• Title/Summary/Keyword: immunophenotype

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A method of isolation and characterization of canine endometrial-derived mesenchymal stem cells

  • Mi Kyung Park;Kun Ho Song
    • Korean Journal of Veterinary Service
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    • v.46 no.2
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    • pp.157-160
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    • 2023
  • Endometrial tissue is a known source of mesenchymal stem cells (MSCs). We isolated canine endometrial stem cells from canine endometrial tissues using an enzymatic method and confirmed the immunophenotype of mesenchymal stem cells and multilineage differentiation. Canine endometrial tissues were obtained from canine ovariohysterectomy surgery and isolated using 0.2% collagenase type I. We measured the immunophenotype of stem cells using flow cytometry. To confirm the differentiation ability, a trilineage differentiation assay was conducted. In this study, canine endometrialderived MSCs (cEM-MSCs) were isolated by enzyme treatment and showed a spindle-shaped morphology under a microscope. Moreover, cEM-MSCs showed a trilineage differentiation ability. In this study, the canine endometrium was a good source of MSCs.

Metastatic Adenocarcinoma of the Skin - Immunohistochemical Study (전이성 피부 샘암종의 면역조직화학적 고찰)

  • Choi, Keum-Ha;Jang, Kyu-Yun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1041-1048
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    • 2009
  • The development of skin metastasis is usually a morbid prognostic feature although they occur infrequently. Adenocarcinomas account for up to about 70% of all metastatic skin cancer. In general, adenocarcinomas are the most difficult metastatic tumor to accurately identify the primary site because they don't have distinctive histological features. For this reason, immunohistochemistry have been used to help identify the origin of metastatic adenocarcinomas. This study performed immunohistochemical staining with metastatic adenocarcinomas of the skin using a variety of antisera to find out characteristic immunohistochemical findings of them. This study was made upon the 29 cases of metastatic adenocarcinomas of the skin, which had been confirmed histopathologically in Chonbuk National University Hospital from January, 1986 to April, 2006, Paraffin blocks were colledted and homemade tissue arrays were made. We performed immunohistochemical staning using 12 antibodies (MUC1, 2, 5AC, 6, cytokeratin (CK) 7, 20, thyroid transcription factor-1 (TTF-1), estrogen receptor (ER), progesterone receptor (PR), beta-catenin, cox-2, claudin-1). The mean age at the time of diagnosis was 60.7 years and the male to female ratio was 1.2:1.0. The most common primary site was lung, followed by stomach and colorectum. MUC1 was expressed by most colorectal, breast and lung adenocarcinoma. MUC2 was expressed infrequently. MUCSAC was expressed by most gastric and colorectal cancer MUC6 was not specific of any primary site in this series. CK7+/CK20+immunophenotype was observed in gastric, lung, colorectal adenocarcinoma. CK7+/CK20- immunophenotype was observed in breast, lung, endometrial, uterine cervical, bile duct adenocarcinoma, while CK7-/CK20+ immunophenotype was observed only in colorectal adenocarcinoma. This results show the utility of TTF-1 to confirm the pulmonary origin. On the other hand ER and PR were not useful markers to assess the origin of primary tumor in this series.

FHIT Gene Expression in Acute Lymphoblastic Leukemia and its Clinical Significance

  • Malak, Camelia A Abdel;Elghanam, Doaa M;Elbossaty, Walaa Fikry
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8197-8201
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    • 2016
  • Background: To investigate the expression of the fragile histidine triad (FHIT) gene in acute lymphoblastic leukemia and its clinical significance. Materials and Methods: The level of expressed FHIT mRNA in peripheral blood from 50 patients with acute lymphoblastic leukemia (ALL) and in 50 peripheral blood samples from healthy volunteers was measured via RT-PCR. Correlation analyses between FHIT gene expression and clinical characteristics (gender, age, white blood count, immunophenotype of acute lymphoblastic leukemia and percentage of blast cells) of the patients were performed. Results: The FHIT gene was expressed at $2.49{\pm}7.37$ of ALL patients against $14.4{\pm}17.9$ in the healthy volunteers. The difference in the expression levels between ALL patients and healthy volunteers was statistically significant. The rate of gene expression did not significantly vary with immunophenotype subtypes. Gene expression was also found to be correlated with increase of total leukocyte and decrease in platelets, but not with age, gender, immunophenotyping or percentage of blast cells. Conclusions: FHIT gene expression is low in acute lymphoblastic leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute lymphoblastic leukemia.

Immunophenotype Characterization for Swine Selected Line, Which is Resistant for the Mycoplasma Pneumonia

  • Katayama, Masafumi;Fukuda, Tomokazu;Okamuara, Toshihiro;Suda, Yoshihito;Suzuki, Eisaku;Uenishi, Hirohide;Suzuki, Keiichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.889-897
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    • 2011
  • Mycoplasma Pneumonia of swine (MPS) decreases the daily growth of pigs, and, co-infection with a virus sometimes causes severe pneumonia. Genetic selection of pigs resistant to the pulmonary MPS lesion might solve the economic loss due to MPS in animal production. Here, we examined the immunophenotype of Landrace line (Miyagino L2), genetically selected to reduce the incidence of pulmonary MPS lesion for 5 generations in Miyagi Prefecture Animal Industry Experiment Station. Although this line is expected to be resistant to the pulmonary MPS lesion, the biological characteristics of its immune function are not clear. We investigated details of the immunorelated phenotype of Miyagino L2 at the hematological and molecular biological level, including cytokine expression, and compared the results with that of non-genetically selected Landrace. Miyagino L2 showed decreased antigen-specific IgG and IgM production and increased CD8-positive T-cell population, and high levels of cortisol concentration, suggesting that the MPS-resistant phenotype is associated these immunological differences. Additionally, T-cell CD4 expression was highly correlated with the MPS expected breeding value. Although the detailed mechanisms underlying this high correlation remain unknown, our result suggested that the genetic selection of the expression level of CD4 might be useful to improve MPS resistance in pig production.

Comparisons of C-kit, DOG1, CD34, PKC-θ and PDGFR-α Expressions in Gastrointestinal Stromal Tumors According to Histopathological Risk Classification

  • Kim, Ki-Sung;Song, Hye-Jung;Shin, Won-Sub;Song, Kang-Won
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.2
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    • pp.48-56
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    • 2011
  • Gastrointestinal stromal tumor (GIST) is a mesenchymal tumor and is associated with a specific immunophenotype index. It is very important to identify the specific immunophenotype and the diagnosis for the treatment GIST patients. Ninety two cases of GIST analyzed in this study were immuno-stained for c-kit, DOG1, CD34, PKC-${\theta}$, PDGFR-${\alpha}$. The rate of positive staining and statistical significance were then compared. In addition, the GISTs were analyzed as followings: very low risk, low risk, intermediate risk and high risk according to tumor size and nuclear division, and later correlated with clinical parameters. The results of the GIST positive stainings were: DOG1 (95.7%), PKC-${\theta}$ (90.2%), PDGFR-${\alpha}$ (88.0%), c-kit (87.0%) and CD34 (71.7%). Only DOG1 staining showed a statistical significance of p<0.05. It was identified in the classification system of histologic risk that staining expression of DOG1, PKC-${\theta}$, PDGFR-${\alpha}$ were significantly increased as histologic risk increases (p<0.05). However, clinical parameters such as age and sex of patients have no correlations with the classification system of histologic risk (p>0.05). Therefore, in this study, the expression of DOG1 showed statistical significance and DOG1, PKC-${\theta}$, PDGFR-${\alpha}$ staining increased significantly as the histologic risk increases in histologic classification system. Taken together, the DOG1 staining should be very effective for the diagnosis of GIST patients.

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Clinical Significance of Co-expression of Aberrant Antigens in Acute Leukemia: A Retrospective Cohort Study in Makah Al Mukaramah, Saudi Arabia

  • Abdulateef, Nahla Ahmad Bahgat;Ismail, Manar Mohammad;Aljedani, Hanadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.221-227
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    • 2014
  • Background: Aberrant phenotypes in acute leukemia have variable frequency and their prognostic and predictive relevance is controversial, despite several reports of clinical significance. Aims: To determine the prevalence of aberrant antigen expression in acute leukemia, assess clinical relevance and demonstrate immunophenotype-karyotype correlations. Materials and Methods: A total of 73 (40 AML and 33 ALL) newly diagnosed acute leukemia cases presenting to KAMC, Kingdom of Saudi Arabia, were included. Diagnosis was based on WHO criteria and FAB classification. Immunophenotyping by flow cytometry, conventional karyotyping and fluorescence in situ hybridization for gene rearrangements were performed. Results: Aberrant antigens were detected in 27/40 (67.5%) of AML and in 14/33 (42.4%) in ALL cases. There were statistically significant higher TLC in Ly+ AML than in Ly-AML (p=0.05) and significant higher blast count in ALL with aberrant antigens at presentation and day 14 (p=0.005, 0.046). There was no significant relation to clinical response, relapse free survival (RFS) or overall survival (p>0.05), but AML cases expressing ${\geq}2$ Ly antigens showed a lower median RFS than those expressing a single Ly antigen. In AML, CD 56 was expressed in 11/40. CD7 was expressed in 7/40, having a significant relation with an unfavorable cytogenetic pattern (p=0.046). CD4 was expressed in 5/40. CD19 was detected in 4/40 AML associated with M2 and t (8; 21). In ALL cases, CD33 was expressed in 7/33 and CD13 in 5/33. Regarding T Ag in B-ALL CD2 was expressed in 2 cases and CD56 in 3 cases. Conclusions: Aberrant antigen expression may be associated with adverse clinical data at presentation. AML cases expressing ${\geq}2$ Ly antigens may have shorter median RFS. No specific cytogenetic pattern is associated with aberrant antigen expression but individual antigens may be related to particular cytogenetic patterns. Immunophenotype-karyotype correlations need larger studies for confirmation.

Comparison of polymerase chain reaction for antigen receptor gene rearrangement and flow cytometric analysis for the diagnosis of canine lymphoma

  • Song, Ru-Hui;Yu, Do-Hyeon;Kim, Jun-Hwan;Lee, Hyun-Seok;Lee, Da-Mi;Park, Chul;Yu, Il-Jung;Park, Jin-Ho
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.265-271
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    • 2011
  • Lymphoma is the most common hematopoietic malignancy in dogs. Diagnosis of lymphoma is classically performed by morphological assessment and immunohistochemistry. But some cases in the early stage are difficult to distinguish and need more objective and accurate methods. So, Polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) and flow cytometric immunophenotype of lymphoma have been developed continuously. In this study, we performed these two methods to classify lymphoma type in 3 cases. According to PARR analysis, B cell origin lymphoma was diagnosed in two of three cases by testing PBMC and lymph node. All fine needle aspiration (FNA) samples of lymph nodes had high expression of CD21 on >88% of total cell population and PBMC samples also showed high expression of CD21 on >30% of total lymphocytes in those two cases, while the expression of CD3, CD4 and CD8 was absent. These results suggest that concurrent use of PARR and flow cytometric immunophenotype is more effective and valuable tool for the diagnosis and monitoring of canine lymphoma patients.

The Expression of Immunomodulation-Related Cytokines and Genes of Adipose- and Bone Marrow-Derived Human Mesenchymal Stromal Cells from Early to Late Passages

  • Mun, Chin Hee;Kang, Mi-Il;Shin, Yong Dae;Kim, Yeseul;Park, Yong-Beom
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.771-779
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    • 2018
  • BACKGROUND: Mesenchymal stromal cells (MSCs) are multipotent stem cells that can differentiate into several cell types. In addition, many studies have shown that MSCs modulate the immune response. However, little information is currently available regarding the maintenance of immunomodulatory characteristics of MSCs through passages. Therefore, we investigated and compared cytokine and gene expression levels from adipose (AD) and bone marrow (BM)-derived MSCs relevant to immune modulation from early to late passages. METHODS: MSC immunophenotype, growth characteristics, cytokine expressions, and gene expressions were analyzed. RESULTS: AD-MSCs and BM-MSCs had similar cell morphologies and surface marker expressions from passage 4 to passage 10. Cytokines secreted by AD-MSCs and BM-MSCs were similar from early to late passages. AD-MSCs and BM-MSCs showed similar immunomodulatory properties in terms of cytokine secretion levels. However, the gene expressions of tumor necrosis factor-stimulated gene (TSG)-6 and human leukocyte antigen (HLA)-G were decreased and gene expressions of galectin-1 and -3 were increased in both AD- and BM-MSCs with repeated passages. CONCLUSION: Our study showed that the immunophenotype and expression of immunomodulation-related cytokines of AD-MSCs and BM-MSCs immunomodulation through the passages were not significantly different, even though the gene expressions of both MSCs were different.

Canine Multicentric Large B Cell Lymphoma with Increased Mott Cells Diagnosed by Flow Cytometry

  • Yang, Yeseul;Jung, Jae-Ha;Hwang, Sung-Hyun;Kim, Yongbaek
    • Journal of Veterinary Clinics
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    • v.38 no.1
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    • pp.36-40
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    • 2021
  • A 12-year-old dog was referred due to multiple superficial lymphadenopathy. On cytology, each lymph node showed different cell populations where some of them consisted of intermediate to large lymphocytes with frequent Mott cells. Presence of Mott cells along with immature lymphocytes made the cytological diagnosis challenging, and therefore, supplementary diagnostic tests including PCR for Antigen Receptor Rearrangement (PARR) assay and flow cytometry were performed. This case report illustrates the value of flow cytometry in the diagnosis of lymphadenopathy with ambiguous cytologic findings.

Differential Potential of Stem Cells Following Their Origin - Subacromial Bursa, Bone Marrow, Umbilical Cord Blood - (줄기세포의 분화능의 기원에 따른 비교 - 견봉하 점액낭, 골수, 탯줄 혈액 -)

  • Sim, Sung Woo;Moon, Young Lae;Kang, Jung Hun
    • Clinics in Shoulder and Elbow
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    • v.15 no.2
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    • pp.65-72
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    • 2012
  • Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.