Michi Hashimoto

Production of soluble ICAM-1 from human endothelial cells induced by IL-1 beta and TNF-alpha.

The present study was designed to establish the effects of cytokines on soluble ICAM-1 (sICAM-1) production by human endothelial cells (EC) and ICAM-1 expression on these cells and the effects of purified sICAM-1 on lympho-cyte-EC adhesion. Expression of ICAM-1 and production of sICAM-1 were measured by a specific ELISA method. ICAM-1 expression was enhanced by IL-1β, TNF-α, and most effectively by IFN-γ. IL-4, IL-6, M-CSF, or GM-CSF showed no effects on ICAM-1 expression. IL-4 (100 units/ml) or IL-6(100 units/ml) abolished the enhancing effect of IL-1β, while TNF-α (1, 10, 100 units/ml) synergized with IL-1β to promote ICAM-1 expression in EC. In contrast with the transient increase of cell-associated ICAM-1 expression after activiation by IL-1β, which peaked 40 h poststimulation and declined thereafter, sICAM-1 continued to accumulate in culture supernatants even after 48 h poststimulation in IL-1β-stimulated EC. IL-1β treatment resulted in an increase in adhesion. sICAM-1, purified from cell-free supernatants obtained after a 48-h culture of EC in IL-1β by affinity chromatography using monoclonal ICAM-1 antibody coupled to Sepharose beads, significantly inhibited lymphocyte EC adhesion. Preincubation of lymphocytes with conditioned medium of EC cultured with 100 units/ml IL-1β for 48 h, which contained a considerable amount of sICAM-1, resulted in a significant inhibition of lymphocyte adhesion to IL-1β-stimulated EC. These results suggest that there is a cumulative increase in sICAM-1 concentration in the vicinity of cytokine-stimulated EC and that this sICAM-1 modulates ICAM-1-mediated cell to cell interaction.

Effect of cytokine-induced soluble ICAM-1 from human synovial cells on synovial cell-lymphocyte adhesion

The present study was designed to establish (i) the effects of cytokines on soluble ICAM‐1 (sICAM‐1) production by human synovial cells (SC) and ICAM‐1 expression on these cells, and (ii) the effects of sICAM‐1 on lymphocyte‐SC adhesion. sICAM‐1 production was enhanced in parallel with ICAM‐1 expression by IL‐1β, TNF‐α and IFN‐γ. IL‐4 showed no effects on ICAM‐1 expression. In contrast with the transient elevation of cell‐associated ICAM‐1 by IL‐1β, which peaked 36 h after stimulation and declined thereafter, sICAM‐1 continued to accumulate in culture supernatants even after 48 h. Purified sICAM‐1 was obtained from a 48h culture synovial cell supernatant by affinity chromatography using ICAM‐1 monoclonal antibody. The purified sICAM‐1 significantly inhibited adhesion of lymphocytes and monocytes to cytokine‐stimulated synovial cells. These results suggest that sICAM‐1 may modulate chronic synovitis by inhibiting ICAM‐1‐mediated cell‐to‐cell adhesion.

PRODUCTION OF SOLUBLE ICAM-1 BY MONONUCLEAR CELLS FROM PATIENTS WITH RHEUMATOID ARTHRITIS PATIENTS

The present study was designed to quantify the level of the soluble form of ICAM-1 (sICAM-1) produced by mononuclear cells (MNC) of rheumatoid arthritis (RA) patients, and to correlate these levels with the disease activity and with the amounts of cytokines or rheumatoid factors (RF) produced by MNC. Unstimulated synovial fluid (SF) MNC produced higher amounts of sICAM-1 than peripheral blood (PB) MNC in RA patients (P<0.01). sICAM-1 production by PHA-stimulated MNC was higher in RA SF MNC than RA or normal PB MNC (P<0.01). The amounts of SICAM-1 produced correlated with the amounts of soluble IL-2 receptor produced (P<0.02) but not with IL-1B or the Lansbury activity index in RA PB MNC. sICAM-1 correlated with the amounts of soluble CD23 and IL-4 produced by normal PB MNC (P<0.01). The amounts of sICAM-1 correlated with IgG-RF (P <0.02) and IgM-RF (P<0.01) produced by unstimulated MNC obtained from the bone marrow (BM) of RA patients. ICAM-1 expression of T-lymphocyte subsets, B lymphocytes, and monocytes obtained from RA PB and RA BM assayed by twocolor flow cytometry ranged from 0.1 to 6%, which was not appreciably different from that of normal controls. The monocyte fraction of RA PB MNC produced significantly higher amounts of sICAM-1 than lymphocyte fraction. These results suggest that sICAM-1 produced by MNC may be a marker of cell activation in T and B lymphocytes, in contrast to the transient increase of ICAM-1 expression.

Human monoclonal rheumatoid factors augment arthritis in mice by the activation of T cells

In order to investigate the in vivo role of rheumatoid factor (RF), the effects of the administration of human monoclonal (m) IgM‐RF and IgG‐RF on the development of arthritis in mice were examined. The administration of human mRFs into mice immunized with type II collagen (CII) markedly enhanced the clinical score and paw swelling. The severity of arthritic joint disease with a marked infiltration of lymphoid cells, proliferation of synovial membrane, pannus formation and destruction of articular cartilage was significantly enhanced in both groups receiving RF (RF‐enhanced arthritis). Skin ulcers were also observed in some of these RF‐enhanced arthritis mice, whereas no such signs were observed in CII‐immunized mice without mRFs. Both IgM‐RF and IgG‐RF increased CII‐specific IgG antibodies in circulation, and the severity of arthritis correlated with the production of high titres of anti‐CII antibodies. In vivo treatment of RF‐enhanced arthritis mice with an anti‐CD4 MoAb or an anti‐CD8 MoAb inhibited the induction and progression of arthritis in these mice. Administration of RF to severe combined immunodeficient (SCID) mice with arthritis developed by the transfer of spleen cells from CII‐immunized mice, prolonged the arthritis and enhanced the severity. This murine model of RF‐enhanced arthritis may provide a useful tool for analysing the pathogenesis of rheumatoid arthritis in RF‐positive patients.

A case of combined primary biliary cirrhosis, ulcerative colitis and chronic myelocytic leukemia

SummaryA rare case of primary biliary cirrhosis, ulcerative colitis and chronic myelocytic leukemia is described in a 49-year-old Japanese diabetic woman. Primary biliary cirrhosis was diagnosed by characteristic liver histology and positive serum mitochondrial antibody test. Ulcerative colitis was diagnosed by typical findings of barium enema and colonoscopy, negative fecal test for pathogens and compatible rectal histology. Chronic myelocytic leukemia was determined by representative hematologic findings and positive result for Ph1 chromosome. This is the first case with combination of primary biliary cirrhosis, ulcerative colitis and chronic myelocytic leukemia.

Abnormality of platelet membrane glycoprotein GPIIb in a myelodysplastic syndrome with 3q inversion presenting with marked dysmegakaryopoiesis.

Platelet membrane glycoproteins were analyzed in a case of myelodysplastic syndrome with inv(3) (q21q26) presenting with prominent dysmegakaryopoiesis by three different labelling techniques for surface proteins. Markedly decreased level of platelet membrane glycoprotein GPIIb was observed in the patients platelets by terminal sialic acid labelling method, whereas no significant changes in the levels of glycoproteins including GPIIb could be detected either by penultimate galactose labelling or by tyrosine/histidine labelling. These results indicate a decreased sialylation of GPIIb in the patients platelets, implying aberrant process in thrombopoiesis in the disease.

AILD-Like Dysplasia Transformed in AILD-Type T Cell Lymphoma in an HTLV-1 Carrier: Usefulness of Interferon-α

A 61-year-old Japanese woman who was an HTLV-1 carrier had angioimmunoblastic lymphadenopathy with dysproteinemia (AILD)-like dysplasia that transformed into AILD-type lymphoma after 3 years. Complete remission was achieved with interferon-alpha (IFN-alpha) treatment following a previous combination of anticancer drugs. IFN-alpha was also effective for maintenance therapy after this complete, albeit transient, remission. From clinical and pathological studies on AILD patients, it appears that AILD may in some cases be the consequence of an immune dysfunction caused by HTLV-1 infection and that AILD-type lymphoma is a malignant disorder of peripheral T cell clones different from those T cells incorporating the HTLV-1 virus.

Missing Y Chromosome in Ph1-Negative Chronic Myeloid Leukemia with bcr Rearrangement

In a case of Philadelphia chromosome (Ph1)-negative chronic myeloid leukemia (CML) without the Y chromosome, we investigated the differences, at the molecular level, from Ph1-positive CML. Using Southern blot analysis and in situ hybridization studies, we could demonstrate a rearrangement within the breakpoint cluster region (bcr), and the location of a bcr-abl fusion gene on chromosome 22. To our knowledge, this is the first case of Ph1-negative CML with a loss of the Y chromosome in which the molecular abnormalities are shown to be identical with those in Ph1-positive CML.

Amyloidosis of the small intestine secondary to rheumatoid arthritis or juvenile rheumatoid arthritis : report of two cases

We report two cases of gastrointestinal amyloidosis, complicated with juvenile rheumatoid arthritis (JRA) in one and rheumatoid arthritis (RA) in the other. A 21-year-old woman, who had been suffering from JRA for the past 12 years, was transferred to our hospital due to intense pain in the epigastrium and back, diarrhea, high fever, and paralytic ileus. Treatment by corticosteroid, antibiotics protease inhibitor and total parenteral nutrition was not effective. Laparoscopic surgery was performed because of repeated melena followed by an episode of hypovolemic shock. The resected specimen of the ileum showed histologically marked amyloid deposition in the arteriolar walls. A 83-year-old man with RA for 14 years was admitted to our hospital with complaints of abdominal pain, nausea and diarrhea. He underwent an emergency operation for perforation of the ileum. The resected specimen revealed amyloid deposition and non-caseating granulomas. The fragility and impaired blood supply caused by amyloid deposition in the vascular walls may have terminated in the severe intestinal lesion. Further clinicopathological studies along this line are keenly desired in order to establish therapeutic modalities for gastrointestinal amyloidosis.

Additive combination therapy of immunomodulators.

Rheumatoid arthritis (RA) is a refractory disease in most cases. Immunomodulators (IMs) such as gold sodium thiomalate (GST) or D-penicillamine (D-Pc) are sometimes very effetive for RA, and can induce a remission occasionally. The remission rate, however, is only 10-20% at most, and the duration of remission is a few years generally. Moreover, we have no more than 10 IMs now. Therefore, we can not induce a remission in most patients with RA, and can not maintain a remission for long time, resulting no significant difference in the long term outcome from the natural course.In order to increase the remission rate and elongate the remission time with the limited number of IMs, we are trying the additive combination therapy of IMs. That is, to RA patients who are treated with an IM effectively but incompletely, another IM is added aiming at complete remission. We have had many cases of successfully treated with this method. For example, a patient controlled incompletely with 200 mg D-Pc, was induced to complete remission by addition of 100 mg tiopronine.According to our multicenter clinical trials, it appears that the combination of GST and bucillamine is fitting, while the combination of GST or D-Pc with lobenzarit is rather unfitting.