Reduced induction of human ß-defensins is involved in the pathological mechanism of cutaneous adverse effects caused by epidermal growth factor receptor monoclonal antibodies.

Epidermal growth factor receptor inhibitors (EGFRIs) frequently cause cutaneous adverse effects such as papulopustular eruptions. However, the mechanism of the reactions remains unclear. To assess the pathological mechanism of cutaneous adverse reactions caused by EGFRIs, we investigated whether EGFRIs have an influence on the innate immune response of the skin. Levels of human ß-defensins (hBDs), which serve as the first line of defence against infection by pathogenic microorganisms, in the stratum corneum samples of patients treated with EGFR. monoclonal antibodies were measured before and after starting therapy. There were no obvious trends in hBD production in patients without eruptions, whereas a significant decrease in hBD1 and hBD3 production and a nonsignficant decrease in hBD2 production were observed in patients who developed papulopustular eruptions. Our results suggest that a reduction in hBD contributes to the increased incidence of papulopustular eruptions.

Essential contribution of germline-encoded lysine residues in Jgamma1.2 segment to the recognition of nonpeptide antigens by human gammadelta T cells.

Human gammadelta T cells display unique repertoires of Ag specificities largely imposed by selective usages of distinct Vgamma and Vdelta genes. Among them, Vgamma2/Vdelta2(+) T cells predominate in the circulation of healthy adults and respond to various microbial small molecular mass nonpeptide Ags. The present results indicate that the primary Vgamma2/Vdelta2(+) T cells stimulated with the distinct groups of nonpeptide Ags, including monoethyl pyrophosphate, isobutyl amine, and aminobisphosphonate, invariably exhibit Jgamma1.2 in the Vgamma2(+) TCR-gamma chains. Gene transfer studies revealed that most of the randomly cloned Vgamma2/Jgamma1.2(+) TCR-gamma genes bearing diverse Vgamma/Jgamma junctional sequences could confer the responsiveness to all these nonpeptide Ags, while none of the Vgamma2/Jgamma1.1(+) or Vgamma2/Jgamma1.3(+) TCR-gamma genes could do so. Furthermore, mutation of the lysine residues encoded by the Jgamma1.2 gene, which are unique in human Jgamma1.2 and absent in other human or mouse Jgamma segments, completely abrogated the responsiveness to all the nonpeptide Ags without affecting the response to anti-CD3 mAb. These results strongly suggested that the positively charged lysine residues in the TCR-gamma chain CDR3 region encoded by the germline Jgamma1.2 gene play a key role in the recognition of diverse small molecular mass nonpeptide Ags.

Targeting of tumor cells for human gammadelta T cells by nonpeptide antigens.

Human Vgamma2/Vdelta2(+) gammadelta T cells respond to low molecular-mass nonpeptide Ags in a gammadelta TCR-dependent manner. Although requirements of Ag presentation have remained controversial, we have indicated that specific responses of the primary gammadelta T cells to pamidronate were dependent on monocytic adherent cells for Ag presentation. Here, we show that human tumor cells can efficiently present aminobisphosphonate and pyrophosphomonoester compounds to gammadelta T cells, inducing specific proliferation and IFN-gamma production. gammadelta TCR dependency of the response to Ag-pulsed tumor cells was confirmed by using a Jurkat line transfected with a Vgamma2/Vdelta2 gammadelta TCR. Furthermore, gammadelta T cells exhibited markedly enhanced cytotoxicity against the Ag-pulsed tumor cells as compared with untreated tumor cells. Survey of a number of human tumor cell lines of different origins revealed that the majority of them became susceptible for gammadelta T cell-mediated cytotoxicity following the Ag pulsing except for breast cancer lines so far examined, while normal PHA blast cells remained resistant. The results not only imply a unique mode of nonpeptide Ag recognition by human gammadelta T cells but also may provide a novel strategic clue for immunotherapy of human malignancy.

Essential requirement of antigen presentation by monocyte lineage cells for the activation of primary human gamma delta T cells by aminobisphosphonate antigen.

Human gammadelta T cells respond to nonpeptide Ags such as pyrophosphomonoesters and alkylamines in a gammadelta TCR-dependent manner in the absence of other APCS: Recently, aminobisphosphonates such as pamidronate have also been shown to activate human gammadelta T cells. In the present study, we indicate that activation of primary gammadelta T cells by pamidronate strictly depends on the presence of monocyte-lineage cells, unlike that by pyrophosphomonoesters. Thus, although pamidronate induced cell clustering, proliferation, and IFN-gamma production of gammadelta T cells in the culture of PBMC, it failed to induce any of these activities in the culture of purified primary gammadelta T cells. By adding back the purified monocytes, however, both cell clustering and IFN-gamma production of gammadelta T cells by pamidronate could be restored. The pamidronate-pulsed, but not untreated, myelomonocytic line, THP-1, was capable of activating the purified gammadelta T cells to produce IFN-gamma, which was associated with the down-regulation of gammadelta TCR. Furthermore, pamidronate-pulsed THP-1 cells were significantly more susceptible to gammadelta T cell-mediated cytotoxicity than untreated THP-1. Also, TCR-defective Jurkat T cells transfected with gammadelta TCR genes produced a significant level of IL-2 in response to the pamidronate-pulsed THP-1 cells. These results have suggested strongly that human gammadelta T cells are functionally activated via gammadelta TCR by aminobisphosphonate Ag presented on the surface of monocyte lineage cells rather than directly by its free form.

4F2 (CD98) heavy chain is associated covalently with an amino acid transporter and controls intracellular trafficking and membrane topology of 4F2 heterodimer.

4F2, also termed CD98, is an integral membrane protein consisting of a heavy chain linked to a light chain by disulfide bond. We have generated a monoclonal antibody to the mouse 4F2 light chain and cloned the cDNA. It encodes a mouse counterpart of rat L-type amino acid transporter-1, and induces system L amino acid transport in Xenopus oocytes in the presence of 4F2 heavy chain. Transfection studies in mammalian cells have indicated that the 4F2 heavy chain is expressed on the plasma membrane on its own, whereas the 4F2 light chain can be transported to the surface only in the presence of 4F2 heavy chain. 4F2 heavy chain is expressed diffusely on the surface of fibroblastic L cells, whereas it is localized selectively to the cell-cell adhesion sites in L cells expressing cadherins. These results indicate that the 4F2 heavy chain is associated covalently with an amino acid transporter and controls the cell surface expression as well as the membrane topology of the 4F2 heterodimer. Although 4F2 heavy and light chains are expressed coordinately in most tissues, the light chain is barely detected by the antibody in kidney and intestine, despite the presence of heavy chain in a complex form. The results predict the presence of multiple 4F2 light chains.

Effects of alcohol on membrane fluidity of human erythrocyte.

Membrane fluidity in human erythrocytes was measured by a spin label method using an electron spin resonance spectrometer in healthy volunteers after ingestion of alcohol (1.5 ml of whisky/kg body weight). Fluidity in the lipid bilayer closer to the hydrophilic face decreased at 30 min and 90 min, and fluidity in the hydrophobic core decreased at 90 min after ingestion of alcohol. In the same experiment, the level of thiobarbituric acid reactive substances in the serum decreased 30 min after ingestion of alcohol, and the triglyceride level increased and free fatty acid level decreased, and serum superoxide dismutase activity increased 150 min after ingestion. Furthermore, membrane fluidity in human erythrocytes was examined in patients with alcohol dependence syndrome who had not any alcohol for about 26 months. Erythrocyte membrane fluidity of patients with alcohol dependence syndrome was not different from that of healthy controls. However, erythrocyte membrane fluidity of the lipid bilayer closer to the hydrophilic face increased in patients who had concomitant liver cirrhosis compared with those who did not. These results suggest that alcohol affects temporal change of membrane fluidity in human erythrocytes.

Changes in free and total catecholamine concentrations in plasma in patients undergoing coronary artery bypass grafting under high-dose fentanyl anesthesia.

We measured free and total catecholamine in ten patients undergoing coronary artery-bypass grafting under high-dose fentanyl (93.9 +/- 2.2 microg.kg(-1), mean +/- SE) anesthesia. Arterial blood samples were obtained: 1) before induction of anesthesia (control), 2) 1 min after intubation, 3) 1 min after skin incision, 4) 1 min after median sternotomy and, 5) just before termination of cardiopulmonary bypass (CPB). The concentrations of free and total catecholamine were measured by HPLC using fully automated analyzer, 8030-TOHSO. Free and total catecholamine concentrations did not change significantly before CPB. At the termination of CPB, however, the levels in free dopamine, norepinephrine and epinephrine all increased several fold as compared with control. Similarly, total norepinephrine and epinephrine also increased at the end of PCB, while total dopamine did not change. Present results indicated that 1) the measurement of free CAs is more significant than the measurement of total CAs for the assessment of sympathoadrenal responses to surgical stimuli, and that 2) high-doses of fentanyl produce hemodynamic stability by suppressing sympathoadrenal responses elicited by the usual surgical procedures. However, stress triggered by CPB could not be suppressed totally by fentanyl even with high dose.

Prevention of the febrile reaction occurring on reinfusion of cell-free and concentrated autogenous ascites.

The febrile reaction that occurs on reinfusion of ascites was studied. Intravenous reinfusion of ascites was performed 213 times in 63 cases of ascites, which were refractory to treatment with various drugs including diuretics. In order to prevent fever on reinfusion of ascites, a screen filter and a depth filter were used; the results were more favorable with the screen filter. Fibrin was considered to be one of the substances removable by the screen filter. HPLC analysis of the filtered and concentrated ascites, after passage through the screen filter, revealed a fraction corresponding to albumin. Intravenous injection of this fraction into rabbits caused fever. Although the screen filter cannot completely prevent fever on reinfusion of ascites, it appears useful to prevent fever in some patients.

[Alpha-atrial natriuretic peptide, antidiuretic hormone and aldosterone levels in patients undergoing aortocoronary grafting under high dose fentanyl anesthesia].

Plasma concentrations of alpha-atrial natriuretic peptide (alpha-ANP), antidiuretic hormone (ADH) and aldosterone (ALDS) were determined by radioimmunoassay in 9 patients undergoing aortocoronary bypass grafting under high dose fentanyl (94.4 micrograms.kg-1) anesthesia. These three levels in pre-anesthetic period (control values) were within normal ranges suggesting the absence of congestion and dehydration. Although these values changed significantly after sternotomy, they all increased at the termination of cardiopulmonary bypass (CPB) reaching 2.5 fold in alpha-ANP, 27.7 fold in ADH and 2.4 fold in ALDS as compared with control (P less than 0.05). Present results indicate that high dose fentanyl anesthesia cannot suppress ADH and ALDS level during CPB as was previously demonstrated and the observed rise in alpha-ANP level is considered to be inadequate not only for diuresis but also for vascular dilatation. Administration of alpha-ANP to ameliorate circulatory insufficiency after CPB should probably be considered.

[Cefmenoxime concentrations in human blood, bile and gallbladder in administration before surgery].

The pharmacokinetics of cefmenoxime (CMX) were studied in 13 patients with cholecystolithiasis, 6 male and 7 female, ranging from 27 to 69 years of age. All patients received a single intravenous dose of CMX 2 g given in a drip infusion over 30 minutes. The level of CMX in gallbladder tissues fell from 350 +/- 101 micrograms/g (mean concentrations, 2.7 hours after administration) to 68 +/- 13 micrograms/g by 7.2 hours. In bile, the mean concentrations of CMX, 2,595 +/- 624 micrograms/ml were reached at 2.7 hours after administration, and at 7.2 hours the mean concentrations were 103 +/- 60 micrograms/ml. In the study reported here, a single 2 g dose of CMX was administered intravenously, and high concentrations of CMX in gallbladder tissue and bile were reached at 2.7 hours after administration. These results suggested that the administration of CMX at about 2.5 hours before surgery might be most effective for prophylaxis.

Overnight sleep EEG and cerebrospinal fluid monoamines in seizures induced by movement.

Clinical, electrical and biochemical studies in an eight years old boy with movement-induced seizures were reported. The attack was usually triggered by sudden initiation of movement, but rarely occurred without any apparent movement. Repeated jumping provoked attacks constantly, which was recorded cinematographically. No abnormality was found either in ictal or interictal EEGs. Haloperidol aggravated the condition, but 1-DOPA had no effect, while DPH (100 mg/day) controlled attacks perfectly with the serum DPH concentration of just 2.0 ug/dl. In overnight sleep analysis, sleep rhythms and characters of REM sleep were not differed significantly from the standard. After DPH therapy, stabilization of sleep in general was noticed; that is, total sleep time prolonged, number of sleep stages decreased and interrupting awakening disappeared. Probenecid loading test revealed that 5-HIAA was normal, HVA high, and large amount of octopamine was detected in CSF.