Lymphocyte modulation by inflammatory bronchoalveolar leukocytes.
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Biomedical subjects
Publications and source records attributed to Y Kusaka.
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Eight asthmatic patients with hard metal asthma due to cobalt underwent bronchial provocation challenge with nickel sulphate. Seven patients developed a fall in FEV1 of 20% or more after inhaling nickel sulphate, four showing an immediate response and three a late response. Eight control subjects, including six asthmatic patients, with no history of hard metal exposure, showed no bronchoconstriction in response to a provocation challenge with nickel sulphate. Specific antibodies to nickel conjugated human serum albumin were present in four of the eight patients with sensitivity to cobalt conjugated human serum albumin but were absent from the serum of 60 unexposed asthmatic patients and 25 exposed symptom free workers. These results suggest that nickel as well as cobalt sensitivity plays a part in hard metal asthma.
Interleukin-1 like activity was produced by neutrophils obtained by bronchoalveolar lavage from experimentally inflamed rat lung. Activity was released spontaneously from neutrophils at high levels but it was enhanced by stimulation with endotoxin in vitro.
A cell line, HuH-33 was cultured in vitro from a patient with hepatocellular carcinoma. This cell line has been in continuous culture over 12 month period with slow growth potential. HuH-33 was composed spindle-or polygonal-shaped cells as a major population. Chromosome number of the cells were widely distributed even in the primary culture. HuH-33 was transplantable into nude mice and secreted alpha-fetoprotein, albumin, beta 2 microglobulin, ferritin and tissue polypeptide antigen.
A serum-free culture system of human hepatoma cell lines (HuH-6 and HuH-7) was used to investigate the activity of bovine serum (BS) and of serum-derived factors on the growth and production of alpha-fetoprotein (AFP) and albumin. At higher concentrations, dialyzed BS was inhibitory to the growth of HuH-6 and caused reduction of the level of AFP production by the cells. AFP and albumin levels in HuH-6 and HuH-7 were reduced or unchanged by fetuin, bovine serum albumin (BSA) and transferrin (TF), although no cytotoxicity was shown by any of them. Commercial preparations of platelet-derived growth factor exhibited cytotoxicity to HuH-6 and HuH-7 and induced a decrease of AFP and albumin levels in a dose-dependent manner. Transforming growth factor (TGF-beta) exhibited no cytotoxicity to HuH-6. AFP levels in HuH-6 were unchanged with 1000 pg/ml TGF-beta, but albumin levels were decreased. TGF-beta at a concentration of 1000 pg/ml was cytotoxic to HuH-7 and AFP levels were a little increased. Albumin levels, however, were unchanged. Following exposure to cycloheximide, AFP and albumin levels in HuH-6 were inhibited.
Eight asthmatic patients who had no history of asthma before starting work in a hard-metal plant and eight control subjects (three atopic, three nonatopic asthmatic, and two normal volunteers) without a history of exposure to hard metal dust were subjected to provocation tests, skin tests, radioallergosorbent tests (RAST) and Farr test with cobalt. Four of the eight patients were atopic, and seven showed bronchial hyperresponsiveness to methacholine (BHR). Patch and intradermal skin tests with cobalt chloride (CoCl2) could not discriminate the patients from control subjects. All patients had positive reactions to CoCl2 in the provocation tests; two developed immediate asthmatic reaction (IAR), four late asthmatic reaction (LAR), and two dual asthmatic reaction (DAR), while the control subjects showed no reaction. Evidence of specific IgE antibodies to cobalt-conjugated human serum albumin (Co-HSA) was presented by four patients (RAST score greater than 2) based on comparison of serum samples from 60 asthmatic patients and 25 asymptomatic workers in the same plant. Positive serum samples selectively bound 57Co, and the test was blocked by nonlabled cobalt sulfate (CoSO4). These findings suggest the development of hard metal-induced asthma from cobalt sensitivity.
A method based on interday fluctuation of contaminant concentrations for evaluating employee's exposure averages (8-h TWAs) was proposed in our previous report. The method was established on the assumption that daily exposure averages of the workers are lognormally distributed in actual workplaces. The study was conducted to elucidate whether the distribution of daily exposure averages is statistically lognormal or not and to examine the relationship between sample geometric standard deviation (sg) of worker's daily exposure averages and its estimate (sg2) calculated by measurements for two consecutive days. These are critical for our proposed method. The data on daily exposure concentrations over five to six weeks were collected from workers exposed to cobalt, acetone, n-hexane, toluene, xylene, ethylbenzene and ethylacetate. The data on organic lead, inorganic lead and mercury exposures reported by Cope et al. and Lindstedt et al. were also used for the study. The result can be summarized as follows: 1. Lognormal distribution of daily exposure averages was confirmed by plotting on normal probability paper and chi 2-test. 2. Median of sg2 on daily exposure averages obtained from individual worker was smaller than sg. 3. sg calculated by a set of measurements for two consecutive days in every worker can be corrected by the equation: (sg2) 1.48 in obtaining a better estimate of sigma g. 4. Statistical analysis on daily exposure averages of all workers showed that median of sg2 was smaller than that of sg, and 88% upper limit of sg2 was equal to that of sg. 5. Therefore, in evaluating TWA obtained by only single day's measurement using the proposed method, median of sg representative of industrial hazardous substance exposure workplaces could be also corrected by the equation described above. However, correction of 90% upper limit of sg2 is not necessary.
The self-administered respiratory questionnaire on persistent cough or sputum prepared and recommended by the Japanese Labour Ministry to detect pneumoconiosis was evaluated and compared with the Japanese version of the standardized interviewer-administered respiratory questionnaire developed by the British Medical Research Council (BMRC). The two questionnaires were given to 800 subjects on the same day. Because the wording of the questions was similar, the two questionnaires yielded very similar results. In terms of the overall prevalence of persistent cough or sputum, no important differences could be demonstrated between the results of the two questionnaires. The results of this study indicate that the self-administered questionnaire has good concordance with the BMRC questionnaire. When compared to the BMRC questionnaire, the specificity of the self-administered questionnaire was found to be more than 99%, but the sensitivity was inferior to the specificity, being less than 80%. When confined to subjects with persistent cough or sputum for two years or more, the sensitivity was up to 84%. These findings suggest the responses of the subjects to the self-administered questionnaire tend to be negative. A validity of the self-administered questionnaire needs to be further studied among dust workers including pneumoconiotics.
Isozyme patterns of glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) in human cell lines derived from primary hepatomas were compared with those in HeLa cells. Some cell lines derived from primary hepatomas having type B G6PD showed one or two isozymes of LDH. On the other hand, HeLa cells having type A G6PD showed four LDH isozymes. These findings suggest that not only G6PD, but also LDH may be useful for the detection of HeLa cell contamination of a culture in some cases.
The effect of various factors and substrates on the growth of a human hepatoblastoma cell line, HuH-6, which was inoculated at low density in a serum-free medium was examined. Several supplements were required to enhance cell growth of HuH-6. These included cholera toxin (CT), glucagon (Glu) and selenium (Se). Type IV collagen (C-IV) provided the most conductive environment tested for cell growth. These results suggest that CT, Glu, Se, and C-IV are important stimulators for the continuous growth of HuH-6 in a serum-free medium at low density.
Lymphocyte transformation tests with cobalt were done in 9 hard metal asthma patients diagnosed on the basis of a positive bronchial provocation test with cobalt. Positive lymphocyte proliferation in response to the metal was defined statistically from the results of 18 controls comprising 12 hard metal-exposed and 6 non-exposed controls. Out of the 4 patients who had been reported to have IgE antibody specific to cobalt, the lymphocytes of 2 proliferated with metal: one showed positive lymphocyte transformation to free cobalt, the other to both free cobalt and cobalt-conjugated human serum albumin. These results suggest that cobalt-sensitized lymphocytes play a role in some hard metal asthmatics. However, the mechanism underlying the hard metal asthma in the other 5 asthmatics who had neither cobalt-specific IgE antibody nor cobalt-sensitized lymphocytes remain to be elucidated.
The effect of various anticancer drugs on alpha-fetoprotein (AFP) secretion and some other properties of human hepatoma cells was investigated in vitro with the following results. (1) There was a high correlation between AFP secretion and cell number after treatment of human hepatoma cells with anticancer drugs and the amounts of AFP secreted per 10(4) cells per 72 hours (AFP-secreting capacity) were not affected within therapeutically achievable concentrations (TAC). (2) The AFP-secreting capacity was affected with some exceptions in the cells treated with higher concentration of drugs than TAC. Furthermore, chromosomal and morphological aberrations in the similarly treated cells, were also observed, suggesting the relationship between the change of AFP-producing capacity and that of some other properties.
An autopsy case of pulmonary asbestosis with synchronous multiprimary lung cancers, gastric cancer and atypical mycobacteriosis of a 64-year-old asbestos textile worker was reported. Chest X-ray film on the first admission showed a coin lesion on the left upper lung field, and on the second admission showed a progressively enlarging shadows in the left hilum. Chemotherapy showed almost no effect and the patient died of respiratory failure. Autopsy revealed moderate asbestosis, with triple lung cancers (squamous cell carcinoma, adenocarcinoma and small cell carcinoma) in the left upper lobe, gastric cancer (signet-ring cell carcinoma) and atypical mycobacteriosis. So far as we know, no case with such complex lesions has ever been reported.
Although on anorectal malignant melanoma is relatively rare, we report two cases, that of a 88-year-old woman, and a 78-year-old man. Both patients were admitted to the hospital due to rectal bleeding and were diagnosed as having rectal carcinoma. Following an abdominoperineal rectal amputation, a pathological examination of the resected specimens revealed a malignant melanoma. Thus, it is necessary to carry out a careful examination of patients with tumors of the anorectal region when there is the least suspicion of a malignancy.
A cell line, HuH-28, was established in vitro from a patient with cholangiocellular carcinoma (CCC). This cell line has grown slowly, revealing a doubling time of approximately 80 h, and the serial passages were carried out 20 times within 10 months. Light microscopy revealed spindle and polygonal morphology of the cells. Chromosome number of the cells were distributed near the hypotriploid region at passages 3 and 14. HuH-28 cells were not transplantable into nude mice, but secreted some tumor markers including alkaline phosphatase (ALP), gamma glutamyltranspeptidase (GGT), beta 2-microglobulin (BMG), ferritin, elastase-1, and tissue polypeptide antigen (TPA). This HuH-28 cell line will represent a good model for the investigation of carcinogenesis, histogenes, and diagnosis of CCC.
The effects on human hepatoblastoma (HuH-6) and hepatoma (HuH-7) cell lines of collagen type I (CI), type IV (CIV), fibronectin (FN) and laminin (LAM) were investigated. CI and CIV promoted almost equally the attachment of both cell lines more strikingly than did FN or LAM. CI and CIV were also superior to FN or LAM in supporting the growth of HuH-6. These substrates, however, had no appreciable effect on the growth of HuH-7. The amount of alpha-fetoprotein (AFP) and albumin secreted in HuH-6 was found to be higher on FN- and LAM- coated substrates than on the other matrix material- coated ones. HuH-7 exhibited increased levels of AFP on LAM- coated substrates 4 days after plating. These results indicate that the ability of extracellular matrix materials to enhance attachment and/or growth is different from that to enhance AFP and albumin production in HuH-6 and probably in HuH-7.
Twelve workers with hard metal asthma diagnosed on the basis of peak flow diaries and positive bronchial reactions to cobalt chloride (CoCl2) were studied for sensitization by detection of specific antibodies to radioactive cobalt (57Co), cobalt-conjugated human serum albumin (Co-HSA) and cobalt-conjugated exchange resin (Co-resin). Their IgE titres ranged from 73 to 1500 IU/ml and eight were atopic individuals. Sixty serum samples from asthmatic patients with IgE titres of 14-4300 IU/ml were studied as controls in all tests. Eleven of twelve subject sera that selectively bound to 57Co after incubation with saturated ammonium sulphate (greater than 232 c.p.m., P less than 0.01) were divided into three groups: (1) six sera showing evidence of specific IgE antibodies to Co-HSA (greater than 673 c.p.m., P less than 0.01) without those to Co-resin; (2) one serum giving a positive radio-allergosorbent test (RAST) only to Co-resin (greater than 417 c.p.m., P less than 0.01), and (3) four sera that were negative for two antigenic agents (Co-HSA, Co-resin). These results suggest that the subjects had occupational asthma due to hard metal exposure from cobalt sensitivity. An immuno-allergic mechanism mediated by specific IgE antibodies to cobalt was confirmed to be responsible for the development of hard metal asthma, with the possibility of some role of the reaction without reagins.
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