Kenji Kaneda

Distribution and morphological characteristics of the pit cells in the liver of the rat

SummaryPit cells, on which almost no further contributions have been presented since the first report by Wisse et al. (1976), are described in detail in the rat liver. These cells show several characteristic features: 1) “rod-cored vesicles”, a new type of vesicular inclusion observed first in our study; 2) electron-dense granules, which we consider to arise from multivesicular bodies by the accumulation of dense material; and 3) well-developed pseudopodia. Although these features clearly differentiate pit cells from conventional lymphocytes, these two cell types display similarities (i) in a number of ultrastructural features, (ii) in the pattern of their intralobular distribution, and (iii) in their presence in the spleen and peripheral blood.

Parallel antitumor, granuloma-forming and tumor-necrosis-factor-priming activities of mycoloyl glycolipids fromNocardia rubra that differ in carbohydrate moiety: Structure—activity relationships

SummaryMultiple intravenous injections (30 µg, ten times) in ICR mice of trehalose dimycolate and glucose monomycolate fromNocardia rubra, containing C36–48 mycolic acids, showed a prominent antitumor effect on a subcutaneously implanted sarcoma-180, an allogeneic sarcoma of mice with a significant granuloma formation in lungs, spleen and liver. On the other hand, mycoloyl glycolipids other than glucose monomycolate and trehalose dimycolate, such as mannose or fructose mycolate, showed no significant activity for tumor regression or granuloma formation in mice.Trehalose dimycolate and glucose monomycolate fromN. rubra, and glucose monomycolate with C56–60 mycolic acids fromRhodococcus terrae also showed a distinctive priming activity for tumor necrosis factor (TNF), when lipopolysaccharide fromEscherichia coli was administered as an eliciting agent. The TNF activity in the sera of mice was abrogated almost completely by anti-(murine TNFα) antibody with protein-A—agarose. Again in contrast, mannose and fructose mycolate fromN. rubra and glucose monomycolate with C30–34 mycolic acids fromRhodococcus equi did not show such activities in mice.Meth-A, a syngeneic fibrosarcoma of BALB/c mice, was less sensitive to administration of glycolipids than sarcoma-180. These results indicated that the existence of a glucose or trehalose molecule was necessary for the expression of immunomodifying activities among various mycoloyl glycolipids differing in carbohydrate structure. However, since the administration of lipopolysaccharide was essentially required as an eliciting agent for the induction of TNF, while no eliciting agent was required for the antitumor activities, TNF does not seem to contribute directly to the antitumor activities of mycoloyl glycolipids in our systems. There was, however, a parallel structure-activity relationship among granuloma-forming, antitumor and TNF-priming activities, indicating that the structures of both the carbohydrate moiety and the mycoloyl residues influenced an initial step, such as macrophage activation, commonly and profoundly.

Three-dimensional structure of endothelial cells in hepatic sinusoids of the rat as revealed by the Golgi method

SummaryThe three-dimensional structure of endothelial cells in the hepatic sinusoids of the rat was studied by application of light- and electron microscopy on Golgi-impregnated specimens. A number of endothelial cells could thus be individually delineated throughout the hepatic lobules. The cytoplasm, showing heavy silver deposits, consists of two distinct areas, a thick and thin portion. The thick portion, issuing from the region of the perikaryon, branches and tapers toward the cell periphery. The thin portion, occupying the remainder of the cytoplasm, consists largely of highly fenestrated sieve plates. Some intralobular variation can be noted; the thick portion of the endothelial cells is well developed in the periportal zone, while the cells in the centrilobular zone are relatively rich in thin portions. In addition, the area of distribution of an individual endothelial cell is larger in the centrilobular sinusoids than in the periportal zone. Some endothelial cells also possess unique cytoplasmic processes projecting into the intercellular space between hepatocytes and connecting the sinusoidal walls of neighboring sinusoids. These processes may anchor the endothelial cells to the hepatic plates.

Localization and ultrastructure of the Kupffer cells in orthotopically transplanted liver grafts in the rat.

Abstract. Kupffer cells play an important role in the acceptance or rejection of liver grafts. We examined the ultrastructure of the Kupffer cells in transplanted rat livers, from an early to a late stage where the graft is accepted, using a DA‐to‐PVG combination. Two days after surgery, endocytic activity of the Kupffer cells had increased, as evidenced by worm‐like structures and many endocytic vacuoles. There was often close apposition to the monocytes or lymphocytes. By day 4, infiltration of mononuclear cells into the sinusoids was readily noticeable. By day 7, several Kupffer cells had migrated into the space of Disse through the openings in endothelial linings. The number of Kupffer cells reached a maximum at 14 days. They were located mostly outside the sinusoid, adhering to the hepatocytes. At this point in time, however, the Kuppfer cells contained few endocytic vacuoles and phagolysosomes, in contrast to those at 2 days. The number and location of Kupffer cells became almost normal at 2 months. The present results indicate that Kupffer cells are highly activated before mononuclear cell infiltration becomes manifest in the sinusoid, and that when a rejecting reaction reaches a peak, they are usually located extrasinusoidally and show a morphologically immature profile.

Light and Electron Microscopic Analysis of Liver Sinusoids during Hepatocarcinogenesis with 2‐Acetylaminofluorene in Rats

To clarify the sequential changes and morphological differences of the sinusoidal structures between hepatocellular carcinoma (HCC) and hepatocellular adenoma (HA), we examined morphological changes of sinusoidal cells and related structures such as basement membrane during hepatocarcinogenesis in the rat. During continuous feeding of carcinogenic diets containing 2‐acetylaminofluorene to rats, HA appeared at the 8th week in the periportal area and then extended toward the centrolobular area. The appearance of HCC was recognized at the 27th week. In the HA lesion, the morphology of sinusoidal cells and related structures was basically the same as that of normal liver except for a slight thickening of the basement membrane and a decreased amount of vitamin A‐lipid droplets of stellate cells. In HCC, the fenestrations of endothelial cells disappeared and the basement membrane became continuous, thick and often multilayered. Stellate cells contained almost no vitamin A‐lipid droplets and were associated with abundant collagen fibers. Kupffer cells and pit cells were not seen inside the sinusoid. All these features of the sinusoids in HCC resembled the morphological characteristics of the capillary. The present study has revealed that HCC possesses sinusoid structures distinct from those of HA. This suggests that HCC may not derive directly from HA but may develop newly within the HA.