[The helper function of monocytes from aged humans in lymphocyte activation by mitogens].
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Biomedical subjects
Publications and source records attributed to M Tsujimoto.
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Heterogeneity of human gamma interferon (IFN-gamma) induced by the combined treatment with OK-432 and Staphylococcal enterotoxin B (SEB) was demonstrated by chromatofocusing. Treatment of IFN-gamma with a mixture of neuraminidase, and beta-galactosidase eliminated the charge heterogeneity. Apparent molecular weight of IFN-gamma was decreased by enzyme treatment. These results suggest that the heterogeneity of IFN-gamma induced in our system was the result of the difference in the content of sialic acids.
Malignant fibrous histiocytoma (MFH) shows a mixed proliferation of both fibroblastic and histiocytic cells. Because of their complex morphologic appearances, the nature of truly neoplastic cells in MFH has been controversial. In the present study, immunoperoxidase method (PAP method) was used to examine the intracytoplasmic lysozyme (LY), alpha-1-antitrypsin (A1AT), fibronectin (FN), and polyvalent immunoglobulin (PI) in the fibroblastic and histiocytic cells. Twenty-three cases with MFH were histologically divided into three groups; predominantly fibroblastic type (Group I; 5 cases), mixed fibroblastic and histiocytic type (Group II; 15 cases), and almost pure histiocytic type (Group III; 3 cases). Fibroblastic cells showed a strong positive reaction for LY and A1AT, suggesting the histiocytic nature, while the proliferating cells in Group II were more intensely stained by each of the antibodies than in Groups I and III. Enzyme histochemical examinations on fresh materials were available in 3 cases. These findings suggest that proliferating cells in MFH possess a histiocyte nature.
A hepatoma comprised of carcinomatous and sarcomatous proliferation has been considered as collision or carcinoma with sarcomatous proliferation. In this report, an autopsy case of primary hepatocellular carcinoma showing almost equal amount of carcinomatous and sarcomatous proliferations is presented. Serial sections from the tumor showed the frequent transition of the figure from carcinoma to sarcoma, justifying the diagnosis of hepatocellular carcinoma with sarcomatous proliferation. When compared with the usual hepatocellular carcinoma, metastasis with sarcomatous figure was more extensive and both ventricles and atriums of the heart, skin, and intestinal tract were involved. A sarcomatous proliferation in hepatocellular carcinoma and manner of metastasis are discussed with a review of the pertinent literature.
Synthetic lipid A analogs which have two amide-bound and two ester-bound (R)-3-hydroxytetradecanoyl groups at the C-2 and -2' and C-3 and -3' positions of beta(1-6)glucosamine disaccharide mono- or diphosphates showed high activities in most in vitro assays, and the lethality of a diphosphate derivative to galactosamine-treated mice was almost comparable to that of natural lipid A. The pyrogenicity and Shwartzman induction activity of the synthetic analogs, however, were much less than those of natural lipid A.
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A 454 base pair fragment of double stranded DNA consisting of a gene for a human immune interferon (hIFN-gamma), initiation and termination signals plus appropriate restriction endonuclease sites, was totally synthesized. The synthesis involved preparation of 62 oligodeoxyribonucleotides by rapid, solid phase procedures, and enzymatic ligation of the oligonucleotides. This synthetic gene was expressed in E. coli under the control of the lac UV5 promoter. The product has antiviral activity which was acid labile and completely neutralized by antiserum to hIFN-gamma but not by antiserum to hIFN-alpha or hIFN-beta. Molecular weight of hIFN-gamma produced by E. coli was estimated to be about 32,000 and 17,000 by gel filtration and SDS-polyacrylamide gel electrophoresis respectively.
The immunopharmacological activities of bacterial cell walls and muramyl peptides were collected in table form with a comprehensive literature. The past and present studies emphasizing the host-defense enhancing activities of muramyl peptides for antitumor immunotherapy were surveyed along three possible approaches: 1) the nonspecific enhancement of natural defense ability of host against tumor cells themselves; 2) the enhancement of nonspecific resistance of host to microbial infections which are frequently encountered and difficult to treat in the advanced stage of tumor patients; and 3) the stimulation of immunity against tumor-specific or tumor-associated immunogens. Finally, the prospects of successful antitumor immunotherapy with muramyl peptides and their derivatives was discussed.
The localization of alkaline phosphatase in a calcifying epithelial odontogenic tumor obtained from a 53-year-old man was examined cytochemically. The majority of enzyme activity was associated with the epithelial cell membranes of the tumor, and faint activity was found in the cell membranes facing the adjacent stromal tissue. The reaction product of ALP was also detected in some membrane-bound vacuoles (lysosomes) and the Golgi apparatus of tumor cells. It is suggested that the appearance of enzyme activity associated mostly with the epithelial cell membrane may be related to transport function of cell membranes.
A case of prolymphocytic leukemia with IgM hypogammaglobulinemia in a 47-year-old man was presented. The leukemic cells possessed Ia-like antigen and receptors for the third component of complement but lacked surface immunoglobulins, cytoplasmic IgM, receptors for sheep red blood cells, or terminal deoxynucleotidyl transferase activity. In vitro immunoglobulin production experiments demonstrated that the leukemic cells did not have the capacity to produce IgM, while patient's T cells were shown to possess helper function on normal B cells to produce immunoglobulins. By these findings, together with the presence of selective IgM hypogammaglobulinemia, it was suggested that the leukemic cells were derived from a B-cell clone of a stage in differentiation and maturation of IgM-forming B-cell spectrum.
A simple and rapid purification method for human serum C-reactive protein (CRP) was developed. CRP was strongly adsorbed on a DEAE-cellulose column and was easily separated from other serum proteins. CRP was purified approximately 1,000-fold with a high yield (50%). The final preparation showed a single band as judged by SDS-polyacrylamide gel electrophoresis, polyacrylamide gel disc electrophoresis, and polyacrylamide gel isoelectric focusing. The fluorescence of the complex of CRP and 8-anilino-1-naphthalene sulfonate (ANS) changed with change of the pH, suggesting that CRP may show pH-dependent conformational change. This finding could account for the peculiar behavior of the protein in isoelectric focusing; it shows an isoelectric point of 7.4 when the starting pH is 7.0, whereas it shows two isoelectric points, 5.3 and 7.4, when the starting pH is 5.5. Ca2+-dependent change of the fluorescence of the complex of CRP and ANS was also detected. These results suggest a pH- and Ca2+-dependent conformational change of CRP.
Interferon (IFN)-gamma was produced with a high yield in cultures of human peripheral mononuclear cells by combined stimulation with OK-432 and staphylococcal enterotoxin B. Human mononuclear cells cultured in serum-free medium produced several times as much IFN as those in RPMI-1640 medium containing 10% fetal bovine serum. A synergistic effect of OK-432 and staphylococcal enterotoxin B on the production of IFN-gamma was demonstrated. Ultrogel AcA54 column chromatography of crude IFN showed a single peak with an apparent molecular weight of 43,000. Our production system for human IFN-gamma offers a feasible approach to preparation of large quantities of purified IFN-gamma for structure studies, antibody production, and clinical applications.
Thirteen acylated and phosphorylated derivatives of beta-1,6-linked glucosamine disaccharide (lipid A analogs), which were synthesized after the structural model of Salmonella-type lipid A, and seven similar derivatives of glucosamine monosaccharide (lipid A-related compounds) were studied for their immunobiological activities. These included mitogenicity and polyclonal B cell activation enhancement of migration of monocytes and polymorphonuclear leukocytes derived from human peripheral blood, stimulation of guinea pig peritoneal macrophages, activation of human complement, and stimulation of serum antibody production and induction of delayed-type hypersensitivity against ovalbumin in guinea pigs. Comparisons were made with lipid A, RE-glycolipid, lipopolysaccharide of natural sources, and a well-known synthetic adjuvant, N-acetylmuramyl-L-alanyl-D-isoglutamine. Some of the lipid A analogs were found to manifest the mitogenic, polyclonal B cell-activating macrophage-stimulating, complement-activating, and immunostimulating activities, although the observed activities were generally far less than those of natural products in intensity and efficiency. Other immunobiological effects exhibited by most of the synthetic lipid A analogs were the enhancement of migration of monocytes and polymorphonuclear leukocytes. It is premature to draw definite conclusions on structure-activity relationships, since a few compounds which were active in some assay systems were scarcely active in other assays. However, an indisputable fact was that beta-1,6-glucosamine disaccharide 1 alpha,4'-diphosphate, which carries two amide-bound (R)-3-hydroxytetradecanoyl and three ester-bound tetradecanoyl residues, and thus has the structure most closely resembling natural lipid A among test compounds in this study, was definitely active in all of the present assay systems. However, its potency was generally much less than natural products. Some of glucosamine monosaccharide derivatives, especially N-(R)-3-[(R)-3-hydroxytetradecanoyloxy]tetradecanoyl glucosamine, also exerted all of the in vitro activities described above. This fact suggests that a glucosamine disaccharide structure may not necessarily be a prerequisite as far as the in vitro immunobiological activities tested are concerned.
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Cell walls isolated from four bacterial species (Streptococcus pyogenes, Lactobacillus plantarum, Streptomyces gardneri, and Nocardia corynebacteriodes), which exhibited the adjuvant effect of stimulating cellular and humoral immune responses against ovalbumin in guinea pigs, caused the slow-starting and long-lasting contraction of guinea pig ileal strips suspended in Tyrode solution. In contrast to these cell walls active in immunoadjuvancy, those isolated from five bacterial species (Micrococcus lysodeikticus, Staphylococcus epidermidis, Arthrobacter atrocyaneus, Corynebacterium insidiosum, and Ampullariella regularis), which lacked immunoadjuvancy at least in intact walls, caused no or very weak contraction of the ileal strips. Further study demonstrated that both a monomer and a polymer of disaccharide-stem peptides, which were obtained by enzymatic degradation of S. epidermis cell wall peptidoglycans, displayed similar contractile effects. It was finally revealed that guinea pig ileum strips showed a definite contractile response to N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP) and 6-O-stearoyl- and 6-O-(2-tetradecylhexadecanoyl)-MDPs, but not to their analogs, whose C-terminal amino acid was L-isoglutamine or D-isoasparagine in place of D-isoglutamine and which lacked adjuvancy. 6-O-(3-Hydroxy-2-docosylhexacosanoyl)-MDP, on the other hand, caused a slow and lasting relaxation of the ileum strips, but its L-isoglutamine and D-isoasparagine analogs did not.
Whole cells of a clinical isolate (strain S-3) of the genus Capnocytophaga were divided into cell envelope (CE) and cytoplasm (CP) fractions by mechanical disintegration followed by differential centrifugation, and a part of the CE fraction was further fractionated by sodium dodecyl sulfate (SDS) treatment into the peptidoglycan and SDS-supernatant fractions. The other part of the CE was extracted with butanol-water or hot phenol-water to isolate butanol-lipopolysaccharide and phenol-lipopolysaccharide, respectively. All of the test fractions except CP exhibited multifold immunomodulating activities, namely, the adjuvant activities to cellular as well as humoral immune responses against ovalbumin in guinea pigs, the mitogenicity on splenocytes of guinea pigs and BALB/c mice (but not on their thymocytes), the stimulation of guinea pig peritoneal macrophages (in terms of increased glucosamine uptake), and the activation of the human complement system through alternative as well as classical pathways. In addition, the test fractions other than the CP evoked dermatoxic reactions on rabbit skin with characteristic variations among them. The immunomodulating activities of SDS-supernatant were noteworthy in view of the fact that this fraction was essentially free of muramic acid and diaminopimelic acid and did not cause the gelation of horseshoe crab amoebocyte lysate except when it was used at the very high dose, suggesting that there was practically no contamination by peptidoglycans and lipopolysaccharides in the SDS-supernatant.
A study was made on the activity of various bacterial cell walls and peptidoglycans to liberate serotonin from rabbit blood platelets. All of the test cell walls or peptidoglycans prepared from 27 strains of 21 bacterial species were shown to cause a marked release of serotonin, regardless of differences in types of peptidoglycan and non-peptidoglycan moieties and in some biological properties. The assay made with the water-soluble "digests" of Staphylococcus epidermidis cell wall peptidoglycans, which were prepared by use of appropriate enzymes, revealed that a polymer of peptidoglycan subunits (a disaccharide-stempeptide) was definitely active in the release of serotonin, but a structural unit monomer was inactive. Among a variety of synthetic muramylpeptides and their 6-O-acyl derivatives, only 6-O-(3-hydroxy-2-docosylhexacosanoyl)-N-acetylmuramyl-L-alanyl-D-isoglutaminyl- L-lysyl-D-alanine was found to hold a strong serotonin-liberating activity.
A 450 base pair fragment of double stranded DNA consisting of a gene for a human immune interferon (146 amino acid residues), initiation and termination signals plus appropriate restriction endonuclease sites was totally synthesized. The synthesis involved preparation of 62 oligodeoxyribonucleotides by rapid, solid phase procedures, and enzymatic ligation of the oligonucleotides. This synthetic gene was expressed in E. coli under the control of the lac UV5 promoter. Molecular weight of IFN-gamma produced by E. coli was estimated to be about 32,000 and 17,000 dalton by gel filtration and SDS-polyacrylamide gel electrophoresis respectively, suggesting that a dimer-form molecule was expressed in E. coli.