Biomedical subjects

Y Nagata

Publications and source records attributed to Y Nagata.

At least 217 records · Page 12Linked to original sources

PubMed1997

Peptides derived from a wild-type murine proto-oncogene c-erbB-2/HER2/neu can induce CTL and tumor suppression in syngeneic hosts.

In this analysis, we examined whether peptides derived from a wild-type murine proto-oncogene, c-erbB-2, function as tumor rejection Ags. Expression of murine c-erbB-2 examined by means of reverse transcription-PCR was observed in several normal adult tissues, such as intestine, kidney, and testis. We then transduced human and murine c-erbB-2 cDNA into two mutually noncross-reactive fibrosarcoma lines of BALB/c origin, CMS7 and CMS17. In BALB/c mice immunized with CMS17HE (CMS17 transduced with human c-erbB-2 cDNA), the growth of subsequently challenged CMS7HE (CMS7 transduced with human c-erbB-2 cDNA) was significantly suppressed. CTL against human c-erbB-2-expressing cells were generated from BALB/c spleen cells in vivo and in vitro sensitized by CMS17HE. The CTL activity was also directed against murine c-erbB-2-expressing cells, CMS7ME and CMS17ME, and was blocked by anti-CD8 or anti-Kd mAbs. A series of peptides of human or murine c-erbB-2 compatible with the Kd binding motif was synthesized. The CTL were reactive with P1.HTR (H-2d) pulsed with three of these peptides, p63-71 (human c-erbB-2 derived), p63-71(A) (murine c-erbB-2 derived), and p780-788 (common for human and murine c-erbB-2). Spleen cells immunized in vivo and in vitro with syngeneic spleen cells pulsed with these peptides became cytotoxic for CMS17HE and/or CMS17ME, but not CMS17neo (CMS17 transduced with control vector). The growth of CMS7ME was suppressed in mice immunized with the murine c-erbB-2-derived peptide, p63-71(A) or p780-788. There was no apparent pathologic change in mice that rejected CMS7ME after vaccination with these peptides.

3T3 Cells↗

PubMed1997

Activation of p38 MAP kinase pathway by erythropoietin and interleukin-3.

Activation of p38 MAP kinase (p38) as well as JNK/SAPK has been described as being induced by a variety of environmental stresses such as osmotic shock, ultraviolet radiation, and heat shock, or the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1 (IL-1). We found that the hematopoietic cytokines erythropoietin (Epo) and IL-3, which regulate growth and differentiation of erythroids and hematopoietic progenitors, respectively, also activate a p38 cascade. Immunoblot analyses and in vitro kinase assay clearly showed that Epo and IL-3 rapidly and transiently phosphorylated and activated p38 in Epo- or IL-3-dependent mouse hematopoietic progenitor cells. p38 can generally be activated by the upstream kinase MKK3 or MKK6. However, in vitro kinase assays in the immunoprecipitates with anti-MKK6 antibody and anti-phosphorylated MKK3/MKK6 antibody showed that activation of neither MKK3 nor MKK6 was detected after Epo or IL-3 stimulation, while osmotic shock clearly induced activation of both MKK3/MKK6 and p38. Together with previous observations, these results suggest that both p38 and JNK cascades play an important role not only in stress and proinflammatory cytokine responses but also in hematopoietic cytokine actions.

Animals↗

PubMed1997

Involvement of T-cell subsets and natural killer (NK) cells in the growth suppression of murine fibrosarcoma cells transfected with interleukin-12 (IL-12) genes.

A 3-methylcholanthrene-induced fibrosarcoma cell line of BALB/c origin, CMS5j, was co-transfected with cDNA for the p40 and p35 subunits of interleukin-12 (IL-12). Injection of transfected cells producing 5 x 10(3) U IL-12/10(6) cells/ml/day in nude mice with an established fibrosarcoma at a contralateral site efficiently eliminated tumor growth in the early phase (injection on day 0 or 4) but not later (day 8). This effect could be abrogated by simultaneous i.v. injection of antibodies against NK1.1 or ASGM1 (asialoGM1 = ganglio-N-tetraosyl-ceramide), which indicates that natural killer (NK) cells play a major role in tumor eradication or suppression when stimulated by IL-12. In wild-type mice, application of IL-12-secreting CMS5j cells abrogated growth of tumors established 8 days before but not earlier. Based on our experiments with antibody blocking in vivo, all CD4+, CD8+ and ASGM1+ cells are involved in tumor rejection. However, in our system, CD4+ cells or CD8+ cells alone, but not ASGM1+ cells alone, also could lead to tumor rejection. IL-12-engineered fibrosarcoma cells may constitute an efficient and safe system for immunotherapy of cancer.

Animals↗

PubMed1997

Function of P-glycoprotein expressed in placenta and mole.

We examined the expression of P-glycoprotein in human placentas and hydatidiform moles. Trophoblasts in all the examined placentas and moles expressed P-glycoprotein, and the size of the P-glycoprotein was smaller than that in multidrug-resistant human epidermoid carcinoma KB cells. The P-glycoprotein in the placenta and mole was photolabeled with [3H]azidopine, and [3H]vincristine was transported in an ATP-dependent manner into membrane vesicles prepared from trophoblasts that expressed P-glycoprotein. These findings indicate that P-glycoprotein expressed in trophoblasts has a drug binding site(s) and the ability to transport vincristine, suggesting that P-glycoprotein in the placenta protects the fetus from xenobiotics and confers drug resistance on moles.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1997

Clinical results of radiofrequency hyperthermia for malignant liver tumors.

PURPOSE: To evaluate thermometry and the clinical results of radiofrequency (RF) hyperthermia for advanced malignant liver tumors. METHODS AND MATERIALS: One hundred seventy-three patients with malignant liver tumors treated between 1983 and 1995 underwent hyperthermia. The 173 tumors consisted of 114 hepatocellular carcinomas (HCCs) and 59 non-HCCs (47 metastatic liver tumors and 12 cholangiocarcinomas). Eight-megahertz RF capacitive heating equipment was used for the hyperthermia. Two opposing 25-cm electrodes were generally used for heating the liver tumors. Our standard protocol was to administer hyperthermia 40-50 min twice a week for a total of eight sessions. The liver tumor temperature was measured by microthermocouples when possible. Transcatheter arterial embolization, radiotherapy, immunotherapy, and chemotherapy were combined with hyperthermia treatment in accordance with each patient's liver function. RESULTS: One hundred forty (81%) of the 173 patients who underwent more than four sessions of hyperthermia were evaluated in this study. Thermometry was performed in 77 (55%) of these 140 patients. The maximum tumor temperature, average tumor temperature, and minimum tumor temperature in the HCC were (mean +/- standard error) 41.2 +/- 0.2 degrees C, 40.3 +/- 1.3 degrees C, and 40.1 +/- 0.2 degrees C, respectively. The same thermometry results for non-HCC were 42.3 +/- 0.2 degrees C, 41.2 +/- 0.2 degrees C, and 40.9 +/- 0.2 degrees C, respectively. The maximum and minimum temperatures (41.8 +/- 0.2 degrees C and 40.3 +/- 0.4 degrees C) in the patients with a complete or partial response (CR or PR) were higher than those in the patients with no response or progressive disease (NR or PD) (41.3 +/- 0.5 degrees C and 39.8 +/- 0.4 degrees C), but the difference was not significant. Of the 73 cases with HCC who were evaluated by computed tomography (CT), CR was achieved in 7 (10%), PR in 15 (21%), NR in 37 (51%), and PD in 14 (19%). Of the 45 cases involving liver metastases evaluated by CT, CR was achieved in 3 (7%), PR in 17 (38%), NR in 12 (27%), and PD in 13 (29%). The 1-year cumulative survival rate for HCC patients was 30.0%, and the 5-year survival rate was 17.5%. The 1-year survival of non-HCC patients was 32.5%, and the longest survival was 30 months. The sequelae of hyperthermia included focal fat necrosis in 20 patients (12%), gastric ulceration in 4 (2%), and liver necrosis in 1 (1%). The sequelae of thermometry were severe peritoneal pain in seven patients (11%), intraperitoneal hematoma in one (1%), and pneumothorax in one (1%). CONCLUSION: Even though the thermometry results for liver tumors were not satisfactory, the treatment results are promising. Further clinical trials of RF capacitive hyperthermia for the treatment of advanced liver tumors should be encouraged.

Bile Duct Neoplasms↗

PubMed1997

Activation of JNK signaling pathway by erythropoietin, thrombopoietin, and interleukin-3.

A variety of environmental stresses, such as osmotic shock, UV radiation, and heat shock, or the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1 reportedly induce activation of c-Jun amino-terminal kinases (JNK), which are usually activated by SEK1/MKK4. We report here that the hematopoietic cytokines interleukin-3 (IL-3), erythropoietin (Epo), and thrombopoietin (Tpo), which regulate growth and differentiation of hematopoietic progenitor cells, erythroids, and megakaryocytes/platelets, respectively, also activate a JNK signaling cascade. In-gel kinase assay as well as in vitro kinase assay clearly showed that IL-3, Epo, and Tpo rapidly and transiently activated both JNK1 and JNK2 in IL-3-, Epo-, or Tpo-dependent mouse hematopoietic progenitor cells. However, immunoblot analysis and in vitro kinase assay showed that neither phosphorylation nor activation of SEK1/MKK4 was induced by IL-3, Epo, or Tpo stimulation. Therefore, we concluded that the JNK signaling cascade plays an important role not only in stress responses and proinflammatory cytokine actions but also in hematopoietic cytokine actions and that hematopoietic cytokines may activate the JNKs through a kinase other than SEK1/MKK4, as previously suggested for stress-activated cells.

Animals↗

PubMed1997

Mutations and DNA diagnoses of classical citrullinemia.

Classical citrullinemia is an autosomal recessive disease caused by a genetic deficiency of argininosuccinate synthetase (ASS). We have previously identified 20 mutations in ASS mRNA of human classical citrullinemia and already established the DNA diagnosis of seven mutations as follows. By Southern blot analysis, each of the alleles with exon 5 or 6 deletion in mRNA appears to involve deletion of genomic DNA from this region. Five mutations involving R304W, G324S, IVS-6(-2) (delta Ex7), IVS-13(+5) (delta Ex13), and delta 13 bp/Ex15&IVS-15 (ins37 b/Ex15&16) are diagnosed by a combination of PCR (or modified PCR) and restriction enzyme digestion. It is important to identify the mutation in genomic DNA for prenatal diagnosis and carrier detection. In the present study, we report a novel missense mutation (R279Q) and a new abnormality in the ASS gene (delta 11 bp/IVS-15). As three missense mutations (R272C, R279Q, and G280R) were found in exon 12, we isolated and sequenced the intron regions surrounding exon 12 to establish a DNA diagnostic test. Although a mutation with a deletion of the first seven bases in exon 16 of mRNA (delta 7 b/Ex16) was found in both Japanese and American patients, the abnormality on the ASS gene was different between the Japanese allele (delta 11 bp/IVS-15) and American allele (IVS-15(-1)). The DNA diagnosis of 47 Japanese alleles with classical citrullinemia showed that the IVS-6(-2) and R304W mutations were found in 49% and 17% of the mutated alleles, respectively. We now have DNA diagnosis systems to detect 14 out of 22 mutations and are performing prenatal diagnosis and carrier detection using genomic DNA on classical citrullinemia.

Amino Acid Metabolism, Inborn Errors↗

PubMed1997

Correlation of body fat distribution with grade of endometrial cancer.

To elucidate whether body fat distribution correlates with the grade of endometrial cancer, we studied 74 postmenopausal women with endometrial cancer (mean age 62.4 +/- 6.6 years, range 49-78 years). The subjects were divided into three groups as follows: well-differentiated adenocarcinoma (G1 group; N = 53), moderately differentiated adenocarcinoma (G2 group; N = 11), and poorly differentiated adenocarcinoma (G3 group; N = 10). Four body fat indices [total fat weight (g), body fat ratio (%), trunk fat weight (g), and weight ratio of trunk fat to leg fat (trunk/leg ratio)] were measured by dual-energy X-ray absorptiometry. Baseline characteristics and body fat indices in the three groups were compared. In all subjects, the correlations of these variables with the grade of adenocarcinoma were investigated using single and stepwise regression analyses. Total fat weight, body fat ratio, and trunk fat weight showed slight increases with the grade of differentiation. The trunk/leg ratio in G1 group was significantly higher than in G2 and G3 groups. The trunk/leg ratio was significantly correlated with the grade in stepwise regression analysis. Body fat distribution in women with endometrial cancer may correlate with the grade of the adenocarcinoma.

Absorptiometry, Photon↗

PubMed1997

Follow-up studies of sphincter-preserving operations for anal fistulas.

PURPOSE: This study was designed to evaluate and to compare the outcome of anal fistula surgery using techniques involving minimal sphincter damage (sphincter-preserving) and conventional laying open techniques. PATIENTS AND METHODS: From January 1987 to December 1993, 2,242 patients with anal fistulas were treated surgically, of whom 1,070 had sphincter-preserving operations. These included 530 patients with low intersphincteric fistulas (II L), 116 patients with high intersphincteric fistulas (II H), 73 patients with combined high and low intersphincteric fistulas, 239 patients with transphincteric fistulas (III), and 112 patients with supralevator fistulas (IV). The open method of fistula surgery was used in 1,172 patients. RESULTS: Of 1,070 sphincter-preserving operations, delayed healing occurred in 49 patients (4.6 percent) and recurrent fistula occurred in 32 patients (3 percent). By using the open method in 1,172 patients, delayed healing occurred in 10 patients (0.9 percent) and recurrence of fistula occurred in 23 patients (2 percent). Failure rates increased with increasing depth and complexity of the fistula. CONCLUSION: Sphincter function is better with sphincter-preserving fistula surgery than with the open method, although healing was delayed more commonly in sphincter-preserving operations that with the open methods, and fistula recurrence was similar with the two methods. Choice of operation depends on depth of fistula, outcome by manometry, and history of former operations.

Anal Canal↗

PubMed1997

Predictors of bone mineral loss in patients with ovarian cancer treated with anticancer agents.

OBJECTIVE: To identify factors predicting bone mineral loss during anticancer chemotherapy. METHODS: Fifteen women (mean age 38.2 +/- 7.8 years; range 30-46 years) with ovarian cancer who had been treated with cisplatin-adriamycin-cyclophosphamide for six cycles every 4 weeks following surgical cytoreductin were studied. Bone mineral density (BMD) of the lumbar spine (L2-L4) was measured by dual-energy x-ray absorptiometry before and after chemotherapy. Fifteen age-matched women whose ovaries had been removed surgically for other reasons. served as controls. None of the patients had received hormonal treatment. The two groups were compared for percentage change of BMD (BMD%) over the same period. In the chemotherapy group, total fat mass, body fat ratio, total lean mass, percent lean, and ration of trunk fat to leg fat were measured by dual-energy x-ray absorptiometry. Lean loss during chemotherapy was also calculated. These variables were compared before and at the end of chemotherapy. Possible correlations of baseline variables with BMD% were determined in univariate and stepwise regression analysis. RESULTS: Mean ( +/- standard deviation) BMD decreased to 87.4 +/- 2.1% after six cycles of chemotherapy and 97.6 +/- 0.4% after 6 months in controls, but the greatest decrease was observed in the chemotherapy group (P < .001). Although baseline lean mass, baseline BMD, body weight, and lean loss during chemotherapy were correlated with BMD% in univariate analysis, baseline lean mass was still significant in stepwise regression analysis. CONCLUSION: Baseline lean mass predicts bone mineral lose with anticancer chemotherapy.

Adult↗

PubMed1997

The effect of body composition on bone density in pre- and postmenopausal women.

OBJECTIVES: To investigate the relative contribution of body composition (lean and fat) to bone mineral density (BMD), 196 premenopausal and 128 postmenopausal Japanese women were enrolled. METHODS: Total fat mass, total lean mass, lumbar BMD (L2-L4), and total body BMD were measured by dual-energy X-ray absorptiometry (DEXA, Hologic QDR 2000, MA. USA). Physical characteristics were also recorded for each subject. Correlation between BMD and variables were calculated for each of the two groups in single and stepwise regression analyses. RESULTS: Total lean mass was significantly higher in premenopausal women than postmenopausal women (P < 0.0001), while body weight, body mass index, and total fat mass were not different between the two groups. In stepwise regression analysis, total lean mass was the most powerful determinant of lumbar BMD and total body BMD in premenopausal women. In postmenopausal women, total fat mass was the most significant determinant of lumbar BMD, while total lean mass was the most significant determinant of total body BMD. CONCLUSIONS: These findings suggest that there is a difference between pre- and postmenopausal women in the relative contribution of lean and fat mass. Total lean mass is the most significant determinant of BMD in premenopausal women. On the other hand, total fat mass may have some advantages in maintaining BMD in postmenopausal women.

Adipose Tissue↗

PubMed1997

In vitro and in vivo antiproliferative effects of simvastatin, an HMG-CoA reductase inhibitor, on human glioma cells.

We analyzed the antiproliferative effect of simvastatin (SV), an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, on human glioma cell lines. Inhibition of cell growth with SV was observed in all cell lines tested. Different culture conditions altered this inhibition of growth: the lower the concentration of fetal bovine serum (FBS) in the medium, the higher the inhibitory effect of SV on glioma cells. On morphological examination, we found that most of the cells exposed to SV became rounded and the proportion of floating cells was increased in a time-dependent manner. Then we examined whether exogenously added mevalonic acid reversed the growth inhibitory effect of SV. Exogenous mevalonic acid suppressed the inhibitory effect of SV on glioma cells in a dose-dependent manner. SV also enhanced the expression of low-density lipoprotein (LDL) receptor on glioma cells. We also found that peroxidized LDL (p-LDL) was cytotoxic to glioma cells and that SV had additive effects on pLDL-induced cytotoxicity. In a mouse model, growth of glioma cells inoculated into nude mice was inhibited by intratumoral injection of both SV and peroxidized LDL. These results indicate that mevalonic acid or a metabolite in the cholesterol synthesis pathway is necessary for the growth of glioma cells and that simvastatin and/or peroxidized LDL should be examined further as potential therapeutic agents for gliomas.

Animals↗

PubMed1997

Advanced imaging of carcinoma of the mandibular gingiva.

OBJECTIVE: To evaluate the validity of MRI in detecting spread of mandibular gingival carcinoma within the bone marrow. SCHEME: Three patients with mandibular gingival carcinoma were examined by panoramic radiography, CT, MRI and radionuclide scanning. The resected mandibles were examined histologically for invasion of bone marrow by the tumor. RESULTS: With MRI the bone marrow in all cases had a low signal intensity on the T1-weighted images, a high signal intensity on the T2-weighted, and high signal intensity on the contrast-enhanced T1-weighted. In one case there was an obvious evidence of tumor invasion of the bone marrow histologically, but in two cases only dilatation of the blood vessels and evidence of hemorrhage associated with tumor. CONCLUSION: MRI is not specific for detecting tumor invasion of bone marrow in mandibular gingival carcinoma.

Aged↗

PubMed1997

Temperature sensitivity of proteoliposomes reconstituted from a mixture of scallop and rabbit sarcoplasmic reticulum Ca2+-ATPases.

We reconstituted proteoliposomes by mixing scallop and rabbit sarcoplasmic reticulum, SR, at different protein weight ratios, and investigated the effects of temperature on their Ca2+-transport activity. When proteoliposomes containing scallop and rabbit SR at a protein ratio of 1:1 were pre-incubated in the presence of Ca2+ at 39 degrees C for 10 min, the Ca2+-transport activity was almost completely lost, whereas the activity of proteoliposomes containing rabbit SR alone decreased only slightly. Essentially the same results were obtained for proteoliposomes reconstituted with Ca2+-ATPases partially purified from scallop and rabbit SR. The susceptibility of the reconstituted proteoliposomes to heat inactivation increased with an increase in the protein weight ratio of scallop to rabbit SR, the maximum being approached at a ratio higher than 1. When the scallop SR was thermally treated before reconstitution, the resulting vesicles showed as high Ca2+-transport activity as that of control vesicles reconstituted from rabbit SR alone. The former vesicles were not inactivated on further treatment at high temperature. In contrast, when the scallop SR was heated in EGTA before reconstitution of vesicles with rabbit SR, the Ca2+-transport activity of the vesicles was strongly inhibited by subsequent treatment at high temperature in the presence of Ca2+. These results can be easily explained if we assume that Ca2+ transport by the reconstituted vesicles can be catalyzed through a dimeric interaction between the scallop and rabbit Ca2+-ATPases in the membrane. Pre-incubation of these vesicles at 39 degrees C for 10 min in the presence of Ca2+ may destroy the dimeric interaction due to denaturation of the scallop Ca2+-ATPase.

Animals↗

PubMed1997

Fibrosarcoma versus cellular fibroma of the ovary: a comparative study of their proliferative activity and chromosome aberrations using MIB-1 immunostaining, DNA flow cytometry, and fluorescence in situ hybridization.

We retrospectively analyzed the proliferative activity and the centromeric copy number of chromosomes 8, 12, and 17 in three cases of fibrosarcoma and eight cases of cellular fibroma of the ovary using MIB-1 immunostaining, DNA flow cytometry, and fluorescence in situ hybridization (FISH) on paraffin-embedded tissue specimens. In our study, both the MIB-1 labeling index (LI) and the proliferative index (% of cells in S + G2 + M phase) in fibrosarcomas were higher than those in cellular fibromas. The FISH analysis demonstrated the sole abnormality of a gain of trisomy 12 cells in all eight cases of cellular fibroma. Both a gain of trisomy 12 cells and a gain of tetrasomy 12 cells were observed in one case of fibrosarcoma. A gain of trisomy 8 cells was observed in all two fibrosarcomas in which signals were detected. By contrast, neither a gain of trisomy 8 cells nor a gain of tetrasomy 12 cells was observed in any of the eight cases of cellular fibroma. Chromosome 17 showed disomy in all eleven cases. On the basis of these findings, a gain of trisomy 8 cells is therefore considered to be an adequately effective marker to distinguish between cellular fibroma and fibrosarcoma of the ovary, and it may also be related to the proliferative activity of fibrosarcoma of the ovary.

Adolescent↗

PubMed1997

Possible association of body-fat distribution with preeclampsia.

OBJECTIVE: To investigate whether preeclampsia is associated with body-fat distribution. METHODS: Twenty-two patients with preeclampsia (mean age: 31.9 +/- 6.0 years) and 126 controls without preeclampsia (mean age: 30.9 +/- 5.2 years) were enrolled in this study. We compared baseline characteristics and 4 body-fat indices measured by DEXA on Day 5 postpartum. The possible correlation between the variables and the development of preeclampsia was evaluated by multivariate analysis. RESULTS: The body-fat ratio, the upper-half body-fat amount, upper-half-body body-fat amount ratio, and upper-/lower-half-body body-fat amounts ratio on Day 5 postpartum were significantly higher in the preeclampsia group. However, the upper-/lower-half-body body-fat amount ratio was best correlated with preeclampsia among variables in multivariate analysis. CONCLUSION: Upper-body-fat distribution might be associated with the development of preeclampsia.

Absorptiometry, Photon↗

PubMed1997

Immunotherapy with husband's leukocytes to primary habitual aborters and autoantibodies in their sera.

OBJECTIVE: This study was performed to determine whether primary habitual aborters have an autoimmunological tendency and whether immunotherapy with leukocytes induces autoantibodies in their sera. METHODS: We measured the levels of antiphospholipid antibodies (APAs) and antinuclear antibodies (ANAs) in the sera of 65 primary habitual aborters. Among them, 8 primary habitual aborters received immunotherapy with their respective husband's leukocytes; these 8 patients were followed up for autoantibodies in their sera before immunotherapy, at the time of pregnancy permission, during the first trimester of pregnancy, at delivery, and postpartum. RESULTS: Sixty-nine percent of the 65 primary habitual aborters were positive for IgG or IgM antibodies to at least 1 of 6 phospholipids, and 29% of them were positive for ANAs. Although the pregnancy outcomes of the 8 primary habitual aborters after immunotherapy were good, their APAs and ANAs converted to positive after the immunotherapy. CONCLUSION: Immunotherapy with leukocytes should be performed after checking and confirming the absence of autoantibodies in the sera of a habitual aborter.

Abortion, Habitual↗

PubMed1997

Characterization of macrophages isolated from the maternal surface of human term placenta by a new method of urokinase treatment.

OBJECTIVES: This study was performed to establish a new method for isolating macrophages from the maternal surface of human term placenta by urokinase treatment and to characterize their immunological functions. METHODS: Macrophages were recovered from the maternal surface of 6 human term placentas by urokinase treatment, adherence to plastic, and density gradient centrifugation using Percoll. The cells retrieved were tested for their surface markers by immunofluorescent staining. Their antigen-presenting capacity was examined by use of a mixed lymphocytes culture (MLC) test. In 2 cases, human leucocyte antigen (HLA) typing was performed. RESULTS: An average of 4 x 10(6) macrophage-like cells were obtained per whole placenta. More than 90% of them were positive for CD14, HLA-DR, and DQ surface antigens. An MLC test revealed that they had antigen-presenting capacity. Of the 6 cases, 2 showed positive MLC test results with maternal peripheral-blood mononuclear cells. In 1 of the 2 cases, macrophages with paternal HLA phenotypes were identified. CONCLUSION: The urokinase treatment is a new useful method for isolating macrophages from the maternal surface of human term placenta. Using this method, we obtained macrophages of paternal HLA phenotypes (fetal origin) that might have migrated to the maternal surface of human term placenta.

Antigen-Presenting Cells↗