Tumor rejection by in vivo administration of anti-CD25 (interleukin-2 receptor alpha) monoclonal antibody.

Immune regulation has been shown to be involved in the progressive growth of some murine tumors. In this study, we demonstrated that a single in vivo administration of an amount less than 0.125 mg of anti-CD25 interleukin 2 receptor alpha monoclonal antibody (mAb; PC61) caused the regression of tumors that grew progressively in syngeneic mice. The tumors used were five leukemias, a myeloma, and two sarcomas derived from four different inbred mouse strains. Anti-CD25 mAb (PC61) showed an effect in six of the eight tumors. Administration of anti-CD25 mAb (PC61) caused a reduction in the number of CD4+ CD25+ cells in the peripheral lymphoid tissues. The findings suggested that CD4+ CD25+ immunoregulatory cells were involved in the growth of those tumors. Kinetic analysis showed that the administration of anti-CD25 mAb (PC61) later than day 2 after tumor inoculation caused no tumor regression, irrespective of depletion of CD4+ CD25+ immunoregulatory cells. Two leukemias, on which the PC61-treatment had no effect, seemed to be incapable of eliciting effective rejection responses in the recipient mice because of low or no antigenicity.

Expression of multiple unique rejection antigens on murine leukemia BALB/c RLmale symbol1 and the role of dominant Akt antigen for tumor escape.

Using the pRL1a Ag-loss RLmale symbol1 tumor variant cell line RM2-1, we demonstrated the presence of tumor Ags other than pRL1a that were recognized by CTLs on RLmale symbol1 cells. Semiallogeneic CB6F1 or syngeneic BALB/c CTLs generated against RM2-1 lysed RM2-1 and RLmale symbol1 cells to a similar extent, but no killing was observed with any other tumor or normal cells examined. Clonal analysis and sensitization with reversed phase-HPLC fractions revealed that there were Dd- and Ld-binding peptides recognized by RM2-1 CTLs. Lysis by bulk CTLs stimulated against RLmale symbol1 and limiting dilution analysis suggested that the pRL1a peptide was dominantly recognized to the RM2-1 peptides by CTLs on RLmale symbol1 cells. The rejection response against the parental RLmale symbol1 tumor was much less than that against RM2-1 cells in either CB6F1 or BALB/c mice, suggesting that the presence of altered Akt molecules from which the dominant pRL1a peptide was derived inhibited the rejection response against RLmale symbol1. Depletion of CD4 T cells caused the regression of RLmale symbol1 at the doses in which the tumor grew in untreated mice. The generation of pRL1a CTLs was inhibited in RLmale symbol1-bearing mice. Thus, immunoregulatory CD4 T cells were most likely activated by the altered Akt molecules and inhibited the efficient generation of CTLs against the dominant pRL1a Ag in RLmale symbol1.

Acute effects of nicotine content in cigarettes on coronary flow velocity and coronary flow reserve in men.

We investigated the acute effects of smoking on coronary flow reserve in terms of the nicotine content of cigarettes in 21 smokers. Coronary flow velocity was measured with a Doppler flow wire. Subjects smoked cigarettes containing >1 mg nicotine (n = 8, group 1) or <1 mg (n = 6, group 2). Subjects in the control group mimicked smoking without a cigarette (n = 7). Coronary flow reserve decreased after smoking in group 1, but not in group 2 or the control group. This reduction may have mediated nicotine or some other unknown substances influenced by smoking.

[Impaired coronary flow reserve due to long-term smoking recovers after quitting].

Cigarette smoking is a major risk factor for coronary artery disease. The present study examined whether impaired coronary flow reserve by smoking can be recovered by quitting. Coronary flow velocity was measured by Doppler guide wire during coronary angiography and coronary flow reserve was determined by injecting 10 mg intracoronary papaverine in 45 patients who were present or former smokers. Twenty-three patients were smoking more than 800 cigarettes/day x years and 22 patients less than 800, and 13 patients had smoked more than 800 but had quit smoking at least for 5 years. None of the patients had any significant coronary stenosis in the left anterior descending artery where the Doppler probe was positioned, nor any coronary risk factors except smoking. Twenty-six non-smokers served as control subjects. There was no difference in the coronary flow reserve between controls and light smokers (3.3 +/- 0.7 vs 3.3 +/- 1.0), but it was significantly reduced in heavy smokers (2.6 +/- 0.8) compared to controls or light smokers (p < 0.05, p < 0.05, respectively), There was no significant difference in coronary flow reserve between controls, light smokers and ex-smokers (3.3 +/- 1.2). These results suggest that the deteriorating effect on the coronary flow reserve by smoking is corrected after its cessation.

Effect of time interval between two balloon inflations on ischemic preconditioning during coronary angioplasty.

Ischemic preconditioning, defined as a reduction in myocardial ischemia caused by repeated brief episodes of coronary occlusions, is observed during percutaneous transluminal balloon angioplasty (PTCA). To elucidate the effects of the length of the interval between consecutive balloon inflations on ischemic preconditioning during PTCA, we examined 62 patients with chronic stable angina (48 males and 14 females; mean age 62 +/- 10 yr). PTCA was performed on the left anterior descending artery lacking in collateral vessels. A 2-min balloon inflation was performed twice and the extent of ST segment elevation in the electrocardiogram and the severity of chest pain (scored from 0 to 10) for each inflation were determined and compared. Patients were divided into three groups according to the interval between the two inflations: 1 min, Group 1; 2 min, Group 2; and 5 min, Group 5. In Groups 2 and 5, ST-segment elevation was significantly decreased during the second balloon inflation, as compared with that during the first inflation (P < 0.01, P < 0.001). A significant decrease was also observed in the severity of chest pain (P < 0.05, P < 0.01). However, Group 1 showed no significant decrease in ST-segment elevation or severity of chest pain between the first and second inflations. ST-segment elevation and chest pain were reduced to a greater extent in Group 5 than in Group 2. Results suggest that an interval of more than 2 min between balloon inflations is required to achieve ischemic preconditioning during PTCA.

[Effects of the time interval between the first and second balloon inflations on ischemic preconditioning during percutaneous transluminal coronary angioplasty].

Ischemic preconditioning is an attenuation of myocardial ischemia by repeated brief coronary occlusions observed during percutaneous transluminal coronary angioplasty (PTCA). The effects of the time delay between balloon inflations during PTCA on ischemic preconditioning were investigated in 48 patients with chronic stable angina but no rich collateral vessels. After successful predilatation, two 2-min balloon inflations were performed and the ST segment elevation in the electrocardiogram and chest pain were measured during each balloon inflation and compared. Patients were divided into three groups according to the interval between balloon inflations; 1 min (I1), 2 min (I2) and 5 min (I5). There were no significant differences in ST elevation (3.4, 3.2 and 3.7 mm) and chest pain during the first balloon inflation between these three groups. ST elevation and chest pain were decreased in groups I2 and I5 (2.6 and 2.8 mm) during the second balloon inflation compared with those during the first balloon inflation. However, there was no significant difference in ST elevation and chest pain during the first and second (3.7 mm) balloon inflations in group I1. ST elevation and chest pain were reduced more in group I5 than in I2. These results suggest that an interval of more than 2 min between balloon inflations is necessary to obtain the effect of ischemic preconditioning during PTCA.

Expression of the catalytic and regulatory subunits of protein phosphatase type 2A may be differentially modulated during retinoic acid-induced granulocytic differentiation of HL-60 cells.

To elucidate the regulation of protein phosphatases types 1 (PP1) and 2A (PP2A) during all-trans retinoic acid (ATRA)-induced granulocytic differentiation of HL-60 cells, the phosphatase activity, proteins, and gene expressions of PP1 and PP2A were examined. Treatment with 1 microM ATRA caused an 85% decrease in the PP2A activity in extracts from HL-60 cells, while the PP1 activity was constant. This reduction in PP2A activity appeared to parallel phenotypic and functional changes of HL-60 cells induced by ATRA. Western blot analysis showed that the level of PP2A catalytic subunit (PP2A-C) decreased during the course of ATRA-induced differentiation, whereas expressions of A and B (M(r) 55,000) regulatory subunits of PP2A were relatively unaltered. Expressions of PP1 catalytic subunit isozymes (PP1 alpha, PP1 gamma, and PP1 delta) were not significantly affected by ATRA treatment. Northern blot analysis revealed that mRNA levels of PP2A-C beta and A alpha regulatory subunits were decreased following treatment with ATRA, while levels of PP2A-C alpha and B (M(r) 55,000) alpha regulatory subunit transcripts were relatively constant. Selective down regulation of PP2A-C beta preceded the granulocytic maturation induced by ATRA. Expressions of PP2A-C isoforms and A and B regulatory subunits may be differentially modulated during ATRA-induced granulocytic differentiation of HL-60 cells.

Purification and characterization of a novel calcium-binding protein, S100C, from porcine heart.

A novel Ca(2+)-binding protein, which we have named S100C (Ohta et al. (1991) FEBS Lett. 295, 93-96), was purified to homogeneity from porcine heart by Ca(2+)-dependent dye-affinity chromatography. S100C possesses some properties of S100 proteins, such as self-association and exposure of a hydrophobic site upon binding of Ca2+ but it differs from S100 proteins in forms of its isoelectric point (pI = 6.2), cross-reactivity with antibodies, staining by Stains-all, and its Ca(2+)-dependent interaction with the immobilized dye. S100C bound to cytoskeletal components at physiological concentrations of Ca2+. Moreover, it was found that 125I-labeled S100C interacted with annexin I in a Ca(2+)-dependent manner. S100C also inhibited the phosphorylation of annexin I by protein kinase C. These data suggest that S100C might act to regulate the cytoskeleton in a Ca(2+)-dependent manner via interactions with annexin I.

Calyculin A and okadiac acid inhibit human platelet aggregation by blocking protein phosphatases types 1 and 2A.

Two potent inhibitors of protein phosphatase type 1 (PP1) and type 2A (PP2A), calyculin A (CAL-A) and okadaic acid (OKA), inhibited human platelet aggregation induced by thrombin, collagen and 9,11-epithio-11,12-methano-thromboxane A2 (STA2). IC50 values of CAL-A and OKA for STA2-induced aggregation were 53 nM and 3.5 microM, respectively. These drugs also inhibited thrombin-induced [14C]serotonin secretion of platelets. CAL-A and OKA elicited phosphorylation of certain proteins with an apparent M(r) (x 10(-3) of 200, 60, 50 and 20 light chain of myosin (MLC). Agonist-induced 47,000 M(r) protein phosphorylation was strongly inhibited by these compounds, whereas phosphorylation of 20,000 M(r) MLC was enhanced. The increase in 50,000 M(r) protein phosphorylation by CAL-A and OKA was observed in the presence of agonists, and the 50,000 M(r) phosphorylation may be involved in the inhibition of platelet activation by these compounds. Subcellular analysis of the phosphatase activity in human platelets showed that MLC phosphatase activity was present mainly (approx. 78%) in the cytosolic fraction. Chromatography of human platelet extract on heparin-Sepharose resolved two peaks of MLC phosphatase activity: PP2A in 0.1 M NaCl eluate and PP1 in 0.5 NaCl eluate. PP2A and PP1 isozymes (PP1 alpha, PP1 gamma and PP1 delta) have also been identified in human platelets, by cross-reactivity with polyclonal antibodies against PP2A and PP1 isozymes, respectively. These results suggest that PP1 and/or PP2A may play an important role in the process of platelet activation by regulating levels of phosphorylation of certain proteins.

Down-regulation by retinoic acid of the catalytic subunit of protein phosphatase type 2A during granulocytic differentiation of HL-60 cells.

Activity of protein phosphatase measured in the absence of divalent cations was decreased by 50% during all-trans retinoic acid (ATRA)-induced HL-60 cell differentiation into the granulocytic phenotype. Treatment of HL-60 cells with ATRA led to a dramatic decrease in the amount of protein phosphatase type 2A (PP2A) protein, whereas that of protein phosphatase type 1 (PP1) protein was relatively constant, as detected by immunoblotting with antibodies specific to PP1 and PP2A. The decreased phosphatase activity may be mainly due to a decrease in the expression of the PP2A protein. The mRNA level of PP2A beta was markedly decreased within 5 h after addition of ATRA, but there was only a slight increase in the mRNA level of PP2A alpha. Selective down-regulation of PP2A beta mRNA clearly preceded the cell differentiation induced by ATRA treatment. Thus, PP2A is down-regulated during ATRA-induced differentiation of HL-60 cells into granulocytes.

Risk factors for the complication of cerebral infarction in Japanese patients with acute myocardial infarction.

The incidence and risk factors for cerebral infarction in Japanese patients with acute myocardial infarction were evaluated. Seven (5.0%) of 140 patients with acute myocardial infarction suffered from cerebral infarction during their initial hospitalization. The incidence was slightly higher than those reported in Western countries. Anterior wall myocardial infarction and a past history of cerebrovascular disease were considered to be probable risk factors for the complication. A beneficial effect of anticoagulant therapy in preventing cerebral infarction in cases of acute myocardial infarction with those risk factors is suggested.

R wave changes in patients with myocardial infarction during treadmill stress testing.

A graded multistage treadmill exercise test was performed on 78 patients with myocardial infarction (65 males and 13 females, ranging in age from 30 to 76). The amplitude of the R wave in CC5 was measured at rest and during the periods of peak exercise. The exercise-induced R wave changes were classified into 3 groups: increased, 39.1%; no changes, 39.7% and decreased, 22.2%. Acute attacks of myocardial infarction were more severe in the patients whose R wave decreased during exercise than in those whose R wave increased. Heart rate, blood pressure and pressure rate products were not different among the groups. However, oxygen consumption was greater in the R-increased group than in the R-decreased group. Therefore, an increase in R wave amplitude during exercise in post-myocardial infarction patients indicates a good efficiency of the myocardium and skeletal muscles. Exercise-induced changes of the mean electrical axes of the transverse plane were significantly related to changes of the R wave amplitude in CC5 (r = 0.50, p less than 0.001). Therefore, change in the transverse plane axis is one of the important determinants of exercise-induced R wave changes.

Exercise capacity of the patients with myocardial infarction.

This study was conducted to investigate the functional capacity in postmyocardial infarction. Eighty-four multistage treadmill exercise tests were performed on 60 patients, none of whom had had any formal rehabilitation or regular exercise. There were 50 men and 10 women, ranging in age from 30 to 81 with an average age of 60. The time interval between the acute attack and the exercise test ranged from one month to 9 years. Even though severe infarction affects the exercise capacity for a long time after an acute attack, its effect on cardiac function was more obvious than that on physical capacity. Age was the most important determinant of physical capacity, and the slope of decreasing physical capacity with age in patients with infarct was the same as that in normal subjects. Cardiac function also decreased with age. However, during the early recovery phase, cardiac function was influenced by the severity of infarction and the influence of age could not be established. There was no significant correlation between early ambulation and physical capacity. The beneficial effects of early ambulation may be lost if physical activity is discontinued for some time after the acute attack. The physical capacity increased 2-3 years after the acute attack, but myocardial function did not change significantly.