Kokichi Kikuchi

Anisakidae and anisakidosis.

During the 1950s, Ishikura noticed in the fishing town of Iwanai, Hokkaido, an intestinal disease that frequently occurred in winter with clinical and histopathologic characteristics different from those of ordinary ileitis terminalis. He reported eight such cases, which occurred within a 2-month period in 1955, as acute ileitis terminalis showing peculiar pathologic features (1). By 1959, thirty cases of this disease had been reported (2). Ishikura, Kikuchi, and their coworkers considered the lesion peculiar because this regional enteritis (or terminal ileitis) caused severe allergic tissue reaction with extensive eosinophilic infiltration. On histologic examination, cross sections of a nematode-like worm were sometimes found, and the specimens were sent to parasitologists for identification. The parasitologists regarded these structures as roundworm larvae, not realizing that they were Anisakis larvae. Meanwhile Otsuru et al. (3), based on a description by Beaver (4), warned that visceral larva migrans might occur in Japan.

Nasal T-cell lymphoma as a type of so-called "lethal midline granuloma".

Six cases were described in which an initial clinical diagnosis of “rhinitis gangrenosa progressiva” or lethal midline granuloma was made. The histological examinations of their surgical and autopsy specimens proved that their nasologic diseases could all be identified as malignant lymphoma arising from the nasal cavity, showing the general histologic characteristics reported for T‐cell lymphomata derived from peripheral T‐cells. This histologic observation was then confirmed by immunofluorescence studies using various antisera directed toward either human T‐ or B‐cell‐surface antigens. These studies clearly demonstrated that their malignant cells bore human Ly‐l‐like antigen but lacked human TL‐like and la‐like antigens as well as surface‐bound immunoglobulins, indicating their peripheral T‐cell origin. These data may suggest that so‐called “rhinitis gangrenosa progressiva” or lethal midline granuloma contains at least two distinct disease categories, one of which is Wegeners granulomatosis, and the other of which is nasal T‐cell lymphoma as described herein.

Non-Hodgkin's lymphoma of Waldeyer's ring and nasal cavity. Clinical and immunologic aspects

Twenty‐nine cases of non‐Hodgkins lymphoma of Waldeyers ring (W‐NHL) and nasal cavity or paranasal sinus (N‐NHL) were studied for tumor‐surface marker phenotype and histopathologic correlation with clinical features. Immunostaining procedures on tissue sections by using xenoantisera and monoclonal antibodies to human B‐ and T‐cells enabled the authors to demonstrate precise surface marker phenotypes of tumor cells and, moreover, the histologic localization of normal or neoplastic B‐ and T‐cells in preserving the original structure of lymphoid organs or tumor tissues. In 22 cases of W‐NHL, 19 (86%) had B‐cell markers and 3 (14%) had T‐cell markers, whereas 6 of 7 cases (86%) of N‐NHL had T‐cell markers. Tumor cells in T‐cell iymphomas in W‐NHL and N‐NHL reacted with antibodies to peripheral T‐cells except one case of W‐NHL. Rappaport “histiocytic” subtype was heterogeneous with respect to both surface marker characteristics and morphologic features, i.e., seven had B‐cell markers and four had T‐cell markers, and they were all subdivided into “large cell” or “large cell, immunoblastic” in Working Formulation and “large cell” or “pleomorpphic” in Lymphoma Study Group classification. The actuarial survival curve for all T‐cell lymphoma patients was characterized by a rapid initial decline and a subsequent plateau, which contained two of the long survivors. In contrast, the B‐cell lymphoma group had a more graded decline. The median and actuarial survivals of the T‐cell lymphoma group were far inferior to those for the lymphoma group that expressed B‐cell markers.

β-Catenin gene mutation in human hair follicle-related tumors

β‐Catenin, a multifunctional protein related to the adherens junction and to signal transduction, is a key molecule of cell proliferation, and it is central to epithelial architecture, regulating the polarity of cells and tissues. β‐Catenin stabilization may play a key role in epidermal signaling leading to hair development, and its aberrant activation may be implicated in formation of hair tumors. Several investigators have shown that pilomatricomas are frequently associated with β‐catenin mutation. In this study, we confirmed β‐catenin gene (CTNNB1) mutation in human pilomatricomas (100% frequency) from which adequate DNA could be obtained for gene analysis. A novel mutation, D32N, was found in one case of pilomatricoma. A preliminary immunohistological study revealed prominent β‐catenin staining in basophilic cells of pilomatricomas, especially in nuclei. Benign tumors which were considered to be derived from hair matrix or hair follicles, and other benign skin tumors, were also investigated. β‐Catenin mutations were not detected in any of the these tumors. These results seem to indicate that hair matrix cells are key players in hair development. Investigation into gene abnormalities of hair‐follicle tumors may elucidate the cause of their neoplastic transformation, and may provide a suggestion for the mechanism of hair development.

Immunohistologic detection of lymphocyte subpopulations infiltrating in human oral cancer with special reference to its clinical significance

Cancer tissues from 30 patients with squamous cell carcinoma of the oral cavity were examined immunohistopathologically as to the responsiveness of the host against its own cancer cells in both biopsy and surgically resected specimens from the same patients. Subpopulations of the infiltrating lymphocytes in cancer tissues were identified on paraffin‐embedded serial sections by a modified indirect immunoperoxidase technique (PAB method) in which it was combined with peroxidase‐antiperoxidase (PAP) complex and avidin‐biotin system with rabbit anti‐human B‐cell, peripheral T‐cell sera. Macrophages were also identified by nonspecific acid esterase staining. T‐cells were predominant over B‐cells in 26 of 30 tissues in biopsy specimens and 23 of 30 in surgically resected specimens with bleomycin treatment. T‐cell infiltration in the peripheral region of the tumor was more prominent than that in the stroma among the cancer nests. T‐cells surrounded the cancer nests, occasionally accumulated around the cancer cells, infiltrated at the marginal part of the cancer nests, and frequently produced perivascular massive accumulations. B‐cells and macrophages, on the other hand, were almost absent or negligible around cancer tissues. The grade of T‐cell infiltration, especially in biopsied specimens, was correlated well to the size of the tumor, and also more marked significantly in patients without cervical lymph node metastasis than in those with lymph node metastasis. Furthermore, there was a significant correlation between the grade of T‐cell infiltration at the peripheral region of the invading cancer mass in initial biopsy specimens and the clinical tumor regression rates with bleomycin treatment, but it did not correlate to the surgically resected specimens. These facts suggest that T‐cells might inhibit the development and spreading of the cancer cells, and that the T‐cell infiltration correlates with the clinical course or prognosis of the oral cancer patients.

Allozyme and morphological identification of shape Anisakis, Contracaecum and Pseudoterranova from Japanese waters (Nematoda, Ascaridoidea)

Allozyme markers were used to identify anisakid nematodes from marine Japanese waters, morphologically assigned to three species complexes: Anisakis simplex (s. l.), Contracaecum osculatum (s. l.) and Pseudoterranova decipiens (s. l.). Samples assigned to A. simplex (s. l.) were found to correspond genetically to A. simplex sensu stricto, those of C. osculatum (s. l.) to C. osculatum A. No morphological characters are yet available to distinguish sibling species of these two complexes. As to the P. decipiens complex, two distinct species were detected: the first corresponded to P. decipiens C, previously recovered in the northern Atlantic, the second to P. decipiens D from Japan. The two species are genetically well differentiated, with five of the 19 loci tested showing distinct fixed alleles. Their reproductive isolation was proved by the lack of hybrids or recombinants in sympatric samples recovered from the same definitive host, Erignathus barbatus. P. decipiens D was found to correspond morphologically to Porrocaecum azarasi, previously considered a synonym of P. decipiens. Accordingly, the name Pseudoterranova azarasi (Yamaguti & Arima, 1942) n. comb. is proposed for P. decipiens D. Similarly, P. decipiens C fits in general morphology, type-locality and host with Ascaris bulbosa, also previously considered a synonym of P. decipiens. The name Pseudoterranova bulbosa (Cobb, 1888) n. comb. is proposed for P. decipiens C.

Establishment and Characterization of Human Gastric Carcinoma Lines with High Metastatic Potential in the Liver: Changes in Integrin Expression Associated with the Ability to Metastasize in the Liver of Nude Mice

There is a need to establish animal models which are suitable for investigation of human gastric cancer metastasis to the liver. To this end, a human gastric carcinoma line, AZ521 was injected into the spleens of nude mice. Cells from the few liver metastatic foci of injected AZ521 were expanded in vitro and subsequently injected into the spleens of nude mice. By repeating these procedures three times, we were able to obtain a cell line, designated as AZ‐M3c, with high metastatic potential in nude mice. Liver metastasis developed in 15 of 21 (71%) animals injected with AZ‐H3c, but in only 14% of those injected with parental AZ521. Further, AZ‐H3c caused faster tumor development than did AZ521. However, the primary AZ‐H3c tumors and liver metastatic AZ‐H3c tumors showed essentially the same histological appearance. We also analyzed the cell surface expression of adhesion molecules. The data showed that the expression of VLA‐1, VLA‐2, VLA‐3, VLA‐4, VLA‐5 was enhanced in AZ‐H3c. In contrast, the expression of VLA‐6, αvβ3, E‐cadherin, ICAM‐1 and LFA‐1 was reduced in this high‐metastatic line. These results suggest that β1 mtegrins play an important role in the liver metastasis of human gastric carcinoma cells. Our high‐metastatic line should be useful for studies aimed at the prevention of liver metastasis.

NKT cells in the rat: organ-specific distribution of NK T cells expressing distinct V alpha 14 chains.

Rat invariant TCR α-chains and NKT cells were investigated to clarify whether CD1d-mediated recognition by NKT cells is conserved further in evolution. Rats had multiple-copies of TRAV14 genes, which can be categorized into two types according to the diversity accumulated in the CDR2 region. Rats retained invariant TCRα forms with the homogeneous junctional region similar to mouse invariant TRAV14-J281. The proportion of invariant TCR among Vα14+ clones was 12.9% in the thymus and increased in the periphery, 31% in the spleen and 95% in hepatic sinusoidal cells. The invariant TRAV14-J281 was expressed by liver sinusoidal and splenic NKT cells with CD8, CD44high, and TCR Vβ8. Type 1 invariant TCRα was expressed more frequently in hepatic lymphocytes, while type 2 invariant TCRα was expressed predominantly in the spleen. Both types of cells cytolyzed to and were stimulated to proliferate by CD1d-expressing cells in a CD1d-restricted manner. These results suggested that rat NKT cells bearing distinct Vα14 chains are distributed in a tissue-specific pattern. NKT cell populations in rats were more variable than those in mice, indicating that they play novel roles in nature. The implication of the molecular interaction between the structurally diverse invariant TCRα and CD1d/ligand complex in different organs is discussed.

70-kDa Heat Shock Cognate Protein Interacts Directly with the N-terminal Region of the Retinoblastoma Gene Product pRb IDENTIFICATION OF A NOVEL REGION OF pRb-MEDIATING PROTEIN INTERACTION

Retinoblastoma protein (pRb) functions as a tumor suppressor, and certain proteins are known to bind to pRb in the C-terminal region. Although the N-terminal region of pRb may also mediate interaction with some proteins, no such protein has been identified yet. We demonstrated previously the in vivo protein association between pRb and 73-kDa heat shock cognate protein (hsc73) in certain human tumor cell lines. In this report we analyzed the interaction between these two proteins in vitro. Our data showed that hsc73 interacts with the novel N-terminal region of pRb; that is, pRb binds directly to hsc73 and dissociates from hsc73 in an ATP-dependent manner. By using deletion mutants of cDNA encoding pRb, the hsc73 binding site of pRb was determined to be located in the region (residues 301-372) outside the so-called A pocket (residues 373-579) of this tumor suppressor protein. This finding was compatible with the fact that the adenovirus E1A oncoprotein, which is known to bind to the E2F binding pocket region of pRb, could not compete with hsc73 for the binding. Furthermore, phosphorylation of pRb by cyclin-dependent kinase inhibited the binding of pRb to hsc73. These data suggest that hsc73 may act exclusively as the molecular chaperone for nonphosphorylated pRb. As a result, hsc73 may function as a molecular stabilizer of nonphosphorylated pRb.

Localization of rat CD1 transcripts and protein in rat tissues— an analysis of rat CD1 expression by in situ hybridization and immunohistochemistry

CD1 molecules are cell surface glycoproteins non‐covalently associated with β2‐microglobulin. Recently, functional features of the CD1 family such as a target ligand and an antigen‐presenting structure for T cells have been reported. In the current study, tissue distribution of rat CD1 was analysed by in situ hybridization (ISH) in conjunction with immunohistochemistry to clarify the precise localization of both transcripts and proteins. CD1 transcripts were detected by ISH in a variety of organs: spleen, thymus, liver, lung, heart, kidney, small intestine and skin. In most organs, CD1 immunoreactivity paralleled the amount of CD1 mRNA expression and was localized in the same regions as its gene transcripts. However, there was a clear difference between the level of rat CD1 transcription and protein expression in the small intestine. CD1 mRNA was detected in the enterocytes of crypts of Lieberkuhn, but not in those of the intestinal villi, while immunoreactivity of CD1 protein was observed in the cells of the intestinal villi but not in those of crypts of Lieberkuhn. This suggests that CD1 gene transcription occurs in the enterocytes of intestinal crypts, and that as the cells of intestinal crypts migrate from the crypts to the intestinal villi, CD1 proteins are synthesized and accumulated in the intestinal villi. Such CD1 expression in the enterocytes appears to be consistent with migration‐associated differentiation, and suggests that rat CD1 may take part in mucosal immunity as a first line of defence. In addition, clear cell membrane CD1 immunoreactivity on lymphoid cells raises the possibility that intercellular interaction via rat CD1 and T cell receptors may be involved in both lymphoid cell differentiation and immunoregulation.