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Maharjan, Anju,Gautam, Ravi,Jo, JiHun,Acharya, Manju,Lee, DaEun,Pramod, Bahadur KC,Gim, Jin,Sin, Sojung,Kim, Hyocher,Kim, ChangYul,Lee, SooYeon,Lee, SooJin,Heo, Yong,Kim, HyoungAh Occupational Safety and Health Research Institute 2022 Safety and health at work Vol.13 No.2
Background: Occupational hazards in crop farms vary diversely based on different field operations as soil management, harvesting processes, pesticide, or fertilizer application. We aimed at evaluating the immunological status of crop farmers, as limited systematic investigations on immune alteration involved with crop farming have been reported yet. Methods: Immunological parameters including plasma immunoglobulin level, major peripheral immune cells distribution, and level of cytokine production from activated T cell were conducted. Nineteen grape orchard, 48 onion open-field, and 21 rose greenhouse farmers were participated. Results: Significantly low proportion of natural killer (NK) cell, a core cell for innate immunity, was revealed in the grape farmers (19.8±3.3%) in comparison to the onion farmers (26.4±3.1%) and the rose farmers (26.9±2.5%), whereas cytotoxic T lymphocyte proportion was lower in the grape and the onion farmers than the rose farmers. The proportion of NKT cell, an immune cell implicated with allergic response, was significantly higher in the grape (2.3±0.3%) and the onion (1.6±0.8%) farmers compared with the rose farmers (1.0±0.4%). A significantly decreased interferon-gamma:interleukin-13 ratio was observed from ex vivo stimulated peripheral blood mononuclear cells of grape farmers compared with the other two groups. The grape farmers revealed the lowest levels of plasma IgG1 and IgG4, and their plasma IgE level was not significantly different from that of the onion or the rose farmers. Conclusion: Our finding suggests the high vulnerability of workplace-mediated allergic immunity in grape orchard farmers followed by open-field onion farmers and then the rose greenhouse farmers.
Maharjan Anju,Gautam Ravi,Acharya Manju,Jo JiHun,Lee DaEun,K C Pramod Bahadur,Lee Young-A,Kwon Jung-Taek,Kim HyoCher,Kim KyungRan,Kim ChangYul,Kim HyoungAh,Heo Yong 한국독성학회 2023 Toxicological Research Vol.39 No.4
Exposure to occupational hazards like dust, pesticides, diesel emission particles, or physical hazards in the agricultural sector is known to cause adverse health effects on farm workers. Our study aimed at addressing the association of immunomodulatory status with plasma levels of lung cancer biomarkers in farming population, attempting to recognition of vulnerable farming group. Blood samples from apparently healthy 51 chicken husbandry, 19 grape orchard, and 21 rose greenhouse workers were subjected to evaluate plasma levels of two representative lung cancer biomarkers, pro-gastrin releasing peptide (Pro-GRP) and cytokeratin fragment 19 (CYFRA 21-1). Peripheral blood mononuclear cells obtained from farmers were used for natural killer (NK) cell phenotyping and cytokines (interferon-gamma, IFN-γ and interleukin-13, IL-13) profiling in the culture supernatant. Compared to the rose greenhouse farmers, the grape orchard and chicken husbandry workers revealed a significantly upregulated plasma Pro-GRP and CYFRA 21-1 level. A low proportion of NK cells was observed among the female grape orchard workers and a lowered IFN- γ:IL-13 ratio was seen in the grape and chicken husbandry workers than the rose workers. Our findings imply that grape orchard and chicken husbandry workers have more disturbed immune homeostasis implicated with augmentation in the levels of lung cancer biomarkers than the rose greenhouse workers.
Anju Maharjan,JiHun Jo,Manju Acharya,SuJeong Yang,Ravi Gautam,Sojung Sin,Hyocher Kim,ChangYul Kim,Yong Heo,HyoungAh Kim 계명대학교 자연과학연구소 2020 Quantitative Bio-Science Vol.39 No.2
The paradigm of immune homeostasis between type-1 helper T cells (Th1) and type-2 helper T cells (Th2) has been extensively examined with respect to infectious diseases, autoimmune diseases, or tumor progression. Such studies have mostly relied on the profiling of cytokines/chemokines using animal studies or cell culture-based in vitro tests. No quantitative analyses have yet been systematically performed to determine the relationship between the various immune parameters in in vivo investigations. Using peripheral blood from 55 chicken husbandry workers or grapevine orchard workers, we obtained data for various immunologic markers as follows: proportion of major immune cell subpopulations, concentrations of plasma immunoglobulin subclasses, and levels of cytokines produced from activated peripheral blood mononuclear cells. Correlational analyses were carried out to examine the association between these parameters. As the IFN-γ:IL-13 ratio was strongly associated with the production levels of IFN-γ, TNF-α, and to the IFN-γ:IL-4 ratio, the IFN-γ:IL-13 ratio could prove to be a valuable monitoring index to examine Th1 or Th2 predominance. IL-17 levels correlated well with those of IFN-γ or TNF-α. Plasma levels of IgG4, a typical IgG subclass expressed upon the occurrence of allergic diseases, strongly correlated with IgG1 levels, suggesting the implication of IgG1 in allergic hyperreactivity. The proportion of cytotoxic T cells correlated significantly with levels of IFN-γ, and the proportion of natural killer T cells correlated well with IL-13 levels. These two cell types are sources for the production of IFN-γ and IL-3 respectively; hence, results of this study might provide potential monitoring markers for cytotoxic T cells or natural killer T cell-mediated immunoreactivity. Overall, the present study provides several putative candidates for the surveillance or prognosis of the in vivo alteration of humoral or cell-mediated immunity.
( Manju Acharya ),( Jihun Jo ),( Anju Maharjan ),( Ravi Gautam ),( Daeun Lee ),( Changyul Kim ),( Yong Heo ) 한국동물실험대체법학회 2020 동물실험대체법학회지 Vol.14 No.1
Although immune suppressive drugs have contributed for the cancer chemotherapy, organ transplant, but their therapeutic efficacy is limited. This study aims to evaluate the immunological modulations induced by four representative immunosuppressive on THP-1 cell line. 75% cell viabilities were determined and four test concentrations, 0.01X, 0.1X, 0.5X, and vehicle control were chosen. Culture supernatants were collected at 24 h after lipopolysaccharide (LPS1 ㎍/㎖) activation in the presence of test substances. 27 target cytokines were measured through luminex system and relative cytokine production levels (RCPLs,%) were calculated. Cytokines with the RCPL below 100% at all the three concentrations were 21 including IL-1beta, -1ra, -2, -4, -5, -6, -7, -9, -10, -12, -13, -15, -17, Eotaxin, FGF-basic, GM-CSF, IFNγ, IP-10, MCP-1α, PDGF-BB, and VEGF in dexamethasone-treated cells. Concerning on cyclophosphamide, the RCPLs on 11 cytokines were less than 100% at all the 3 concentrations. Treatment of cyclosporine demonstrated 10 cytokines with below 100% RCPL at all the 3 concentrations. The RCPLs below 100% were observed with 22 cytokines including IL-1β, -1ra, -2, -4, -5, -6, -7, -9, -12, -13, -15, -17, Eotaxin, GFG-basic, G-CSF, GM-CSF, IFNγ, MCP-1, MIP-1α, PDGF-BB, TNFα, and VEGF from THP-1 cells treated with tacrolimus at all the 3 concentrations. The present study indicates that tacrolimus and dexamethasone are stronger immunosuppressants than cyclophosphamide and cyclosporine on THP-1 cell line.
Ravi Gautam,DaEun Lee,Anju Maharjan,Manju Acharya,JiHun Jo,HyunJi Park,YuBin Kim,Pramod Bahadur KC,Chang Yul Kim,HyoungAh Kim,JiMin Choi,SooMin Lim,YoungWoo Shin,SuBin Yang,Yong Heo 한국실험동물학회 2021 한국실험동물학회 학술발표대회 논문집 Vol.2021 No.7
Nanoparticles (NPs), fine particles ranging from 1 to 100 nm, are being used extensively in various fields including industry, agriculture, engineering. Nanoparticles also have become the imperative parts of cosmetic and dermal products owing to the higher benefits over their counter bulk particles. Manganese oxide (Mn2O3) and copper oxide (CuO) are used in cosmetics and dermal products owing to their UV protection and antimicrobial properties and also to make products transparent and aesthetically acceptable. Although Mn2O3 and CuO are used in consumer products that get exposed to the skin, their skin sensitization potential has not been well investigated. In this study we employed, Local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method (LLNA: BrdU-FCM) to predict the skin sensitization (SS) potential of nanoparticles. LLNA: BrdU-FCM is a method for screening of SS potential that addressed the activation and proliferation of T-lymphocytes, key event-4 (KE-4), on adverse outcome pathway (AOP) for SS. Mn2O3 (5%, 10%, and 25%), and CuO (5%, 10%, and 25 %) NPs were suspended in dimethyl sulfoxide (DMSO) for experimentation. AOO (acetone: olive oil = 4:1) and α-hexyl cinnamaldehyde were used as negative and positive control respectively. Mn2O3 and CuO NPs were both classified as non-sensitizer with maximum stimulation indices of 1.4 and 0.99 respectively. Although both NPs were classified as non-sensitizer using LLNA: BrdU-FCM, further assays addressing other KEs of AOP for SS could be carried out as confirmation and cautious use of products containing these nanoparticles is recommended as various studies have revealed their toxicity in vitro . This study was supported by the Korea National Research Foundation (grant number: 2020R1I1A3A0403650911) and the Ministry of Environment-Educational training program for the management of information on the hazards and risk of chemical substances.
화학물질들의 면역독성 모니터링을 위한 THP-1 수지상세포주 활용
( Manju Acharya ),( Ravi Gautam ),( Anju Maharjan ),조지훈 ( Ji-hun Jo ),이다은 ( Daeun Lee ),박현지 ( Hyunji Park ),김유빈 ( Yubin Kim ),박성민 ( Sungmin Park ),허용 ( Yong Heo ) 대구가톨릭대학교 자연과학연구소 2020 자연과학연구논문집 Vol.18 No.1
Alternative test methods have been developed for various areas of toxicologic evaluation including skin sensitization or irritation, eye irritation, genotoxicity, or carcinogenicity. Whereas, alternative test method, especially based on in vitro approach, for screening immunotoxic substances was not actively challenged and therein, no test guideline has not been officialized internationally. THP-1 dendritic cell line was chosen for developing in vitro alternative test method for screening immunotoxic chemical or medicinal substances. Regarding major pathway for immune suppression or non-specific immune enhancement, antigen presenting cell is considered as most important target immune cells. Two test substances representing immune-enhancement(ribavirin) or immune-suppression (cyclophosphamide) were tested whether production of cytokines or chemokines is differentially modulated through supplementation of 0.01x, 0.1x, and 0.5x 75% cell viability(CV75) concentration of those compounds. Cell culture supernatants were collected at 24 h after the activation with 1 ㎍/㎖ lipopolysaccharide. Relative cytokine production levels (% versus each vehicle control, RCPL) were calculated. Production of all 27 cytokines evaluated was confirmed, which included IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7,IL-8, IL-9,IL-10, IL-12, IL-13, IL-15, IL-17, Eatoxin, FGF basic, G-CSF, GM-CSF, IFNγ, IP-10, MCP-1, PDGF-BB, MIP-1α, MIP-1β, RANTES, TNFα, VEGF. Treatment of ribavirin demonstrated all the RCPL values over 100% at 0.01x and 0.1x CV75 concentration. Treatment of cyclosphosphamide demonstrated the RCPL values below 100%except IL-12(0.01x), IL-13(0.1x), MCP-1(0.01x), TNFα (0.01x, 0.1x, and 0.5x), and VEGF(0.01x, 0.1x, and 0.5x). This findings suggest that THP-1 cell line is a valuable cell line to develop an in vitro alternative test method for screening immunotoxic chemical substances.