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Biomedical subjects

S Sakamaki

Publications and source records attributed to S Sakamaki.

87 records · Page 5Linked to original sources
PubMed1992

Effect of Krestin (PSK) on the induction of IL-2 activated killer cells.

The effects of Krestin (PSK) on the generation of lymphokine-activated killer (LAK) cells were examined in tumor-bearing mice. BALB/c mice were inoculated subcutaneously with methylcholanthrene-induced fibrosarcoma (Meth A) cells, and PSK was administered intraperitoneally every other day. The reduced LAK activity in tumor-bearing mice was restored by the administration of PSK. Since involvement of the humoral immunosuppressive factor in the impairment of LAK activity has been suggested, the effect of PSK on the impaired LAK activity in the presence of an immunosuppressive factor isolated from the ascites of X5563 (plasmacytoma)-inoculated mice was examined. The activity reduced by the immunosuppressive factor in an in vitro induction of LAK was restored by incubation with PSK. The antimetastatic effect of IL-2 was also augmented by its combined use with PSK. The data provide a rational basis for using PSK in combination with recombinant IL-2 in cancer immunotherapy.

Adjuvants, Immunologic↗
PubMed1991

Human tumor-infiltrating lymphocytes transfected with tumor necrosis factor gene could augment cytotoxicity to autologous tumor cells.

Human tumor-infiltrating lymphocytes (TILs) derived from pleural or ascitic fluid were incubated with recombinant interleukin 2 and transfected with human tumor necrosis factor (TNF) alpha gene by the lipofection procedure. The resulting TILs secreted significant amounts of TNF in the culture supernatant and exhibited cytotoxicity against established cell lines, such as K562 and Daudi, and autologous tumor cells. The TNF gene-transfected TILs exhibited an augmented killing of autologous tumor cells.

Ascites↗
PubMed1991

Cytomegalovirus DNA in the sera of patients with cytomegalovirus pneumonia.

We attempted to detect cytomegalovirus DNA (CMV-DNA) in the sera of four leukaemia patients who underwent an allogeneic bone marrow transplant (BMT), in six leukaemia patients who suffered from pneumonia and in 16 healthy subjects, using the polymerase chain reaction (PCR). Three of the four BMT patients subsequently developed CMV pneumonia. In two cases, CMV-DNA was detected in the sera at about the time the pneumonia occurred, and the amount of DNA increased with disease progression. The serum of the third patient became positive for CMV-DNA before he developed pneumonia. The fourth patient did not develop CMV pneumonia, but his urine became persistently positive for CMV-DNA soon after the BMT, whereas the serum was negative. A relationship was found between the occurrence of pneumonia and the serum level of CMV-DNA. CMV-DNA was also detected in three of six pneumonia patients whose anti-CMV IgM antibodies were elevated in the circulation. Sera from the 16 normal subjects were negative for CMV-DNA, regardless of their being seropositive or seronegative for CMV. While it had been previously thought that CMV did not exist in serum, we detected CMV-DNA in serum by PCR in the active disease stage. Our results suggest that PCR would be useful for the early diagnosis of CMV pneumonia and in monitoring its course.

Adolescent↗
PubMed1990

A shared and unique epitope(s) on human colon, skin, and biliary epithelium detected by a monoclonal antibody.

An immune-complex-mediated inflammation has been suggested in the pathogenesis of skin and biliary complications in ulcerative colitis, although specific antigen-antibody complexes have not been found. We recently identified an Mr 40,000 colonic protein that reacts with tissue-bound immunoglobulin G eluted from colon specimens of patients with ulcerative colitis and not with immunoglobulin G eluted from colon specimens of patients with other diseases (J Clin Invest 1985;7:311). Murine monoclonal antibodies to the purified Mr 40,000 colonic protein are developed. The presence of the Mr 40,000 protein was examined with a monoclonal antibody (7E12H12, immunoglobulin M isotype) by an immunoperoxidase assay against various human epithelial tissues (74 specimens) including colon, ileum, jejunum, duodenum, stomach, esophagus, pancreaas, liver, respiratory, urinary, and genital tracts. In addition, gall bladder, bile duct, hepatic ducts, skin, synovial membrane, and eye tissue were also examined for any cross-reactivity with 7E12H12 and another anti-Mr 40,000 monoclonal antibody (7E6A5, immunoglobulin G1 isotype). 7E12H12 reacted only with colonic-epithelial cells mainly along the basolateral domains of plasma membrane and apical areas. The reactivity was present in both adult and fetal colon including the appendix. Among all the noncolonic tissue specimens, 7E12H12 reacted only with the gall bladder, the bile duct, the hepatic ducts, and the skin. All other epithelial tissues from 15 different organs, including the small intestine, synovium, and eye tissue, did not react. In the skin and biliary tract the immunoreactivity was exclusively present in the epidermal epithelial cells and mucosal epithelium respectively. 7E6A5 stained colonic mucosal cells but did not react with skin, biliary epithelium, synoyium and eye tissue. These results indicate the presence of a unique epitope or epitopes on Mr 40,000 colonic epithelial protein that is shared by the skin and biliary tract epithelial cells. Future studies of this shared epitope or epitopes and its immune recognition may provide further understanding in the pathogenesis of extraintestinal complications involving these organs in patients with ulcerative colitis.

Antibodies, Monoclonal↗
PubMed1989

Alteration in autonomic function and cardiovascular regulation in amyotrophic lateral sclerosis.

Autonomic function mediating cardiovascular regulation was evaluated in amyotrophic lateral sclerosis (ALS) in comparison with Shy-Drager syndrome. The subjects were 14 normal controls and 9 patients each with ALS and Shy-Drager syndrome. To evaluate the autonomic function in detail, a new series of quantitative autonomic function tests was conducted, in conjunction with conventional tests. In patients with ALS, the data indicated subclinical sympathetic hyperfunction and parasympathetic (vagal) hypofunction, probably resulting in cardiovascular dysfunction.

Aged↗
PubMed1988

Somatosensory evoked potentials in chronic alcoholics with spasticity.

Somatosensory evoked potentials to median and bilateral tibial nerve stimulation were investigated in eight chronic alcoholics with spasticity, 12 patients with alcoholic polyneuropathy, and 11 normal subjects. Central conduction velocities from the third lumbar vertebra to the fifth cervical vertebra and from the 12th thoracic vertebra to the fifth cervical vertebra were significantly lower in the chronic alcoholics with spasticity than in the alcoholic polyneuropathy group and in the healthy nonalcoholic group. The result indicates that chronic alcoholics with spasticity have conduction disturbance in the posterior column and/or the medial lemniscus, which is considered to be due to alcoholic myelopathy and/or a brainstem lesion.

Alcoholism↗
PubMed1988

[Induction of LAK cells by high density dialyzing culture device and its cytotoxicity].

Lymphokine activated killer (LAK) cells are generated by culture of lymphocytes with interleukin 2 (IL-2) in short term culture (3 to 5 days) and are used for adoptive immunotherapy for advanced cancer patients. The culture condition hitherto reported are essentially based on the rotating culture system, in which the maximum cell density was at 2 X 10(6) cell/ml and the cell recovery was usually less than 100%. The inability to induce LAK cells efficiently in vitro made the culturing of cells for therapy rather difficult and costly work because the mean infusion dose of LAK cells of one patient requires more than 1 X 10(10)/ml. We have therefore attempted to culture lymphocytes in 10 times higher concentration comparing with conventional methods. By using a new dialyzing culture system under continuous regulation of the amount of infused IL-2, nutrition medium, and pO2 and pCO2, we could culture cells at 2 X 10(7)/ml for more than 21 days and the resulted LAK cells showed a 100 times increase of activity on a per cell basis. By limiting dilution procedure, these killer cells mostly express T cell markers such as CD3 and CD8 but dose not express CD16.

Cell Division↗
PubMed1987

The production and characterization of monoclonal antibodies to a human colonic antigen associated with ulcerative colitis: cellular localization of the antigen by using the monoclonal antibody.

We detected in human colon extracts a 40 kDa protein(s) that specifically reacts with tissue-bound IgG obtained from the colon of patients with ulcerative colitis or CCA-IgG. Using the hybridoma technology, we developed monoclonal antibodies to this 40 kDa protein. The specific immunoreactivity of one of the monoclonal antibodies (7E12H12, IgM isotype) against the 40 kDa protein was demonstrated both by ELISA and by immunotransblot. Competitive binding experiments showed that CCA-IgG inhibits the binding of 7E12H12 to the 40 kDa protein, suggesting the recognition of common epitope(s) on the 40 kDa protein by the monoclonal antibody and CCA-IgG. 7E12H12 was used to determine cellular localization of the 40 kDa protein. Biopsy tissue specimens from colon, esophagus, stomach, duodenum, jejunum, ileum, liver, pancreas, lungs, kidneys, salivary, and mammary glands were obtained. Tissue specimens were fixed in 4% paraformaldehyde or in 10% formalin. Sections were sequentially incubated with the hybridoma supernatant, biotinylated anti-mouse IgM, avidin-biotin-peroxidase complex, and 3,3'-diaminobenzidine. An unrelated hybridoma supernatant was used as control. The monoclonal antibody exclusively recognized colonic epithelial cells both in the crypt and on the luminal surface. Immunoreactivity was present on the plasma membrane chiefly along the basolateral areas of the cells. Plasma membrane localization of the 40 kDa protein was confirmed by immunoelectron microscopy. All colonic mucosal biopsy specimens from both adult and fetal colon reacted with the monoclonal antibody. None of the biopsy specimens from stomach, duodenum, jejunum, ileum, liver, pancreas, or non-gastrointestinal tissue reacted with the antibody, confirming the organ specificity of the 40 kDa protein. The interaction between this colonic epithelial membrane protein and the CCA-IgG may play an important role in the pathogenesis of ulcerative colitis.

Antibodies, Monoclonal↗
PubMed1987

Development of a monoclonal antibody specifically reactive to gastrointestinal goblet cells.

A mouse monoclonal antibody (7E6A5) of IgG isotype, reacting specifically with mucin-producing goblet cells of the human gastrointestinal tract, has been developed. 7E6A5 reacts by an ELISA with colonic protein eluted from a DEAE column. A screening by immunoperoxidase assay of 76 specimens from 19 different human tissues showed that the immunoreactivity of 7E6A5 was confined exclusively in the globules of goblet cells in the colon, the appendix, and the small intestine. Nongoblet small and large intestinal epithelial cells did not react. Immunoelectron microscopy demonstrated the reactivity with mucin droplets in a homogeneous granular pattern inside the globules of goblet cells. Mucus-secreting cells from remaining parts of the gastrointestinal tract and other mucus-secreting organs such as respiratory, genitourinary tracts, salivary and mammary glands did not show any reactivity to 7E6A5. These findings indicate that the antigen recognized by 7E6A5 is shared by the goblet cells of both the small and large intestines and is unique to them. The monoclonal antibody may be useful in the study of function of mucus-secreting goblet cells and may represent an important tool in the evaluation of diseases such as ulcerative colitis, colon cancer, and intestinal metaplasia in gastric mucosa that are associated with quantitative changes in goblet cell numbers or with qualitative differences in mucin secretion.

Antibodies, Monoclonal↗
PubMedDate not supplied

Immunochemotherapy in B-16-melanoma-cell-transplanted mice with combinations of interleukin-2, cyclophosphamide, and PSK.

The effect of combination immunochemotherapy using interleukin-2 (IL-2), PSK and cyclophosphamide (CY) was evaluated in a pulmonary metastasis model in BDF1 mice. B-16 melanoma cells were inoculated into a hind limb. On day 3 after inoculation, 20 mg/kg of CY was administered intraperitoneally, and IL-2 (3.75 x 10(4) BRM units/head) was injected into the tail vein on days 7, 8 and 9. PSK (1,000 mg/kg) was administered orally every day from day 1 to day 10 using a stomach tube. This treatment cycle was repeated three times. Using this combination therapy, the cytotoxicity of lymphokine-activated killer cells and tumor-infiltrating lymphocytes was enhanced. Pulmonary metastasis was remarkably suppressed and a prolongation of survival was obtained compared with the nontreated group and an IL-2+CY group. The effect was augmented by repeating the therapy protocol. By analyzing the killer activity and surface markers of tumor-infiltrating lymphocytes, it was recognized that increased numbers of Lyt-2-positive T cells with augmented cytotoxicity were obtained. This treatment modality should have clinical significance.

Animals↗
PubMedDate not supplied

Ulcerative colitis: specific antibodies against a colonic epithelial Mr 40,000 protein.

A colon tissue-bound disease specific IgG (CCA-IgG) antibody can be eluted from the colonic tissue of patients with ulcerative colitis (UC). CCA-IgG recognizes an Mr 40,000 protein present in both normal and diseased colon. Using an anti-Mr 40,000 murine monoclonal antibody this protein has been localized exclusively in colonic epithelial cells mainly along the baso-lateral domains of plasma membrane. Non-colonic epithelial tissues from 12 different organs including small intestine did not react with the anti-Mr 40,000 monoclonal antibody. The Mr 40,000 protein is present in the RPMI-4788 colon cancer cells but not in HeLa. The antibody dependent cell-mediated cytolysis by UC sera against RPMI-4788 cells was inhibited by anti-Mr 40,000 polyclonal antibody. These findings suggest that CCA-IgG and the Mr 40,000 colonic epithelial protein are involved in an autoimmune reaction and may be important in the pathogenesis of UC.

Antibodies, Monoclonal↗