The presence of calcium (Ca2+) in the culture medium is a requirement for the NK cytotoxic reacti... more The presence of calcium (Ca2+) in the culture medium is a requirement for the NK cytotoxic reaction. To further explore the role of Ca2+ and calmodulin (a cytoplasmic protein that mediates most of the biological effects of Ca2+) in this process, we evaluated the effects of nifedipine (a Ca2+ channel antagonist), BAY-K-8644 (a Ca2+ channel agonist), and haloperidol (an inhibitor of calmodulin) on the NK activity of human peripheral blood mononuclear cells (PBMC), and the augmentation of this activity by recombinant interleukin 2 (r-IL 2) and interferon-gamma (r-gamma-IFN). We found that all of these drugs inhibit NK activity in a dose-dependent fashion. This appears to result from interference with the programming for lysis stage of the lytic process. In contrast, the presence of these agents during the incubation of PBMC with r-IL 2 or r-gamma-IFN did not induce any change in the enhancement of NK activity. These data suggest that Ca2+ exerts its effect at the intracellular level du...
Chromosomes were studied in cells from 15 japanese patients with adult T-cell leukemia (ATL). Mit... more Chromosomes were studied in cells from 15 japanese patients with adult T-cell leukemia (ATL). Mitoses were obtained from unstimulated peripheral blood in 12 patients and a lymph node in one patient. In two other patients, mitotic cells were obtained solely from peripheral blood stimulated with phytohemagglutinin (PHA). Chromsomally abnormal cells were seen in 14 of the 15 patients. The abnormal cells had a modal number of chromosomes in near diploid range in 12 patients, and in near triploid and tetraploid range in the remaining 2 patients, respectively. Eight of the nine patients analyzed by Q-banding had clonal chromosome abnormalities. The most common abnormality was trisomy no.7 or 7q, which was seen in 5 cases and has been primarily observed in lymphoid neoplasms. A 14q+ marker chromosome was found in two patients and a Dq+ in one patient; loss of a sex chromosome was found in three patients. Most chromosomes were involved in gains, losses, or structural rearrangements, but abn...
Leukemic cells from nine patients with adult T-cell leukemia were examined for Fey and Fcp recept... more Leukemic cells from nine patients with adult T-cell leukemia were examined for Fey and Fcp receptors, human B-lymphocyte antigens (HBLA), and effect on PWM-induced B-cell differentiation. Fey receptors were detected in one and Fcp receptors in another. Leukemic cells in both cases bore HBLA and expressed a suppressive effect on PWMinduced B-cell differentiation. However, leukemic cells from the remaining seven cases did not bear Fc receptors. In four, leukemic cells possessed HBLA and all expressed a suppressive effect on the PWM system. Some leukemic cells which did not express HBLA had the same effect on the PWM system. The origin of adult T-cell leukemia was discussed.
European Journal of Cancer and Clinical Oncology, 1984
The phototoxicity of acridine orange and argon laser irradiation on Walker carcinosarcoma 256 sto... more The phototoxicity of acridine orange and argon laser irradiation on Walker carcinosarcoma 256 stomach tumors was studied. Wistar strain rats bearing stomach tumors 4-6 mm in diameter 5-10 days after their implantation were injected intraperitoneally with 40 mg/kg of acridine orange 2 hr before irradiation. Then the forestomach was opened and the tumors were exposed to the argon laser at 488 nm at an intensity of 15 mW/cm2 for 20 min. Tumors in rats treated with acridine orange were brightly fluorescent during irradiation. No marked temperature rise was detected during irradiation. Argon irradiation significantly prolonged the survival of rats treated with acridine orange. Histologically, complete or partial tumor necrosis was observed, with sparing of surrounding mucosa, in rats treated by the combination therapy. Phase-contrast and electron microscopy showed that cytotoxicity was mediated by changes in the cell, nuclear and lysosomal membranes. Neither the dye nor laser alone had any effect.
The pathogenesis of airway inflammation in diffuse panbronchiolitis (DPB) is unknown. Neutrophil ... more The pathogenesis of airway inflammation in diffuse panbronchiolitis (DPB) is unknown. Neutrophil survival-enhancing activity, partially mediated by granulocyte-macrophage colony-stimulating factor (GM-CSF), has been shown in the sputum from DPB patients. This study investigated the mechanisms of GM-CSF expression in the airways of DPB patients. This involved examining the effects of mucoid Pseudomonas aeruginosa strains derived from chronically colonized patients with DPB on neutrophil survival and GM-CSF expression. Neutrophils from healthy subjects were cultured with the culture supernatants of P. aeruginosa isolated from sputum of DPB patients in the presence or absence of anti-GM-CSF and anti-GM-CSF receptor (alpha chain) antibodies, and viable neutrophils were counted daily. GM-CSF gene expression in neutrophils was evaluated by RT-PCR. Neutrophils cultured with the culture supernatants showed significantly prolonged survival, compared with neutrophils cultured with the control broth. The neutrophil survival-enhancing activity in the culture supernatants was lost by heating. The enhanced survival of neutrophils was abolished in the presence of anti-GM-CSF and anti-GM-CSF receptor (alpha chain) antibodies. GM-CSF mRNA was detected in neutrophils cultured with the bacterial supernatants, but not in those with the control broth. P. aeruginosa-derived factors (rich in proteins) stimulated neutrophils to synthesize GM-CSF, which enhanced neutrophil survival in an autocrine/paracrine fashion.
We examined the effects of extracellular adenosine 5&... more We examined the effects of extracellular adenosine 5'-triphosphate (ATP) on single airway smooth muscle (ASM) cells from porcine trachea using a patch-clamp technique. ATP induced a sustained inward current. Phospholipase C inhibitor U-73122 failed to inhibit the current, suggesting the involvement of P2X receptor. A specific effecter of P2X(4), ivermectin, augmented the current indicating the existence of P2X(4) receptors. Immunohistochemistry and reverse transcription/polymerase chain reaction analysis and Western blot analysis also showed the distribution of the P2X(4) receptors. The inward current was reduced by SKF-96365, an inhibitor of both voltage-dependent Ca(2+) channels (VDCCs) and voltage-independent Ca(2+) channels, although a VDCC antagonist, verapamil, did not affect the current. SKF-96365 caused complete suppression of both the increase in the intracellular Ca(2+) concentration and the contraction of ASM cells induced by ATP. Our results demonstrate that P2X(4) receptors exist on ASM and that the receptors are responsible for Ca(2+) influx. These findings suggest that the Ca(2+) influx regulated by P2X(4) receptors plays an important role in ASM contraction by a pathway distinct from VDCC.
An autopsy case of a 37‐year‐old Japanese man, confirmed as an AIDS patient infected by an undete... more An autopsy case of a 37‐year‐old Japanese man, confirmed as an AIDS patient infected by an undetermined route of transmission, is presented. The initial symptoms of full‐blown AIDS in this case were neurological, and the patient died of severe pneumonia 9 months after onset. The main histo‐ and immu‐nopathological features were a marked depletion of helper‐inducer T cells and dendritic reticulum cells in the lymphoid tissues, opportunistic infections, and some neuropathology changes. Very few cells, possibly macrophages, immu‐noreactive with a monoclonal antibody (VAK‐5) against HIV‐gag protein P24 were found in the mediastinal lymph nodes. Numerous pathogens had induced opportunistic infections in many organs: severe and generalized cytomegalovirus infection, Pneumocystis carinii pneumonia, bronchopneumonia (possibly due to Pseudomonas aeruginosa), candidiasis in the tongue and oral cavity, and atypical mycobacteriosis in the pulmonic hilar lymph nodes. Vascular proliferation was f...
We have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) fr... more We have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) from the culture supernatants of CHO‐Sec cells that secreted truncated 140‐kDa precursor and mature 120‐kDa Env glycoproteins. The concentrated culture supernatants were applied to a column coupled with cibacron blue 3GA (CB3GA) to separate albumin from the Env proteins because CB3GA, a triazine dye, has been known to have a high affinity to albumin. Unexpectedly, Env proteins as well as albumin bound to the column, and the bound Env proteins were eluted by increasing the ionic strength using KC1. Gp120 was eluted at 0.5–0.9 m of KC1, while a higher concentration (0.9–1.5 m) was necessary for the elution of gp140. The agarose gel coupled with reactive red 120 (RR120), another triazine dye with similar characteristics, also retained both Env proteins, and the bound Env proteins could be eluted in a similar manner. In addition, these agents inhibited syncytium formation caused by HTLV‐IIIB a...
The presence of calcium (Ca2+) in the culture medium is a requirement for the NK cytotoxic reacti... more The presence of calcium (Ca2+) in the culture medium is a requirement for the NK cytotoxic reaction. To further explore the role of Ca2+ and calmodulin (a cytoplasmic protein that mediates most of the biological effects of Ca2+) in this process, we evaluated the effects of nifedipine (a Ca2+ channel antagonist), BAY-K-8644 (a Ca2+ channel agonist), and haloperidol (an inhibitor of calmodulin) on the NK activity of human peripheral blood mononuclear cells (PBMC), and the augmentation of this activity by recombinant interleukin 2 (r-IL 2) and interferon-gamma (r-gamma-IFN). We found that all of these drugs inhibit NK activity in a dose-dependent fashion. This appears to result from interference with the programming for lysis stage of the lytic process. In contrast, the presence of these agents during the incubation of PBMC with r-IL 2 or r-gamma-IFN did not induce any change in the enhancement of NK activity. These data suggest that Ca2+ exerts its effect at the intracellular level du...
Chromosomes were studied in cells from 15 japanese patients with adult T-cell leukemia (ATL). Mit... more Chromosomes were studied in cells from 15 japanese patients with adult T-cell leukemia (ATL). Mitoses were obtained from unstimulated peripheral blood in 12 patients and a lymph node in one patient. In two other patients, mitotic cells were obtained solely from peripheral blood stimulated with phytohemagglutinin (PHA). Chromsomally abnormal cells were seen in 14 of the 15 patients. The abnormal cells had a modal number of chromosomes in near diploid range in 12 patients, and in near triploid and tetraploid range in the remaining 2 patients, respectively. Eight of the nine patients analyzed by Q-banding had clonal chromosome abnormalities. The most common abnormality was trisomy no.7 or 7q, which was seen in 5 cases and has been primarily observed in lymphoid neoplasms. A 14q+ marker chromosome was found in two patients and a Dq+ in one patient; loss of a sex chromosome was found in three patients. Most chromosomes were involved in gains, losses, or structural rearrangements, but abn...
Leukemic cells from nine patients with adult T-cell leukemia were examined for Fey and Fcp recept... more Leukemic cells from nine patients with adult T-cell leukemia were examined for Fey and Fcp receptors, human B-lymphocyte antigens (HBLA), and effect on PWM-induced B-cell differentiation. Fey receptors were detected in one and Fcp receptors in another. Leukemic cells in both cases bore HBLA and expressed a suppressive effect on PWMinduced B-cell differentiation. However, leukemic cells from the remaining seven cases did not bear Fc receptors. In four, leukemic cells possessed HBLA and all expressed a suppressive effect on the PWM system. Some leukemic cells which did not express HBLA had the same effect on the PWM system. The origin of adult T-cell leukemia was discussed.
European Journal of Cancer and Clinical Oncology, 1984
The phototoxicity of acridine orange and argon laser irradiation on Walker carcinosarcoma 256 sto... more The phototoxicity of acridine orange and argon laser irradiation on Walker carcinosarcoma 256 stomach tumors was studied. Wistar strain rats bearing stomach tumors 4-6 mm in diameter 5-10 days after their implantation were injected intraperitoneally with 40 mg/kg of acridine orange 2 hr before irradiation. Then the forestomach was opened and the tumors were exposed to the argon laser at 488 nm at an intensity of 15 mW/cm2 for 20 min. Tumors in rats treated with acridine orange were brightly fluorescent during irradiation. No marked temperature rise was detected during irradiation. Argon irradiation significantly prolonged the survival of rats treated with acridine orange. Histologically, complete or partial tumor necrosis was observed, with sparing of surrounding mucosa, in rats treated by the combination therapy. Phase-contrast and electron microscopy showed that cytotoxicity was mediated by changes in the cell, nuclear and lysosomal membranes. Neither the dye nor laser alone had any effect.
The pathogenesis of airway inflammation in diffuse panbronchiolitis (DPB) is unknown. Neutrophil ... more The pathogenesis of airway inflammation in diffuse panbronchiolitis (DPB) is unknown. Neutrophil survival-enhancing activity, partially mediated by granulocyte-macrophage colony-stimulating factor (GM-CSF), has been shown in the sputum from DPB patients. This study investigated the mechanisms of GM-CSF expression in the airways of DPB patients. This involved examining the effects of mucoid Pseudomonas aeruginosa strains derived from chronically colonized patients with DPB on neutrophil survival and GM-CSF expression. Neutrophils from healthy subjects were cultured with the culture supernatants of P. aeruginosa isolated from sputum of DPB patients in the presence or absence of anti-GM-CSF and anti-GM-CSF receptor (alpha chain) antibodies, and viable neutrophils were counted daily. GM-CSF gene expression in neutrophils was evaluated by RT-PCR. Neutrophils cultured with the culture supernatants showed significantly prolonged survival, compared with neutrophils cultured with the control broth. The neutrophil survival-enhancing activity in the culture supernatants was lost by heating. The enhanced survival of neutrophils was abolished in the presence of anti-GM-CSF and anti-GM-CSF receptor (alpha chain) antibodies. GM-CSF mRNA was detected in neutrophils cultured with the bacterial supernatants, but not in those with the control broth. P. aeruginosa-derived factors (rich in proteins) stimulated neutrophils to synthesize GM-CSF, which enhanced neutrophil survival in an autocrine/paracrine fashion.
We examined the effects of extracellular adenosine 5&... more We examined the effects of extracellular adenosine 5'-triphosphate (ATP) on single airway smooth muscle (ASM) cells from porcine trachea using a patch-clamp technique. ATP induced a sustained inward current. Phospholipase C inhibitor U-73122 failed to inhibit the current, suggesting the involvement of P2X receptor. A specific effecter of P2X(4), ivermectin, augmented the current indicating the existence of P2X(4) receptors. Immunohistochemistry and reverse transcription/polymerase chain reaction analysis and Western blot analysis also showed the distribution of the P2X(4) receptors. The inward current was reduced by SKF-96365, an inhibitor of both voltage-dependent Ca(2+) channels (VDCCs) and voltage-independent Ca(2+) channels, although a VDCC antagonist, verapamil, did not affect the current. SKF-96365 caused complete suppression of both the increase in the intracellular Ca(2+) concentration and the contraction of ASM cells induced by ATP. Our results demonstrate that P2X(4) receptors exist on ASM and that the receptors are responsible for Ca(2+) influx. These findings suggest that the Ca(2+) influx regulated by P2X(4) receptors plays an important role in ASM contraction by a pathway distinct from VDCC.
An autopsy case of a 37‐year‐old Japanese man, confirmed as an AIDS patient infected by an undete... more An autopsy case of a 37‐year‐old Japanese man, confirmed as an AIDS patient infected by an undetermined route of transmission, is presented. The initial symptoms of full‐blown AIDS in this case were neurological, and the patient died of severe pneumonia 9 months after onset. The main histo‐ and immu‐nopathological features were a marked depletion of helper‐inducer T cells and dendritic reticulum cells in the lymphoid tissues, opportunistic infections, and some neuropathology changes. Very few cells, possibly macrophages, immu‐noreactive with a monoclonal antibody (VAK‐5) against HIV‐gag protein P24 were found in the mediastinal lymph nodes. Numerous pathogens had induced opportunistic infections in many organs: severe and generalized cytomegalovirus infection, Pneumocystis carinii pneumonia, bronchopneumonia (possibly due to Pseudomonas aeruginosa), candidiasis in the tongue and oral cavity, and atypical mycobacteriosis in the pulmonic hilar lymph nodes. Vascular proliferation was f...
We have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) fr... more We have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) from the culture supernatants of CHO‐Sec cells that secreted truncated 140‐kDa precursor and mature 120‐kDa Env glycoproteins. The concentrated culture supernatants were applied to a column coupled with cibacron blue 3GA (CB3GA) to separate albumin from the Env proteins because CB3GA, a triazine dye, has been known to have a high affinity to albumin. Unexpectedly, Env proteins as well as albumin bound to the column, and the bound Env proteins were eluted by increasing the ionic strength using KC1. Gp120 was eluted at 0.5–0.9 m of KC1, while a higher concentration (0.9–1.5 m) was necessary for the elution of gp140. The agarose gel coupled with reactive red 120 (RR120), another triazine dye with similar characteristics, also retained both Env proteins, and the bound Env proteins could be eluted in a similar manner. In addition, these agents inhibited syncytium formation caused by HTLV‐IIIB a...
Uploads
Papers by 俊夫 服部