A triploid partial mole placenta from paternal isodisomy with a diploid fetus derived from one sperm and one oocyte may have caused angiogenic imbalance leading to preeclampsia-like symptoms at 19 weeks of gestation.

Partial hydatidiform mole with a normal fetus is extremely rare. A 30-year-old woman presented at 19 weeks gestation with clinical manifestations of severe preeclampsia. The fetus revealed a normal 46,XX karyotype and the placenta revealed triploid 69,XXX from paternal isodisomy. Microsatellite analysis revealed that the fetus and the triploid partial mole were derived from one sperm and one oocyte, followed by duplication of paternal chromosomes in only a trophectodermal cell. The maternal serum levels of angiogenic factors were extremely high compared with those reported in preeclampsia, suggesting an angiogenic imbalance may have caused preeclampsia-like symptoms before 20 weeks of gestation.

Down-regulation of Smad3 accelerates palatal wound repair.

Smad3-deficient mice exhibit accelerated re-epithelialization and tissue remodeling during palatal wound repair. In addition, transforming growth factor beta 1 (TGF-ß1) and other inflammatory factors are down-regulated compared with those in wild-type mice. The aim of this study was to examine whether targeting of Smad3 with small interfering RNA (siRNA) accelerates wound-healing and inhibits wound contraction in palatal mucoperiosteal wounds. An initial histological examination of wound closure in mouse palates treated with Smad3-targeted siRNA vs. a scrambled siRNA found that wound-healing was accelerated when levels of Smad3 were decreased. Furthermore, with real-time PCR, mRNA levels of Smad3, TGF-ß1, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) were found to be significantly down-regulated in palatal tissue treated with Smad3-targeted siRNA vs. a control siRNA. Protein and mRNA levels of α-smooth-muscle actin (α-SMA), type I collagen, and fibronectin were also lower in palates treated with Smad3-targeted siRNA vs. control siRNA. Taken together, these results indicate that down-regulation of Smad3 expression by siRNA can accelerate wound-healing and may inhibit wound contraction. Therefore, siRNA-targeted inhibition of Smad3 may represent a valuable therapeutic tool for palatal mucoperiosteal wound-healing.

Discovery of a potent, metabolically stabilized resorcylic lactone as an anti-inflammatory lead.

With bioactivity-guided phenotype screenings, a potent anti-inflammatory compound f152A1 has been isolated, characterized and identified as the known natural product LL-Z1640-2. Metabolic instability precluded its use for the study on animal disease models. Via total synthesis, a potent, metabolically stabilized analog ER-803064 has been created; addition of the (S)-Me group at C4 onto f152A1 has resulted in a dramatic improvement on its metabolic stability, while preserving the anti-inflammatory activities.

[Recent donation and clinical results of homograft tissue].

Between August 1995 and July 1999, we have experienced 14 donors for allografts (mean age: 39.8 +/- 15.8, M/F = 10/4, mean warm ischemic time: 359 minutes). Donated tissues were included 12 aortic valves and 12 pulmonary valves, respectively. Since February 1994, clinical diagnoses of 14 patients included 7 congenital heart disease, 5 infective heart disease, 1 artificial graft infection, and 1 thrombosed valve. There was no graft-transmitted disease. In congenital heart disease, 3 patients (HLHS: 1, Truncus: 1, TOF + PA: 1) died (early mortality, 42%) and 1 with TGA had residual conduit stenosis. However, in infective heart disease, all patients survived without recurrent infection and did not need reoperation (early mortality, 0%). Our clinical results of homograft implantation for infective heart disease were excellent, but more careful consideration will be needed for congenital heart disease in neonates and/or patients with poor preoperative condition.

Close correlation between estrogen treatment and renal phosphate reabsorption capacity.

To determine the influence of estrogen on the activity of renal proximal tubular reabsorption of inorganic phosphate (Pi) in women, we examined the changes of the renal threshold phosphate concentration (also denoted as TmP/GFR), as well as the changes in the concentrations of mineral components in the circulation in two groups of women--one receiving hormone replacement therapy (HRT) and one receiving gonadotropin-releasing hormone agonists (GnRH-a) therapy. We also examined the changes in the concentrations of serum PTH in the GnRH-a group. The patients in the HRT group were continuously treated with 0.625 mg conjugated equine estrogens plus 2.5 mg medroxyprogesterone acetate per day. The patients in the GnRH-a group were treated with a monthly injection of 3.75 mg leuprolide acetate depot for 6 months. The values of TmP/GFR decreased in all of the patients who received HRT. The mean percentage change in TmP/GFR was -14.5% (range, -24.3% to -9.6%). In contrast, in all of the patients treated with GnRH-a, the values of TmP/GFR increased after 6 months of treatment (the mean percentage change was 28.5%; range, 18.2-78.3%) and returned to the preadministration level at 12 weeks after stopping therapy. In these patients, both the values of TmP/GFR and the concentrations of serum Pi correlated significantly with circulating estradiol levels (r = -0.767, P < 0.01 and r = -0.797, P < 0.01, respectively), but the concentrations of serum corrected calcium did not correlate. Moreover, in the same patients, the levels of serum intact PTH decreased significantly (P < 0.05) after 6 months of treatment, but at 12 weeks after stopping therapy the trends of these levels varied among individual patients. These results suggest that estrogen could act directly to suppress sodium-dependent Pi reabsorption in the renal proximal tubules.

Elevated remnant-like lipoprotein particles in impaired glucose tolerance and type 2 diabetic patients.

OBJECTIVE: Impaired glucose tolerance (IGT) in association with insulin resistance is considered to be a risk factor for atherosclerosis. Thus, patients with IGT may have abnormal lipid and lipoprotein profiles. The purpose of this study was to investigate presence of remnant-type hyperlipoproteinemia in patients with IGT. RESEARCH DESIGN AND METHODS: Serum levels of remnant-like lipoprotein particles (RLP) were measured in 541 subjects (362 men and 179 women, age 53 +/- 7.9 years) who visited our health center for routine medical examinations. We measured RLP cholesterol (RLP-C) and RLP triglycerides (RLP-TG) using immunoaffinity gel containing monoclonal anti-human apoproteins A-I (H-12) and B-100 (JI-H) antibodies. After a 75-g oral glucose tolerance test, subjects were divided into three groups: normal, IGT, and type 2 diabetic. RESULTS: After matching for sex, age, and body weight, serum RLP-C in normal, IGT, and diabetic groups were 4.2 +/- 1.7, 6.2 +/- 3.4, and 6.2 +/- 4.2 mg/dl, respectively. The corresponding RLP-TG values were 16.7 +/- 9.2, 28.0 +/- 19.1, and 29.0 +/- 27.2 mg/dl. We found that RLP-C and RLP-TG values were significantly higher in the IGT and diabetic groups compared with the normal group (P < 0.001). In the same order, total serum cholesterol levels were 206 +/- 29, 205 +/- 34, and 206 +/- 34 mg/dl and LDL cholesterol levels were 127 +/- 27, 124 +/- 34, and 123 +/- 34 mg/dl, showing no marked difference in these groups. However, serum levels of triglyceride were higher in the IGT and diabetes groups (155 +/- 76 and 151 +/- 81 mg/dl vs. 106 +/- 41 mg/dl; P < 0.0001). Further, the incidence of remnant hyperlipoproteinemia in normocholesterolemic subjects was up to four times higher in IGT and diabetic groups compared with the normal group. CONCLUSIONS: High serum RLP-C and RLP-TG levels in IGT and diabetic patients may represent an increased risk of atherosclerosis in these patients.

Combination of automatic HPLC-RIA method for determination of estrone and estradiol in serum.

We developed a highly sensitive assay for estrone and 17 beta-estradiol in serum. Estrone and 17 beta-estradiol, obtained by solid-phase extraction using a Sep pak tC18 cartridge, were purified by high-performance liquid chromatography (HPLC). Quantitation of estrone and 17 beta-estradiol were carried out by radioimmunoassay. Not insignificantly, this automatic system of extraction and HPLC succeeded in analyzing 80 samples a week. Intra-assay coefficients of variation (CV) for estrone and 17 beta-estradiol ranged from 19.5 to 28.7%, and from 8.5 to 13.7%, respectively. The minimum detectable dose for estrone and 17 beta-estradiol were 1.04 pg/ml and 0.64 pg/ml, respectively. The serum levels of 17 beta-estradiol using our method strongly correlated with those by Gas chromatography mass spectrometry (GC-MS). The serum levels of estrone and 17 beta-estradiol in 154 peri- and postmenopausal women were estimated to be between 15 and 27 pg/ml and between 3.5 and 24.0 pg/ml, respectively, while the serum level of 17 beta-estradiol in postmenopausal women, in particular, was estimated to be from 3.5 to 6.3 pg/ml. For postmenopausal women who suffered from vasomotor symptoms, the mean levels of estrone and 17 beta-estradiol at 12 to 18 hours after treatment with daily 0.625 mg conjugated equine estrogen (CEE) and 2.5 mg medroxyprogesterone acetate (MPA) were 135.0 and 21.3 pg/ml at 12 months, respectively. On the other hand, levels of estrone and 17 beta-estradiol at 12 to 18 hours after treatment with CEE and MPA every other day, were 73.4 and 15.3 pg/ml, respectively. These highly sensitive assays for estrone and 17 beta-estradiol are useful in measuring low levels of estrogen in postmenopausal women, and monitoring estrogen levels in women receiving CEE as hormone replacement therapy.

The influence of ovariectomy on ob gene expression in rats.

Ovarian steroid hormones exert major influences on eating behaviour and body weight regulation of female rats. Ovariectomy (OVX) results in an increase in food intake and a concomitant increase in body weight, while estradiol (E2) replacement reverses these effects. In this study, we examined the influence of OVX on obese (ob) gene expression in rat adipose tissues and serum leptin concentration. Female Wistar rats, 10 weeks old, were divided into three groups: sham-operated control rats receiving corn oil (group 1, n = 4), ovariectomized rats receiving corn oil (group 2, n = 5), and ovariectomized rats receiving 17beta-E2 (10 microg/kg/day) replacement (group 3, n = 4). After 4 weeks, the rats and food consumption were weighed and serum E2 and leptin levels were measured by radioimmunoassays. Furthermore, the expression levels of ob mRNA obtained from the bilateral perimetric fat pads were estimated by Northern blot analysis. The mean weight and food consumption in group 2 were significantly (p < 0.01) heavier than those in group 1. But there were no significant differences between group 1 and group 3. The expression levels of ob mRNA in group 2 were lower than those in group 1, however, the levels of group 3 were restored to the level of group 1. On the other hand, no significant differences among the 3 groups as to serum levels of leptin were observed. The data herein clearly indicate that ovarian steroid hormones may be one of the factors involved in the regulation of ob gene.

Measurement of N- and C-terminal-region fragments of parathyroid hormone-related peptide in milk from lactating women and investigation of the relationship of their concentrations to calcium in milk.

Parathyroid hormone-related peptide (PTHrP) is found in very high concentrations in the milk of various mammals. However, little is known about its physiological role in this fluid. To obtain detailed profiles of PTHrP in milk, we measured the concentrations of PTHrP in human milk by two different region-specific assays, PTHrP(1-87) (N-PTHrP) and PTHrP(109-141) (C-PTHrP). We also examined the correlations between PTHrP and Ca concentrations in milk as well as the correlations between PTHrP and secreted milk volume. The levels of N-PTHrP and C-PTHrP were relatively low after delivery and gradually increased to 13.87 +/- 2.40 nmol/l (mean +/- S.E.M.) and 56.39 +/- 11.31 nmol/l respectively on the 10th day postpartum. N-PTHrP concentration remained steady until the 6th month postpartum when weaning starts, at which point it decreased slightly. C-PTHrP levels changed in a similar way to N-PTHrP levels but were 2- to 5-fold higher. Milk Ca concentration, and content, correlated with C-PTHrP concentration, and content (r = 0.422 and r = 0.769 respectively; P < 0.0001) but not with N-PTHrP. N-PTHrP concentration in the milk samples on the 4th day postpartum correlated with the volume of milk secreted during the 24 h before the samples were taken (r = 0.524, P < 0.01), but C-PTHrP concentration did not. These results suggest that PTHrP in human milk may play some role in the maintenance of lactation through the N-terminal region and in promoting Ca transfer into milk via the C-terminal region.

Usefulness of recombinant human prolactin for treatment of poor puerperal lactation in a rat model.

Recombinant human prolactin (r-hPRL) was produced by a line of murine C127 cells transfected with human PRL gene. To assess the biological efficacy of r-hPRL in vivo, we studied its influence on milk secretion using a rat model in which lactation was reduced by bromocriptine treatment. Puerperal rats were injected daily for 9 days after delivery with bromocriptine or bromocriptine plus r-hPRL, and lactational performance was assessed by weighing the pups. The concentrations of rat and human PRL in rat serum were measured by specific radioimmunoassays and the mammary glands were examined on postpartum day 10. Daily injection of bromocriptine (0.1 mg/rat) significantly reduced the endogenous level of rat PRL and impaired the weight gain of the pups. Administration of r-hPRL increased the serum level of human PRL. Daily injections of r-hPRL (50 micrograms/rat, twice a day) restored lactational performance and significantly increased the weight of the pups. The detrimental effect of bromocriptine on the mammary glands, assessed by both weight and histological appearance, was reversed by administration of r-hPRL. These results demonstrate that r-hPRL is biologically active in vivo and replacement therapy of r-hPRL is effective in improving the lactational performance in bromocriptine-treated rats, and also that r-hPRL may be useful for the treatment of women with poor lactation.

[Assessment of serum estrogen levels by RIA with HPLC during hormone replacement therapy].

We studied the serum estradiol and estrone levels in 146 peri and postmenopausal women, and in 38 women who had complained of various climacteric disturbance symptoms during daily hormone replacement therapy (HRT) with conjugated equine estrogen (CEE) 0.625 mg and medroxyprogesterone acetate (MPA) 2.5 mg. Serum estradiol and estrone were measured before treatment, and at 6 months, and after one year of the HRT therapy by HPLC-radioimmunoassay. In 146 peri and postmenopausal women, the serum level of estradiol was from 3 to 6pg/ml. The serum level of estradiol in 38 women after HRT significantly increased (p < 0.01) from 3.34 to 23.6 pg/ml at 6 months, and 21.5 pg/ml at 12 months. The serum level of estrone significantly increased (p < 0.01) from 26.6pg/ml to 156.7pg/ml at 6 months, and 137.2pg/ml at 12 month. These results are very useful for deciding on the doses of hormones and the expected serum estradiol level in HRT for Japanese women.

[Usefulness of the combined transfusion of autologous blood as pre-deposited and preoperatively diluted in gynecological surgery].

To eliminate the risks involved in homologous blood transfusion in gynecological surgery, we studied the effect of the combined transfusion of autologous blood as predeposited and preoperatively diluted. Thirty-nine cases were operated on and studied. In 27 cases, a total of 695 ml/person of autoblood was taken during the 15 preoperative days. In 39 cases, an average of 1,038 ml/person of autoblood was collected at the beginning of surgery. In 37 out of 39 cases, we were able to avoid homologous blood transfusion. In the remaining two cases with homologous blood transfusion, although more than 3,000 ml of operative blood loss was observed, only about 700 ml of homologous blood was needed. We did not find any serious side effect with autoblood transfusion. These results demonstrate the usefulness of the combined transfusion of autologous blood as predeposited and preoperatively diluted in gynecological surgery.

Expression pattern of CD45 RA/RO isoformic antigens in T-lineage neoplasms.

The expression of CD45 RA/RO antigen was investigated in neoplasms including cases expressing CD7 antigen as the sole pan-T antigen (n = 8), T-lineage acute lymphoblastic leukemia (ALL)/lymphoblastic lymphoma (LBL) at various stages of differentiation (n = 32), peripheral stage T-lineage leukemia (n = 10) and adult T-cell leukemia (ATL) (n = 14). The p56lck gene expression was also investigated in selected cases. The expression pattern of CD45 RA/RO antigen was defined as of RA, mixed, or RO type. All but one CD7+ CD5- CD2- case were of the RA type. The CD7+ CD5+ CD2- prothymic stage included seven RA and one mixed type cases. One CD7+ CD5- CD2+ case was of the RA type, but the other was of the RO type. The CD7+ CD5+ CD2+ prothymic stage included three RA and four mixed type cases. All seven CD3- CD4+ CD8+ (double-positive) thymic cases were of the RO type. The CD3+ CD4+ CD8+ (triple-positive) stage included two RO and three mixed-type cases. One CD3+ CD4+ CD8- late thymic case was of the mixed type. The peripheral stage cases included five RA, three RO, and two mixed type cases. All ATL cases were of the RO type. The expression of p56lck gene in the prothymic stage was less marked than that in the thymic stage. On the basis of these results, the following sequence of pattern of the CD45 RA/RO antigen expression along with T-lineage differentiation was reconstructed: prothymic stage [RA and mixed type]-->double-positive thymic stage [RO type]-->triple-positive thymic stage [RO and mixed type]-->peripheral stage [RA, mixed, and RO type]. While one RO-type CD7+ CD5- CD2- and one RO-type CD7+ CD5- CD2+ cases were not in accord with this sequence, the pattern of CD45 RA/RO antigen expression in most of T-lineage neoplasms could be determined by the respective stage of differentiation. The poor expression of the p56lck gene by the prothymic blasts compared with the thymic blasts may be related to the expression pattern of the CD45 RA/RO molecules, which exhibits phosphatase activity. The consistent RO-type expression in the ATL cases may reflect the activated status of the neoplastic T cells due to the presence of the HTLV-I gene. Alternatively, the target cells for HTLV-I-induced neoplastic transformation may possible be of the RO type.

Novel leukemic lymphoma with probable derivation from immature stage of natural killer (NK) lineage in an aged patient.

A 66-year-old male patient was admitted with dyspnea; physical examination revealed petechiae and systemic lymphadenopathy. Laboratory findings showed leukemia. The blasts in the peripheral blood were negative for cytochemical myeloperoxidase, and had condensed nuclear chromatin with a nucleolus. The histological diagnosis of the biopsied neck lymph node was lymphoblastic lymphoma. The leukemia cells expressed CD2, CD6, CD7, CD13low, CD56, beta chain of IL-2 receptorlow (IL-2R beta), and HLA-DR antigens, but not other pan-T (CD5, CD3, CD4, and CD8); pan-B (CD10, CD19, CD20, and CD24); natural killer (NK) (CD16, CD57); or myeloid (CD33) antigens. Electronmicroscopy revealed convoluted nuclei with conspicuous nucleoli and peripherally condensed heterochromatin. Membrane-bound granules containing an electron dense matrix were observed in the cytoplasm, indicating the NK cell nature of the neoplastic cells. While terminal deoxynucleotidyl transferase (TdT) and cytoplasmic CD3 were not detected by immunofluorescence on fixed smears, Northern blot analysis revealed the gene expression of CD3 epsilon, CD3 zeta, and TdT. Gene rearrangement analysis revealed that the beta, gamma, and delta chains of T-cell receptor (TCR) and immunoglobulin heavy chain (IgH) were of germline genotype. While the overall interpretation of the phenotype and genotype was difficult, the derivation of an immature stage of NK lineage was strongly suggested, based predominantly on the electronmicroscopic features. Despite initially successful chemotherapy, the patient died 14 months after initial presentation.

Lineage determination of CD7+ CD5- CD2- and CD7+ CD5+ CD2- lymphoblasts: studies on phenotype, genotype, and gene expression of myeloperoxidase, CD3 epsilon, and CD3 delta.

The gene expression of myeloperoxidase (MPO), CD3 epsilon, and CD3 delta molecules, the gene rearrangement of T-cell receptor (TCR) delta, gamma, and beta and immunoglobulin heavy (IgH) chain, and the expression of cell-surface antigens were investigated in seven cases of CD7+ CD5- CD2- and four cases of CD7+ CD5+ CD2- acute lymphoblastic leukemia or lymphoblastic lymphoma (ALL/LBL) blasts, which were negative for cytochemical myeloperoxidase (cyMPO). More mature T-lineage blasts were also investigated in a comparative manner. In conclusion, the CD7+ CD5- CD2- blasts included four categories: undifferentiated blasts without lineage commitment, T-lineage blasts, T-/myeloid lineage blasts, and cyMPO-negative myeloblasts. The CD7+ CD5+ CD2- blasts included two categories; T-lineage and T-/myeloid lineage blasts. The 11 cases were of the germ-line gene (G) for TCR beta and IgH. Four cases were G for TCR delta and TCR gamma. The others were of the monoclonally rearranged gene (R) for TCR delta and G for TCR gamma or R for both TCR delta and TCR gamma. The expression or in vitro induction of CD13 and/or CD33 antigens correlated with the immaturity of these neoplastic T cells, since it was observed in all 11 CD7+ CD5- CD2- and CD7+ CD5+ CD2-, and some CD7+ CD5+ CD2+ (CD3- CD4- CD8-) cases, but not in CD3 +/- CD4+ CD8+ or CD3+ CD4+ CD8- cases. CD3 epsilon mRNA, but not CD3 delta mRNA, was detected in two CD7+ CD5- CD2- cases, while mRNA of neither of the two CD3 molecules was detected in the other tested CD7+ CD5- CD2- cases. In contrast, mRNA of both CD3 epsilon and CD3 delta were detected in all CD7+ CD5+ CD2- cases, indicating that CD7+ CD5- CD2- blasts at least belong to T-lineage. The blasts of two CD7+ CD5- CD2- cases with entire germ-line genes and without mRNA of the three molecules (MPO, CD3 epsilon, and CD3 delta) were regarded as being at an undifferentiated stage prior to their commitment to either T- or myeloid-lineage. The co-expression of the genes of MPO and CD3 epsilon in a CD7+ CD5- CD2- case MPO, CD3 epsilon, and CD3 delta in a CD7+ CD5+ CD2- case suggested the presence of some overlapping phase for T- and myeloid-lineage commitment during immature stages of differentiation. This helps understand the conversion of some T-ALL/LBL cases to acute myeloblastic leukemia (AML).(ABSTRACT TRUNCATED AT 400 WORDS)

CD21 antigen in T-lineage neoplastic lymphoid cells: characteristic expression at thymic stage.

The expression of CD21 antigen, a receptor for the C3d fragment of complement and Epstein-Barr virus (EBV), was investigated in a total of 85 cases of neoplastic lymphoid cells including 39 cases of T-lineage acute lymphoblastic leukemia (ALL)/lymphoblastic lymphoma (LBL), although CD21 antigen is usually regarded as a pan-B antigen. The CD21 antigen was expressed by one of the eight cases of neoplastic lymphoid cells expressing the CD7 antigen as a sole pan-T antigen, by three of the 20 cases of pro- or early thymic stage (CD7+ CD5+ CD2-, CD7+ CD5- CD2+, or CD7+ CD5+ CD2+, and ten of 11 cases of thymic stage (CD3+/- CD4+ CD8+), but not by one case of late thymic stage (CD3+ CD4+ CD8-) T-ALL/LBL cells. The CD21 antigen was not expressed by any of the 11 cases of adult T-cell leukemia (ATL) or two cases of chronic T-lineage leukemia. At most 4% of the normal thymocytes obtained from seven infants or children expressed the CD21 antigen. While only a very limited population of normal thymocytes expresses CD21 antigen, T-ALL/LBL cells at the thymic stage characteristically express CD21 antigen in contrast to pro- or early thymic ALL/LBL or peripheral-stage neoplastic T cells. The estimation of the expression of CD21 antigen is useful for delineating stages of differentiation in T-ALL/LBL. Furthermore, these observation are notable, considering the possibility that the reported EBV-carrying T-cell lymphomas result from the penetration of EBV into EBV-negative neoplastic T cells.

Frequency analysis of QRS complex with bundle branch block in patients with and without sustained ventricular tachycardia.

To distinguish patients with bundle branch block (BBB) and sustained ventricular tachycardia (s-VT) from patients with BBB but without s-VT, a frequency analysis of the QRS complex was performed in 71 patients. Frequency analysis of the QRS complex of patients with left bundle branch block (LBBB) showed that patients with s-VT had significantly larger areas and area ratios between 50 and 100 Hz in the X lead than patients without s-VT (area: -0.905 +/- 0.231 vs -1.195 +/- 0.286. area ratio: -0.783 +/- 0.230 vs -1.125 +/- 0.310; P < 0.05). The area and area ratios from 100 to 200 Hz in the Z lead were also larger in patients with s-VT. The highest predictive accuracy using the area ratio from 50 to 100 Hz in the X lead was 86%, with a sensitivity and specificity of 83% and 88%, respectively. In cases with LBBB, time domain analysis showed no significant difference between patients with s-VT and those without s-VT. Frequency analysis of the QRS complex may be useful for distinguishing LBBB patients with s-VT from those without s-VT.

Pattern of expression of CD45 RA/RO isoformic antigens in acute myeloblastic leukemia cells.

The authors used cell surface immunofluorescence to investigate the expression of CD45 RA (4KB5)/RO (UCHL1) antigen by acute myeloblastic leukemia (AML) cells from 78 patients. Four types--RA+/RO- (RO < 15%), RA-/RO+ (RA < 15%), mixed (20% RA and 20% RO), and RA-/RO- (RA < 10% and RO < 10%)--were observed. The number of cases with RA+/RO-, RA-/RO+, mixed, and RA-/RO- types in each French-American-British subclass of AML were as follows: M1 (n = 22): 18, 4, 0, 0; M2 (n = 21): 16, 0, 5, 0; M3 (n = 14): 11, 0, 0, 3; M4: 2, 2, 1, 0; and M5: 5, 5, 6, 0, respectively. The M1 RA-/RO+ type, which was always CD7- CD34- HLA-DR-, constituted a rare and distinct M1 subtype because there was no mixed type among the M1 cases. All CD7+ AML cells were the RA+/RO- type and HLA-DR+ except one. The expression of CD45 RO antigen, found in patients with M2, M4, and M5 subtypes of AML, was thought to be associated with the maturity of blasts as monocyte or granulocyte lineage cells. CD45 antigen has tyrosine phosphatase activity in association with nonreceptor-type tyrosine kinases. It was speculated that the functional status and stage of differentiation of the granulocyte/monocyte lineage determine which type of nonreceptor-type tyrosine kinases will operate, which then select the pattern of expression of the CD45 isoform. Thus, the determination of tyrosine kinases associated with CD45 isoforms seems to be important in understanding the AML subsets defined by the pattern of CD45 RA/RO expression.

[Phenotypic analysis of immature T-lineage cells].

The characteristics of very immature T-lineage blasts including co-expression of myeloid properties and the distinctions between the phenotypes of pro-thymic (CD7+ CD5+ CD2+ CD3- CD4- CD8- or more immature) and thymic (CD3- CD4+ CD8+ or more mature) blasts are shown. The lineage-derivation of CD7+ CD5- CD2- and CD7+ CD5+ CD2- lymphoid blasts was investigated based on the gene expression of CD3 epsilon, CD3 delta and myeloperoxidase (MPO). The former group included 4 categories; undifferentiated blasts without commitment to myeloid or T-lineage, T-lineage blasts with mRNA of CD3 epsilon, T/myeloid blasts with mRNA of CD3 epsilon and MPO and blasts with mRNA of MPO. The latter included 2 categories; T-lineage blasts with CD3 epsilon and CD3 delta mRNA and T/myeloid blasts with CD3 epsilon, CD3 delta and MPO mRNA. Thus, CD3 epsilon was expressed at a more immature stage than CD3 delta. The blasts at the pro-thymic stage were different from those at the thymic stage in several respects. The T-cell receptor (TCR) beta-chain gene showed a germ-line configuration in most pro-thymic blasts. The CD13/CD33, CD11b and class II MHC antigens were expressed very frequently in the pro-thymic stage. The CD21 antigen was most selectively expressed at the thymic stage. Pro-thymic blasts were of RA type of CD45 antigen, while thymic blasts were of RO type. Recombination activation gene-1 (RAG-1) was only limitedly expressed at the pro-thymic stage, but was highly expressed at the thymic stage. These findings indicate that there are distinctive differences before and after the initiation of clonal selection which is unique and thymus-specific, and that the neoplastic cells represent each stage of T-lineage differentiation involving the unique thymic function.