• The Korean Journal of Physiology and Pharmacology
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• 제24권6호
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• pp.463-472
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• 2020

Direct reprogramming, also known as a trans-differentiation, is a technique to allow mature cells to be converted into other types of cells without inducing a pluripotent stage. It has been suggested as a major strategy to acquire the desired type of cells in cell-based therapies to repair damaged tissues. Studies related to switching the fate of cells through epigenetic modification have been progressing and they can bypass safety issues raised by the virus-based transfection methods. In this study, a protocol was established to directly convert fully differentiated fibroblasts into diverse mesenchymal-lineage cells, such as osteoblasts, adipocytes, chondrocytes, and ectodermal cells, including neurons, by means of DNA demethylation, immediately followed by culturing in various differentiating media. First, 24 h exposure of 5-azacytidine (5-aza-CN), a well-characterized DNA methyl transferase inhibitor, to NIH-3T3 murine fibroblast cells induced the expression of stem-cell markers, that is, increasing cell plasticity. Next, 5-aza-CN treated fibroblasts were cultured in osteogenic, adipogenic, chondrogenic, and neurogenic media with or without bone morphogenetic protein 2 for a designated period. Differentiation of each desired type of cell was verified by quantitative reverse transcriptase-polymerase chain reaction/western blot assays for appropriate marker expression and by various staining methods, such as alkaline phosphatase/alizarin red S/oil red O/alcian blue. These proposed procedures allowed easier acquisition of the desired cells without any transgenic modification, using direct reprogramming technology, and thus may help make it more available in the clinical fields of regenerative medicine.

• Molecules and Cells
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• 제42권3호
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• pp.228-236
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• 2019

CD4 T cells differentiate into $ROR{\gamma}t/IL$-17A-expressing cells in the small intestine following colonization by segmented filamentous bacteria (SFB). However, it remains unclear whether SFB-specific CD4 T cells can differentiate directly from naïve precursors, and whether their effector differentiation is solely directed towards the Th17 lineage. In this study, we used adoptive T cell transfer experiments and showed that naïve CD4 T cells can migrate to the small intestinal lamina propria (sLP) and differentiate into effector T cells that synthesize IL-17A in response to SFB colonization. Using single cell RT-PCR analysis, we showed that the progenies of SFB responding T cells are not uniform but composed of transcriptionally divergent populations including Th1, Th17 and follicular helper T cells. We further confirmed this finding using in vitro culture of SFB specific intestinal CD4 T cells in the presence of cognate antigens, which also generated heterogeneous population with similar features. Collectively, these findings indicate that a single species of intestinal bacteria can generate a divergent population of antigen-specific effector CD4 T cells, rather than it provides a cytokine milieu for the development of a particular effector T cell subset.

• IMMUNE NETWORK
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• 제7권1호
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• pp.1-9
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• 2007

[ $CD4^+CD25^+$ ] regulatory T cells (Tregs) expressing the lineage-specific marker Foxp3 represent an important regulatory T cell that is essential for maintaining peripheral tolerance. Although it was believed that Treg development is solely dependent on the thymus, accumulating evidence demonstrates that Tregs can also be induced in the periphery. Considering the various origins of peripherally developed $CD4^+CD25^+Foxp3^+$ regulatory T cells, it seems likely that multiple factors are involved in the peripheral generation of Tregs.

• Molecules and Cells
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• 제44권5호
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• pp.318-327
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• 2021

CD4+ T helper (Th) cells play a crucial role in the modulation of innate and adaptive immune responses through the differentiation of Th precursor cells into several subsets, including Th1, Th2, Th17, and regulatory T (Treg) cells. Effector Th and Treg cells are distinguished by the production of signature cytokines and are important for eliminating intracellular and extracellular pathogens and maintaining immune homeostasis. Stimulation of naive Th cells by T cell receptor and specific cytokines activates master transcription factors and induces lineage specification during the differentiation of Th cells. The master transcription factors directly activate the transcription of signature cytokine genes and also undergo post-translational modifications to fine-tune cytokine production and maintain immune balance through cross-regulation with each other. This review highlights the post-translational modifications of master transcription factors that control the differentiation of effector Th and Treg cells and provides additional insights on the immune regulation mediated by protein argininemodifying enzymes in effector Th cells.

• IMMUNE NETWORK
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• 제4권1호
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• pp.44-52
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• 2004

Background: As a potent antigen presenting cell and a powerful inducer of antigen specific immunity, dendritic cells (DCs) are being considered as a promising anti-tumor therapeutic module. The expected therapeutic effect of DCs in renal cell carcinoma was tested in the mouse model. Established late-stage tumor therapeutic (E-T) and minimal residual disease (MRD) model was considered in the in vivo experiments. Methods: Syngeneic renal cell carcinoma cells (RENCA) were inoculated either subcutaneously (E-T) or intravenously (MRD) into the Balb/c mouse. Tumor cell lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started 3 week (E-T model) or one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, the tumor growth and the systemic immunity were observed. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with RENCA cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor growth (E-T model) or formation (MRD model) was suppressed in pulsed-DC treated group. RENCA specific lymphocyte proliferation was observed in the RENCA tumor-bearing mice treated with pulsed-DCs. Primary cytotoxic T cell activity against RENCA cells was increased in pulsed-DC treated group. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs in established or minimal residual disease/metastasis state of renal cell carcinoma. Systemic tumor specific immunity including cytotoxic T cell activity was modulated also in pulsed-DC treated group.

• Genomics & Informatics
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• 제14권4호
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• pp.205-210
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• 2016

Interleukin-10 (IL10) plays an important role in initiating and maintaining an appropriate immune response to non-Hodgkin lymphoma (NHL). Previous studies have revealed that the transcription of IL10 mRNA and its protein expression may be infl uenced by several single-nucleotide polymorphisms in the promoter and intron regions, including rs1800896, rs1800871, and rs1800872. However, the impact of polymorphisms of the IL10 gene on NHL prognosis has not been fully elucidated. Here, we investigated the association between IL10 polymorphisms and NHL prognosis. This study involved 112 NHL patients treated at the National Cancer Center, Korea. The median age was 57 years, and 70 patients (62.5%) were men. Clinical characteristics, including age, performance status, stage, and extra-nodal involvement, as well as cell lineage and International Prognostic Index (IPI), were evaluated. A total of four polymorphisms in IL10 with heterozygous alleles were analyzed for hazard ratios of overall survival (OS) and progression-free survival (PFS) using Cox proportional hazards regression analysis. Diffuse large B-cell lymphoma was the most common histologic type (n = 83), followed by T-cell lymphoma (n = 18), mantle cell lymphoma (n = 6), and others (n = 5). Cell lineage, IPI, and extra-nodal involvement were predictors of prognosis. In the additive genetic model results for each IL10 polymorphism, the rs1800871 and rs1800872 polymorphisms represented a marginal association with OS (p = 0.09 and p = 0.06) and PFS (p = 0.05 and p = 0.08) in B-cell lymphoma patients treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). These findings suggest that IL10 polymorphisms might be prognostic indicators for patients with B-cell NHL treated with R-CHOP.

• 대한수의학회지
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• 제47권1호
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• pp.85-89
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• 2007

A case of multicentric high grade T cell lymphoma is reported in a 5-year-old male Maltese dog with generalized lymphadenopathy. The dog showed depression, anorexia, blindness, jaundice, arrhythmia, and hematuria for 8 months. Complete blood count and chemistry profile revealed anemia and increased alanine transferase, alkaline phosphatase, total bilirubin, and total cholesterol. Grossly, most of lymph nodes, spleen, and liver were enlarged and neoplastic masses were occupied in these tissues. Histologically, massive accumulation of small noncleaved neoplastic lymphocytes with high mitotic figures was observed in all lymph nodes and spleen. Infiltration of neoplastic lymphocytes was also noted in the lung, liver, kidney, eye, skin, muscle, and bone marrow of femur. Immunohistochemistry revealed that tumor cells were CD3-positive and but CD79a-negative, consistent with T-cell lineage. In our best knowledge, this is the first report of multicentric lymphoma clarified the origin of tumor cells in Korea.

• Journal of Yeungnam Medical Science
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• 제14권2호
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• pp.359-369
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• 1997

저자들은 1993년 3월에서 1997년 9월까지 영남대학교 의과대학 부속병원 임상병리과에 면역학적 표지자 검사가 의뢰된 153명의 백혈병 환자의 결과를 분석하였다. 여기에는 AML 61례, ALL 46례, FAB 아형이 확정되지 않았던 12례 등이 포함되어 있었다. EDTA 항응고처리한 골수나 말초혈액에서 단핵세포층을 분리한후 일련의 단클론항체와 FITC 표지 이차항체를 이용하여 면역형광염색법을 실시한 후 형광현미경으로 관찰하였다. AML 61례에서의 비전형적인 표지자 표현 양상을 보면 CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6%, CD4 4.9%, CD19 1.6%의 양성률을 보였고, TdT 양성은 3.9%였다. 혼성 백혈병으로 최종 진단을 내린 경우는 13.1%로 그 중 단구성 계열이 대부분(6/8)이었다. ALL 46례에서의 면역학적 검사에 따른 분류에 따르면 CALLA(+) B precursor형이 65.2%를 차지하였고, CALLA(-) B precursor형이 10.9%, T 세포형 8.7%, B 세포형 2.2%, 혼성림프구형(B&T) 4.3%, 미분화성 백혈병 2.2%였고, 혼성 백혈병으로 진단된 경우는 6.5%였다. 골수성 표지자 양성인 ALL은 CD13이나 CD33 양성인 례가 각각 2.2%였다. 급성 백혈병에서 골수형태와 세포화학검사로 FAB 아형을 확정하지 못한 12례에서 혼성 백혈병이 5례(41.7%)였고, 림프구 계열이 7례(58.3%)였으며 그 중 B 세포계열이 3례, T 세포계열이 3례, 혼성 림프구형(B&T)이 1례로 나타났다. 결론적으로는, 면역학적 표지지 검사로 더 정확하게 AML과 ALL을 감별할 수 있었으며, M0, M7 아형진단에도 도움이 되며, 형태학적, 세포화학적검사로 정확히 진단할 수 없었던 급성 백혈병의 분류에 도움이 되었다. 따라서 급성 백혈병의 진단을 위하여 FAB 분류 뿐 만 아니라 면역학적 표지자 검사를 함께 시행한다면 보다 정확한 진단에 도움을 줄 수 있을 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권6호
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• pp.2909-2916
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• 2016

Background: Canine malignant lymphoma is classified into B- or T-cell origin, as in the human case. Due to differences in prognosis, a suitable method needs to be developed for lineage identification. Aims: To determine the accuracy of immunophenotypic and molecular information between three methods: immunocytochemistry (ICC), immunohistochemistry (IHC) and heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) in spontaneous canine lymphomas. Materials and Methods: Peripheral blood, fine needle aspiration and tissue biopsies from enlarged peripheral lymph nodes prior to treatment of 28 multicentric lymphoma patients were collected. Cytopathology and histopathology were examined and classified using the updated Kiel and WHO classifications, respectively. Anti-Pax5 and anti-CD3 antibodies as B- and T-cell markers were applied for immunophenotyping by ICC and IHC. Neoplastic lymphocytes from lymph node and white blood cell pellets from peripheral blood were evaluated by hPARR. Results: In this study, low grade B-cell lymphoma accounted for 25% (7/28), high grade B-cell lymphoma for 64.3% (18/28) and high grade T-cell lymphoma for 10.7% (3/28). According to the WHO classification, 50% of all cases were classified as diffuse large B-cell lymphoma. In addition, ICC showed concordant results with IHC; all B-cell lymphomas showed Pax5+/CD3, and all T-cell lymphomas exhibited Pax5-/CD3+. In contrast to hPARR, 12 B-cell lymphomas featured the IgH gene; seven presented the $TCR{\gamma}$ gene; five cases showed both IgH and $TCR{\gamma}$ genes, and one case were indeterminate. Three T-cell lymphomas showed the $TCR{\gamma}$ gene. The percentage agreement between hPARR and ICC/IHC was 60%. Conclusions: Immunophenotyping should not rely on a single method. ICC or IHC with hPARR should be used concurrently for immunophenotypic diagnosis in canine lymphomas.

• IMMUNE NETWORK
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• 제3권2호
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• pp.89-95
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• 2003

Background: Unusually high amounts of N region addition and CDR3 length diversity were found in immunoglobulin (Ig) light chain Vk and Jk joins in patients with rheumatoid arthritis (RA). We sought to determine whether this finding is due to excessive activity of the enzyme responsible for N region addition (terminal deoxynucleotidyl transferase [TdT]) in B lineage cells in bone marrow or from positive antigenic selection of B cells with long CDR3 lengths. Methods: We used FACS to isolate $IgM^+/IgD^+$ B cells (predominantly naive) and $IgM^-/IgD^-$ B cells (predominantly class-switched) B cells from peripheral blood of a patient with RA known to have enrichment for long Vk CDR3s and from that of two normal controls. RT-PCR of VkIII transcripts was performed, followed by sequencing of individual cDNA clones. We analyzed the CDR3 lengths and N region additions in 97 clones. Results: There was enrichment for long CDR3 lengths (11 or 12 amino acids) in both $IgM^+/IgD^+$ and $IgM^-/IgD^-$ B cells in RA compared to B cell subsets in the normal controls. The $IgM^+/IgD^+$ B cell subset in RA was markedly enriched for N region addition and was similar to that seen in the $IgM^-/IgD^-$ subset. Conclusion: These data suggest that enrichment for N region addition and long CDR3 lengths in RA may result from unusually high or prolonged activity of TdT in bone marrow.