• Title/Summary/Keyword: Immunophenotyping

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ZAP-70 Protein Expression in B-cell Chronic Lymphoid Leukemia: a Single Center Experience from Pakistan

  • Zeeshan, Rozina;Irfan, Syed Mohammed;Sultan, Sadia;Bhimani, Sanjana
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1587-1590
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    • 2015
  • Background: Chronic lymphoid leukemia (CLL) is the most frequent type of adult leukemia. The Rai and Binet staging systems have been well recognized as standards for assessing the treatment requirements and overall survival in CLL patients. However, there is a need to seek newer prognostic markers to identify stable or progressive forms of CLL that will facilitate risk-adapted treatment strategies. Currently a molecular biomarker ZAP-70 has attracted interest as providing prognostic information in CLL patients. Objective: To determine the frequency of ZAP-70 positivity in B-CLL patients at disease presentation. Materials and Methods: From January 2011 to September 2014, 89 patients were diagnosed to have chronic lymphoid leukemia. Complete blood count was done on an automated analyzer (Cell Dyne, Abott Architect, USA), while immunophenotyping was conducted for each patient to establish the diagnosis of the disease. ZAP-70 expression was evaluated by flow cytometry. Data were compiled and analyzed by SPSS version 21. Results: Out of the total of 89 B-CLL patients, 62 (69.7%) were male and 27 (30.3%) were females with a male to female ratio of 2:1. The mean age was $57.5{\pm}12.1years$. The frequency of ZAP-70 positivity in our B-CLL patients was found to be 13.5%. ZAP-70 positivity was significantly correlated with stage III disease and high absolute lymphocytic count (P<0.05). No correlation of ZAP-70 could be established with age and gender (p>0.05). Conclusions: The frequency of ZAP-70 in our patients appears low. It is approximately half that in international data. We would recommend to screen all the newly diagnosed patients with CLL for ZAP-70 protein expression for risk stratification, family counseling and to predict overall survival.

A Case of MALT Lymphoma in Parotid Gland Duct (이하선관에 발생한 MALT 림프종 1예)

  • Kim, Ki Yup;Yang, Won Yong;Kwon, Seok Min;Kang, Sang Yoon
    • Archives of Plastic Surgery
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    • v.36 no.5
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    • pp.663-666
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    • 2009
  • Purpose: Primary malignant lymphomas of the salivary glands are uncommon. The parotid gland was most frequently involved, followed by the submandibular gland, minor salivary gland and sublingual gland. The most common subtype is mucosa - associated lymphoid tissue(MALT) lymphoma. We experienced a case of salivary MALT lymphoma involving parotid gland duct, so report a case with a review of the literature. Methods: A 65 year old female presented with a palpable mass on the left side of her cheek. There was no clinical or laboratory evidence of pre - existing autoimmune disease. Preoperative facial and neck CT with contrast showed $2.1{\times}1.7cm$ sized, ill defined, homogeneous low density mass near left masseter muscle, and no evidence of other enlarged lymph nodes. Results: At operation, a yellowish oval shaped mass was found slightly adhered to middle portion of the parotid gland duct, meaduring $2{\times}1.5{\times}0.7cm$. Microscopic finding showed that centrocyte - like cells, monocyte B cells and plasma cells were diffusely infiltrated. Immunophenotyping was preformed on fixed section. The majority of the small cells were immunoreactive for the B cell marker CD20. Based on the typical histological findings supported by immunostaining, the mass was defined as MALT lymphoma. Conclusion: We report that very rare case of MALT lymphoma involving parotid gland duct in 65 year old female patient was experienced with clinical characteristics, histologic features and references.

A Case of Canine Mammary Comedocarcinoma with Regulatory T Cell Infiltration

  • Siwon Jeong;Jiwoong Yoon;Woo-Jin Song;Jongtae Cheong;Young-min Yun;Gee Euhn Choi;Myung-Chul Kim
    • Journal of Veterinary Clinics
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    • v.41 no.4
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    • pp.215-222
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    • 2024
  • An adult female dog was presented for evaluation of rapid growth of mammary gland masses. Complete blood count, serum biochemistry, and diagnostic imaging results were unremarkable. Fine needle aspirates of the mammary masses indicated mammary carcinoma characterized by large globoid cells with finely granular eosinophilic globules or Melamed-Wolinska-like bodies. A regional mastectomy was performed on the masses. Subsequent histopathologic examination of the surgically resected masses resulted in a diagnosis of mammary comedocarcinoma with nodal metastasis and distinct perivascular immune infiltrates, which were subject to immunohistochemical and flow cytometric immunophenotyping. Immunohistochemical examination confirmed the infiltration of CD3+ T and PAX5+ B lymphocytes. Flow cytometric analysis demonstrated tumor-infiltrating CD4+CD25+FOXP3+ regulatory T, CD8+ T, CD11b+ myeloid, and CD21+ B cells. Of note, paired flow cytometric analysis of peripheral blood and tumor tissues showed a preferential tumor infiltration of regulatory T and B cells. Approximately two months after the mastectomy, the tumor reoccurred at the surgery site. The dog died due to deteriorating conditions. We report a rare case of canine mammary comedocarcinoma, providing clinical, clinicopathologic, histologic, and immunophenotypic characteristics. Our case is valuable in providing a rationale for basic research that maps the immune landscape of mammary comedocarcinoma to identify key immune subsets for cancer progression.

A Case of Canine Colorectal Carcinoma In Situ with Regulatory T Cell Infiltration

  • Yunhee Joung;Jiwoong Yoon;Dong Ju Lee;Woo-Jin Song;Jongtae Cheong;Hyunjung Park;Young-min Yun;Gee Euhn Choi;Myung-Chul Kim
    • Journal of Veterinary Clinics
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    • v.41 no.4
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    • pp.207-214
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    • 2024
  • An adult castrated male dog was presented with persistent hematochezia. Digital rectal examination and endoscopy found multiple colorectal masses. Complete blood count and serum biochemical results were within the reference interval. Fine needle aspirate of the masses indicated a diagnosis of inflamed polyps with a primary differential of malignancy. Histopathologic examination using endoscopy-guided incisional biopsy of the masses revealed an inflamed neoplasm with ossification. A colectomy was performed to remove the tumor. Subsequent histopathologic examination of the surgically resected masses resulted in a diagnosis of colorectal carcinoma in situ (CiS) with immune infiltrates, which were subject to immunohistochemical and flow cytometric immunophenotyping. The immunohistochemistry confirmed intraepithelial CD3+ T cells within CiS. The flow cytometric analysis indicated tumor-infiltrating CD4+ T, CD8+ T, and CD11b+ myeloid subsets. The flow cytometric analysis of circulating and tumor-infiltrating leukocytes demonstrated a preferential expansion of CD25+FOXP3+ regulatory T cells within CiS. To the author's knowledge, this is the first report to show clinical evidence emphasizing the immunogenicity and immune-suppressive environment of canine colorectal CiS. Our case will be valuable in providing a rationale for basic research that dissects the immune environment for canine colorectal cancers for the future development of cancer immunotherapy.

Clinical Features and Prognostic Factors in 41 Patients of Primary Nasal/Nasopharyngeal Non-Hodgkin's Lymphoma: Prognostic Significance of Immunophenotype (비/비인강 비호즈킨 림프종의 임상양상과 예후 인자 : 면역 표현형의 임상적 의의)

  • Park Soon-Seo;Park Jong-Beom;Suh Cheol-Won;Park Ji-Woon;Lim Soo-Duk;Huh Joo-Ryung;Nam Soon-Yuhl;Kim Sang-Yoon;Lee Ho-Gyu;Chang Hye-Sook;Kim Tae-Won;Lee Je-Hwan;Kim Sung-Bae
    • Korean Journal of Head & Neck Oncology
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    • v.15 no.2
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    • pp.149-155
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    • 1999
  • Objectives: To study the clinical features of the primary nasal/nasopharyngeal non-Hodgkin's lymphomas and to evaluate the implication of immunophenotyping as a prognostic factor. Patients and Methods: From January 1990 to December 1997,41 patients(median age, 41 years) of primary nasal/nasopharyngeal non-Hodgkin's lymphoma were studied. The clinical records and paraffin-embedded tissue blocks were reviewed retrospectively. The histologic features, immunophenotypic findings(pan-T, pan-B, CD3, CD56) and Epstein-Barr virus in situ hybridizatios were examined. The prognostic factors for clinical outcome were evaluated in these patients. According to Ann-Arbor system, there were 30 patiets(73%) with stage IE, 4(10%) with stage IIE, 3(7%) with stage IIIE, 4(10%) with stage IVE lymphoma. Among the patients with stage IE/IIE, 4 patients received local radiation alone, 4 received chemotherapy alone, 25 received combination chemotherapy and radiotherapy and 1 refused treatment. The patients with stage IIIE/IVE were given combination chemotherapy and radiotherapy. Results: Immunophenotyping were performed in 40 patients and staining results were as follows: 3(7%) patients with B cell, 17(42%) with T cell, 18(44%) with NK/T cell(CD56 positive), and two patients with unclassifiable result. Epstein-Barr(EB) virus in situ hybridization were performed in 28 patients and 23(82%) patients had positive EBV-encoded RNAs(EBERs). 21(55%) patients achieved a complete remission. There was no difference in complete remission between radiation alone and combination therapy. With median follow-up of 30 months, 5-years disease free survival of complete responders was 60% and 5-years overall survival rate was 36%. Multivariate analysis showed that better overall survival was related with absence of B symptoms, ECOG performance${\leq}1$ and non-NK cells. Conclusion: Most of all cases were positive for EBER. Since NK/T phenotype carried the worst prognosis, analysis for CD56 expression should be done. Further prospective studies were warranted to evaluate the role of chemotherapy in stage IE/IIE.

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Isolation and Culture of Human Embryonic Stem-like Cells from Abnormal Blastocysts (비정상 포배기 배아에서 인간 배아줄기 유사 세포의 분리 및 배양에 관한 연구)

  • Lim, Chun-Kyu;Sung, Ji-Hye;Park, Jong-Hyuk;Kim, Sun-Jong;Yoon, Hyun-Soo;Koong, Mi-Kyoung;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.4
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    • pp.293-298
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    • 2003
  • 목 적: 인간의 배아줄기세포는 전분화능과 영속성을 가지고 있어 발생 및 분화에 관련된 기초 연구 뿐 만 아니라 재생의학, 약물검색 등에서도 매우 유용한 재료로 이용될 수 있다.본 연구에서는 유전체의 변형이 배아줄기세포주의 확립 효율에 미치는 영향을 살펴보고자 비정상적인 포배기 배아에서 내세포괴를 분리하여 배양하였다. 연구 방법: 인간의 체외수정 및 배아이식술에서 공여 받은1개 또는3개의 전핵이 관찰되는 비정상 수정란 (n=20)과 착상전 유전진단에서 이수성이 확인된 배아 (n=27)를 대상으로 하였다. 일반적인 immunosurgery 방법으로 영양배엽세포들을 제거하고 내세포괴를 분리한 후 PMEF 혹은 STO feeder 세포위에서 배양하였다. 배아줄기세포의 배양시스템을 검증하기 위해서 이미 확립된 Miz-hES1 cell line을 동시에 같은 조건 하에서 계대배양하였다. 결 과: 비정상 수정란에서 발생된 포배기 배아에서 분리한 1개의 내세포괴가 배아줄기세포와 유사한 colony를 형성하였으나, 계대배양에는 실패하였다. 이수성 배아에서 발생된 포배기 배아의 내세포괴 배양에서는 두개의 colony가 계대배양 중에 영양배엽세포의 형태로 분화되어 미분화 상태를 유지하지 못하였다. 동일한 시기와 조건 하에서 계대배양된 Miz-hES1 cell line이 미분화상태로 유지됨을 karyotyping (46, XY)과 immunophenotyping (positive in SSEA-3 and -4)으로 확인하였다. 결 론: 본 연구의 결과에서 비정상 수정란과 이수성 배아에서 발생된 포배기 배아에서 유래한 내세포괴는 배아줄기세포주 확립 및 미분화 상태 유지 능력이 매우 저조한 것으로 여겨진다. 따라서, 인간의 배아줄기세포주를 확립하는데 있어 배아의 정상여부가 중요한 요소로 작용할 것으로 생각된다.

The Effect of Gefitinib on Immune Response of Human Peripheral Blood Monocyte-Derived Dendritic Cells (인간 말초혈액 단핵구 유래 수지상세포의 면역반응에 미치는 Gefitinib의 영향)

  • Cho, Jin-Hoon;Kim, Mi-Hyun;Lee, Kwang-Ha;Kim, Ki-Uk;Jeon, Doo-Soo;Park, Hye-Kyung;Kim, Yun-Seong;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.6
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    • pp.456-464
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    • 2010
  • Background: Synergistic antitumor effects of the combined chemoimmunotherapy based on dendritic cells have been reported recently. The aim of this study is to search new applicability of gefitinib into the combination treatment through the confirmation of gefitinib effects on the monocyte derived dendritic cells (moDCs); most potent antigen presenting cell (APC). Methods: Immature and mature monocyte-derived dendritic cell (im, mMoDC)s were generated from peripheral blood monocyte (PBMC) in Opti-MEM culture medium supplemented with IL-4, GM-CSF and cocktail, consisting of TNF-${\alpha}$ (10 ng/mL), IL-$1{\beta}$ (10 ng/mL), IL-6 (1,000 U/mL) and $PGE_2$ ($1{\mu}/mL$). Various concentrations of gefitinib also added on day 6 to see the influence on immature and mature MoDCs. Immunophenotyping of DCs under the gefitinib was performed by using monoclonal antibodies (CD14, CD80, CD83, CD86, HLA-ABC, HLA-DR). Supernatant IL-12 production and apoptosis of DCs was evaluated. And MLR assay with $[^3H]$-thymidine uptake assay was done. Results: Expression of CD83, MHC I were decreased in mMoDCs and MHC I was decreased in imMoDCs under gefitinib. IL-12 production from mMoDCs was decreased under $10{\mu}M$ of gefitinib sinificantly. Differences of T cell proliferation capacity were not observed in each concentration of geftinib. Conclusion: In spite of decreased expressions of some dendritic cell surface molecules and IL-12 production under $10{\mu}M$ of gefitinib, significant negative influences of gefitinib in antigen presenting capacity and T cell stimulation were not observed.

Morphological aspects of small intestinal mucosal injury and repair after electron irradiation

  • Grigory Demyashkin;Elza Karakaeva;Siuzanna Saakian;Vladimir Shchekin;Emir Elbuzdukaev;Umar Bamatgiraev;Daniia Ashgalieva;Makka Evsultanova;Daniil Kovalev;Darya Kabanova;Oleg Shatunov;Dmitrii Atiakshin
    • Anatomy and Cell Biology
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    • v.57 no.3
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    • pp.384-391
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    • 2024
  • Morphological evaluation of the small intestine mucosa and apoptosis activity (caspase-3) is necessary to assess the severity of damage to the small intestine. At the same time, proliferative index based on Ki-67 can be used to assess the regenerative potential of the small intestine. Fragments of small intestine of Wistar rats (n=60) of three groups: I) control (n=20); II) experimental group (n=20; local single electron irradiation at a dose of 2 Gy), III) experimental group (n=20; local single electron irradiation at a dose of 8 Gy) were studied by light microscopy using hematoxylin and eosin staining and immunohistochemical reactions with antibodies to Ki-67 and caspase-3. In all samples of the experimental groups, a decrease in all morphometric indices was observed on day 1 with a tendency to recover on day 3. Small intestinal electron irradiation led to disturbances in the histoarchitecture of varying severity, and an increase in cell apoptosis was observed (increased expression of caspase-3 and decrease in Ki-67). In addition, modulation of the PI3K/AKT and MAPK/ERK signaling pathways was detected. The most pronounced destructive changes were observed in the group of 8 Gy single electron irradiation. Local irradiation of the small intestine with electrons at a dose of 2 and 8 Gy results in a decrease in the number of enterocytes, mainly stem cells of the intestinal crypts.