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Biomedical subjects

K Nomoto

Publications and source records attributed to K Nomoto.

At least 361 records · Page 20Linked to original sources
PubMed1993

T cells expressing both L-selectin and CD44 molecules increase in number in peritoneal exudate cells and in vitro-stimulated spleen cells from mice immunized intraperitoneally with Listeria monocytogenes.

L-selectin, which was first reported as MEL-14 antigen in mice, is a type of animal lectin and expressed on lymphocytes, neutrophils and macrophages. L-selectin has been reported to be a homing receptor of lymphocytes to peripheral lymph nodes and to have an important role in initial adhesion of lymphocytes and neutrophils to endothelial cells activated by inflammatory cytokines. On the other hand, it has been reported that naive T cells express L-selectin while memory T cells and in vitro antigen-stimulated T cells lose its expression. If all memory T cells lack L-selectin, trafficking of memory T cells into inflammatory sites would be difficult. To know whether all memory T cells lack L-selectin expression, kinetics of expression of L-selectin was analysed on memory T-cell subsets, which are detected by expression of CD44, in mice after intraperitoneal immunization with a sublethal dose of viable Listeria monocytogenes. T cells expressing both L-selectin and CD44 were detected in splenocytes and peritoneal exudate cells (PEC) from untreated mice, though at low levels. L-selectin+ CD44+ T cells increased in PEC, which are known to be highly enriched in antigen-primed T cells, and reached maximum level on day 14 after immunization. Furthermore, we found increases not only of L-selectin- CD44+ but also of L-selectin+ CD44+ T cells by in vitro Listeria antigen stimulation of Listeria-immune spleen cells on day 14. These results showed that T cells expressing both L-selectin and CD44 increase after antigen stimulation in vivo and in vitro. The L-selectin+ CD44+ T cells may be a subset of memory T cells which retain their capacity of trafficking to inflammatory sites.

Animals↗
PubMed1993

Structure and function of the molluscan myoactive tetradecapeptides.

Effects of myoactive tetradecapeptides, Achatina excitatory peptide 2 and 3 (AEP2 and AEP3) and Fusinus excitatory peptide 4 (FEP4), on several molluscan muscles and neurons were investigated. In the penis retractor and radula retractor muscles of Achatina fulica (pulmonate), the three peptides enhanced the tetanic contraction elicited by nerve stimulations. The order of potency was AEP2 > AEP3 > FEP4, although the effects of AEP3 and FEP4 on the radula retractor were somewhat irregular. AEP2 also induced rhythmic bursts of activity in the buccal ganglionic neuron B4 known as a cholinergic motoneuron of the radula retractor. In the radula protractor and retractor muscles of Fusinus ferrugineus (prosobranch), FEP4 was most potent in enhancing the contraction. The enhancement was greater in the protractor than in the retractor. It was suggested that myoactive tetradecapeptides modulate mainly the cholinergic transmission in molluscan muscles.

Amino Acid Sequence↗
PubMed1993

Characterization of T-cell receptor gamma delta T cells appearing at the early phase of murine Listeria monocytogenes infection.

It has been shown that T-cell receptor (TcR) gamma delta + CD4- CD8- T cells increase in number and have an important role in early protection in murine Listeria monocytogenes infection. In this report, to characterize further the phenotype of the gamma delta T cells in listeriosis, we analysed V region gene usage and in vitro antigen recognition of the TcR gamma delta T cells in the peritoneal cavity of mice at the early phase after i.p. infection with a sublethal dose of L. monocytogenes. The gamma delta T cells predominantly expressed V delta 6 which has been reported to be expressed by TcR gamma delta-bearing foetal thymocyte hybridomas specific to mycobacterial and self heat-shock protein (hsp) 60. These early appearing CD3+ CD4- CD8- T cells in Listeria-infected mice, which were reported to be TcR gamma delta T cells, increased in proportion and in size by in vitro stimulation with recombinant hsp 60 from Mycobacterium bovis and purified protein derivative from M. tuberculosis but not by stimulation with heat-killed L. monocytogenes. A 65,000 MW molecule was detected in the lysate of viable L. monocytogenes but not in the lysate of heat-killed L. monocytogenes by a monoclonal antibody (mAb) raised against mycobacterial hsp 60. These results suggest that the V delta 6-bearing peripheral gamma delta T cells are activated by recognizing listerial hsp 60 expressed by viable L. monocytogenes. The hsp 60-reactive V delta 6-bearing T cells may have an important role in protection against L. monocytogenes and other parasites that express hsp 60 at high level.

Animals↗
PubMed1993

Increase of Thy-1 antigen on the thymocytes accompanied with their augmented adhesion capacity to thymic epithelial cells in the mice infected with Listeria monocytogenes.

It becomes increasingly clear that adhesion systems such as CD2/LFA-3 (lymphocyte function-associated antigen-3), LFA-1/ICAM-1 (intercellular adhesion molecule-1) and Thy-1/putative Thy-1 ligand participate in the association between murine thymocytes and thymic epithelial cells. In the present study, thymocytes showed an increase in surface Thy-1 levels in mice infected with Listeria monocytogenes, but no significant changes in the levels of CD2 or LFA-1. No alteration was found either in the ratio of CD3high/CD3low/CD3- or in that of CD4/CD8 subsets in these thymocytes compared with uninfected control thymocytes which excluded the possibility of enrichment of 'cortical thymocytes' with Thy-1high/CD3low/CD4+ CD8+ in the thymocyte population of infected mice. Moreover such Thy-1high thymocytes exhibited a highly augmented ability of adhesion to a thymic epithelial cell line due to the increase of surface Thy-1 antigens as an adhesion molecule. At such intervals after infection, the total number of thymocytes was found to be reduced. These results suggest that the expression level of surface Thy-1 on thymocytes is regulated in response to in vivo stimulation and may play a role in the intrathymic development of thymocytes by affecting the adhesion of thymocytes with thymic stromal cells. The implication of the enhanced ability of adhesion in the decrease in the number of thymocytes is discussed.

Animals↗
PubMed1993

Phenotypic and functional modulation of interleukin-2-activated peripheral blood mononuclear cells by anti-CD3 and anti-CD28 antibody.

For effective adoptive cancer immunotherapy, not only killer lymphocytes (cytotoxic T cells or NK cells) but also helper T cells are considered to be beneficial. The conventional culture of human lymphocytes with high-dose of recombinant interleukin -2 (rIL-2) yields a high proportion of killer cells (CD8+ or CD56+ cells) but is usually accompanied by a decrease in the proportion of CD4+ cells. Peripheral blood mononuclear cells (PBMC) stimulated with anti-CD3 monoclonal antibody (10 ng/ml) in the absence of rIL-2 during the initial 48-hr period of cultivation followed by the addition of rIL-2 (200 Japan reference units/ml) (referred to as anti-CD3-rIL-2) resulted in CD4+ cells expanding 8.0- to 63.3-fold at day 12-14. When PBMC were cultivated with rIL-2 alone, or with rIL-2 together with anti-CD3 initially, CD4+ cells expanded 1.7-6.2- or 5.7-36.9-fold, respectively. The expansion of CD4+ cells from PBMC in the anti-CD3-rIL-2 cultures was enhanced by the addition of anti-CD28 monoclonal antibody at the initiation of cultures (500 ng/ml) (referred to as anti-CD3/anti-CD28-rIL-2) (12.1- to 127.8-fold expansion). The anti-CD3-rIL-2 and anti-CD3/anti-CD28-rIL-2 cultures also demonstrated a lesser expansion of CD3-CD56+ cells (NK cells). The PBMC in the anti-CD3/anti-CD28-rIL-2 cultures showed low cytotoxic activity per cell; however, the cytotoxic activity per total culture was comparable to that of PBMC expanded with rIL-2 alone because of the larger total expansion of the former culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗
PubMed1993

Cancer-specific binding of a mouse MAb vs. Candida krusei cytochrome c: an antigen recognized by a cancer-associated human MAb HB4C5.

More than 60 mouse monoclonal antibodies directed to cytochrome c from Candida krusei with different specificities were raised. Most of these monoclonal antibodies, except for three of them, did not cross-react with bovine cytochrome c. By the immunoblotting method, the monoclonal antibodies of clones HCC 5-13, 9-2, and 10-5 reacted with the Candida cytochrome c, which had been transferred onto nitrocellulose membrane, but those of clones HCC 1-22, 6-3, and 17-3 did not, although all these monoclonal antibodies strongly reacted with coated Candida cytochrome c on plastic immunoplates when examined by ELISA. On the contrary, monoclonal antibody activities of clones HCC 1-22, 6-3, and 17-3 in binding to the coated cytochrome c in ELISA were inhibited competitively by the addition of extra Candida cytochrome c, whereas those of clones HCC 5-13, 9-2, and 10-5 were not inhibited. Among these monoclonal antibodies, the antibody of clone HCC 6-3, which showed a good reactivity to added cytochrome c in inhibiting ELISA reaction but was not reactive with the transblotted cytochrome c on nitrocellulose, was found to be reactive with human lung cancer tissues specifically with no reactivity to normal tissues. The immunostaining of lung cancer tissue showed that this mouse monoclonal antibody to Candida cytochrome c reacted to the cytoplasmic fraction of the cancer cells specifically.

Animals↗
PubMed1993

Direct evidence for clonal destruction of allo-reactive T cells in the mice treated with cyclophosphamide after allo-priming.

It has previously been reported that a single i.p. injection of 200 mg/kg cyclophosphamide (CP) 2 days after priming with 10(8) donor spleen cells (SC) leads to donor-specific skin allograft tolerance in H-2 compatible, multiminor antigen incompatible murine strain combinations. It is speculated that the i.v. injection of donor cells may result in synchronized proliferation of donor-reactive host T cells and subsequently administered CP may specifically destroy these proliferating T cells in the periphery. Although this unique action of CP is considered to be a principal mechanism in this method, direct evidence has not yet been obtained. In the present article, this in vivo destructive effect of CP is clearly demonstrated by assessing detailed kinetics of host-derived blastoid T cells and donor (Mls-1a)-reactive V beta 6+ T cells in the model system of C3H mice rendered tolerant to AKR. Frequencies of the blastoid cells and V beta 6+ cells, which increased as a result of AKR priming, decreased rapidly with the administration of CP. C3H mice, which received AKR SC alone, also exhibited partial deletion of V beta 6+ T cells, but both tempo and magnitude of decrease in the frequency of V beta 6+ cells were quite different from those of the C3H mice given AKR SC and CP, which showed more rapid and profound elimination of V beta 6+ T cells. In accordance with these kinetic studies, in vitro proliferative response to Mls-1a antigens was greatly impaired in mice treated with SC and CP, whereas a low but appreciable response was detected in mice given SC alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

A lymphocyte-specific protein tyrosine kinase, p56lck, regulates the PMA-induced internalization of CD4.

p56lck, a member of the src family of non-receptor protein tyrosine kinases (PTKs), is expressed predominantly in T-lymphocytes. Association of p56lck with CD4 and CD8 T-cell receptor (TcR) accessory molecules suggests that p56lck may play a specialized role in antigen-induced T-cell activation. CD4 and CD8 molecules are known to stabilize the interaction between TcR and the major histocompatibility complex during T-cell activation. To examine the role of p56lck in the dynamics of the CD4 molecule, p56lck-expressing transfectant cell clones were prepared by the transfection of an lck-gene plasmid containing an inducible promoter into a CD4+lck- human monocytoid cell line. When these transfectant cells were stimulated with phorbol ester, CD4 internalization on these p56lck-expressing cell lines was selectively and markedly retarded, as compared to p56lck-negative control cell lines. When cell-surface CD4 and intracellular CD4 were selectively precipitated after stimulation, the intracellular CD4 molecules were dissociated from p56lck whereas the surface-retained CD4 molecules were still associated with p56lck. Moreover, the dissociation of p56lck from CD4 appeared to occur prior to the PMA-induced internalization of CD4. These data indicate that p56lck regulates the PMA-induced internalization of CD4 possibly via its association with CD4. Treatment with genistein, a PTK inhibitor, revealed that the PTK activity of p56lck might not be involved in this regulatory effect of p56lck on CD4 internalization.

Blotting, Western↗
PubMed1992

Interference with cyclophosphamide-induced skin allograft tolerance by cyclosporin A.

In a murine strain combination identical in H-2 Ag but disparate in minor histocompatibility (H) Ag consisting of C3H/He (C3H; H-2k, Mls-1b) mice as recipients and AKR/J (AKR; H-2k, Mls-1a) mice as donors, a permanent skin allograft tolerance can be achieved by the cyclophosphamide (CP)-induced tolerance system that consists of i.v. injection of donor spleen cells (day -2) and i.p. injection of CP 2 days later (day 0). Such permanent take of allografts in CP-induced tolerant mice was interfered with by intramuscular injection of cyclosporin A (CsA) from day -5 to day -1 and their grafts were rejected by 21 days after grafting. Mls-1a-reactive CD4+V beta 6+ T cells in the periphery, as the indicator to follow the kinetics of donor-reactive T cells, increased on day 0 and day 3 in the C3H mice treated with AKR spleen cells alone, whereas they disappeared rapidly from day 0 to day 3 in CP-induced tolerant mice. When CsA capable of interfering with IL-2 production and T cell proliferation was administered before CP treatment in CP-induced tolerance system, the number of CD4+V beta 6+ T cells in periphery did not increase on day 0 and 3, but increased on day 7 in contrast to the decreased number of those in CP-induced tolerant mice. On day 7, MLR against donor cells was decreased in CP-induced tolerant mice, but maintained in CsA-interfered tolerant mice. These result may indicate that the destruction of donor-Ag-stimulated, proliferating T cells by CP is interfered with by CsA, probably because CsA inhibits the proliferation of donor-reactive T cells at the time of CP treatment. Furthermore, these results also implicate that the protocol for immunosuppression with CsA and antimetabolites has to be designed carefully in clinical transplantation.

Animals↗
PubMed1992

Conformational feature of neuroactive domoic acid: X-ray structural comparison with isodomoic acid A and alpha-kainic acid.

As an aid for developing a new type of potent insecticide acting on the neuromuscular junction, conformational characteristics of domoic acid and isodomoic acid A, the naturally occurring glutamate agonists, were investigated by X-ray crystal analyses. Conformational comparison with a neuroactive alpha-kainic acid provides information concerning the stereochemical feature responsible for the biological activity.

Insecticides↗
PubMed1992

Effect of the D-Phe2 residue on molecular conformation of an endogenous neuropeptide achatin-I. Comparison of X-ray crystal structures of achatin-I (H-Gly-D-Phe-Ala-Asp-OH) and achatin-II (H-Gly-Phe-Ala-Asp-OH).

The molecular conformation of achatin-II neutral form (H-Gly-Phe-Ala-Asp-OH), an endogenous peptide from the Achatina fulica ganglia, was elucidated by X-ray crystal analysis. The molecule takes an extended beta-pleated structure stabilized by 5 intermolecular hydrogen bonds with the antiparallely arranged molecules. This is in contrast with the turn conformation of a neuroactive achatin-I (H-Gly-D-Phe-Ala-Asp-OH) [(1992) FEBS Lett. 276,95-97]. The conformational comparison of both of the molecules makes clear the structural role which D-Phe residue of achatin-I plays in forming a definite active form.

Amino Acid Sequence↗
PubMed1992

Sequential analysis of T cells in the liver during murine listerial infection.

Phenotypes and functions of T cells in the liver were studied after an i.p. inoculation with viable Listeria monocytogenes in mice. T cells in the liver of untreated C3H/HeN mice (C3H; H-2k, Mls-2a) contain Thy-1.2+TCR-alpha beta + cells as a majority and Thy-1.2+TCR-gamma delta + cells and Thy-1.2-TCR-gamma delta + cells as minorities. The liver of untreated C3H mice did not contain T cells expressing V beta 3 and V beta 11, which are potentially autoreactive against self-superantigens of Mls-2a and Dvbl, respectively. On days 3 to 6 after infection, Thy-1.2-CD4lowTCR-alpha beta + T cells or Thy-1.2-TCR-gamma delta + T cells increased significantly in number and proportion in the liver whereas T cells with these phenotypes were hardly detected in the spleen, lymph nodes, peripheral blood, and peritoneal cavity during the course of the infection. The Thy-1.2-CD4lowTCR-alpha beta T cells contained V beta 3 or V beta 11-bearing cells in high frequencies. The potentially autoreactive V beta 3- or V beta 11-bearing T cells disappeared from the liver on day 7 after infection. Furthermore, the V beta 3+ and V beta 11+ cells but not V beta 8+ cells disappeared after culture for 24 h at 37 degrees C. In vitro stimulation of liver T cells using anti-V beta 11 mAb showed no proliferative response. These results suggest that the potentially autoreactive clones with Thy-1.2-CD4low phenotypes, which increased in number after listerial infection, may be anergized after interaction with self-Ag and may be programmed to die. These potentially autoreactive clones induced in the liver of Listeria-infected mice may not be functionally relevant to the host defense against Listeria.

Animals↗
PubMed1992

IFN-gamma-producing ability as a possible marker for the protective T cells against Mycobacterium bovis BCG in mice.

We searched for a functional marker with which T cells mediating acquired cellular resistance (ACR) can be discriminated from those mediating delayed-type hypersensitivity (DTH) in mice immunized with Mycobacterium bovis BCG. Four wk after injection of the mice with 10(5) viable BCG (vBCG) cells emulsified in IFA, a passive transfer experiment revealed that T cells mediating DTH as well as ACR, which we designated TACR, were generated in the spleen. In contrast, T cells mediating only DTH (TDTH) were generated by immunization with 10(7) killed BCG cells along with IFA. The transferring ability of both TACR and TDTH was substantially reduced by treatment with anti-Thy-1.2 or anti-CD4 mAb plus complement, whereas anti-CD8 treatment had no effect. To determine the functional differences between TACR and TDTH, we assessed IL-2 and IFN-gamma production from TACR and TDTH after stimulation with PPD in vitro. Similar levels of enhanced IL-2 activity were detected in the culture supernatants of both groups of T cells. However, augmented production of IFN-gamma was observed only in TACR. This finding was confirmed by Northern blot analysis for detection of IFN-gamma-specific mRNA. In addition, a significant increase in the number of IFN-gamma-producing cells was observed only in T cells from mice immunized with vBCG. The production of IFN-gamma was also totally abolished by treatment with anti-Thy-1.2 and anti-CD4 mAb plus complement in vitro, whereas anti-CD8 mAb treatment had no effect. These results suggest that CD4+ protective T cells generated by immunization with vBCG are characterized by the ability to produce IFN-gamma after stimulation with specific Ag.

Animals↗
PubMed1992

Trypsin inhibitors from bottle gourd (Lagenaria leucantha Rusby var. Depressa Makino) seeds. Purification and amino acid sequences.

Two almost identical trypsin isoinhibitors, LLDTI-I and LLDTI-II, from bottle gourd (Lagenaria leucantha Rusby var. Depressa Makino) seeds were purified by acetone precipitation, gel filtration and reversed phase chromatography. LLDTI-I and LLDTI-II consist of 30 and 29 amino acid residues, respectively, and have identical sequences, except that LLDTI-I has one additional pyroglutamic acid residue at N-terminus. Both proteins are strong inhibitors of bovine trypsin, with Ki values of 2.4.10(-10) M (LLDTI-I) and 9.6.10(-11) M (LLDTI-II). Amino acid sequences are as follows: [sequence: see text]

Amino Acid Sequence↗