebral murine cytomegalovirus infections in immunocompetent and immunodeficient mice.

The efficacy of HPMPC and DHPG against systemic and intracerebral murine cytomegalovirus (MCMV) infections was examined in immunocompetent NMRI mice and in mice with severe combined immunodeficiency (SCID). HPMPC proved to be far superior to DHPG in preventing mortality and growth retardation in MCMV-infected NMRI mice even when given as a single dose on the day of infection. In intraperitoneally infected SCID mice, HPMPC administered as a single dose of 2, 10, 20 or 50 mg/kg per week increased the survival period of the mice by 22, 49, 77 and 156 days, respectively. In contrast, DHPG at daily doses of 10, 20 or 50 mg/kg for five consecutive days every week did not delay death by more than 13, 17 and 21 days, respectively. About one week before the MCMV-infected SCID mice (treated with either DHPG or HPMPC) died, they developed signs of neurological disease and intranuclear inclusion-bearing cells were found in their brains. The virus that was recovered from the brains of these mice did not prove to be resistant to HPMPC or DHPG. Only the virus recovered from the brains of mice treated with HPMPC at a dosage of 50 mg/kg/week had a slightly decreased susceptibility to HPMPC. When HPMPC (50 mg/kg for 4 consecutive days) was administered to SCID mice at the time when neurological symptoms became apparent, death of the animals could be delayed by another 35 to 40 days.

Animals↗

Angiogenic activity of peritoneal fluid from women with endometriosis.

OBJECTIVE: To investigate the presence of angiogenic factors in peritoneal fluid (PF) and follicular fluid (FF). DESIGN: The PF samples of 48 women with (n = 24) or without (n = 24) endometriosis were investigated. Angiogenesis was assayed using the chorioallantoic membrane of 11-day-old fertilized chicken embryos. Glass fiber prefilters impregnated with the fluids were placed on the chorioallantoic membrane. MAIN OUTCOME MEASURE: The vascular reaction was analyzed by an independent observer after 72 hours. RESULTS: There was a positive reaction in 58.3% of the PF from women with endometriosis and in 12.5% of the women without endometriosis. No correlation was found between the angiogenic response and the severity of endometriosis. The reaction remained after charcoal treatment of the PF. Positive reaction was found in three of six FF samples. CONCLUSION: The PF of women with endometriosis contain more angiogenic factors than PF from women without endometriosis. This angiogenic activity could be important for the further outgrowth and progression of the lesions.

Angiogenesis Inducing Agents↗

Influence of different growth factors on a rat choriocarcinoma cell line.

The influence of epidermal growth factor, insulin-like growth factors I and II, insulin, transforming growth factor beta 1 and transferrin on the growth of a postgestational rat choriocarcinoma was examined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The cell line was cultured in RPMI 1640 medium supplemented with fetal calf serum, beta-mercaptoethanol, glucose, sodium pyruvate and antibiotics. The experiments were done in media supplemented with 10% (optimal) or 3% (suboptimal) fetal calf serum. Among the different growth factors tested, only epidermal growth factor and to a certain extent insulin had a growth-promoting effect by themselves. The other growth factors had either an additive effect in the presence of epidermal growth factor or no effect at all. The cytotrophoblast cells expressed both epidermal growth factor and transferrin receptors whereas the more differentiated giant cells expressed only transferrin receptors.

Animals↗

Immunosuppressive activity of peritoneal fluid in women with endometriosis.

OBJECTIVE: To investigate the immunosuppressive effect of peritoneal fluid and follicular fluid on both natural killer-mediated cytotoxicity and phytohemagglutinin-induced lymphocyte proliferation. METHODS: The peritoneal fluid of women with endometriosis was compared to both fertile and infertile control fluids. Lymphocytes were pretreated for 2 or 20 hours with peritoneal or follicular fluids, and their cytotoxicity toward K562 tumor cells was measured. We also investigated the phytohemagglutinin-induced stimulation of lymphocytes cocultured with peritoneal or follicular fluid. RESULTS: Peritoneal fluid from women with endometriosis had a significantly greater immunosuppressive effect on natural killer-mediated cytotoxicity and on phytohemagglutinin stimulation of lymphocytes compared to peritoneal fluid of fertile women without endometriosis (P < .01 and P < .05, respectively). Using the peritoneal fluid of infertile women without endometriosis, these differences were significant only when compared to women with severe endometriosis. Inhibition of the natural killer activity increased when the incubation period was prolonged from 2 to 20 hours (P < .04). There was no correlation between the immunosuppressive effect of peritoneal fluid and the volume of peritoneal fluid, the day of the menstrual period, or estradiol, progesterone, prostaglandin E2, or prostaglandin F2 alpha levels. In peritoneal fluid, the factor responsible for inhibition of natural killer activity was not removed with charcoal treatment. In follicular fluid, on the other hand, the inhibition of natural killer activity decreased significantly after treatment with charcoal. CONCLUSION: Natural killer activity is suppressed by the peritoneal fluid of women with severe endometriosis; this may be important in the pathogenesis of endometriosis. The factor responsible for the inhibition of natural killer activity in peritoneal fluid is different from that in follicular fluid.

Ascitic Fluid↗

Visceral yolk sac-derived tumors.

Externalization of the visceral yolk sac, after fetectomy, induces the development of extra-embryonal fetal tumors in rodents. These tumors are either benign teratomas that appear 3 to 4 weeks after the displacement of the yolk sac or malignant tumors, i.e. yolk sac carcinomas. The latter appear 4 to 8 months after the surgery. If however, Mouse Sarcoma Virus (MSV) is injected in the placentas at the time of fetectomy (day 12 of pregnancy) the malignant tumors develop much earlier (2 to 3 months after surgery) and some display characteristics of embryonal carcinoma. Whether virus induced or not, the yolk sac carcinomas that develop from the displaced visceral yolk sac possess the same morphological and biological characteristics. They are composed of both parietal and visceral yolk sac structures and sometimes trophoblast. The tumors metastasize, grow in ascites form and kill their host. They are readily transplantable in syngeneic rats and grow in tissue culture as an epithelial-like sheet of cells. On the other hand, the benign teratomas are composed of various well differentiated adult tissues. In these tissues, derivatives of all three germ layers are observed. Numerous experiments prove that the stem cells for these various adult tissues are not germ cells. Instead the stem cells are multipotential cells that arise in the displaced yolk sac by a process of dedifferentiation. These poorly differentiated cells originate from the endoderm of the displaced visceral yolk sac. By redifferentiation they give rise to the various adult tissues characteristic for benign teratomas. The multipotential poorly differentiated cells are also likely to be the target cells for malignant transformation. Malignant transformation of these cells, whether induced by a virus or spontaneously occurring in the displaced yolk sac, leads not only to the development of yolk sac carcinomas and eventually embryonal carcinoma but also, although rarely, to choriocarcinoma. The latter tumor is transplantable in allogeneic hosts. It is hormonally active since it secretes lactogen and progesterone. The extra-embryonal fetal tumors and in particular the rat yolk sac carcinomas and choriocarcinoma proved to be a good source for the detection of oncofetal antigens. At least two different oncofetal endodermal antigens were detected with monoclonal antibodies (mab) made after immunization with yolk sac carcinoma. Another mab, made against choriocarcinoma, was found to react specifically with the cytotrophoblast both in the normal placenta and in the tumor. No other placental cells showed a positive reaction.

Animals↗

Factors involved in peripheral T cell tolerance: the extent of clonal deletion or clonal anergy depends on the age of the tolerized lymphocytes.

After injection of SEB (staphylococcus enterotoxin B), normal adult mice, or thymectomized irradiated mice (TX irr.) reconstituted with lymphocytes taken from normal adult mice became specifically tolerant of SEB. At the same time the percentage of Vbeta8 positive CD4 lymphocytes known to be responsive to SEB was almost 50% decreased, indicating that a high level of clonal deletion was realized. In contrast, mice with an exclusively old T cell compartment (old thymectomized mice, TX irr. mice reconstituted several months previously) became tolerant of SEB without deleting their Vbeta8 + CD4 + cells, indicating that clonal anergy was the major mechanism in play in the induction of tolerance. Finally, TX irr. mice reconstituted with single positive thymocytes known to become recent thymic emigrants developed tolerance for SEB together with a high level (70%) of clonal deletion. Altogether these results indicated that the mechanism involved in peripheral tolerance depended on the age of the lymphocyte: very young lymphocytes underwent mainly clonal deletion whereas long lived lymphocytes underwent predominantly clonal anergy.

Animals↗

The influence of neonatal thymectomy on the development of radiation myelopathy in rats.

To investigate the possible contribution of cellular immunity in the development of radiation injury of the central nervous system, Wag/Rij rats were thymectomized at birth and irradiated to the cervical spinal cord at the age of 3 months. At the time of paralysis or at the end of the follow-up period (when rats were 1-year-old) the animals were sacrificed and the mediastinum was examined histologically. In 95% of the neonatally thymectomized animals no thymus was left. These rats showed a firm impairment of the cellular immunity, as they had a 40% reduction of the T-lymphocytes in the spleen, and a 70% reduction of the mixed lymphocyte reaction, compared to age-matched controls. Both single dose and two-fraction irradiation experiments were performed. No modification of the latency time to develop paralysis was observed comparing thymectomized and age-matched controls. The incidence of foreleg paralysis after cervical spine irradiation (single dose or two-fraction) was identically distributed in the follow-up period for both neonatally thymectomized and control Wag/Rij rats. The ED50 value derived in the single dose experiments was 20.3 Gy for the control animals, and 20.9 Gy for thymectomized rats, and in the two fraction experiments 29 Gy for controls and 29.6 Gy for thymectomized rats. None of these differences are significant. It appears that neonatal thymectomy, in spite of its firm suppression of the cellular immunity, has no major influence on the development of radiation myelopathy in rats.

Animals↗

Changes of lymphocyte subsets after local irradiation for early stage breast cancer and seminoma testis: long-term increase of activated (HLA-DR+) T cells and decrease of "naive" (CD4-CD45R) T lymphocytes.

Blood lymphocyte subsets of early breast cancer patients and of men with stage I seminoma of the testis were studied up to 6 years after radiotherapy. Similar results were obtained in the two patient groups. After a temporary decrease, the CD4-w29 or "memory" T cells recovered completely, while the CD4-45R or "naive" T cells remained decreased up to 6 years after irradiation. The number of CD8 T lymphocytes did not change during or after treatment. Because of the decrease of a subset of CD4 cells, and the unchanged values of CD8 cells, the CD4/CD8 ratio decreased significantly after irradiation, and remained lower than before treatment up to 5-6 years after radiotherapy. The number of both HLA-DR positive CD4 and HLA-DR positive CD8 T cells ("activated" T cells) increased significantly after irradiation. The natural killer (NK) cells were not affected by treatment. We propose that the recovery of the CD4 cells is limited to the CD4-w29 ("memory") population because of thymic dysfunction in older humans. The impact of the observed immune modulation on the low susceptibility for infections after local irradiation, and on putative antitumour immune responses is discussed.

Adult↗

The natural killer activity of peritoneal fluid lymphocytes is decreased in women with endometriosis.

OBJECTIVE: To investigate the local natural killer (NK) activity of the peritoneal fluid mononuclear cells (PFMC). DESIGN PATIENTS: In a prospective way, the NK activity (K562-assay) was measured in peripheral blood (PB) and peritoneal fluid (PF) of 44 women who underwent a laparoscopy for infertility and/or pain at the University Hospital of Gasthuisberg, Leuven, Belgium. MAIN OUTCOME MEASURE: The NK activity of peripheral blood mononuclear cells and PFMC, the number and concentration of PFMC, the percentage of lymphocytes versus macrophages by May-Grünwald-Giemsa staining and the estradiol and progesterone concentration of the PF were correlated together and with the severity of endometriosis. RESULTS: We demonstrated that there is a significant NK activity in PF and that this activity is decreased in women with endometriosis. This defect was more pronounced in the follicular phase of the cycle compared with the postovulatory phase. In PB of the same 44 women, the decreased NK activity correlated with the severity of the disease. This confirms our previous report on another 34 women. CONCLUSIONS: The NK activity is decreased in women with endometriosis and correlated significantly with the severity of the disease in both the PB and PF of women with endometriosis.

Ascitic Fluid↗

A rat cytotrophoblast antigen defined by a monoclonal antibody.

A monoclonal antibody that recognizes specifically a cytotrophoblast antigen was obtained. The monoclonal antibody 22H6 was tested on rat choriocarcinoma (in vivo, in vitro), normal placenta, ectoplacental cone, blastocysts, and several normal organs. The antigen was detected on frozen sections and on tissue culture by indirect immunofluorescence. The monoclonal antibody 22H6 reacts with the cytotrophoblasts of rat choriocarcinoma. The giant cells do not display a positive reaction. It is not expressed on other tumors than choriocarcinoma. In adult rats the only cells revealing a positive reaction are the hepatocytes and the epithelial cells lining the small intestine. In the pregnant rat, the antigen is expressed on the cytotrophoblasts of the junctional zone in the placenta, but not on the giant cells. The mab reacts only with the small trophoblast cells of the ectoplacental cone, but not with trophectoderm of blastocyst. The mab has an IgG2b isotype and is not cytotoxic for choriocarcinoma cells in a complement-dependent cytotoxicity test. The described monoclonal antibody is to our knowledge the only known marker of rat benign and malignant cytotrophoblast.

Animals↗

1,25-Dihydroxyvitamin D3 prevents insulitis in NOD mice.

The active form of vitamin D, 1,25(OH)2D3, can prevent various forms of experimentally induced autoimmune disorders. The aim of this study was to confirm these findings in NOD mice that spontaneously develop an autoimmune type of diabetes mellitus. Therefore, the effect of a long-term 1,25(OH)2D3 treatment on the incidence of insulitis, the histological lesion preceding diabetes, was studied. Forty-three NOD mice were treated with 1,25(OH)2D3 (5 micrograms/kg) i.p. every other day from age 21 days on, when no insulitis was present yet. At day 100, 16 control mice receiving the treatment vehicle (arachis oil) had an incidence of insulitis of 75%, whereas only 41% of the 1,25(OH)2D3-treated animals developed insulitis (P < 0.025). Calcemia, determined 24 h after the last 1,25(OH)2D3 injection was 2.5 +/- 0.1 mM, which was higher than in control animals (2.3 +/- 0.1 mM), but was well tolerated. Cellular immunity, as assessed with the mixed lymphocyte reaction performed at day 100, was not impaired significantly. This study demonstrates that long-term treatment with high doses of 1,25(OH)2D3 is able to decrease the incidence of insulitis in spontaneous autoimmune diabetes without major side effects.

Animals↗