Biomedical subjects

K Nomoto

Publications and source records attributed to K Nomoto.

At least 253 records · Page 14Linked to original sources

PubMed1995

The antitumor effect of tumor-draining lymph node cells activated by both anti-CD3 monoclonal antibody and activated B cells as costimulatory-signal-providing cells.

To establish an efficient cell-culture system for adoptive immunotherapy, we attempted to use lipopolysaccharide(LPS)-activated B cells (LPS blasts) as costimulatory-signal-providing cells in the in vitro induction of antitumor effector cells. Both normal and tumor-draining lymph node cells were efficiently activated by both anti-CD3 monoclonal antibody (mAb) and LPS blasts, and subsequently expanded by a low dose of interleukin-2 (IL-2; anti-CD3 mAb and LPS blasts/IL-2). The expanded cells were predominantly CD8+ T cells and showed a low level of tumor-specific cytotoxic T lymphocyte (CTL) activity. The adoptive transfer of B16-melanoma-draining lymph node cells expanded by anti-CD3 mAb and LPS blasts/IL-2 showed significant antitumor effect against the established metastases of B16 in combination with intraperitoneal injections of IL-2. This treatment cured all B16-bearing mice. In addition, these mice also showed tumor-specific protective immunity against B16 at the rechallenge. Considering that activated B cells express several kinds of costimulatory molecules, these findings thus indicate an efficacy of costimulation that is derived from activated B cells for the in vitro induction of tumor-specific CTL, in co-operation with anti-CD3 mAb. The culture system presented here may thus be therapeutically useful, providing potent effectors for adoptive immunotherapy against various types of cancer.

Animals↗

PubMed1995

Specific antitumor activity of tumor-infiltrating lymphocytes expanded first in a culture with both anti-CD3 monoclonal antibody and activated B cells and then in a culture with interleukin-2.

In order to expand tumor-infiltrating lymphocytes (TIL) efficiently and in order to use them for immunotherapy, we utilized lipopolysaccharide-activated B cells (LPS blasts) as costimulatory-signal-providing cells in an in vitro culture system. TIL, prepared from subcutaneously inoculated B16 melanoma, failed to expand when cultured with anti-CD3 monoclonal antibody (mAb) alone followed by a low dose of interleukin(IL)-2. In contrast, such TIL did expand efficiently in culture with both anti-CD3 mAb and LPS blasts followed by culture with IL-2. These findings suggest that the presence of LPS blasts in the initial culture was essential for the cell expansion. The expansion of TIL was partially blocked by the addition of CTLA4 Ig, which is an inhibitor of costimulatory molecules such as CD80 and CD86, and was almost blocked by the addition of anti-(Fc receptor gamma II)mAb. These findings thus indicate that such molecules, in conjunction with the receptor on the LPS blasts, participate in the efficient expansion of TIL. The B16-derived TIL, which expanded in our culture system, were predominantly CD8+ T cells and showed a higher level of cytolytic activity against B16 melanoma than either lymphokine-activated killer cells or TIL cultured with a high dose of IL-2. In addition, the in vitro expanded B16-derived TIL produced interferon gamma, but not IL-4, in response to B16 melanoma. What is more important, the adoptive transfer of such TIL had a significant antitumor effect against pulmonary metastasis in B16 melanoma, even without the concurrent administration of IL-2. Collectively, our results thus indicate the therapeutic efficacy of the protocol presented here for antitumor immunotherapy with TIL.

Abatacept↗

PubMed1995

Cardiac dystrophin abnormalities in Becker muscular dystrophy assessed by endomyocardial biopsy.

Duchenne and Becker muscular dystrophy (DMD/BMD) are allelic variants caused by mutations in gene-encoding dystrophin. Abnormal expression of dystrophin in skeletal muscle has been shown to correlate with severity of disease. However, in BMD the severity of skeletal and cardiac involvement are not well correlated. We studied the immunostaining pattern of cardiac dystrophin in endomyocardial biopsy specimens from 83 patients with heart disease. Immunohistochemical assessment of dystrophin in four patients with BMD and cardiomyopathy showed a variable distributions of myocytes with continuous, discontinuous, or absent membrane immunostaining patterns. These patterns were obviously different from patterns of other heart diseases. We conclude that the discontinuous immunostaining pattern of cardiac dystrophin is characteristic of BMD and that an absent pattern may be associated with more severe cardiac dysfunction. Because genetic analysis cannot determine the correct diagnosis in 35% of DMD/BMD cases, we recommend routine examination of immunostaining patterns of dystrophin in endomyocardial biopsy specimens in patients with cardiomyopathy suspected to be the result of BMD.

Adult↗

PubMed1995

In vitro generation of IFN-gamma-producing Listeria-specific T cells is dependent on IFN-gamma production by non-NK cells.

In vitro 5-day cultures of naive spleen cells with viable Listeria monocytogenes (VLM), but not heat-killed L. monocytogenes, induced CD4+ T cells that produced IFN-gamma upon secondary antigen stimulation. The VLM-induced Listeria-specific T cells produced IFN-gamma but lacked expression of IL-2 and IL-4. To study the role of IFN-gamma in the induction of the IFN-gamma-producing T cells, we added anti-IFN-gamma mAb to the primary culture and analyzed IFN-gamma production upon secondary antigen stimulation. Addition of anti-IFN-gamma mAb to the culture suppressed generation of IFN-gamma-producing CD4+ T cells, suggesting that IFN-gamma is important in the induction of IFN-gamma-producing CD4+ T cells. Furthermore, our results showed that depletion of NK cells from spleen cells by anti-asialo GM1 antibody plus complement before culture enhanced induction of IFN-gamma-producing CD4+ T cells. Although NK cells are known to produce IFN-gamma, the results indicate that NK cell-derived IFN-gamma may not be important in induction of the Listeria-specific IFN-gamma-producing CD4+ T cells in the culture system. In addition, we demonstrated that IFN-gamma expression was high in CD4+ T cells from cultures of spleen cells with VLM at the primary culture level. These results suggest that IFN-gamma derived from T cells may enhance production of IFN-gamma by CD4+ T cells, while NK cells rather suppress the induction of IFN-gamma producing CD4+ T cells.

Animals↗

PubMed1995

Age-associated changes in the proliferative response of rat intestinal intraepithelial leukocytes to bacterial antigens.

BACKGROUND & AIMS: Intraepithelial leukocytes (IELs) residing in the intestinal epithelium are thought to recognize bacterial antigens and play an important role in providing protection against the invasion of microorganisms through the intestinal wall. This report studies the effect of various bacterial antigens on the proliferation of cultured IELs. METHODS: Bacterial antigens mycobacterial heat shock protein (hsp) 60 or 70, staphylococcal enteroxin A (SEA), or purified protein derivative (PPD) from Mycobacterium tuberculosis were added to mixed cultures of IELs and irradiated syngeneic or allogeneic spleen cells, and the proliferative response was evaluated. Monoclonal antibodies to CD4, CD8, and major histocompatibility complex class I or class II molecules were added to assess the role of these proteins in the proliferative response. RESULTS: The proliferation of IELs isolated from mature rats in mixed culture was significantly enhanced by the addition of PPD, hsp 60, hsp 70, or SEA. The proliferation of mixed cultures of IELs isolated from young rats was enhanced only by the addition of SEA. In mixed cultures of mature rat IELs and syngeneic spleen cells containing PPD, proliferating populations were of the CD4-CD8+ and CD4+CD8+ phenotypes. Addition of anti-CD4 monoclonal antibody suppressed the proliferative response to PPD to control levels. CONCLUSIONS: IELs isolated from mature rats contain mycobacterial antigen-specific T cells, and these cells may be selected by long-term exposure to common intestinal bacterial antigens such as hsp. These cells may contain populations that recognize bacterial antigens in a CD4-dependent manner.

Age Factors↗

PubMed1995

Analysis of the role of CD4+ T-cells during murine cytomegalovirus infection in different strains of mice.

We examined the role of CD4+ T-cells in peritoneal exudate cells (PECs) during the course of acute murine cytomegalovirus (MCMV) infection in two strains of mice. Cell counts of PECs and cytofluorometric analysis showed that C57BL/6, a resistant strain, had more CD4+ T-cells than BALB/c, a susceptible strain, after intraperitoneal infection of 3 x 10(3) PFU of the Smith strain of MCMV, though both strains had an equivalent number of CD8+ T-cells. CD4+ T-cells of both strains expressed mRNA of IFN-gamma, IL-2, and IL-4 on days 5 and 7 after infection, with much higher expression of these cytokines in C57BL/6 than in BALB/c. At the same time point after infection, macrophages were shown to express mRNA of IL-1 alpha and TNF-alpha with higher expression of IL-1 alpha in C57BL/6 than in BALB/c. Production of nitric oxide, recently shown to be one of the antiviral effector mechanisms of macrophages, by macrophages of both strains was examined showing more production of nitric oxide on day 7 after infection in C57BL/6 than in BALB/c. From these findings, we suggest the possibility that CD4+ T-cells contribute to the protection against MCMV infection via the secretion of cytokines and the resultant activation of macrophages to produce nitric oxide.

Animals↗

PubMed1995

Changes in carnitine metabolism with ketone body production in obese glucose-intolerant patients.

To elucidate the relationship between carnitine metabolism and plasma ketone body concentrations in moderately obese patients with mild glucose intolerance, the ketone body and carnitine levels in the basal state were determined in 72 obese patients: 20 with normal glucose tolerance (NGT), 29 with impaired glucose tolerance (IGT), and 23 with non-insulin-dependent diabetes mellitus (NIDDM) having a fasting plasma glucose (FPG) level of less than 200 mg/dl. Total ketone body (TKB) levels significantly (P < 0.05) increased in the order of NGT, IGT, NIDDM, while the FPG and free fatty acid (FFA) concentrations were significantly (P < 0.05) higher in the NIDDM group than in the other two groups. In contrast, the insulin, glucagon and glycerol levels were comparable in the three groups. The plasma short-chain acylcarnitine (SCAC) concentration and the acylcarnitine/free carnitine (AC/FC) ratio were similar in the IGT and NIDDM groups, and significantly (P < 0.05) greater than those in the NGT group. The AC/FC ratio correlated significantly with the FPG and FFA, but not with the TKB. These results suggest that the combination of IGT with simple obesity may trigger the acceleration of hepatic ketogenesis in conjunction with an elevated SCAC and an increased AC/FC ratio. In addition, the data also imply that, in patients with mild NIDDM, factors other than the carnitines may play a greater role in enhancing ketonemia.

Acylation↗

PubMed1995

Combined treatment with ren-shen-yang-rong-tang (Japanese name: ninjin-youei-to) plus prednisolone on adjuvant-induced arthritis in Lewis rat.

The effects of combined treatment with 250 mg/kg Ren-shen-yang-rong-tang (Japanese name: Ninjinyouei-to, NYT) plus 4 mg/kg, an average dosage, or 0.2 mg/kg, a suboptimal dosage, of prednisolone (PSL) on adjuvant-induced arthritis in Lewis rats were investigated using two treatment schedules. The agents were administered orally every day from day 0 to 21 (schedule A), or from day -7 to 21 (schedule B) after adjuvant injection. PSL treatment (4 mg/kg) obviously inhibited paw swelling due to non-immune inflammation, diminished the weights of the thymus, spleen, adrenals and iliac lymph nodes, and suppressed the increment of serum interleukin (IL)-6 concentration in both schedules compared to controls. NYT treatment alone inhibited paw swelling due to immune inflammation, diminished the weight of the adrenals and decreased IL-6 concentration only in schedule A. Combined treatment with NYT plus PSL (4 mg/kg) showed: (1) a superior effect to that of PSL on paw swelling in the uninjected hind foot, especially in schedule B, (2) a tendency to diminish adrenal weight in schedule A and the weights of all four organs in schedule B compared with PSL treatment alone, and (3) a suppressive effect on IL-6 concentration weaker than that of PSL alone in schedule B. The suppressive effect of combined treatment with NYT plus PSL (0.2 mg/kg) on paw swelling was significantly stronger compared with either NYT or PSL treatment alone in schedule B. Although this dose of PSL had no influence upon the IL-6 concentration, the combined treatment or NYT alone increased the IL-6 concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1995

Hot water extracts of Chlorella vulgaris reduce opportunistic infection with Listeria monocytogenes in C57BL/6 mice infected with LP-BM5 murine leukemia viruses.

The bacterial elimination after infection with Listeria monocytogenes was impaired in mice with murine acquired immunodeficiency syndrome (MAIDS) by infection with LP-BM5 murine leukemia virus. Oral administration of hot water extracts of Chlorella vulgaris (CVE) restored the capacity of MAIDS mice to eliminate L. monocytogenes in association with improvement of the deteriorated immune response to L. monocytogenes. DTH response to Listeria in CVE-treated MAIDS mice was significantly higher than that of MAIDS mice after Listeria infection in association with increases in number of CD4+CD8- and CD4-CD8+ alpha beta T-cells in the infected sites. CVE might be effective in the treatment of opportunistic infection in retrovirus-induced immunodeficient patients.

Animals↗

PubMed1995

Structure-activity relations of fulicin, a peptide containing a D-amino acid residue.

Structure-activity relations of fulicin (H-Phe-D-Asn-Glu-Phe-Val-NH2) isolated from the ganglia of a pulmonate mollusc Achatina fulica were investigated on the contraction of penis retractor muscle of this snail. The contraction-enhancing activity of fulicin was reduced to about 1/3000 when D-Asn2 was replaced by L-Asn, whereas substitution of the other D-amino acid for D-Asn2 showed no significant reduction of the activity. On the contrary, if a D-amino acid was introduced into any position other than position 2 the activity was reduced remarkably. The presence of Glu3 and five amino acid residues as a whole were also required for activity.

Amino Acid Sequence↗

PubMed1995

A novel gut tetradecapeptide isolated from the earthworm, Eisenia foetida.

A novel bioactive tetradecapeptide, GFKDGAADRISHGFamide, was isolated from the gut of the oligochaete annelid, Eisenia foetida, using the isolated anterior gut (crop-gizzard) as a bioassay system. A highly homologous peptide, GFRDGSADRISHGFamide, was also purified from the whole body of another species of earthworm, Pheretima vittata. These peptides were termed Eisenia tetradecapeptide (ETP) and Pheretima tetradecapeptide (PTP), respectively. Both the peptides showed a potent excitatory action on spontaneous contractions of the anterior gut with a threshold as low as 10(-10)-10(-9) M. These peptides were significantly homologous to molluscan tetradecapeptides and, to a lesser extent, to arthropodan tridecapeptides that have been reported to date. All these peptides seem to be evolutionally related to each other.

Amino Acid Sequence↗

PubMed1995

Isolation and characterization of four novel bioactive peptides from a polychaete annelid, Perinereis vancaurica.

Four bioactive peptides were purified from an extract of the polycheate annelid Perinereis vancaurica using an HPLC system for fractionation of the extract and the esophagus of the worm as bioassay system. The sequences of the peptides were determined to be H-Trp-Val-Val-Gly-Asp-Val-Gln-OH, H-Ala-Thr-Trp-Leu-Asp-Thr-OH, H-Trp-Met-Val-Gly-Asp-Val-Gln-OH and H-Phe-Tyr-Glu-Gly-Asp-Val-Pro-Tyr-OH. These peptides showed an excitatory activity on the esophagus of P. vancaurica. The excitatory effect of the second and fourth peptides was marked. They may be neuropeptides involved in regulation of the esophagus.

Amino Acid Sequence↗

PubMed1995

Partial hepatic grafting: porcine study on critical volume reduction.

BACKGROUND: The safe limit of volume reduction in partial hepatic transplantation, including extracorporeal hepatic resection, remains to be clarified. This study evaluated such a limit and pathologic features associated with transplantation of a less than critical volume. METHODS: Partial hepatic grafting was performed in a porcine orthotopic autotransplantation model. According to the remnant liver volume, animals were classified into three groups: group 1, 73.8% +/- 4.2% (SD); group 2, 52.6% +/- 6.7%; and group 3, 29.4% +/- 6.7% of the whole liver (n = 5 each). RESULTS: Three-day survival was achieved in five (100%), four (80%), and zero animals, respectively. All animals in group 3 died of graft nonfunction; their intraoperative clearance of the total bile acids was significantly worse than the other groups (p < 0.01). After operation the clearance of the total bile acids and hyaluronic acid, which is selectively cleared by hepatic endothelial cells, was significantly better in group 1 than group 2 (p < 0.01 and < 0.05, respectively). On histologic examination postperfusion biopsy specimens of group 3 exhibited severe ischemic changes and portosinusoidal hyperemia, whereas that of groups 1 and 2 exhibited only mild ischemic damages. CONCLUSIONS: Transplantation of less than 30% of expected full liver volume could lead to primary graft nonfunction after partial hepatic grafting.

Animals↗

PubMed1995

The induction of skin xenograft tolerance in rat-to-mouse combination could be affected by DFR mediating cells and antibodies against rat bone marrow cells as well as NK cells in the cyclophosphamide-induced tolerance system.

We investigated whether the prolongation of skin xenograft survival was obtained by a tolerance-inducing method using cyclophosphamide (CY), by which long-lasting skin allograft tolerance could be induced. The long-lasting skin allograft survival could be obtained in the recipient C3H/HeN (C3H) mice which were given 100 micrograms of anti-CD4 mAb on day -3, 1 x 10(8) spleen cells (SC) plus 3 x 10(7) bone marrow cells (BMC) derived from C57BL/6 (B6) mice on day -2,200 mg/kg CY on day 0, and which were grafted with allogeneic B6 skin on day 14. When the C3H mice were treated with anti-CD4 mAb, 1 x 10(8) s.c. plus 5 x 10(7) BMC derived from F344 rat and CY, the F344 skin grafts survived slightly longer (about 15 days) than those in untreated recipients (about 8.4 days). Such a prolongation of skin xenograft survival was considered donor-specific because rejection of 3rd party skin grafts from BN rats occurred significantly earlier than that of F344 skin grafts. In the recipient C3H mice treated with anti-CD4 mAb, F344 s.c. plus BMC and CY, mixed chimerism in the periphery was detected for a few days after CY administration, although intrathymic chimerism was not detected throughout this study. In these recipient C3H mice, cytotoxic T lymphocytes (CTL) against F344 antigens were completely abrogated through the delayed footpad reaction (DFR) remained at a low but significant level. Moreover, though antibody (Ab) activity against F344 s.c. was completely abrogated, neither Ab activity against F344 BMC, which seemed to have a background common to natural Ab activity, nor NK activity were abrogated by this treatment. These results suggested that DFR mediating cells directly mediated skin xenograft rejection in the recipient mice treated with anti-CD4 mAb, F344 cells, and CY. Such cells may interfere with establishment of mixed chimerism and long-lasting skin xenograft tolerance, presumably in cooperation with CY-resistant Ab activity and NK cells.

Animals↗

PubMed1995

Anti-metastatic activity induced by the in vivo activation of purified protein derivative (PPD)-recognizing Th1 type CD4+ T cells.

The Th1 type Cd4+ T cell clone (MH2), which is capable of recognizing purified protein derivative from Mycobacterium tuberculosis (PPD), was examined for its anti-metastatic activity against melanoma. In using an in vitro proliferative assay, MH2 was able to recognize PPD-derived antigen in a major histocompatibility complex class II-restricted manner. MH2 showed neither any natural killer (NK) activity nor cytolytic activity against syngeneic B16 melanoma. This clone produced interferon-gamma, tumor necrosis factor and interleukin-2, but not interleukin-4, when co-cultured with PPD and irradiated syngeneic C57BL/6 spleen cells, suggesting that this clone could thus be assigned to the Th1 subset. An intraperitoneal (i.p.) co-injection of 2 x 10(6) MH2 and 50 micrograms PPD increased the NK activity of the peritoneal exudate cells (PEC) and the percentage of NK1.1+ cells in the PEC. These activated NK cells showed a low but significantly cytolytic activity against B16 melanoma. The augmented NK activity induced by the co-injection of MH2 and PPD was maintained by the weekly additional i.p. injections of PPD alone. Using a murine metastatic model, and i.p. co-injection of MH2 and PPD-induced anti-metastatic activity against B16 melanoma. This anti-metastatic activity was then abrogated by the in vivo administration of anti-asialo GM1 serum. In addition, the NK activity in both peripheral blood and metastatic lungs was significantly augmented in the mice which were co-injected with MH2 and PPD. Taken together, these findings indicate that the in vivo activation of Th1 type CD4+ T cells augmented the NK activity in vivo and thus could potentially be an efficient immunotherapeutic weapon against metastasis of melanoma. These results also imply that adoptive immunotherapy could induce anti-metastatic activity through cytokine production but not through any direct cytolytic activity.

Adjuvants, Immunologic↗

PubMed1995

T-cell receptor V alpha and V beta gene use by infiltrating T cells in labial glands of patients with Sjögren's syndrome.

Sjögren's syndrome is an autoimmune disease affecting the exocrine glands; it is thought to result from T-cell-mediated damage. In the labial glands of 20 patients with Sjögren's syndrome, infiltrating T cells were immunohistochemically characterized, and T-cell receptor (TCR) gene expression was examined with a method based on polymerase chain reaction. Most of these lymphocytes expressed CD3, CD4, CD45RO, and TCR alpha beta, whereas less than 5% of them expressed CD25 and CD69. The TCR V alpha and V beta genes expressed in peripheral blood mononuclear cells were diverse, whereas the TCR V alpha and V beta repertoires in the labial glands were more restricted but were still heterogeneous. The predominantly used V alpha and V beta families in the labial glands, when compared with those in peripheral blood mononuclear cells, were found to vary in individual patients and also to differ from patient to patient. Thus the T-cell population in the labial glands was polyclonal but showed a more restricted pattern than that seen in peripheral blood mononuclear cells.

Adult↗

PubMed1995

Extrathymic and thymic origin of murine IEL: are most IEL in euthymic mice derived from the thymus?

Although an abundance of literature supports the hypothesis that murine intestinal intraepithelial lymphocytes (IEL) can develop extrathymically, whether most IEL in euthymic mice ar extrathymically derived is unknown. Recent evidence, however, suggests that the development of most IEL in euthymic mice is influenced either directly or indirectly by the thymus. While some evidence suggests that the thymus can influence the extrathymic development of IEL through time-derived factors, clearly a substantial portion (if not most) of the IEL in euthymic mice are derived directly from the thymus. The ability of the thymus to generate IEL depends on the age of the thymus, as fetal/neonatal thymus up to approximately 2 weeks of age generates a totally different subset of IEL than the adult thymus. These results suggest that the IEL generated from the fetal early neonatal thymus recognize a different antigen than the IEL generated from the adult thymus. The former probably recognizes self-antigen and the later recognizes bacterial antigen.

Aging↗

PubMed1995

A novel cDNA sequence encoding the precursor of the D-amino acid-containing neuropeptide fulicin and multiple alpha-amidated neuropeptides from Achatina fulica.

Fulicin (Phe-D-Asn-Glu-Phe-Val-NH2) is an endogenous neuropeptide containing a D-amino acid from ganglia of the African giant snail Achatina fulica. We have cloned a novel cDNA (1,995 nucleotides) encoding a fulicin precursor from the snail cerebral and subesophageal ganglia. The fulicin precursor protein (357 amino acids) contains one copy of fulicin and at least nine other putative alpha-amidated neuropeptides composed of four to six amino acid residues. Seven of the nine neuropeptides were novel, and the other two had the same structures as Mytilus inhibitory peptide-related peptides previously isolated from the ganglia of Helix pomatia. All sequences of 10 peptides are flanked by Lys-Arg(Lys) at the N-terminus and by Gly-Lys-Arg(Lys) at the C-terminus. Nucleotide sequence analysis revealed that D-Asn present in fulicin is encoded by the usual L-Asn codon (AAT). Although fulicin has as yet only been isolated from the central ganglia. RNA blot analysis revealed that single transcripts of approximately 2.0 kb in size also exist in the ventricles and atria. These results suggest that fulicin and related peptides are produced in neurons and the heart by the processing of a ribosomally made precursor, although the mechanism of in-chain epimerization remains unclear.

Amino Acid Sequence↗