Production of heavy-chain class-switch variants of human monoclonal antibody by recombinant DNA technology

We have previously established a human-mouse heterohybridoma (H6-3C4), which produced a human sperm-immobilizing antibody (mu, lambda of human type). The human rearranged immunoglobulin mu-chain and lambda-chain genes were cloned from the hybridoma H6-3C4. The cloned V region of the heavy chain (VH) gene was ligated to human immunoglobulin gamma 1-heavy chain constant region (C gamma 1) genes. This resulted in the heavy-chain class-switch from mu-chain to gamma 1-chain of H6-3C4 antibody. The class-switched heavy-chain gene as well as the cloned lambda-chain gene were introduced into mouse myeloma cell line X63Ag8.653 by protoplast fusion and electroporation. The stable transformants produced the human IgG monoclonal antibody, which fully retained specificity to human sperm cells and sperm-immobilizing activity.     

Acute pulmonary toxicity of inhaledβ-1,3-glucan and endotoxin

The number of inflammatory cells was studied in lung walls and airways after inhalation of endotoxin or -1,3-glucan. In the water unsoluble form, -1,3-glucan caused a delayed response in terms of a decrease in macrophages and lymphocytes in the lung wall, 1 to 7 days after exposure but no invasion of neutrophils into the airways. When solubilized in 0.02 N NaOH, the cell response was the same as that observed after exposure to endotoxin.     

Comparative anatomical studies of thyroid and thymic arteries: I. Rat (Rattus norvegicus albinus)

The thyroid and thymic arteries were investigated in 50 male and 50 female rats. In more than 70% of the animals, on both sides the cranial thyroid artery forms a common trunk with the ascending pharyngeal artery. The caudal thyroid artery arises not from the deep cervical but from the pericardiacophrenic artery. It may be replaced, however, by a branch of some other artery, such as the brachiocephalic, subclavian, vertebral, or ascending cervical, suggesting a shift of its origin from the internal thoracic artery to the thyrocervical trunk as in man. All the thoracic lobes of the thymus are supplied directly by a thymic branch of the internal thoracic artery or indirectly by a branch of the pericardiacophrenic artery. More than half of the specimens have a cervical thymic lobe of variable size, which is supplied by a branch of the cranial thyroid, external carotid, and/or occipital arteries. Some of these thymic arteries, except those from the external carotid and occipital arteries, reach the thoracic lobe. The thoracic lobes lacking a cervical lobe may be supplied by the thymic branch arising only from the cranial thyroid artery. Other anomalous arteries supplying the thoracic lobe are derived from the superficial cervical and/or the right common carotid arteries. These results show that the thymic arteries of rats are basically similar to those of man, although they display a clear difference in their frequency and origin.     

Characteristics of the antihistamine effect of TAK-427, a novel imidazopyridazine derivative

OBJECTIVE AND DESIGN: The characteristics of the antihistamine effect of the new antiallergic compound TAK-427 were investigated. MATERIALS AND METHODS: In vitro binding assay of [(3)H] pyrilamine was performed using recombinant human histamine H(1) receptors (rhH(1)R). In vivo studies were performed in male ICR mice or Hartley guinea pigs. Drugs were administered orally 1 h before examinations. Determinations were made of histamine-induced skin reaction, ex vivo measured radioligand binding to brain and lung H(1) receptors, pentobarbital-induced sleeping time, passive cutaneous anaphylaxis (PCA) reaction, and antigen-induced itch-scratch responses (ISRs). RESULTS: TAK-427 inhibited ligand binding to rhH(1)R with an IC(50) value of 17.3 nmol/l. TAK-427 inhibited histamine-induced skin reactions in guinea pigs and mice with an ID(50) value of 0.884 and 0.450 mg/kg, p.o., respectively; significant inhibition associated with 10 mg/kg of TAK-427 was still observed 24 h after dosing in guinea pigs. TAK-427 showed as high selectivity for peripheral H(1) receptors as terfenadine and epinastine did, which was evaluated by ex vivo measured radioligand binding. Even at 300 mg/kg, TAK-427 did not affect pentobarbital-induced sleeping time in mice. TAK-427 significantly inhibited PCA in mice and guinea pigs, and also inhibited antigen-induced ISRs in guinea pigs. CONCLUSIONS: These results suggest that TAK-427 may have a long-lasting antihistamine activity with minimum sedative side effect and suppress acute phase allergic reactions.     

IgE-mediated14C-serotonin release from passively sensitized rat mast cells: Comparative kinetic study with formaldehyde and ice-cold methods

The kinetics of IgE-mediated release of serotonin from passively sensitized rat mast cellsin vitro was studied by stopping¹⁴C-serotonin release with the application of formaldehyde fixative or ice-cold mast cell medium (MCM). Antigen dose-release curves of¹⁴C-serotonin and/or histamine were comparable when mediator release was terminated with either formaldehyde at a final concentration of 1% or ice-cold MCM 15 min after antigen challenge. However, the kinetic study of immunological mediator release stopped by formaldehyde showed that the addition of antigen resulted in a progressive increase of released¹⁴C-serotonin for 7 min, the release curve being sigmoidal, whereas the application of ice-cold MCM artificially enhanced¹⁴C-serotonin and histamine release in the first 2 min. The results suggest that stopping IgE-mediated release of¹⁴C-serotonin with formaldehyde is a simple, rapid and accurate method of studying the kinetics of mediator release from mast cells.     

Hypercomplex models of insulin and glucose dynamics: Do they predict experimental results?

A hypercomplex circulation and organs model of glucose and insulin dynamics is presented. The model is based on physiological parameters, incorporating blood and plasma flow rates, circulatory paths, intra- and extravascular glucose and insulin spaces, as well as the specific organs and tissues involved both with insulin disappearance and with glucose production or uptake. Its simulations readily lend themselves to physiological interpretation. To explore its validity, the model was assigned parameters typical of a 12 kg dog and was arranged to accept known glucose and insulin infusions from two different experiments on pancreatectomized diabetic animals. It predicted the observed glucose and insulin concentrations as well as uptake rates for both moieties in the individual organs and tissues. This confirmed the ability of the model to predict with consistency the group mean outcomes of both experiments when differing routes (portal or peripheral) of infusion were applied. Excellent agreement was achieved for the studies. The model isolates glucose uptake in the periphery, the liver, the brain, and the gut and allows a direct comparison of glucose disposal along various routes. Thus, the total amount of glucose uptake by peripheral, insulin-dependent tissues is directly calculated to be 30% of an intravenous glucose load, with peripheral infusion, which is higher than that with portal infusion (18%). This investigation was conducted as a project of the Artificial Pancreas Programme and is supported in part by a grant MA5767 from the Medical Research Council of Canada and a negotiated contract No. NO1-AM-9-2201 from The National Institutes of Health, Bethesda, Maryland.     

Duodenal wall cysts may be derived from a ductal component of ectopic pancreatic tissue

AIMS: To clarify the mechanism of origin of duodenal wall cysts in patients with chronic pancreatitis, developing into duodenal stenosis. METHODS AND RESULTS: Specimens from 12 pancreatoduodenectomized patients with chronic pancreatitis and 51 controls were studied histopathologically and immunohistochemically. Variously shaped cystic lesions, averaging about 15 mm in diameter, were found in the duodenum in six of the 12 patients with chronic pancreatitis, but were not observed in the controls. Each case had an average of two cysts, which were located mainly in the muscularis propria of the duodenum with or without submucosal or extraduodenal-peripancreatic extensions. The inner part of the cyst wall consisted of a moderate rim of granulation tissue, with both myofibroblasts and smooth muscle proliferation in the tissue surrounding the cyst and the submucosal layer of the duodenum, occasionally accompanied by an epithelial lining. A ductal structure in the muscularis propria of the duodenum, possibly a ductal component of ectopic pancreatic tissue, was found in five of the six cases. Some of these structures showed cystic changes. Three of the six patients had accompanying duodenal stenosis. CONCLUSIONS: Duodenal wall cysts occur mainly in the muscularis propria of the duodenum associated with both myofibroblasts and smooth muscle proliferation, and may result in duodenal stenosis. These cysts may be derived from a ductal component of ectopic pancreatic tissue.     

Injection-site granulomas resulting from the administration of both leuprorelin acetate and goserelin acetate for the treatment of prostatic cancer.

Although injection-site granulomas caused by leuprorelin acetate have been reported, there have been no reports of granulomas caused by both leuprorelin acetate and goserelin acetate. An 81-year-old man presented with subcutaneous nodules of the abdominal wall and upper arm, where 11.25 mg of leuprorelin acetate had been injected for the treatment of prostate cancer. Because of these nodules, treatment was changed to goserelin acetate. Nevertheless, he presented with another subcutaneous nodule at the injection site. Histological examination showed that these nodules consisted of numerous giant cells that were CD3-positive T lymphocytes and CD68-positive histiocytes associated with granulomatous changes. The granulomas had likely been caused by delayed-type hypersensitivity to leuprorelin acetate injection. The granuloma that formed after goserelin acetate injection might thus have developed owing to the immunogenicity of the previous leuprorelin acetate injections. The patient underwent surgical castration. The present case suggests that both leuprorelin acetate and goserelin acetate can cause injection-site disorders.