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M Kotani

Publications and source records attributed to M Kotani.

At least 19 recordsLinked to original sources
PubMed1992

Generation of one set of monoclonal antibodies specific for a-pathway ganglio-series gangliosides.

We established five murine monoclonal antibodies (MAbs) specific for a-pathway ganglio-series gangliosides by immunizing C3H/HeN mice with these purified gangliosides adsorbed to Salmonella minnesota, followed by fusion with mouse myeloma cells. The binding specificities of these MAbs were determined by enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatogram. These five MAbs, designated GMR6, GMB28, GMB16, GMR17, and GMR11 reacted strongly with the gangliosides GM3, GM2, GM1, GD1a, and GT1a, respectively, that were used as immunogens. Three MAbs, GMB28 (anti-GM2), GMB16 (anti-GM1), and GMR11 (anti-GT1a) showed highly restricted binding specificities, reacting only with the immunizing ganglioside. None of the other various authentic gangliosides or neutral glycolipids was recognized. On the other hand, the other two MAbs, GMR6 (anti-GM3) and GMR17 (anti-GD1a) exhibited broader specificities. MAb GMR6 cross-reacted with GM4, GM1b, GD1a, GT1b, and IV3NeuAc alpha-nLc4Cer. MAb GMR17 also reacted with GM1b and GT1b. Neither GMR6 nor GMR17 reacted with other gangliosides or neutral glycolipids tested. Using these MAbs, we determined the expression of these gangliosides, especially GM1, GD1a, and GT1a on mouse, rat and human leukemia cells. GM1 and GD1a were expressed on some leukemia cells, whereas GT1a was not detected in these cells.

Animals↗
PubMed1992

Generation of one set of monoclonal antibodies specific for b-pathway ganglio-series gangliosides.

We established six murine monoclonal antibodies (MAbs) specific for b-pathway ganglio-series gangliosides by immunizing C3H/HeN mice with these purified gangliosides adsorbed to Salmonella minnesota mutant R595. The binding specificities of these MAbs were determined by an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatogram. These six MAbs, designated GGB19, GMR2, GMR7, GGR12, GMR5, and GGR13 reacted strongly with the gangliosides GD3, O-Ac-GD3, GD2, GD1b, GT1b, and GQ1b, respectively, that were used as immunogens. All these MAbs except GGB19 showed highly restricted binding specificities, reacting only with the immunizing ganglioside. None of other various authentic gangliosides or neutral glycolipids were recognized. On the other hand, MAb GGB19 exhibited a broader specificity, cross-reacting weakly with O-Ac-GD3, GQ1b, and GT1a, but not with other gangliosides or neutral glycolipids. Using these MAbs, we determined the expression of these gangliosides, especially GD1b, GT1b, and GQ1b on mouse, rat, and human leukemia cells. GD1b was expressed on rat leukemia cells, but not on mouse and human leukemia cells tested. Neither GT1b nor GQ1b was detected in these cell lines.

Animals↗
PubMed1992

Oestrogen retards the development of spontaneous thymomas in BUF/Mna rats.

BUF/Mna rats develop spontaneous thymomas with nearly 100% incidence in both sexes. While the thymomas in males develop from around 9 months of age, those in females start from 13-15 months of age. To clarify the mechanism of the delay of thymomagenesis in females, the effect of sex hormones on the development of thymomas was examined after either gonadectomy or oestrogen treatment. Prepubertal ovariectomy accelerated the thymoma development in females, whereas orchiectomy did not affect it. An intraperitoneal injection of oestriol (20 mg) into males at 2 months of age remarkably diminished the thymic weight to about one-tenth of age-matched controls at 16 months of age. These results suggest that oestrogen can actually retard the onset of thymoma in spite of genetic control of its incidence. However, oestrogen did not cause thymic involution when it was injected into rats over 9 months of age. Immunohistochemically, there seemed to be no distinct difference in distribution of oestrogen-receptor-bearing epithelial cells between thymomas and 2- to 3-month-old thymuses. The oestrogen sensitivity of the thymus might be destined to be lost, as the thymic epithelial cells start neoplastic changes with the impairment of oestrogen-receptor function.

Age Factors↗
PubMed1992

A study of mast cells in autoimmune NZB/W F1 mice: possible relationship between mast cells and increased vascular permeability in the thymus of NZB/W F1 mice.

We examined the possible relationship between thymic mast cells and increased vascular permeability in the thymus of autoimmune NZB/W F1 mice. Light-microscopic observation of tissue sections showed that non-autoimmune BDF1 mast cells increased with age. In contrast, autoimmune NZB/W F1 mast cells did not increase in the thymic parenchyma at the age of 9 weeks. However, NZB/W F1 mast cells resumed the age-associated increase from the age of 12 weeks and exceeded the number of BDF1 mast cells at the age of 30 weeks. Blood histamine levels of 9-week-old NZB/W F1 mice were higher than those of BDF1 mice of comparable age. Furthermore, peritoneal mast cells of NZB/W F1 mice were more sensitive to compound 48/80 than those of BDF1 mice. Increased blood histamine levels of NZB/W F1 mice seem to be due to the enhanced histamine release from mast cells. These results suggest a possible correlation between the high histamine levels by degranulation of mast cells and increased vascular permeability in the thymus of NZB/W F1 mice.

Age Factors↗
PubMed1991

Characterization of the rat leukocyte integrin, CD11/CD18, by the use of LFA-1 subunit-specific monoclonal antibodies.

We have attempted to characterize the rat leukocyte integrin, CD11/CD18, by the use of newly generated monoclonal antibodies (mAb) WT.1 (anti-CD11a) and WT.3 (anti-CD18) in conjunction with an mAb, OX42, reactive with a rat integrin-like molecule, with respect to the biochemistry, cellular distribution and function. The conclusion that the mAb WT.1 and WT.3 specifically recognize the rat CD11a and CD18, respectively, was based on: (a) their ability to inhibit homotypic aggregation of splenic concanavalin A (Con A) blasts; (b) sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the antigens recognized; (c) their ability to inhibit binding of Con A blasts to the purified ligand, namely the ICAM-1 antigen and (d) their blocking abilities in mixed leukocyte reaction. In the rat, CD18 has an apparent molecular mass of 95-100 kDa and can associate with at least three distinct alpha subunits of 160-170 kDa (CD11a), 140-150 kDa and 120-130 kDa. The latter two are precipitated by OX42 from M phi but not from unstimulated lymphocytes. They presumably represent the rat CD11b and CD11c, respectively. Rat thymocytes, PBL, thoracic duct lymphocytes, monocytes and neutrophils expressed differential levels of CD11a and CD18. Peritoneal M phi showed virtually no CD11a expression, although CD18 was expressed at levels similar to those seen on blood monocytes, showing an interesting pattern of LFA-1 expression regulation in this cell lineage. Both WT.1 (anti-CD11a) and WT.3 (anti-CD18) apparently recognize a "low-affinity" as well as a "high-affinity" form of LFA-1 and do not discriminate between the two.

Animals↗
PubMed1991

Molecular mechanisms underlying lymphocyte recirculation. II. Differential regulation of LFA-1 in the interaction between lymphocytes and high endothelial cells.

Although it has been suggested that LFA-1 is one of the important molecules mediating interaction between lymphocytes and high endothelial (HE) cells, the implication was based on the observation that lymphocyte binding to high endothelial venules in frozen lymph node sections was partially inhibited by anti-LFA-1 monoclonal antibody at 4 degrees C. However, it has previously been unequivocally demonstrated that LFA-1 molecule is unable to function at this low temperature. To assess the actual involvement of LFA-1 in lymphocyte-HE cell interaction at body temperature, we examined effects of newly developed anti-rat LFA-1 and anti-rat ICAM-1 monoclonal antibody on binding of resting or activated lymphocytes to a rat HE cell line at 37 degrees C. We found that (a) LFA-1/ICAM-1-independent pathway was predominant in the interaction between resting lymphocytes and HE cells, indicating that LFA-1 functions little, if any, in the interaction and (b) lymphocyte triggering through CD3 significantly increased binding inducing a LFA-1/ICAM-1-dependent pathway, whereas phorbol 12-myristate 13-acetate stimulation also enhanced lymphocyte binding but inducing a LFA-1-dependent/ICAM-1-independent pathway. In both cases no apparent alteration in LFA-1 expression was observed, suggesting that the increase in lymphocyte adhesion was not due to quantitative but a qualitative change induced in LFA-1 molecule upon lymphocyte stimulation. These findings suggest that LFA-1 is normally inert but can be "switched" upon lymphocyte stimulation to participate in lymphocyte-HE cell interaction, and that the LFA-1's ligand specificity can be differentially regulated at the lymphocyte-HE venule interface.

Animals↗
PubMed1991

Infection of human CD4+ rabbit cells with HIV-1: the possibility of the rabbit as a model for HIV-1 infection.

Although human T cell surface glycoprotein CD4 is the cellular receptor for human immunodeficiency virus 1 (HIV-1), the introduction of the human CD4 gene into murine cells does not render them susceptible to HIV-1 infection. Here we have established rabbit transfectant cell lines expressing human CD4 on the cell surface and demonstrated that the CD4+ rabbit transfectants could be readily infected by HIV-1 by co-cultivating with a HIV-1-infected human MOLT-4 T cell line (MOLT-4/HIV). Avid syncytia formation was observed upon co-cultivation and the syncytia abundantly produced HIV-1 mature particles, as revealed by electron microscopy. A significant increase of HIV-1 p24 antigen was also detected in the culture supernatant. The syncytia formation was blocked by pretreating the transfectant with anti-human CD4 or by pretreating the MOLT-4/HIV with anti-HIV-1 serum obtained from an infected individual, indicating that the syncytia formed as a result of the interaction of human CD4 on the rabbit transfectant with the HIV-1 envelope protein expressed on MOLT-4/HIV. In contrast, only a very small proportion of the rabbit transfectants expressed HIV-1-specific antigens upon infection with an HIV-1 stock. This may indicate that, although rabbit cells have partially acquired susceptibility to HIV-1 by transfection of human CD4 gene, rabbit cells may further require such a molecule as might be provided by MOLT-4 to become fully susceptible to HIV-1 infection. The possibility of the rabbit as a model for HIV-1 infection is also discussed.

Animals↗
PubMed1991

Clastic cells of Hassall's corpuscles during acute involution of the thymus induced by cyclophosphamide in guinea pigs.

General and histochemical observations of the thymus were carried out in guinea pigs after injection of cyclophosphamide (280 mg/kg). Acute involution of the thymus induced by cyclophosphamide was accompanied by marked enlargement of Hassall's corpuscles in the first week after injection. However, the markedly enlarged Hassall's corpuscles disappeared entirely by the fourth week. Large cells characterized by pale nuclei with one or two prominent nucleoli became aggregated in the enlarged Hassall's corpuscles by the second week. Their cytoplasm frequently was foamy or vesicular in appearance. Histochemical observations revealed strong activities of nonspecific esterase, acid phosphatase and beta-glucuronidase in these cells. Staining for these lysosomal hydrolytic enzymes was evident not only intracellularly but also extracellularly, indicating the dissolution of Hassall's corpuscles by intensive extracellular enzyme release.

Acid Phosphatase↗
PubMed1991

Increased vascular permeability of Brucella abortus bacilli in the thymus of NZB/W F1 mice.

Various amounts of the bacterium, Brucella abortus (BA) were injected intravenously into autoimmune NZB/W F1 mice and non-autoimmune BDF1 mice and then the localization of BA in the thymus was traced using an immunohistochemical method at 30 min and 3 h after injection. The results showed that a greater amount of BA became consistently localized in the thymic parenchyma in a free form or in a phagocytized form in NZB/W F1 mice in comparison with BDF1 mice, indicating a marked increase of vascular permeability in the thymus of NZB/W F1 mice. The extravascular leakage of BA was clearly dose-dependent. The significance of invasion of bacterial antigens from the general circulation into the thymic parenchyma is discussed in relation to autoimmune states.

Animals↗
PubMed1991

Thymic nurse cells (TNC) in spontaneous thymoma BUF/Mna rats as a model to study their roles in T-cell development.

In order to elucidate possible roles of thymic nurse cells (TNC) we isolated them as lympho-epithelial cell complexes from spontaneous thymomas of BUF/Mna rats and characterized them using immuno- and enzyme-histochemical techniques. A remarkable increase in the number of TNC was seen at 8 months of age, immediately before or in accordance with the onset of thymomas. No structural abnormality in the TNC was detected by light-microscopy compared with those from normal control thymi. Phenotypically, the TNC per se were positive for major histocompatibility complex (MHC) class I, class II, cytokeratin and thymulin, but lacked macrophage markers, indicating their epithelial origin. They also expressed some of the markers for non-epithelial components, such as Thy-1, S100 and peanut agglutinin (PNA). The majority of intra-TNC cells were MHC class 1+, Thy-1+, CD5+, CD4+ CD8+ (double positive) and PNA+, but with some heterogeneity in their phenotype. The intra-TNC cells from thymomas revealed higher proliferation indices than those from control thymi, as assessed by 5-bromo-2'-deoxyuridine (BrdU)-uptake. It was also demonstrated for the first time that, not only in thymoma rats but also in normal control rats, about 15-30% of TNC enclosed macrophage populations within them. These results may suggest that the TNC in BUF/Mna thymomas represent typical TNC populations, but they are hyperactive particularly in their number and nursing capacity, resulting in the unusual increment of thymocytes in the thymomas. This animal model lends itself greatly to studies on the regulatory roles of TNC in T-cell development.

Aging↗
PubMed1990

Localization in the rat spleen of carbon-laden macrophages introduced into the splenic artery: a subpopulation of macrophages entering the white pulp.

Heavily carbon-laden (HC) macrophages, largely derived from the red pulp of the donor spleen, were injected into the splenic artery of recipient rats. Immediately after injection, HC macrophages were found only in the marginal sinus and in the splenic cords. With time after injection, they appeared successively at the periphery of the white pulp, in the deeper white pulp, and finally in and near the germinal centers, suggesting migration of HC macrophages from the marginal sinus towards the germinal centers. The number of HC macrophages in and near the germinal centers reached a peak at 12 h. Most of the HC macrophages in the white pulp were spherical or ovoid in shape with a diameter of 7-11 microns in sections, having an eccentric round or oval nucleus often with a distinct nucleolus and a cap-like or horseshoe-like cytoplasm filled with carbon. When immunostained with monoclonal antibodies against rat macrophage subpopulations, more than 90% of HC macrophages in the white pulp were found to be ED1+2-3-. A population of the same type of macrophages, both in morphology and phenotype, were found in the red pulp of the donor spleen. They were different from the major residents, red pulp scavenger macrophages, which were ED1+2+3- and larger in size and irregular in shape. These results suggest the presence of a distinct subpopulation of macrophages which actively migrate into the splenic white pulp including the germinal centers. A discharge of transferred macrophages from the red pulp to the general circulation is also suggested.

Alkaline Phosphatase↗
PubMed1990

The liver lymphatics as a migratory pathway of macrophages from the sinusoids to the celiac lymph nodes in the rat.

A migratory pathway of macrophages as well as lymphatic communications from the liver to the celiac lymph nodes were studied both macroscopically and histologically. The injection of gelatinized carbon into the porta hepatis revealed a new pathway of the liver lymphatics running independently of the portal vein in addition to the ordinary periportal lymphatics. By obstruction of the efferent lymph flow of the celiac nodes and immunostaining with a monoclonal antibody to lymphatics, perilobular lymphatic vessels in the portal tract was readily demonstrated. It was suggested that heavily carbon-laden (HC) macrophages had migrated from the sinusoid into the interlobular connective tissue by 6 h after an intravenous injection of carbon, and then entered the lymphatic vessels of the portal tract. By 9 h to 12 h after carbon injection, HC macrophages started to migrate into the celiac nodes via the two lymphatic pathways. From the marginal sinus in the celiac nodes, they moved into the interfollicular area of the superficial cortex, then accumulated in the paracortex by 12 h to 24 h. They finally ended up in the corticomedullary junction. Migrating HC macrophages showed morphological homogeneity. The liver lymphatic pathway in the rat and a significance for the translocation and function of migrating macrophages were discussed.

Animals↗
PubMed1990

A new approach for identification of rat lymphatic capillaries using a monoclonal antibody.

In order to discriminate rat lymphatics from blood vessels on tissue cryosections by immunohistochemistry, a mouse monoclonal antibody (B27) was produced by immunization of mice with a homogenate of rat thoracic duct. B27 strongly recognized endothelial cells of almost all rat lymphatics, such as lacteals, lymphatic capillaries in the diaphragm at sites of absorption from the peritoneal cavity, collecting lymphatics and the thoracic duct. Besides the lymphatics, B27 reacted with the endothelium of some types of blood vessel, the mesothelium of the peritoneal cavity and substances between smooth muscle fibers. A new double immunostaining technique was then developed to distinguish the lymphatic capillaries, having no or only scanty basement membranes more clearly from the blood vessels. Cryosections were stained first with an anti-type IV collagen antibody for basement membranes, secondly with B27 for the endothelium, and then examined by either light microscopy or fluorescence microscopy. With this technique, the lymphatic capillaries were easily distinguished from other vessels by their positive reaction with B27 alone. B27 appears to be very useful for the simple and reliable identification of rat lymphatics, particularly lymphatic capillaries, in various tissues when applied for double immunostaining.

Animals↗
PubMed1990

A novel monoclonal antibody, Mar 1, directed specifically against mononuclear phagocyte system cells in rats.

Three different monoclonal antibodies (mAb), designated Mar 1, Mar 2, and Mar 3, recognizing three distinct novel antigen molecules expressed preferentially in rat macrophages, were produced by the hybridoma technique. Binding of these mAb to isolated cells or fixed cells was detected by radioactive binding assay, immunohistochemical technique and flow cytometry. Mar 1 binds specifically to the cells constituting the mononuclear phagocyte system (MPS), but not to granulocytes nor endocytosis-positive cells from non-lymphoid tissues. Mar 2 and Mar 3 recognize both the former and the latter. The isotypes of Mar 1, Mar 2 and Mar 3 were defined as IgG1, IgG1 and IgG2b, respectively. These mAb were species specific, allo-non-specific and not cytotoxic for rat peritoneal macrophages. Immunoelectron microscopic observation demonstrated that Mar 1-3 antigens are located on both surface membrane and cytoplasmic membrane structures of peritoneal macrophages, particularly on the limiting membrane of phagocytic small vesicles and large phagosomes. Immunoprecipitation experiments demonstrated that the apparent molecular weights (MW) of the reactive antigens of Mar 1, Mar 2 and Mar 3 are 95,000, 100,000 and 55,000 and 27,000, respectively. These findings indicate that all of Mar 1-3 mAb have considerable value in the identification of rat phagocytes and that, of the three kinds of antigens detected with Mar 1-3, Mar 1 antigen is a specific marker for identification of the cells constituting the MPS and may offer the means to assess the functional capability and differentiation process of the macrophage populations.

Animals↗
PubMed1990

Characterization of intrathymic and extrathymic T cell development in spontaneous thymoma Buffalo/Mna rats.

The developmental status of T cell lineages at prethymic, intrathymic and postthymic stages in spontaneous thymoma Buffalo/Mna (BUF/Mna) rats was characterized on the basis of surface phenotypes and some immune responses. The proportion of bone marrow cell populations was shown to be normal in thymoma rats by immunofluorescence flow cytometry (FACS). In spite of a cortex-predominant appearance of thymomas, the proportion of thymocyte populations was phenotypically normal as assessed by FACS. Double immunohistochemical stainings revealed that a substantial number of single-possible (CD4+CD8- or CD4-CD8+) thymocytes existed not only in the narrowed medullary areas but also in the enlarged cortical areas of the thymomas. In lymph nodes, the proportion of T cells increased with age, reaching 88% after 20 months of age. This increase was due mainly to an increase in the CD8+ population but not the CD4+ population, resulting in low CD4/CD8 ratios. An abnormal increase of Thy1+ immature T cells was also observed in the lymph nodes. However, these phenotypic changes in the T cell lineages in the thymoma rats were not so influential as to alter their immunological reactivities, such as the primary antibody response to a T-dependent antigen, the graft-versus-host reaction and the mixed lymphocyte reaction to allo-antigens. These results suggest the possible presence of some altered differentiation pathways for intrathymic and postthymic T cell development in BUF/Mna rats.

Animals↗
PubMed1990

IL-2 receptors on rabbit T-cell lines and their transfectants expressing the human IL-2 receptor alpha chain.

Low-affinity (dissociation constant: Kd = 7 nM) and high-affinity (Kd = 27 pM) interleukin-2 receptors (IL-2R) were detected on rabbit T-cell lines by IL-2 binding studies. Chemical cross-linking studies using 125I-labelled IL-2 showed that rabbit low-affinity IL-2R was singly expressed alpha-chain (MW 55,000) and that high-affinity IL-2R was composed of at least alpha- and beta- (MW 75,000) chains, similar to the human and murine counterparts. The existence of an additional chain (MW 25,000) was suggested in the rabbit IL-2R. Rabbit T-cell transfectant lines were established by human IL-2R alpha-chain (IL-2R alpha) cDNA transfection. These transfectant lines possessed not only extremely large numbers of human IL-2R alpha (over 10 times more than endogenous rabbit alpha-chain) but also twice as many high-affinity sites as their parental lines. The number of high-affinity sites on the transfectants significantly decreased when human alpha-chains were blocked, indicating that these transfectants expressed high-affinity receptor consisting of the exogenous human alpha-chain and rabbit beta-chain. This was confirmed by cross-linking experiments. The observation that expression of extremely large numbers of exogenous alpha-chains lead to an increase of the total number of high-affinity sites in the apparent absence of an increase of beta-chain expression raises the possibility that not only the beta-chain but also the alpha-chain may play an important role in regulating the number of high-affinity receptors.

Animals↗