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Biomedical subjects

M Adachi

Publications and source records attributed to M Adachi.

At least 721 records · Page 40Linked to original sources

T-cell hybridoma co-expressing Fc receptors for different isotypes. I. Reciprocal regulation of Fc alpha R and Fc gamma R expression by IgA and interferon.

To clarify the co-expression phenomenon of T-cell Fc receptors (FcR) specific for different isotypes on the clonal level, a murine hybridoma clone T2D4 was studied. T2D4 cells originally reported to bear FcR for IgG (Fc gamma R) and to release a Fc gamma R-related T-cell factor binding to IgG (immunoglobulin binding factor; IBF) proved to have also the receptor for IgA. The binding of IgA was detected by rosette formation with trinitrophenylated ox red blood cells (TNP-ORBC) after preincubation of T2D4 cells with MOPC 315 IgA having anti-TNP activity, or directly with TNP-ORBC sensitized with MOPC 315 IgA. While the binding of MOPC 315 IgA was competed for by IgA but not by IgG2A nor IgG2B, IgA failed to inhibit the rosette formation of the cells with ORBC sensitized with rabbit IgG antibody (EA ox gamma), proving that T2D4 cells express FcR specific for IgA (Fc alpha R) in addition to Fc gamma R. Co-expression of both receptors on the same cell surface was demonstrated by a double rosette technique using TNP-quail red blood cells (TNP-QRBC) and EAox gamma. Fc alpha R activity of the cells was completely abrogated by 15 min. incubation with 0.1 mg/ml trypsin, whereas Fc gamma R was resistant even to 1 mg/ml trypsin. The expression of Fc alpha R was augmented (up-regulation) by IgA at the concentration above 300 micrograms/ml and inhibited (down-regulation) by 1000 u./ml of murine beta-interferon (beta-IFN). Conversely, the expression of Fc gamma R was down-regulated by IgA and up-regulated by alpha-IFN. Thus, Fc gamma R and Fc alpha R are co-expressed and reciprocally regulated on these cell lines. The possible co-production of IBF and the Fc alpha R-related binding factor specific for IgA is discussed.

Animals↗

T cell hybridomas coexpressing Fc receptors (FcR) for different isotypes. II. IgA-induced formation of suppressive IgA binding factor(s) by a murine T hybridoma bearing Fc gamma R and Fc alpha R.

T2D4 murine T hybridoma cells have previously been shown to express Fc receptors (FcR) for IgG (Fc gamma R) and for IgA (Fc alpha R) and to produce an IgG binding factor (IgGBF) that suppresses IgG and IgM responses. In the present work we report on the behavior of IgA bound to T2D4 cells and on the production of IgA binding factor (IgABF) and its ability to suppress IgA antibody production. A dose-dependent binding of MOPC315 IgA with anti-TNP activity by T2D4 cells was demonstrated by rosette formation with trinitrophenylated ox red blood cells (TNP-ORBC) and fixation of iodinated DNP-BSA. IgA bound to the cells disappeared after a short-term culture of 3 hr at 37 degrees C, but not at 4 degrees C. Because this phenomenon was inhibited by 0.1% sodium azide and 100 microM dansylcadaverine, a transglutaminase inhibitor, Fc alpha R-IgA complexes seemed to be released by an active process involving receptor movement. In the culture supernatant of IgA-treated T2D4 cells, we detected a factor(s) that binds to IgA-Sepharose and competitively inhibits the binding of IgA to T2D4 cells. The factor (IgABF) failed to inhibit the rosette formation of Fc gamma R(+) cells with IgG-sensitized ORBC (EAox gamma), indicating that it binds specifically to IgA. IgABF was undetectable in the culture supernatants of untreated T2D4 cells of Fc alpha R(-) BW5147 T lymphoma cells used as parent cells for the establishment of the hybridoma. To study the effect of IgABF on antibody formation, culture filtrates of IgA-treated or untreated T2D4 cells were fractionated on IgA-Sepharose beads and were added to BALB/c spleen cells cultured with pokeweed mitogen. By use of a reverse plaque assay, it was shown that the IgA plaque-forming cell (PFC) response was suppressed by the acid eluate but not by the effluent of IgA-Sepharose beads incubated with the filtrates of IgA-treated T2D4 cell cultures. The suppression was IgA specific, because neither IgG nor IgM responses were suppressed by the eluate. As expected, there was no significant IgA suppressive activity in the acid eluates of the beads incubated with the culture filtrate of untreated T2D4 cells or IgA-treated BW5147 cells. IgA-specific suppressive activity proved to be due to IgA binding factor(s), because suppressive activity in the eluate was completely adsorbed by IgA-Sepharose but not by IgG- nor BSA-Sepharose.

Animals↗

Altered expression of lymphocyte Fc alpha receptor in selective IgA deficiency and IgA nephropathy.

To study the expression of FcR specific for IgA (Fc alpha R) on human peripheral lymphocytes (PBL), PBL from normal donors were incubated with 300 to 500 micrograms/ml MOPC 315 IgA having anti-trinitrophenyl (TNP) antibody activity at 4 degrees C or 37 degrees C for 60 min. Under this condition, less than 2% of total cells could form rosettes with TNP-coated ox red blood cells (TNP-ORBC). When cultured with MOPC 315 IgA at 37 degrees C for 18 hr, however, there was a dose-dependent increase of the rosette-forming cells (RFC) binding TNP-ORBC. Because 15 to 20% of the total cells bound TNP-ORBC but not unsensitized ORBC, the rosette formation appeared to be due to the cytophilic binding of IgA to the cells. The binding of MOPC 315 IgA was competed by TEPC 15 IgA and human myeloma IgA, but not by murine myeloma proteins of other classes, indicating that the receptor is specific for IgA. Fc alpha R was induced on 15 to 20% of fractionated T and B cells, as well as on 15 to 18% of concanavalin A-(Con A) activated lymphocytes when cultured with IgA. The induction of the receptor was dependent on protein and RNA synthesis, but not on DNA synthesis as suggested by the sensitivity to metabolic inhibitors, such as mitomycin C, actinomycin D, puromycin, and cycloheximide. In five patients with selective IgA deficiency (serum IgA, 0 to 4 mg/dl), only 5.1% +/- 1.7 of PBL formed rosettes with TNP-ORBC after culture with MOPC 315 IgA, whereas 12.5% +/- 2.5 of PBL from normal donors (serum IgA, 90 to 330 mg/dl) formed rosettes. Fc alpha R was induced on more than 15% of the cells from these patients, however, when cultured with IgA in the presence of a conditioned medium obtained from mixed lymphocyte culture from two normal donors. The results suggested that the abnormality in the patients' PBL might be in the induction mechanism rather than in the number of precursor cells that could express Fc alpha R in the presence of IgA. On the other hand, Fc alpha R was induced on 10.4% +/- 1.5 of PBL from the patients with IgA nephropathy (serum IgA, 382 +/- 11 mg/dl) when they were incubated with IgA for 1 hr at 37 degrees C. Because Fc alpha R on normal PBL was not induced by 1 hr of incubation with IgA, it appeared that the receptor was already expressed in vivo on the cells of these patients.

Animals↗

[Evaluation of autopsy cases in pancreatic cancer].

The incidence of pancreatic cancer has increased recently. The average age of pancreatic cancer patients in our series was 60.6 years; the male to female ratio was 2.3:1. As for tumor location, the head (45.5%) was the most frequent followed by the body (32.7%) and the tail (20%). Pancreatic cancers were histologically classified as duct carcinoma (88.2%), acinar cell carcinoma (0.9%), islet cell carcinoma (1.8%) and undifferentiated carcinoma (9.1%). In 17 cases (15.5%), diabetes mellitus was an accompanying complication.

Adult↗

[Role of the spleen in tumor immunity--with special reference to anti-tumor immunity induced by solubilized and purified tumor-associated antigen].

The effects of post-operative immunization with purified tumor-associated antigen on the development of lung metastases following resection of the Lewis lung carcinoma (3LL) were studied. The soluble tumor-associated antigens were isolated from 3LL cells by detergent solubilization and purified by affinity chromatography on peanut agglutinin (PNA)-agarose. The immunization with subcutaneous injections of PNA-binding glycoproteins at early phase following resection of primary footpad tumors of 3LL resulted in a significant decrease in incidence and size of lung metastases. The delayed immunization, however, failed to suppress the development and growth of metastatic lung tumors after surgery. The suppression of postoperative development of lung metastases was found to depend on timing of immunization following local tumor resection. Splenectomy performed simultaneously with amputation of tumor-bearing leg was completely abolished the decrease in the development and growth of postoperative lung metastases in immunized-mice. Evidence of lymphoid system involvement in the control of metastatic tumor growth was also supported by the observation of in vivo tumor neutralization assay utilizing the spleen cells.

Animals↗

Enteric coated polymyxin B in the treatment of hyperammonemia and endotoxemia in liver cirrhosis.

Effects of enteric coated polymyxin B capsules on hyperammonemia and endotoxemia in liver cirrhosis were investigated. Six million units of polymyxin B were orally administered daily to 21 patients with liver cirrhosis and 3 patients with hepatoma cum liver cirrhosis, whose plasma ammonia was higher than normal limit and/or whose plasma endotoxin was positive, for 5-32 days, and serum polymyxin B concentration (in 5 cases), changes of plasma ammonia level (in 19 cases) and plasma endotoxin (in all cases) were observed. Serum polymyxin B concentration was below the detectable limit (0.5 unit/ml) in all cases observed. In the patients with liver cirrhosis, plasma endotoxin and ammonia levels decreased rapidly after polymyxin B treatment, and the decreases in endotoxin levels were kept throughout the treatment. Twelve patients with liver cirrhosis (10 among them were treated with lactulose) were served as controls. All patients who were treated with lactulose alone showed rapid decrease in plasma ammonia, but the decrease in endotoxin in these patients was slower than that in those treated with polymyxin B. From these results, oral administration of polymyxin B is concluded to be useful in the treatment of hyperammonemia and endotoxemia in liver cirrhosis, as a poorly absorbed antibiotic and as an antiendotoxin agent.

Administration, Oral↗

The relationships of free to conjugated normetanephrine in plasma and spinal fluid of hypertensive patients.

The concentrations of free and total normetanephrine (NMN) were determined in the plasma of normotensives and patients with primary hypertension and pheochromocytoma. NMN values were measured in the cerebrospinal fluid (CSF) of patients. Free and conjugated NMN, the latter after acid hydrolysis, were assayed using S-adenosylmethionine in the presence of phenylethanolamine-N-methyltransferase to form labeled metanephrine. The conjugates of NMN were present in plasma as sulfates principally, as they were also liberated with arylsulfatase. Free and conjugated NMN levels were 117 +/- 10 and 1417 +/- 109 ng/liter, respectively in plasma of normotensives. The mean ratio of the content of conjugated to free NMN was 14.9 +/- 1.8 (mean +/- SEM). The contents of free and conjugated NMN were 155 +/- 33 and 1670 +/- 320 ng/liter in primary hypertensives, respectively, and the ratio of conjugated to free NMN was 18.5 +/- 3.3. These values did not differ significantly from those in normotensives. The contents of free and total NMN in the plasma of patients with pheochromocytoma were 50- to 60-fold greater than values in normotensive and primary hypertensives. The mean ratio of conjugated to free NMN in the plasma of patients with pheochromocytoma was similar to those in normotensives and primary hypertensives. The contents of free and conjugated NMN in the CSF of patients with pheochromocytoma exceeded those in primary hypertensives (P less than 0.01 and P less than 0.001). Further, the ratio of conjugated to free NMN in CSF was increased in patients with pheochromocytoma (33.9 +/- 8.1) compared to that primary hypertensives (8.3 +/- 2.3; P less than 0.001). The measurement of NMN in plasma and CSF may help characterize sympathetic nerve tone in patients with primary hypertension to elucidate the pathophysiology of the elevated blood pressure.

Adolescent↗