Immunological cell situation in the skin of atopic model mice.

BACKGROUND: We observed nishikinezumi, cinnamon-coloured (NC)/Fujita (F) mice aged between 5 and 28 weeks. These NC mice have skin eruptions that resemble human atopic dermatitis (AD) under conventional circumstances. OBJECT: We investigated the skin of eruptive and non-eruptive lesions in NC/F mice by using haematoxylin-eosin (H&E) staining, toluidine blue staining and immunohistopathological study with immunoglobulin (Ig)EepsilonRI, CD23, interleukin (IL)-4, IL-5, interferon (INF)-gamma and Ia antigen. RESULTS: Histological examination of the eruptive lesions revealed the perivascular infiltration of many lymphocytes and mast cells into the upper dermis. Intracellular oedema of the epidermis, lymphocyte infiltration into the epidermis and liquefaction degeneration of the basal layer were also observed. The numbers of IL-4 and IL-5 positive cells in the eruptive lesions were larger than those of the non-eruptive lesions. IL-4 and IL-5 positive cells in the eruptive lesions increased weekly. Some IFN-gamma positive cells were observed in the eruptive lesions after 21 weeks. IFN-gamma positive cells were scarce in the skin of both the non-eruptive and eruptive lesions before 21 weeks. Serum IgE increased from 7 weeks to 21 weeks. DISCUSSION: We confirmed that these findings indicated that T helper (Th)2-dominant immunological activation transformed to a Th1-dominant situation. Many IgEepsilonRI positive cells were recognized in the dermis of the eruptive lesions by the time IgE had decreased. We assumed that the dermatitis before 21 weeks was an IgE-mediated allergy. We have previously reported that older NC/F mice had positive patch-test reactions to mites. Because serum IgE decreased after 21 weeks, dermatitis after 21 weeks might be associated more with cell-mediated delayed hypersensitivity than with IgE-mediated immediate allergy.

Male siblings with dyserythropoiesis, microcephaly and intrauterine growth retardation.

Male siblings with intrauterine growth retardation, hydrops, mild liver dysfunction, chronic diarrhoea, failure to thrive and microcephaly are reported. In both patients, the intrauterine growth retardation was detected in the second trimester of pregnancy. Relatively severe early onset neonatal jaundice, microcytosis, anisocytosis and abnormal iron metabolism were also seen. Bone marrow examination in the second sibling showed marked ringed sideroblasts and multilobulated erythroblasts in late developmental stages. The brain was very small with enlarged cerebrospinal fluid space, a reduced number of gyri and a thin cortex. The clinical and laboratory findings in these patients appear to be unique.

A case of AFP-producing early gastric carcinoma with rapid growth liver metastasis.

An 84-year-old man presented with complaints of epigastric discomfort. Upper gastrointestinal series and endoscopy showed an elevated lesion at the posterior wall of greater curvature on the gastric fundus. Diagnosed as moderately differentiated tubular adenocarcinoma by biopsy, wedge resection and 4sa regional lymph node dissection were carried out. The tumor morphology showed type I with slight elevation, 2.5 x 1.7 cm in size; histological showed papillary, tubular, and solid formations having clear cytoplasm and large bizarre nuclei invading the deep submucosal layer (sm2). This case was evaluated as T1(sm) N0 M0 stage Ia early gastric cancer. In the 5th month after operation, multiple liver metastases were detected. He died of liver failure by rapid growth of metastatic tumors in the 6th month after operation. The serum alpha-fetoprotein level at recurrence was 1,900 ng/mL, and alpha-fetoprotein-positive cells were immunohistochemically detected in operative and liver biopsy specimens.

Primary malignant melanoma arising from the base of the tongue.

A rare case of malignant melanoma of the base of the tongue in a 65-year-old Japanese woman is described. The primary and metastatic tumors were treated by surgery, chemohormone and immunotherapy. There was no recurrence for 5 years after the initial diagnosis. Electron microscopy suggested that the melanosomes of the melanoma cells in the cervical lymph nodes were less mature than those in the primary melanoma cells; melanosome immaturity was consistent with a more aggressive quality.

Increased activity and intranuclear expression of phospholipase D2 in human renal cancer.

We examined the PLD activities of human renal cancers and found that the PLD2 activity was greatly elevated in almost all cases examined as compared with the adjacent normal region. Western blot analysis showed the increased levels of PLD2 protein, but the PLD1 was not discernible. The oleate-dependent PU) activity was very low but appeared to increase in most cases. Interestingly, the immunohistochemical observations indicated the high expression of PLD2 in the nuclei of clear carcinoma cells. This is the first demonstration which suggests the possible involvement of PLD2 in tumorigenesis of renal cancer.

Basic study on gene therapy of human malignant glioma by use of the cationic multilamellar liposome-entrapped human interferon beta gene.

For gene therapy of human malignant glioma, we adopted positively charged multilamellar liposomes entrapping the human interferon beta (hIFN-beta) gene. One week after the transplantation of human malignant glioma U251-SP cells to produce glioma in nude mouse brain, the liposomes entrapping the gene (500 ng of DNA per 25 nmol of lipids per 2 microl) were injected into the same site of the cell transplantation once every second day for a total of five injections; and by this means the tumor completely disappeared. To confirm the antiproliferative effect of hIFN-beta, we performed an in vitro study using a plasmid containing a secretion signal sequence-deleted hIFN-beta gene and one containing the hIFN-beta gene inserted in reverse. In both cases, there was no hIFN-beta release into the medium and no growth inhibition effect. On addition of anti-hIFN-beta antibody to the medium, the growth inhibition effect was abolished. As this cell line expresses IFN-alpha/beta receptor, the hIFN-beta produced in the transfected cells could be released and acted in a paracrine manner. For 120 days the body weight change of normal mice treated by the same procedure as used in the curing experiment was not significant among the groups injected with empty liposomes, plasmid only, and liposomes entrapping the gene. In all of these three groups, death, abnormal behavior, and significant histological changes were not observed.

Paracortical hyperplasia of superficial lymph nodes in a new mutant strain of hairless rats (ISh): a lesion similar to human dermatopathic lymphadenopathy.

The dermal histology and the regional draining superficial lymph node of a new mutant strain of hairless rats (ISh) were investigated. The homozygote ISh rat was characterized as having naked and wrinkled skin. The comedo-like cysts in the skin resembled human acne and vulgaris. Histopathologically, many Langerhans' cells positive for anti-protein kinase C type II antibody (PKC-II) were recognized in the epidermis. The superficial lymph nodes were significantly larger than those of the heterozygote. The paracortex of these lymph nodes was expanded and the number of Langerhans' cells mainly increased in these areas. These lymph node lesions were similar to those of human dermatopathic lymphadenopathy. The ISh rats should be a very useful animal model for studying dermatopathic lymphadenopathy in humans. Furthermore, they may be a valuable animal model for investigating the mechanisms of not only maturation, movement and migration of Langerhans' cells, but also of their function for antigen presentation.

Acute leukemia with the phenotype of a natural killer/T cell bipotential precursor.

An acute leukemia with an unusual immunophenotype developed in a 17-year-old girl. At the initial presentation, extramedullary involvement was not evident, but with advancing disease, massive splenomegaly and an osteolytic rib tumor developed. The disease was aggressive and refractory to intensive chemotherapeutic regimens for myeloid and lymphoid malignancies, and the patient died 3 months after the initial presentation. The leukemic cells were of irregular shape and variable size; they had deeply indented or bi-lobed nuclei and relatively fine, azurophilic granules in their cytoplasm. They were positive for acid phosphatase and beta-glucuronidase in granular staining, but they were negative for myeloperoxidase. The leukemic cells had a unique immunophenotype: it was positive for T-cell antigens (CD1a, CD2, cytoplasmic CD3, CD4), myeloid antigens (CD13 and CD33), NK-cell antigen (CD56), CD19 and CD30. DNA analysis revealed no gene rearrangement in the T-cell receptor beta, gamma and delta, or immunoglobulin heavy chain genes. The leukemic cells of our patient are thought to have arisen from the transformation of a putative precursor cell common to both the T- and NK-cell lineage in the bone marrow. The current literature on precursor NK-cell malignancy is reviewed, and its clinicopathological feature is discussed.

An effective method for suspicious cases in urinary cytodiagnosis by the combined use of scanning and transmission electron microscopy following light microscopy.

In urinary cytodiagnosis, pathological characterization of atypical cells is sometimes difficult because cells in urine samples tend to be degenerated. To overcome this problem, we adopted serial examination under light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Five patients with transitional cell carcinoma primarily developed in the bladder, three patients with dysplastic cells, and one patient with metastatic and infiltrative colorectal carcinoma were subjects of the present study. Sample cells were smeared on a film slide especially suitable for serial use (a film slide used in X-ray diagnosis), fixed in 1% glutaraldehyde (GA) for 30 min, stained with Papanicolaou (Pap) staining, and immediately processed for fixation in 2% paraformal-dehyde (PA) and 2.5% GA. Slides were postfixed in 0.5% osmium tetroxide. Then, cell samples were subjected to a series of observations under LM, SEM, and TEM. One of three patients in whom dysplastic cells were suggested under LM was finally diagnosed as transitional cell carcinoma by subsequent examination under SEM followed by TEM. Thus, serial use of LM, SEM, and TEM proved useful in discriminating transitional cell carcinoma (grade 1) from dysplastic cells and also in distinguishing metastatic tumor cells from primary tumor cells in urine samples.

Presence of nuclear grooves in endometrial cytology.

Although the presence of nuclear grooving in papillary carcinomas of the thyroid has been well-described, so far the attention paid to similar structures in endometrial cell samples has been limited. In order to investigate the occurrence of nuclear grooves in endometrial specimens from patients with various pathologic conditions of the endometrium, we compared their appearance with papillary thyroid cancers. A total of 10 cases was studied (age range, 40-72 yr), all cases demonstrating nuclear grooves. In each case, 10 random high-power fields (HPFs) were investigated, and the numbers of fields in which nuclear grooving could be seen were recorded. Nuclear grooves were observed in 3-36 of each HPFs and were more often observed in atypical endometrial cells than in their normal-appearing counterparts; however, these nuclear alterations were thought to be nonspecific findings.

Isolation and expression of rat interferon beta gene and growth-inhibitory effect of its expression on rat glioma cells.

Rat interferon beta (IFN beta) was cloned from a part of rat genomic DNA by the polymerase chain reaction (PCR) with reference to the nucleotide sequence data of a mouse IFN beta. The sequence result showed that the clone had 555 bp of open reading frame (ORF) which encoded a 184-amino acid polypeptide. Comparison of our data with those of mouse and human IFN beta revealed 86.7 and 67.6% homology at the nucleotide level, and 76.2 and 48.9% homology at the amino acid level, respectively. This ORF was inserted into an expression vector, pCAGGS, and COS-1 cells were then transfected with the plasmid encapsulated in cationic multilamellar liposomes. From the results of reverse transcription-PCR and cytopathic activity assay, this ORF expressed IFN beta in COS-1 cells. When the cultured cells of rat glioma cell line T9 were transfected with the plasmid, their growth was markedly suppressed due to the expression of rat IFN beta.

Studies on intranuclear inclusions and nuclear grooves in papillary thyroid cancer by light, scanning electron and transmission electron microscopy.

OBJECTIVE: To successively examine intranuclear inclusions and nuclear grooves in the same papillary thyroid cancer specimens using a light microscope (LM), scanning electron microscope (SEM) and transmission electron microscope (TEM). STUDY DESIGN: We stained cells by the Papanicolaou method after fixation in 1.25% glutaraldehyde for LM and then attempted to observe them successively by SEM-TEM after fixation in 2% paraformaldehyde and 2% osmium tetroxide. RESULTS: On SEM, intranuclear inclusions were observed as elevated parts, like hills, and nuclear grooves were observed as deep fissures or shallow cracks, sometimes with a few in one cell. On TEM, both intranuclear inclusions and nuclear grooves seemed formed by the nuclear membranes. Intranuclear inclusions also possessed cytoplasm and/or cytoplasmic organelles within some expanded areas in the nuclear grooves. CONCLUSION: It was evident from our three-step technique that intranuclear inclusions and nuclear grooves were essentially the same structures.

Do epidermal Langerhans cells, migrating from skin lesions, induce the paracortical hyperplasia of dermatopathic lymphadenopathy?

In the present study, immunohistochemical and immuno-electron microscopic techniques were used to differentiate Langerhans cells (LC) from interdigitating cells (IDC) in the lymph nodes (LN) of dermatopathic lymphadenopathy. The majority of the dendritic cells that existed in the LN of dermatopathic lymphadenopathy were positive for OKT-6 (CD 1a) antibody. It was concluded that these dendritic cells were not IDC, but LC. Electron microscopically, LC in these LN contained a few Birbeck granules (BG). In order to prove the fact that these dendritic cells were LC, the existence of BG was investigated ultrastructurally by examining serial sections, and immunoelectron microscopically for CD 1a positive cells. Most of the LC in the lymph nodes we examined were negative for the anti-proliferating nuclear antigen (PCNA) antibody. This finding may mean that LC in the LN are fully developed cells and do not divide in the LN. Langerhans cells may migrate from the skin lesions to the paracortical areas in the LN, which then may become enlarged.

Cellular differentiation and development of pyloric mucosal metaplasia in the human gall-bladder.

The development of pyloric mucosal metaplasia (PMM) with regard to cellular differentiation in the human gallbladder was studied by mucin staining (paradoxical concanavalin A (Con A), galactose oxidase-Schiff (GOS) and alcian blue (pH 2.5) PAS (AB-PAS) and immunohistochemistry (pepsinogen II (PgII) and SH-9, and proliferating cell nuclear antigen (PCNA). PMM was divided into three stages of development by three-dimensional (3D) computer graphic reconstruction analysis. In the early stage, a transitional zone of PCNA positive cells was observed between areas of SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells in flat monolayered epithelium. In the middle stage, shallow pits became apparent as areas enlarged, with these becoming deeper in the advanced stage, whereby SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells were observed at the upper and lower portion of the pits, respectively, with PCNA-positive cells forming a narrow zone between the two cell populations. Consequently, the structure of PMM gradually resembles that of the normal gastric pyloric mucosa.

Familial macrothrombocytopenia with unusually elongated mitochondria.

We report a familial case of macrothrombocytopenia without inclusion bodies in polymorphonuclear cells or any congenital abnormalities. The results of the hemostatic and platelet function tests were all normal except for the platelet retention rate. The number of megakaryocytes increased slightly and some were relatively small. Electron microscopic studies revealed a unique morphological abnormality of the platelets' mitochondria.

Differentiation of Langerhans cells from interdigitating cells using CD1a and S-100 protein antibodies.

The present study shows that Langerhans cells can be differentiated from interdigitating cells at the light microscopic level. Superficial lymph nodes and skin taken from necropsies and the lymph nodes of dermatopathic lymphadenopathy (DPL) were used for this experiment. Sections of lymph node and skin were embedded using the acetone, methyl benzoate and xylene (AMeX) method and dendritic cells were immunostained with anti S-100 protein antibody (S-100, and OKT-6 (CD1a) using the restaining method. Langerhans cells in the skin were positive for both CD1a and S-100. Dendritic cells positive for both CD1a and S-100, and dendritic cells positive for S-100, but not for CD1a were observed in superficial lymph nodes. In normal superficial lymph nodes, there were more interdigitating cells than Langerhans cells. The majority of the dendritic cells in the DPL were Langerhans cells. We conclude that the S-100 and CD1a positive cells are Langerhans cells, and the S-100 positive-CD1a negative cells are interdigitating cells.

A comparative study on Langerhans cells in lymph nodes with dermatopathic lymphadenopathy and histiocytosis X cells.

It was elucidated that the majority of DC were LC which were positive for CD 1a, but negative for PCNA, and possessed BG in the lymph nodes with DPL. On the other hand, HCX cells were almost always positive for PCNA. From this point of view, it can be speculated that LC in the lymph nodes of the DPL are non dividing mature cells and migrate from the skin lesion. HCX cells which were positive for CD 4 may be more immature cells than LC in DPL, and may be pathological cells which can divide in the foci.

Comparative studies of the same adenocarcinoma cells, macrophages, and mesothelial cells by light microscopy, scanning electron microscopy, and transmission electron microscopy.

The identification of cells in body cavities of cancer patients is sometimes difficult to make. In order to make a definite cytological diagnosis, we observed the same cells by using light microscopy (LM)-scanning electron microscopy (SEM)-transmission electron microscopy (TEM). In this study we first stained cells by the Papanicolaou method after fixation in 1% glutaraldehyde for LM, and then attempted to observe them successively by SEM-TEM after fixation in 1% paraformaldehyde and 1.25% OsO4. Our method and procedures in examining successively one and the same cells in body cavity fluids by using LM, SEM, and TEM ensured accurate discrimination among adenocarcinoma cells, mesothelial cells, and macrophages. The results of this study suggest that LM-SEM-TEM may be of diagnostic value in distinguishing among mesothelial cells, macrophages, and adenocarcinoma cells. This method also succeeded in disclosing differences between the ultrastructure of the cell surfaces, and those of the cytoplasm, and of the nuclei. It is desirable that LM-SEM-TEM observation can be introduced into various aspects in order to obtain an improvement in the diagnosis by cytologic examination, the judgment of therapeutical effects, drug selection, and prognostic presumption.

Migration and maturation of Langerhans cells in squamous metaplasia of the rat trachea induced by vitamin A deficiency.

The migration and maturation of Langerhans cells (LCs) in rat tracheal squamous metaplasia due to vitamin A deficiency were investigated immunohistochemically and electron microscopically. In the early stage of metaplasia, i.e. basal cell hyperplasia, no LCs with Birbeck granules (BGs) could be found, but there were desmosome-free cells which had the morphological characteristic of immature LCs. They were clearly different from inflammatory cells such as macrophages and lymphocytes, and were, therefore, considered to be precursors of LCs. In the stage of stratification, small numbers of Ia- and protein kinase C type II (PKCII)-positive cells were recognized. Ultrastructurally they were immature LCs with ovoid nuclei, many free ribosomes and few dendrites. The cytoplasm was dark and a few BGs and atypical granules (AGs) could be seen in the Golgi area. In the early stage of cornification, LCs with partially intended nuclei, prominent nucleoli and well-developed Golgi complexes were found. There were many BGs and AGs and structures transitional between them in the Golgi areas. In epithelium showing mature squamous metaplasia, many Ia- and PKCII-positive dendritic cells could be seen. Most of these were typical mature LCs with lobulated nuclei, clear cytoplasm and prominent dendritic processes. The number of BGs and AGs were fewer than in the LCs found in the early stage of cornification, and these granules were distributed throughout the cytoplasm. In the final stage, where the basal cells had differentiated into a flatter epithelium, few LCs could be seen.(ABSTRACT TRUNCATED AT 250 WORDS)