• Proceedings of the Microbiological Society of Korea Conference
• /
• 2008.05a
• /
• pp.62-64
• /
• 2008

A major hurdle to the development of RNA interference as therapy for HIV infection is the delivery of siRNA to T lymphocytes which are difficult cells to transfect even in vitro. We have employed a single chain antibody to the pan T cell surface antigen CD7 was conjugated to an oligo-9-arginine peptide (scFvCD7-9R) for T cell-specific siRNA delivery in NOD/SCIDIL2${\gamma}$-/- mice reconstituted with human peripheral blood lymphocytes (Hu-PBL). Using a novel delivery, we first show that scFvCD7-9R efficiently delivered CD4 siRNA into human T cells in vitro. In vivo administration to Hu-PBL mice resulted in reduced levels of surface CD4 expression on T cells. Mice infected with HIV-1 and treated on a weekly basis with scFvCD7-9R-siRNA complexes targeting a combination of viral genes and the host coreceptor molecule CCR5 successfully maintained CD4/CD3 T cell ratios up to 4 weeks after infection in contrast to control mice that displayed a marked reduction in CD4 T cell numbers. p24 antigen levels were undetectable in 3 of the 4 protected mice. scFvCD7-9R/antiviral siRNA treatment also helped maintain CD4 T cell numbers with reduced plasma viral loads in Hu-PBL mice reconstituted with PBMC from donors seropositive for HIV, indicating that this method can contain viral replication even in established HIV infections. Our results show that scFvCD7-9R could be further developed as a potential therapeutic for HIV-1 infection.

• Korean Journal of Clinical Laboratory Science
• /
• v.38 no.1
• /
• pp.1-8
• /
• 2006

The purposes of this study were to evaluate the changes in hematologic indices after plateletpheresis and to identify the preapheresis platelet count and clinical factors (age, gender, height, and weight) that showed some influence on the percentage of platelet decrement, yield and efficiency. Plateletpheresis was performed on 101 healthy donors in Bundang CHA general hospital. The data was analyzed using the SAS program with t-test, ANOVA test and Multiple regression. The mean percentage decrease after plateletpheresis was 2.0% in hemoglobin, 1.8% in hematocrit, and 29.7% in the platelet count, while a WBC count showed an increase of 2.6%. The mean percentage decrease of hemoglobin and hematocrit were 1.7% and 1.4%, in males and 3.6% and 3.7% in females, respectively. Particularly the percentage decrease of platelet count was significantly higher in females (40.0%) than in males (27.2%). The platelet decrementage and yield were significantly higher in females, but the efficiency did not differ significantly between males and females. The yield showed the lowest levels in subjects who were 40 years old or over but the platelet decrement and efficiency did not change according to age. The platelet decrement increased as height and weight increased. Also, the platelet decrement and yield increased as the initial platelet counts increased, but the efficiency did not. From multiple regression analysis, the platelet decrement was associated with gender, weight, and initial platelet count. The yield was related to the initial platelet count, but the efficiency was not related to gender, age, weight, height or initial platelet counts. This study has a limitation of the generality of the study results since this study was conducted only in a single university hospital. Further study would be necessary to find out a subpopulation that is sensitive to the hematologic change after plateletpheresis, and to determine the standard criteria for blood donation based on the subpopulation.

• Toxicological Research
• /
• v.12 no.2
• /
• pp.195-201
• /
• 1996

Endotoxic shock causes death in humans and animals via extreme hypoperfusion of peripheral organs. A massive production of nitric oxide (NO) both from the endothelical cells and smooth muscle cells has been proposed as a possible mechanism in this process. Since NO attenuated the contractility to vasoconstricting agents such as norepinephrine (NE) by directly acting on the smooth muscle cells, this mechanism was considered mainly as a postsynaptic mechanism. In this research it was investigated whether NO, thus released, also participates in the presynaptic events for the regulation of vascular tone in endotoxic shock. The role of NO was studied by adding NO donors or NO synthase inhibitor $N^\omega $methyl-L-arginine (NMA) in stimulated sympathetic nerves of the mesenteric vascular bed and the Langendorff heart of rats. Sodium nitroprusside (SNP), an NO donor, reduced the pressor responses of isolated mesenteric artery either to electrical stimulation or exogenously administered phenylephrine (PE). In this mesentery, although neither agent influenced NE release, in the presence of the adrenergic $\alpha_2$-receptor antagonist yohimbine, elecrical stimulation-evoked NE release was augumented by SNP. In the heart SNP facilitated the NE release induced by electrical stimulation, while NMA had no effect. From these results it is proposed that there exists a local reflex phenomenon in the junction between the sympathetic nerve terminals and the smooth muscle of resistance blood vessels; by which sympathetic responses are reduced by NO at the postjunctional level while NO facilitates NE release contributing to augumentation of sympathetic tone. All these facts suggest that NO produced during endotoxic shock has dual effects: whereas NO blunts the vasoconstrictive activity of NE at the postsynaptic level, NO presynaptically facilitates the release of NE from sympathetic nerve terminals.

• Korean Journal of Transplantation
• /
• v.32 no.4
• /
• pp.108-112
• /
• 2018

Antibody-mediated rejection (AMR) is a major complication after ABO-incompatible liver transplantation. According to the 2016 Banff Working Group on Liver Allograft Criteria for the diagnosis of acute AMR, a positive serum donor specific antibody (DSA) is needed. On the other hand, the clinical significance of the histological findings of AMR in the absence of DSA is unclear. This paper describes a 57-year-old man (blood type, O+) who suffered from hepatitis B virus cirrhosis with hepatocellular carcinoma. Pre-operative DSA and cross-matching were negative. After transplantation, despite the improvement of the liver function, acute AMR was observed in the protocol biopsy on postoperative day 7; the cluster of differentiation 19+ (CD19+) count was 0% and anti-ABO antibody titers were 1:2. This paper presents the allograft injury like AMR in the absence of DSA after ABOi living donor liver transplantation with low titers of anti-ABO antibody and depleted serum CD19+ B cells.

• Genomics & Informatics
• /
• v.21 no.2
• /
• pp.18.1-18.11
• /
• 2023

Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell-derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55⁺) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

• The Journal of Korean Society for Radiation Therapy
• /
• v.19 no.2
• /
• pp.83-90
• /
• 2007

Purpose: In this study, it was investigated whether commercially produced beer is able to prevent a lymphocyte from radiation induced apoptosis. Materials and Methods: Whole blood samples were acquired from 5 healthy volunteers (male, 26$\sim$38 years old) and the lymphocyte were isolated by density gradient centrifugation. Radiation induced apoptosis of the lymphocyte were investigated by 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy to 5.0 Gy irradiation. In some experiments, the donor drunk beer and then blood samples were collected. In other experiments, melatonin or glycine betain was added to lymphocyte culture medium. Treated or untreated lymphocytes were cultured for 60 hours and radiation induced apoptosis of the lymphocyte was analyzed by annexin-V staining through flow cytometery. Results: Relative radiation induced apoptosis ratio of the untreated lymphocytes is 1.22$\pm$1.1, 1.22$\pm$1.1, 1.38$\pm$1.0, 1.47$\pm$1.1, 1.50$\pm$1.2 by radiation dose of 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy and 5.0 Gy respectively. Relative radiation induced apoptosis ratio of lymphocytes is isolated from beer drunken donors is 0.97$\pm$1.0, 0.99$\pm$1.0, 1.11$\pm$0.9, 1.29$\pm$1.1, 1.15$\pm$1.1 by radiation doses respectively which are reduced 21.5% compared with untreated lymphocyte. Relative radiation induced apoptosis ratio of the lymphocytes is isolated from non-alcohol beer drunken donors is 1.22$\pm$1.1, 1.17$\pm$1.1, 1.13$\pm$1.3, 1.38$\pm$1.2, 1.32$\pm$1.1 by radiation dose of 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy and 5.0 Gy respectively which are reduced 10.8% compared with the untreated lymphocyte. Conclusion: As a result, it is suggested that beer may protect the lymphocyte from radiation damage and inhibit apoptosis.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.11
• /
• pp.6403-6409
• /
• 2013

Purpose: Biallelic germline variants of the 8-hydroxyguanine (8-OG) repair gene MYH have been associated with colorectal neoplasms that display somatic $G:C{\rightarrow}T:A$ transversions. However, the effect of single germline variants has not been widely studied, prompting the present investigation of monoallelic MYH variants and susceptibility to sporadic colorectal cancer (CRC) in a Chinese population. Patients and Methods: Between January 2006 and December 2012, 400 cases of sporadic CRC and 600 age- and sex-matched normal blood donors were screened randomly for 7 potentially pathogenic germline MYH exons using genetic testing technology. Variants of heterozygosity at the MYH locus were assessed in both sporadic cancer patients and healthy controls. Univariate and multivariate analyses were performed to determine risk factors for cancer onset. Results: Five monoallelic single nucleotide polymorphisms (SNPs) were identified in the 7 exon regions of MYH, which were detected in 75 (18.75%) of 400 CRC patients as well as 42 (7%) of 600 normal controls. The region of exon 1 proved to be a linked polymorphic region for the first time, a triple linked variant including exon 1-316 $G{\rightarrow}A$, exon 1-292 $G{\rightarrow}A$ and intron 1+11 $C{\rightarrow}T$, being identified in 13 CRC patients and 2 normal blood donors. A variant of base replacement, intron 10-2 $A{\rightarrow}G$, was identified in the exon 10 region in 21 cases and 7 controls, while a similar type of variant in the exon 13 region, intron 13+12 $C{\rightarrow}T$, was identified in 8 cases and 6 controls. Not the only but a newly missense variant in the present study, p. V463E (Exon 14+74 $T{\rightarrow}A$), was identified in exon 14 in 6 patients and 1 normal control. In exon 16, nt. 1678-80 del GTT with loss of heterozygosity (LOH) was identified in 27 CRC cases and 26 controls. There was no Y165C in exon 7 or G382D in exon 14, the hot-spot variants which have been reported most frequently in Caucasian studies. After univariate analysis and multivariate analysis, the linked variant in exon 1 region (p=0.002), intron 10-2 $A{\rightarrow}G$ (p=0.004) and p. V463E (p=0.036) in the MYH gene were selected as 3 independent risk factors for CRC. Conclusions: According to these results, the linked variant in Exon 1 region, Intron 10-2 $A{\rightarrow}G$ of base replacement and p. V463E of missense variant, the 3 heterozygosity variants of MYH gene in a Chinese population, may relate to the susceptibility to sporadic CRC. Lack of the hot-spot variants of Caucasians in the present study may due to the ethnic difference in MYH gene.

• Journal of Preventive Medicine and Public Health
• /
• v.30 no.1 s.56
• /
• pp.17-29
• /
• 1997

To estimate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and to determine associated risk factors, a population-based seroepidemiologic study was carried out. In 1993, a health examination survey of the population was carried out in rural area known to have a high incidence of liver cancer. The study population were those who volunteered to participate in a health survey over 10 years of age. Examinees were interviewed by specially trained staffs. Sera from 1,033 study subjects were tested for hepatitis B surface antigen (HBsAg) by .everse passive hemagglutinin (RPHA) estimation and for hepatitis C virus antibody (anti-HCV) by 2nd generation passive hemagglutinin (PHA) estimation. The age and sex standardized prevalence of HBsAg was 6.3% which was similar to national average, but that of anti-HCV was 5.1% which was 4 to 5 times higher than that of blood donors or other health examinees in Korea. In a multivariate analysis, transfusion history, surgical operative history, and acupuncture history were not associated with HBsAg positivity. In contrast, acupuncture history (adjusted odds ratio[OR]=2.2 : 95% Confidence interval[CI] 1.0-4.7) and surgical operative history(adjusted OR=2.0 : 95% CI 1.0-4.1) were associated with anti-HCV positivity. The present study suggest that there is an highly endemic area of HCV infection in Korea and probably this endemicity is associated with a parenteral source of HCV infection other than blood transfusion.

• Childhood Kidney Diseases
• /
• v.5 no.2
• /
• pp.87-99
• /
• 2001

Purpose: We studied to find out apo-E genotype polymorphism in minimal change nephrotic syndrome(MCNS) and IgA nephropathy(IgAN) and to determine the relationship between apo-E genotype and clinical course of MCNS. Materials and Method: 43 MCNS patients and 15 IgAN patients were examined for apo-E polymorphism. 50 healthy blood donors were examined for apo-E genotype as control. Genomic DNA was prepared front peripheral blood leukocytes according to standard procedures. Results: As compared with control group, e4 allele frequency was significantly increased in MCNS (p<0.01). However, in IgAN e2 allele frequency, however, was 2.6 times higher than normal control (P<0.01). The frequency of e4 allele of frequent relapser group was 4.6 times higher than normal control and was 2 times higher than infrequent relapser group. Conclusion: We think that apo-E typing might be one of the parameters, which should be considered to predict the course of MCNS in children. MCNS with risky HLA profile and E4/4 genotype could indicate the need for a longer steroid administration. And apo-E genotype needs to be considered for the evaluation of therapeutic responses to other drugs. (J, Korean Soc Pediatr Nephrol 2001 ; 5 : 87-99)

• Journal of Life Science
• /
• v.24 no.9
• /
• pp.1030-1038
• /
• 2014

Platelet products are used to treat hemorrhagic or platelet dysfunction diseases. Plateletpheresis involves collecting the platelet components of blood using an apheresis blood-collection system. Various indicators are available for evaluating the qualities of the apheresis platelets. The productivity of platelet collection is evaluated through both the collection efficiency and collection rates. Platelet storage quality can be evaluated in vitro using several indicators, including visual appearance, metabolic activities, volume, platelet count, white blood cell count, microparticles, and various platelet activation markers. Platelet activation markers have been used as indicators of storage quality in various studies. Post-transfusion platelet quality can be evaluated based on the corrected count increment and the percentage of platelet recovery. Although various studies have investigated the aspects of plateletpheresis, no article has systemically presented assessments of the platelet products obtained from different plateletpheresis devices. The present study provides a review of plateletpheresis, including the specifics of the process, the types of devices employed, the platelet quality, the overall efficacy, and the evaluation indicator qualities. Furthermore, the differences in functionality among the different apheresis devices are discussed. Although adverse reactions to the citrate anti-coagulant have been reported, apheresis processing may provide a safer option for donors who are at a high risk for presyncopal or syncopal reactions related to whole blood collection.