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Biomedical subjects

M Harada

Publications and source records attributed to M Harada.

At least 1,027 records · Page 57Linked to original sources
PubMed1987

Stimulatory effect of forphenicinol on normal human bone marrow granulocyte-macrophage progenitor cells mediated by T-lymphocytes.

Forphenicinol, L-2-(4-formyl-3-hydroxymethylphenyl) glycine, is a newly discovered low molecular weight immunomodifier. Its effects on normal human bone marrow granulocyte-macrophage progenitor cells (CFU-C) were studied in vitro. Addition of forphenicinol to cell cultures resulted in a significant (p less than 0.05) increase in the number of, and preservation of the ability to form CFU-C colonies per fixed number of human non-adherent bone marrow cells. This effect was not observed in T-lymphocyte depleted fractions of human non-adherent bone marrow cells. Furthermore, colony stimulating factor was released when T-lymphocytes were incubated with forphenicinol. These data suggested that T-lymphocytes mediated the stimulatory effect of forphenicol on human bone marrow CFU-C.

Adjuvants, Immunologic↗
PubMed1987

Mouse anti-benzylpenicilloyl IgE monoclonal antibody: preparation, characterization and cross-reactivity.

Anti-benzylpenicilloyl (BPO-) monoclonal antibody of the IgE class was prepared from spleens of immune C57BL/6 mice whose sera reacted with BPO-hapten, penicillin G(PCG) polymer, cephalothin (CET)-hapten and CET polymer. Affinity chromatography experiments showed that the haptenic specificity of the IgE monoclonal antibody (designated BIE-13CE) was directed mainly to phenylacetyl portion of BPO group. BIE-13CE antibody reacted on passive cutaneous anaphylaxis (PCA) assay with BPO-hapten, CET-hapten, cephaloridine-hapten and CET polymer, but did not react with PCG polymer, ampicillin-hapten, or cefazolin-hapten. These results indicated that the sera of the immune C57BL/6 mice contained IgE antibodies capable of cross-reacting at the monoclonal antibody level with various forms of eliciting antigens and that the cross-reactivity of the antibody could be ascribed essentially to the structural similarity of acyl side chains of the antibiotics. The structure of the CET polymer is also discussed in terms of its PCA reactivity with the monoclonal antibody and analytical and spectral data of the polymer.

Animals↗
PubMed1986

Nonspecific suppressor T cells cause decreased mixed lymphocyte culture reactivity in bone marrow transplant patients.

Decreased reactivity in mixed lymphocyte culture (MLC) was observed in patients within 1 yr after allogeneic and autologous bone marrow transplantation. Suppressor activity of peripheral blood mononuclear cells (PBMC) from transplant patients was studied by adding these cells as modulator cells to a bidirectional MLC with cells from normal individuals. PBMC from transplant patients markedly suppressed MLC reactivity in a dose-dependent manner. Suppressor activity was present in cells forming rosettes with sheep erythrocytes. Treatment of modulator cells with monoclonal antibodies against T cell differentiation antigens (OKT8, OKIa1) and complement completely abolished suppression of MLC. Suppressor activity was unaffected by 30 Gy irradiation. Suppressor activity declined gradually after transplantation and was inversely correlated with MLC reactivity of each patient at a significant level (p less than 0.01). These observations suggest that OKT8+ Ia+ radioresistant suppressor T cells play a role in the development of decreased MLC reactivity observed during the early post-transplant period.

Adolescent↗
PubMed1986

Effect of recombinant interleukin 2 (R-IL2) on in vivo growth of murine myeloma X5563.

The present study deals with the effect of recombinant interleukin 2 (R-IL2) on in vivo growth of murine myeloma X5563. Administration of R-IL2 (5 X 10(4) J.U./mouse per day) s. c. starting 1 day after X5563 inoculation i.d. had a marginal effect on the growth of X5563, and all the mice repeatedly given R-IL2 from day 1 to day 17 died. However, daily administration of R-IL2 starting 7 days after the tumor inoculation was highly effective and significantly lengthened survival time compared with the control mice injected with vehicle alone. About 50% of the treated mice were completely cured, and survived for more than a month after the therapy ceased. In a representative experiment, where the growth of X5563 was slow because of the small number of inoculated tumor cells, all the mice (n = 6) given R-IL2 from day 11 to day 23 showed complete cure of the established X5563 solid tumor. These mice showed in vivo protective immunity and in vitro cytotoxic T cell responses to X5563 tumor antigens. Histologically, a large number of macrophages and lymphocytes had infiltrated the area around the necrotic X5563 tumor mass in the mice which had received R-IL2 therapy. These results suggest that repeated injections of R-IL2 at the local site after tumor development can augment antitumor immunological responses and subsequently induce tumor regression.

Animals↗
PubMed1986

Suppressive and augmentative effects of recombinant human interleukin-2 upon delayed type of hypersensitivity in the mouse.

The in vivo action of recombinant human interleukin-2 (IL-2) was investigated against the delayed type of footpad swelling to sheep red blood cells (SRBC) in B6D2F1 mice. A single intravenous injection of 1 X 10(5) of SRBC induced transient cell-mediated immunity for delayed type of hypersensitivity (DTH) which peaked four days after the immunization and diminished rapidly thereafter. IL-2 exerted both suppressive and augmentative effects, depending on the timing of its administration in relation to the immunization. Daily intraperitoneal injections of as much as 5 X 10(4) Jurkat units of IL-2 from day 0 through day 3 caused marked suppression of DTH elicited on day 4. On the other hand, three or four consecutive injections of IL-2 from day 4 or day 3 through day 6 produced potent DTH on day 7, when DTH was faint, if there was any, without IL-2 administration. Potentiation of DTH was also observed when three injections of IL-2 were given from day 7 through day 9 and the antigen was challenged on day 10. These results suggest that IL-2 acts as an immunomodulator against cell-mediated immunity to SRBC in the mouse.

Animals↗
PubMed1986

Two-dimensional electrophoresis in the analysis of a mixture of human sublingual and submandibular salivary proteins.

Complex protein mixtures of unstimulated human sublingual and submandibular saliva were fractionated by two-dimensional (2-D) gel electrophoresis, visualized by silver staining and then analysed by immunostaining. Specific proteins were identified by incubation with specific antibody and peroxidase-conjugated second antibody (Western blot). Electrophoresis and silver staining revealed over 50 protein components in 2 microliter of unconcentrated mixture. The Western-blot technique allowed detection of protein spots of plasma origin when an antibody against whole serum was used, but only the albumin spot could be found. Albumin, secretory IgA, acid phosphatase and alpha-amylase were identified with specific antibodies.

Acid Phosphatase↗
PubMed1986

Analysis and identification of human parotid salivary proteins by micro two-dimensional electrophoresis and Western-blot techniques.

Using this method, without denaturing agents, over 60 protein components in 2 microliter (about 4 micrograms of protein) of unconcentrated parotid saliva could be detected. After two-dimensional separation, the spots of albumin, secretory IgA, IgG, acid phosphatase, amylase and plasma-originating protein were identified by antibodies with the Western-blot technique.

Adolescent↗
PubMed1986

Site of alkylation of the major ribonuclease from Aspergillus saitoi with iodoacetate.

A base non-specific and adenylic acid preferential ribonuclease from Aspergillus saitoi (RNase M) was modified by [14C]iodoacetic acid. RNase M was inactivated with concomitant incorporation of about 1 mol equivalent of carboxymethyl group. Carboxymethylated RNase M (CM RNase M) thus obtained was reduced and carboxymethylated (RCM CM RNase M). From tryptic and chymotryptic digests of RCM CM RNase M, two carboxymethylated histidine-containing peptides labeled with radioactivity were isolated. The amino acid sequences of these two peptides were determined to be Thr-Ile-His-Gly-Leu-Trp-Pro-Asp-Asn-Cys-Asp-Gly-Ser-Tyr... and His-Gly-Thr-Cys-Ile-Asn-Thr-Ile-Asp-Pro-Ser-Cys-Tyr-Pro-Asp-Asp-Tyr-Ala. .... The distribution of the radioactivity on the former and latter peptides was 43% and 57%, respectively. The results indicated that two histidine residues are involved in the active site of RNase M, and the modification of either one of the two histidine residues inactivates RNase M. The CD spectrum of carboxymethylated RNase M indicated that some tryptophan residue(s) with a CD band at 287 nm is in the proximity of the active site histidine residues of RNase M.

Alkylation↗
PubMed1986

Enhancement of in vitro erythropoiesis by peripheral blood mononuclear cells from allogeneic marrow recipients in the early post-transplant period.

The effect of peripheral blood mononuclear cells (PBMCs) from bone marrow recipients on in vitro growth of erythroid burst-forming units (BFU-E) was studied. PBMCs were obtained from 5 allogeneic, 1 syngeneic and 1 autologous bone marrow recipient(s) at different intervals after transplantation. The number of BFU-E was significantly increased when donor bone marrow cells were co-cultured with PBMCs obtained from allogeneic marrow recipients in the early post-transplant period. No effect was observed using PBMCs obtained in the later post-transplant period, PBMCs from a syngeneic marrow recipient, or PBMCs from an autologous marrow recipient. The BFU-E enhancing activity was present among T cells and was abolished by treating them with OKT3 or OKT4 antibody and complement. These observations suggest that chimeric T lymphocytes, probably of the helper/inducer subset, from allogeneic marrow recipients in the early post-transplant period have a potent enhancing effect on in vitro erythropoiesis.

Adolescent↗
PubMed1986

Inorganic pyrophosphatase activity of purified bovine pulp alkaline phosphatase at physiological pH.

At physiological pH, the hydrolytic activity of purified bovine pulp alkaline phosphatase toward phosphorus compounds was observed to be in the order of inorganic pyrophosphate greater than beta-glycerophosphate greater than phosphorylcholine greater than p-nitrophenylphosphate greater than glucose-6-phosphate. Optimum pH of the enzyme toward inorganic pyrophosphate was shown to be 8.5, with around 60% of the activity at pH 7.5. The activity was increased by the addition of Mg2+, but a different pattern of activation was observed between pH 7.5 and 8.5.

Alkaline Phosphatase↗
PubMed1986

Cephalothin and penicillin G polymers as elicitors of rat PCA mediated by mouse IgE antibodies.

PCA-eliciting activities of cephalothin- and penicillin G-polymers were examined in rats sensitized with homologous IgE antibodies of mouse origin. Cephalothin-polymers elicited PCA regardless of the source of antibodies and the methods to raise them, the minimal effective dose being 2 to 20 micrograms/animal. Penicillin G-polymers provoked PCA only when anti-benzylpenicilloyl IgE antibody of C57BL/6J mouse raised early after immunization was used. The minimal effective dose in this case was 5 micrograms/animal, being comparable to that of cephalothin polymers.

Animals↗
PubMed1986

Heterogeneity of cross-reactivity of mouse anti-benzylpenicilloyl antibodies against cephalothin haptens or polymers.

The cross-reactivity of mouse anti-benzylpenicilloyl antibodies against cephalothin-haptens and cephalothin-polymers was divergent depending on the source and isotype of antibodies and the methods to raise them. High cross-reactivity nearly equal to the reactivity to homologous haptens and polymers was obtained only with the IgE antibody of C57BL/6J mouse raised shortly after an immunizing injection of the emulsion containing Freund's complete adjuvant and benzylpenicilloyl-bovine gamma-globulin conjugate. With the other antibody preparations, cross-reactivity was moderate to negligible.

Animals↗