Generation of monoclonal antibodies to the rabbit interleukin-2 receptor alpha chain (CD25) and its distribution in HTLV-1-transformed rabbit T cells

Rabbits can be infected with human retroviruses such as human T-cell leukemia virus-1 (HTLV-1) and human immunodeficiency virus (HIV), and provide useful animal models to study retroviral diseases such as adult T-cell leukemia and HIV. Previously we have succeeded in generating monoclonal antibodies (mAbs) against rabbit CD4, CD5 and CD11a antigens. To make this animal species more amenable to cellular and molecular studies, we have attempted to extend the panel of mAbs against rabbit CD antigens. Here we report on the generation of three neutralizing mAbs against interleukin-2 receptor alpha chain (IL-2R alpha) (CD25), Kei-alpha 1 (IgG2b), Kei-alpha 2 (IgG2a) and Kei-alpha 3 (IgG1). They specifically recognize the rabbit Mr 55,000 IL-2 binding protein, IL-2R alpha, and completely inhibit both high- and low-affinity IL-2 binding to F648b cells that express IL-2R alpha as well as IL-2R beta. The use of mAb Kei-alpha 1 confirmed that the rabbit IL-2R alpha is not only a low-affinity IL-2R on its own but also an essential component of high-affinity IL-2R as found in other animal species, and that rabbit activated T cells including HTLV-1-transformed cell lines express high levels of the IL-2R alpha. Together with mAbs against various rabbit CD antigens that we reported previously, these neutralizing mAbs to IL-2R alpha will be valuable for studies of human retrovirus infections, such as those induced by HTLV-1 or HIV, in rabbits.     

Oxidation of methane to formaldehyde over Fe/SiO2 and Sn---W mixed oxides

In an attempt to develop new catalysts for the formation of formaldehyde from methane, the promotion effect of Fe on SiO₂ and that of Sn on WO₃ have been studied. The formation of formaldehyde on silica can be appreciably enhanced by the impregnation of Fe, as far as iron loadings are kept below 0.1 atom.% (Fe/Si × 100). In the case of Sn---W---Ox catalysts, both the addition of Sn to WO₃, and that of W on SnO₂ were effective to the selective formaldehyde formation. Absorption spectra (UV-Vis) and ESR measurements revealed that tetrahedrally coordinated Fe³⁺ in the silica network plays an important role in the formation of formaldehyde. A thin surface layer consisting of W and Sn oxides can account for the selective formaldehyde formation on the Sn---W---Ox catalysts.     

Minor role of fractographic features in basic fatigue crack propagation properties

The fatigue crack propagation rate and the fatigue threshold were investigated for transverse-butt-welded joints of the austenitic stainless steel SUS304. Specimens were of the center-cracked type. In three sets of tests the fatigue crack passed through the weld metal, and in the other two sets, through the base metal. The fatigue crack propagation properties coincided with each other at different stress ratios for the weld metal, and at the higher stress ratios for the base metal. The propagation properties improved at the stress ratio of zero for the base metal. The absence of a stress ratio effect means that the coincided properties are basic ones in which fatigue crack closure does not occur. However, fractographic appearance and surface roughness were quite different between weld metal and base metal. The coincidence of fatigue crack propagation properties in spite of the difference in fractographic features reveals that the fractographic appearance and surface roughness only have a minor effect on basic fatigue crack propagation behaviour.     

A monocyte chemotactic factor, S19 ribosomal protein dimer, in phagocytic clearance of apoptotic cells

HL-60 cells derived from a human promyelocytic leukemia underwent apoptosis by heat treatment. When the heat-treated HL-60 cells were injected into guinea pig skin, monocyte/macrophage infiltration was observed 24 or 36 hours later, and the apoptotic cells were phagocytically cleared by 48 hours after their injection. The infiltration and clearance patterns were quite different from those observed in injection of necrotic or boil-fixed HL-60 cells. The apoptotic cells released a monocyte chemotactic factor in vitro 24 hours after the heat treatment. The chemotactic factor generated was identified as the cross-linked homodimer of S19 ribosomal protein by its immunologic and physicochemical properties. A serine protease that inactivates the monocyte chemotactic factor was also released from the apoptotic cells 30 hours after the heat treatment. A super infusion of this protease into the skin where the apoptotic cells had been injected diminished the number of infiltrated monocytes. The present results indicate an important role of the S19 ribosomal protein dimer in the phagocytic clearance of apoptotic cells.     

Identification of osteopontin in human dental calculus matrix

Osteopontin is a prominent non-collagenous component of bone matrix, although it is expressed in several other tissues. Recently, osteopontin was reported to be involved in urinary stone formation and atherosclerotic lesions of the aorta, suggesting that it may be a key protein associated with these types of pathological mineralization. In this study, whether or not human dental calculus contains osteopontin was investigated by immunoblotting and immunohistochemical analyses. After extraction of calculus proteins with EDTA and separation of the proteins by electrophoresis, immunoblotting analysis revealed the presence of osteopontin. Two forms of osteopontin appeared at 61 and 68 kDa on 10% polyacrylamide gel and the proteins were digested with thrombin, a highly specific protease. Moreover, immunohistochemical analysis revealed that osteopontin was localized in dental calculus adherent to tooth roots. These findings indicate that osteopontin is, in fact, present in human dental calculus and may be involved in calculus formation as the stone matrix.     

RANTES-induced T cell activation correlates with CD3 expression

The chemokine RANTES induces a unique biphasic cytoplasmic Ca2+ signal in T cells. The first phase of this signal, similar to that of other chemokines, is G-protein mediated and chemotaxis associated. The second phase of this signal, unique to RANTES and evident at concentrations greater than 100 nM, is tyrosine kinase linked and results in a spectrum of responses similar to those seen with antigenic stimulation of T cells. We show here that certain jurkat T cells responded to RANTES solely through this latter pathway. A direct correlation between the RANTES-induced second phase response and CD3 expression was demonstrated in these cells. Sorting the Jurkat cells into CD3(high) and CD3(low) populations revealed that only the CD3(high) cells were responsive to RANTES. Furthermore, stimulation of these Jurkat cells with anti-CD3 mAb significantly depresses their subsequent response to RANTES. While a RANTES-specific chemokine receptor is expressed at a low level on these Jurkat cells, the RANTES-induced activation is dependent on the presence of the TCR. Thus, stimulation through TCR may partially account for RANTES' unique pattern of signaling in T cells.     

Estimating the 3D skeletons and transverse areas of coronary arteries from biplane angiograms

A method for estimating the three-dimensional (3D) skeletons and transverse areas of the lumens of coronary arteries from digital X-ray angiograms is described. The method is based on the use of a 3D generalized cylinder (GC) consisting of a series of 3D elliptical disks transverse to and centered on a 3D skeleton (medial axis) of the coronary arteries. The estimates of the transverse areas are based on a nonlinear least-squares-error estimation technique described by D.W. Marquardt (1963). This method exploits densitometric profiles, boundary estimates, and the orientation of the arterial skeleton in 3-space and includes an automatic artery tracking procedure. It applies an adaptive window to the densitometric profile data that are used in the parameter estimation. Preliminary experimental tests of the procedure on angiograms of in vivo human coronaries and on synthetic images yield encouraging results.     

Effect of oxidized oil on lipogenic enzymes

Male Wistar rats were fed for 4 wk on diets containing 2% oxidized corn oil. Liver tissue was then studied to determine the effect of feeding peroxidized oil on lipogenic enzymes. Although substances which reacted with thiobarbituric acid increased in liver microsomes and mitochondria with increasing peroxide values of the dietary corn oil fed, the activities of glucose-6-phosphate dehydrogenase, malic enzyme and acetyl-CoA carboxylase in liver were unchanged. However, when rats were fed for 2 wk on diets containing 10% fat, of which 0.5, 5 or 10% was unoxidized corn oil and the remainder was hydrogenated beef tallow filler, the lipogenic enzyme activities and also the liver triglyceride levels were observed to decrease with increasing amounts of dietary corn oil. Therefore, although a synthetic diet containing corn oil was easy to oxidize spontaneously, the reductions of lipogenic enzymes in rats fed the diet would not have been caused by lipid peroxides but by unsaturated fatty acids themselves.