Yatsuki Aratake

Coexistent anaplastic and differentiated thyroid carcinoma : An immunohistochemical study

The aim of the present study was to clarify the underlying molecules that might contribute to the highly aggressive behavior of anaplastic thyroid carcinoma. We selected 5 cases of anaplastic thyroid carcinoma that had a differentiated area to determine differences in the molecules of undifferentiated and differentiated cancer cells. We immunohistochemically examined the localization of nuclear antigen (Ki-67), proliferating cell nuclear antigen (PCNA), p53, apoptotic protease-activating factor-1 (Apaf-1), CD26, galectin-3, E-cadherin, and CD147. We found increased Ki-67, PCNA, and p53 labeling indices; decreased levels of Apaf-1, CD26, galectin-3, and E-cadherin; and overexpression of CD147 in the undifferentiated area compared with the differentiated area. These findings indicate high proliferative properties, suppression of apoptosis, disruption of cell-cell interaction, and induction of matrix metalloproteinases in the undifferentiated areas. Thus the molecules examined might be useful for evaluating the aggressive nature of this tumor and the prognosis.

Dipeptidyl peptidase IV (DPP IV/CD26) staining predicts distant metastasis of ‘benign’ thyroid tumor

Because follicular thyroid carcinoma is extremely difficult to diagnose, several cases were encountered which have been rediagnosed as carcinoma due to distant metastasis. In the search for a method of correctly diagnosing ‘benign’ thyroid tumor, dipeptidyl peptidase (DPP) IV immunostaining was applied to 10 cases whose diagnoses had been corrected to follicular thyroid carcinoma because of distant metastases. The positive rate of immunostaining using paraffin sections in the rediagnosed follicular thyroid carcinoma group (7/10) was much higher than that of the control group (1/29), which consisted of 15 cases of follicular thyroid adenoma and 14 cases of nodular hyperplasia. These results suggested that pre‐ or postoperative DPP IV staining is useful for predicting distant metastasis of ‘benign’ thyroid tumor.

Expression of functional Fas antigen on adult T-cell leukemia

The expression of Fas antigen was analyzed in the peripheral blood mononuclear cells of 12 patients with adult T-cell leukemia (ATL) by flow cytometry. The induction of apoptosis in these cells by adding an anti-Fas antibody and the expression of activated T-cell surface antigens, CD25 and CD26, were also studied. It appears that the cells in ATL expressed a significantly larger number of Fas antigens than those in normal subjects (p < 0.01) and their fluorescent intensity was also shown to be much stronger in ATL (p < 0.01). The large number of ATL cells showed apoptosis in a short-term culture in the presence of the anti-Fas antibody. There was no difference in the expression of Fas antigen among ATL cells with different phenotypes of CD4+/CD8-, CD4-/CD8- and CD4+/CD8+ as well as with clinical subtypes of ATL. Interestingly, the expression of Fas and CD26 antigens showed a negative correlation (p < 0.01, r = 0.78). The strong expression of functional Fas antigen in ATL leads to the impression that anti-Fas antibody could be one of the treatment modalities for ATL which is known to be a very difficult disease to cope with.

Identification of biclonal (duplex) leukaemic cells expressing either CD4+/CD8‐ or CD4‐/CD8+ from a patient with adult T‐cell leukaemia/lymphoma

A 24‐year‐old Japanese woman was admitted to our hospital in 1987 with a chief complaint of skin eruptions, and was diagnosed as having chronic ATLL. In 1993 the leucocyte count increased gradually to 126.0x109/l with 91.5% abnormal lymphocytes expressing two different types of antigenicity, either CD4+/CD8‐ or CD4‐/CD8+. Monoclonal integration of human T‐cell lymphotropic virus type‐I proviral DNA was detected at different sites of the genomic DNA in each cell type. These studies clearly indicate that CD4+/CD8‐ and CD4‐/CD8‐ leukaemic cells originated from two independent clones.

A case of CD4+/CD8- adult T-cell leukemia with good response to interferon-beta terminating as a CD4+/CD8+ adult T-cell lymphoma.

The leukemic cells of adult T-cell leukemia (ATL) usually express the helper/inducer associated antigen reactive with anti-CD4 antibodies but not with anti-CD8. We present a 63-yr-old woman with ATL characterized by circulating leukemic cells with CD4+/CD8- phenotype, hepatosplenomegaly with no lymphadenopathy, and the presence of proviral DNA of human T-cell leukemia virus I in the leukemic cells. She was successfully treated with interferon beta and the remission lasted for 12 months. She then relapsed in the lymph nodes with minimal peripheral blood involvement. The neoplastic cells of the lymph node now co-expressed CD4 and CD8 antigens indicating that the change in clinical manifestation was accompanied by a phenotypic change of the leukemic cells.

Coexistent Anaplastic and Differentiated Thyroid Carcinoma.

The aim of the present study was to clarify the underlying molecules that might contribute to the highly aggressive behavior of anaplastic thyroid carcinoma. We selected 5 cases of anaplastic thyroid carcinoma that had a differentiated area to determine differences in the molecules of undifferentiated and differentiated cancer cells. We immunohistochemically examined the localization of nuclear antigen (Ki-67), proliferating cell nuclear antigen (PCNA), p53, apoptotic protease-activating factor-1 (Apaf-1), CD26, galectin-3, E-cadherin, and CD147. We found an increased Ki-67, PCNA, and p53 labeling indices; decreased levels of Apaf-1, CD26, galectin-3, and E-cadherin; and overexpression of CD147 in the undifferentiated area compared with the differentiated area. These findings indicate high proliferative properties, suppression of apoptosis, disruption of cell-cell interaction, and induction of matrix metalloproteinases in the undifferentiated areas. Thus the molecules examined might be useful for evaluating the aggressive nature of this tumor and the prognosis.

Clinical significance of autorosette-forming cells in T-cell lymphoma

The capability to form rosettes with sheep erythrocytes (E), antibody-complement-sensitized ox erythrocytes (EAC) and autologous erythrocytes (ARFC), and surface immunoglobulin determinations were studied using 21 lymph nodes and one tonsil with pathologically-proven non-Hodgkins lymphoma and 10 lymph nodes with benign pathology. Fourteen of 22 non-Hodgkins lymphoma patients (64%) had a high incidence of E-rosette formation and they were further differentiated into ARFC-positive and ARFC-negative lymphomas. The clinicopathological findings of the latter were compatible with those of adult T-cell leukemia. ARFC-positive lymphoma was regarded as non-Hodgkins lymphoma of T-cell type and one patient showed lymphoblastic lymphoma with high ARFC counts. ARFC counts were very low in B-cell and non-T, non-B lymphomas. The results from benign lymph nodes were too variable to draw any conclusion, although ARFC counts were relatively high in lymphadenitis and hyperplasia.

Cytology of primary cutaneous Langerhans cell histiocytosis with a malignant phenotype. A case report.

BACKGROUND Langerhans cell histiocytosis (LCH) is a proliferative disorder of Langerhans cells, but the nature of LCH, whether reactive, benign, or malignant and neoplastic, is controversial. We encountered a case of LCH showing a malignant phenotype initially localized in the skin of an elderly woman. Since there is no other report on the cytologic appearance of primary cutaneous LCH or on LCH with a malignant phenotype, we compared the cytologic features of this case with those of benign cases at other sites reported in the literature. CASE A 74-year-old woman presented with a gradually enlarging and partially ulcerated skin lesion expanding both sides of her right hand. On histologic and ultrastructural analyses of surgically resected tissue, we diagnosed the lesion as Langerhans cell histiocytosis originating in the skin. Although the patient had no recurrence or metastases for six months after surgical resection of the primary skin lesion and radiation therapy, the tumor extended multisystemically, and the patient died of multiple organ failure 14 months after the initial diagnosis. CONCLUSION Imprint and scrape cytology of multiple skin lesions six months after surgery was useful in immediately diagnosing the recurrent LCH. The tumor cells had indented, twisted or grooved nuclei, and some had intranuclear inclusions. Immunocytochemically the cells were positive for CD1a and S-100 protein. Numerous eosinophils were seen in the background.

Comparative analysis and characterization of mutated thyroid peroxidases with disturbance expressed on the cell surface

Five mutated thyroid peroxidases (TPO) with varying degrees of disturbance in cell surface expression, probably owing to misfolding, were comparatively analyzed. CHO-K1 cells transfected with these mutated mRNAs expressed TPO protein in 65.6-82.1% of cells in antibody staining, and the TPOs were located in intracellular structures like the nuclear envelope and ER as well as cytoplasmically like wild-type TPO. When cell surface expression was examined, three mutated TPOs, G533C-, D574/L575del-, and G771R-TPOs, were expressed to varying degrees. In contrast, R175Q- and R665W-TPOs were thought not to be expressed on the cell surface, although a vague increment in R175Q-TPO was observed with increasing amounts of mRNA. In the kinetic study, three mutated TPOs having insufficient expression on the cell surface showed delays in decrease at 4 and 8 h after chase, although between 8 and 24 h after chase they decreased rapidly, as did the two other mutated TPOs. In immunoprecipitation by anti-TPO antibody, G533C-, D574/L575del-, and G771R-TPOs exhibited increasing interaction with calnexin. The combined evidence suggested that some of the mutated TPOs with disturbance in cell surface expression, probably owing to misfolding, exhibited the delay in kinetics of newly synthesized protein as a result of increasing interaction with calnexin and that such TPOs could be expressed to some extent on the cell surface.

Increased plasma lactoferrin levels in leucocytapheresis therapy in patients with rheumatoid arthritis

OBJECTIVE The aim of this study was to clarify the mechanism of leucocytapheresis (LCAP) in patients with RA. METHODS Protein profiles of blood samples from two patients with RA obtained via LCAP column inlet and outlet lines were analysed by two-dimensional fluorescence difference gel electrophoresis and mass spectrometry. The lactoferrin (LTF) levels of peripheral and circulating blood samples from seven patients obtained via the LCAP column blood circuit were then determined by ELISA. Peripheral blood samples from 14 patients with RA were exposed to unwoven polyester fibre filters and the LTF level was determined. In addition, morphological changes in neutrophils after exposure to the filter were examined by optical microscopy, electronic microscopy and LTF immunostaining. RESULTS LTF levels were increased in both samples from the LCAP column outlet and peripheral blood at the end of LCAP treatment. Furthermore, peripheral blood samples exposed to the filter revealed a decreased number of neutrophils and an increased level of LTF. Morphological analysis of the exposed neutrophils showed vacuolization of the cytoplasm and degranulation of LTF-positive granules. These data suggest that LTF stored in the granules of neutrophils is released from the neutrophils caught in the LCAP column. CONCLUSION Because LTF has been reported to have multiple anti-inflammatory properties, increased levels of LTF may contribute to the clinical effect of LCAP in patients with RA.