Experimental arthritis induced by continuous infusion of IL-8 into rabbit knee joints.

To determine the roles of IL-8 in inflammatory synovitis, examination was made of the results of continuously injecting human recombinant IL-8 into the knee joints of New Zealand while rabbits. Recombinant human IL-8 was infused continuously into the joint cavity at 75 ng/h for 14 days by a polypropylene catheter connected to a mini-osmotic pump implanted in each rabbit. Infiltration of inflammatory cells into joint cavity and histopathological changes in synovial tissue were examined at 7 and 14 days following the start of infusion. The continuous infusion of IL-8 for 14 days led to severe arthritis characterized by apparent erythema and joint pain, the accumulation of leucocytes, infiltration of mononuclear cells in synovial tissue, and marked hypervascularization in the synovial lining layer. IL-8 may be a factor which can contribute to the inflammatory process of chronic arthritis by mediating leucocyte recruitment and hypervascularization in inflamed joints.     

A solid-phase blocking ELISA for detection of antibodies to Nipah virus

A monoclonal antibody (MAb) based solid-phase blocking ELISA was developed for detection of antibodies to Nipah virus. The ELISA was designed to detect remaining antigens on the plate with anti-Nipah MAb conjugate after the reaction with sample serum, and enabled simple procedure, detection of neutralizing antibody to Nipah virus, and application of samples from different animal species. Forty of 200 swine reference sera examined were positive by the ELISA, of which thirty seven were found positive by serum neutralization test. Sera from a total of 131 fruit bats captured in Malaysia were also tested and all found negative by the both tests. It is considered that the solid-phase blocking ELISA can be used as a screening test for Nipah virus infection followed by the serum neutralization test as confirmatory test.     

Major organ function under mechanical support: comparative studies of pulsatile and nonpulsatile circulation

We examined a major organ function during 3 h biventricular assisted circulation after acute myocardial infarction model in the pig. In left ventricular circulation, the outflow cannula was placed in the ascending aorta and an inflow cannula through the mitral valve in the left ventricle. A pump (pulsatile group, Zeon Medical, Inc., Tokyo, Japan and nonpulsatile group, Nikkiso HPM-15, Nikkiso, Inc., Tokyo, Japan) was connected to each cannula. In right ventricular circulation, the outflow cannula was placed in the pulmonary artery and an inflow cannula in the right ventricle. The right ventricular circulation was supported by a nonpulsatile pump (Nikkiso HPM-15). The items measured were the regional blood flows of the cortex and medulla in the kidney, white matter and gray mater in brain, and liver; renal arterial flow; carotid arterial flow; portal vein flow; common hepatic arterial flow; arterial ketone body ratio (AKBR); and lactate/pyrubic acid (L/P). In the pulsatile group, the renal cortical blood flow increased, and the medulla blood flow decreased. On the other hand, in the nonpulsatile group, both regional blood flows decreased. That means that in the pulsatile assisted group intrarenal redistribution improved rather than in the nonpulsatile assisted group. In addition the liver regional blood flow, AKBR, and L/P showed significant differences between the pulsatile and nonpulsatile groups. On the other hand, the white matter and gray matter regional blood flows and carotid arterial flow did not show significant differences between the groups. The results of our study indicated that pulsatile circulation produced superior circulation in the kidney and liver, and microcirculation on the cell level was superior as well in early treatment of acute heart failure.     

Algogenic mediator-induced nociceptive response in diabetic mice

The duration of the somatostatin-, bradykinin- or prostaglandin F2alpha-induced nociceptive response was significantly less in diabetic mice than in non-diabetic mice. Subcutaneous injection of 7-benzylidenenaltrexone (0.1, 0.3 and 1 mg/kg), an antagonist of delta1-opioid receptors, had no significant effect on either somatostatin-, bradykinin- or prostaglandin F2alpha-induced nociceptive responses in non-diabetic mice. 7-Benzylidenenaltrexone (0.1 and 0.3 mg/kg, s.c.) also had no significant effect on somatostatin- or prostaglandin F2alpha-induced nociceptive responses in diabetic mice. However, the bradykinin-induced nociceptive response in diabetic mice was dose-dependently and significantly increased when 7-benzylidenenaltrexone (0.1, 0.3 and 1 mg/kg, s.c.) was injected 10 min before the injection of bradykinin. These results suggest that a spinal delta1-opioid receptor-mediated endogenous antinociceptive system may inhibit the bradykinin-mediated nociceptive responses in the second phase of the formalin-induced nociceptive response in diabetic mice.     

Stimulation of interstitial collagenase in co-cultures of rat hepatocytes and sinusoidal cells

Although the fibrosis observed during chronic liver injury is the result of a complex process, the striking accumulation of collagen in end stage liver disease has provoked interest in the mechanisms that regulate both collagen production and degradation in the diseased liver. The present studies have examined the cell interactions that may be important in the regulation of collagen degradation. Although minimal amounts of interstitial collagenase activity were noted in cultures of normal hepatocytes and sinusoidal cells, the co-cultures of these cells in the presence of lipopolysaccharide showed a substantial increase in collagenase activity. When the hepatocytes were obtained from rats that had been treated with carbon tetrachloride in vivo, the enhanced activity seen in the co-cultures did not require the addition of lipopolysaccharide. Further characterization of this interaction suggested that the increase in collagenolytic activity was partially due to the elaboration of soluble factors by the hepatocyte, which stimulated collagenase production by the sinusoidal cell population. Elaboration of collagenase activity by the sinusoidal cells was inhibited by cycloheximide, suggesting that protein synthesis was required. The proteolytic activity was abrogated by inhibitors of metalloproteinases but not by serine or thiol proteinase inhibitors. The degradation products of type I collagen were typical of the expected products seen with vertebrate collagenases. Thus, it appears that the increased collagenolytic activity detected in this co-culture system is attributable to the production of interstitial collagenase by the sinusoidal cell population. Such cell-cell interactions may play an important role in the maintenance of normal connective tissue structure of the liver during disease processes.     

The role of Ia positive T cells in patients with rheumatoid arthritis

Summary In peripheral blood and synovial fluid of patients with rheumatoid arthritis (RA), increased levels of Ia antigen-positive (Ia+) T cells have been demonstrated. Therefore, we examined these Ia+ T cells in vitro to identify their role in the production of rheumatoid factor (RF) and to study the immunologic abnormalities of RA. When Ia+ T cells from peripheral blood of RA patients were added to pokeweed mitogen (PWM)-non-stimulated autologous B cells, the amount of IgM-RF production was 25.8±6.4 (mean±SE) (p<0.001) as compared to 16.0±4.6 ng/ml (mean±SE) in the presence of Ia^– T cells. When Ia^– OKT4+ cell fractions, obtained by excluding Ia+ T cells from OKT4+ cells, were added to B cells, the increase in IgM-RF production was markedly lower than that obtained with the OKT4+ cell fraction. These results indicate that the helper T cells which induce the production of IgM-RF may derive from the Ia+ OKT4+ cell fraction. B cells from rheumatoid synovial fluid produced IgM-RF levels as high as 102.7±19.2 ng/ml (mean±SE) even without stimulation. When T cells from autologous synovial fluid were added, IgM-RF production was not increased. These data suggest that B cells from RA synovial fluid had already been activated. When synovial fluid T cells were added to B cells from autologous peripheral blood, larger amounts of IgM-RF were produced as compared to experiments in which T cells from peripheral blood were added, suggesting that T cells from synovial fluid induce an enhanced IgM-RF production by B cells. The presence or absence of Ia antigen on the surface of synovial fluid T cells did not affect the level of IgM-RF production. Our results indicate that Ia+ T cells from the peripheral blood of RA patients induce the production of IgM-RF by autologous B cells.     

Adhesion molecule expression on peripheral blood mononuclear cells in rheumatoid arthritis: Positive correlation between the proportion of L-selectin and disease activity

Summary The expression of a series of adhesion molecules (the integrin family, the immunoglobulin superfamily, CD44 and the selection family) on peripheral blood mononuclear cells (PBMC) from RA patients was investigated by flow-cytometry. L-Selectin (CD62L) in the selectin family was more significantly expressed on PBMC from RA patients as compared to those from normal controls. Further, RA patients exhibited a slight but significant increase of VLA-2 (CDw49b), p150,95 (CD11c), and VNR- (CD61) in the integrin family. The enhanced expression of L-selectin was positively correlated with disease activity.An increase in the proportion of L-selectin⁺CD4⁺ cells in PBMC from patients with RA and its clinical significance is reported.     

Suppressed serum erythropoietin response to anemia and the efficacy of recombinant erythropoietin in the anemia of rheumatoid arthritis

Serum erythropoietin (EPO) was measured by radioimmunoassay in 67 patients with rheumatoid arthritis (RA). Twenty of these patients judged to have iron deficiency anemia, based on reduced serum ferritin levels, had higher serum EPO levels than did the 24 other anemic patients with normal or elevated serum ferritin levels. A significant negative correlation between serum EPO and hemoglobin concentrations was noted in the former group, but not in the latter. Human recombinant erythropoietin (r-EPO) was administered to 6 anemic patients with RA, resulting in improvement of anemia in 4 patients, 2 of whom showed no change in RA activity. These findings suggest a suppressed serum EPO response ot anemia and the effectiveness of r-EPO in treating anemia associated with RA.