scholarly journals Cytokine and Chemokine mRNA Expressions after Mycobacterium tuberculosis-Specific Antigen Stimulation in Whole Blood from Hemodialysis Patients with Latent Tuberculosis Infection

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 595
Author(s):  
Ji Young Park ◽  
Sung-Bae Park ◽  
Heechul Park ◽  
Jungho Kim ◽  
Ye Na Kim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mrna Expression ◽  
Whole Blood ◽  
Latent Tuberculosis Infection ◽  
Latent Tuberculosis ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Healthy Individuals ◽  
Relative Mrna Expression ◽  
Mrna Expression Levels

There have been few reports on the kinetics of hemodialyzed (HD) patients’ immune responses in latent tuberculosis infection (LTBI). Therefore, in the present study, messenger ribonucleic acid (mRNA) expression levels of nine immune markers were analyzed to discriminate between HD patients with LTBI and healthy individuals. Nine cytokines and chemokines were screened through relative mRNA expression levels in whole blood samples after stimulation with Mycobacterium tuberculosis (MTB)-specific antigens from HD patients with LTBI (HD/LTBI), HD patients without LTBI, and healthy individuals, and results were compared with the QuantiFERON-TB Gold In-Tube (QFT-GIT) test. We confirmed that the C-C motif chemokine 11 (CCL11) mRNA expression level of the HD/LTBI group was significantly higher than the other two groups. Especially, the CCL11 mRNA expression level of the >0.7 IU/mL group in the QFT-GIT test was significantly higher than the <0.2 IU/mL group in the QFT-GIT test and the 0.2–0.7 IU/mL group in the QFT-GIT test (p = 0.0043). The present study reveals that the relative mRNA expression of CCL11 was statistically different in LTBI based on the current cut-off value (i.e., ≥0.35 IU/mL) and in the >0.7 IU/mL group. These results suggest that CCL11 mRNA expression might be an alternative biomarker for LTBI diagnosis in HD patients.

scholarly journals Cytokine and Chemokine mRNA Expressions After Mycobacterium Tuberculosis-Specific Antigen Stimulation in Whole Blood From Hemodialysis Patients with Latent Tuberculosis Infection

2020 ◽  
Author(s):  
Ji Young Park ◽  
Sung-Bae Park ◽  
Heechul Park ◽  
Jungho Kim ◽  
Ye-Na Kim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mrna Expression ◽  
Whole Blood ◽  
Latent Tuberculosis Infection ◽  
Specific Antigen ◽  
Latent Tuberculosis ◽  
Tuberculosis Infection ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Relative Mrna Expression

Abstract There have been few reports on the kinetics of hemodialyzed (HD) patients’ immune responses in latent tuberculosis infection (LTBI). Nine cytokines and chemokines were screened through relative mRNA expression levels in whole blood samples after stimulation with Mycobacterium tuberculosis (MTB)-specific antigens from HD patients with LTBI (HD/LTBI), HD patients without LTBI, and healthy individuals and results were compared with QuantiFERON-TB Gold In-Tube (QFT-GIT) test.Only the CCL11 mRNA expression level of the HD/LTBI group was significantly higher than the other two groups (P = 0.028). CCL11 mRNA expression level of the > 0.7 IU/mL group in QFT-GIT test was significantly higher than the < 0.2 IU/mL group in QFT-GIT test and the 0.2-0.7 IU/mL group in QFT-GIT test (P = 0.0043). The present study reveals that the relative mRNA expression of CCL11 was statistically different in LTBI based on the current cut-off value (i.e., ≥ 0.35 IU/mL) and in the > 0.7 IU/mL group. These results suggest that CCL11 might be an alternative biomarker for LTBI diagnosis in HD patients.

scholarly journals Th2 cytokine bias induced by silver nanoparticles in peripheral blood mononuclear cells of common bottlenose dolphins (Tursiops truncatus)

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5432 ◽  
Author(s):  
Wen-Ta Li ◽  
Lei-Ya Wang ◽  
Hui-Wen Chang ◽  
Wei-Cheng Yang ◽  
Chieh Lo ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Mononuclear Cells ◽  
Bottlenose Dolphins ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Expression Levels ◽  
Th2 Cytokine ◽  
Tnf Α ◽  
Ifn Γ ◽  
Mrna Expression Levels

Background Silver nanoparticles (AgNPs) have been widely used in many commercial products due to their excellent antibacterial ability. The AgNPs are released into the environment, gradually accumulate in the ocean, and may affect animals at high trophic levels, such as cetaceans and humans, via the food chain. Hence, the negative health impacts caused by AgNPs in cetaceans are of concern. Cytokines play a major role in the modulation of immune system and can be classified into two types: Th1 and Th2. Th1/Th2 balance can be evaluated by the ratios of their polarizing cytokines (i.e., interferon [IFN]-γ/Interleukin [IL]-4), and animals with imbalanced Th1/Th2 response may become more susceptible to certain kinds of infection. Therefore, the present study evaluated the in vitro cytokine responses of cetacean peripheral blood mononuclear cells (cPBMCs) to 20 nm citrate-AgNPs (C-AgNP20) by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Methods Blood samples were collected from six captive common bottlenose dolphins (Tursiops truncatus). The cPBMCs were isolated and utilized for evaluating the in vitro cytokine responses. The cytokines evaluated included IL-2, IL-4, IL-10, IL-12, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α. The geometric means of two housekeeping genes (HKGs), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and β2-microglobulin (B2M), of each sample were determined and used to normalize the mRNA expression levels of target genes. Results The ratio of late apoptotic/necrotic cells of cPBMCs significantly increased with or without concanavalin A (ConA) stimulation after 24 h of 10 µg/ml C-AgNP20 treatment. At 4 h of culture, the mRNA expression level of IL-10 was significantly decreased with 1 µg/ml C-AgNP20 treatment. At 24 h of culture with 1 µg/ml C-AgNP20, the mRNA expression levels of all cytokines were significantly decreased, with the exceptions of IL-4 and IL-10. The IFN-γ/IL-4 ratio was significantly decreased at 24 h of culture with 1 µg/ml C-AgNP20 treatment, and the IL-12/IL-4 ratio was significantly decreased at 4 or 24 h of culture with 0.1 or 1 µg/ml C-AgNP20 treatment, respectively. Furthermore, the mRNA expression level of TNF-α was significantly decreased by 1 µg/ml C-AgNP20 after 24 h of culture. Discussion The present study demonstrated that the sublethal dose of C-AgNP20 (≤1 µg/ml) had an inhibitory effect on the cytokine mRNA expression levels of cPBMCs with the evidence of Th2 cytokine bias and significantly decreased the mRNA expression level of TNF-α. Th2 cytokine bias is associated with enhanced immunity against parasites but decreased immunity to intracellular microorganisms. TNF-α is a contributing factor for the inflammatory response against the infection of intracellular pathogens. In summary, our data indicate that C-AgNP20 suppresses the cellular immune response and thereby increases the susceptibility of cetaceans to infection by intracellular microorganisms.

scholarly journals Angiogenesis system, as a part of endothelial dysfunction in patients with diabetes mellitus type 2: relationship with obesity

2020 ◽  
Vol 92 (10) ◽  
pp. 23-28
Author(s):  
A. S. Severina ◽  
M. V. Shestakova
Keyword(s):  
Diabetes Mellitus ◽  
Mrna Expression ◽  
Body Mass ◽  
Diabetes Mellitus Type 2 ◽  
Diabetes Mellitus Type ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Patients With Diabetes ◽  
Mrna Expression Levels

Aim.To investigate parameters of angiogenesis system in patients with diabetes mellitus and their relationship with obesity. Materials and methods.104 patients with diabetes mellitus type 2 were included in the study. Patients were divided in 2 groups: Obesity+ (body mass index30 kg/m2;n=63) and Obesity- (body mass index 30 kg/m2;n=41). In all patients was performed clinico-diagnostical examination. mRNA expression levels of vascular endothelial growth factor (VEGF), its receptors flt-1 (fms-like tyrosine kinase 1), KDR (human kinase insert domain receptor) were determined in blood mononuclear cells. Results.There were no statistically significant differences in investigated parameters between study groups. mRNA expression level of VEGF was slightly lower in men compared to women: 0.19 (0.14; 0.32)vs0.28 (0.12; 0.4) respectively,р=0.2236. MRNA expression level of flt-1 was lower in men compared to women: 0.14 (0.04; 0.3)vs0.25 (0.12; 0.38),р=0.0321 (statistically significant). We found statistically significant correlations of mRNA expression level of VEGF with mRNA expression level of flt-1 and KDR. Also we found strong positive correlations of BMI and mRNA expression levels VEGF, flt-1, KDR (r=0.86107,r=0.86125,r=0.86112, respectively,p0.001). Conclusion.Results of the study displayed relationship of obesity and angiogenesis system condition in patients with diabetes mellitus type 2. Further investigations are perspective for the future as a way to new therapeutical approach of obesity and its complications treatment.

The Feature of CD3-zeta Chain Gene Expression in Mononuclear Cells without and with Stimulation by Different Factors from Umbilical Cord Blood.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3870-3870
Author(s):  
Shaohua Chen ◽  
Yangqiu Li ◽  
Lijian Yang ◽  
Si Chen
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Cord Blood ◽  
Mrna Expression ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Healthy Adults ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Relative Mrna Expression

Abstract CD3 zeta gene is required for efficient TCR expression and plays a central role in the signal-transducing events leading to T and NK-cell activation and proliferation. Umbilical cord blood (UCB) has been used successfully as a source of allogeneic transplantation and UCB T cells have showed capacity for production the specific CTL by tumor associated antigen ex vivo. In order to investigate the feature of CD3 zeta gene expression pattern in cord blood, UCB T cell without or with stimulation by different stimulators, including PHA, IL-2, CD3 monoclonal antibody (McAb), CD28 McAb + IL-2 and PML-RARα peptide) were used to analyze. By using Real-Time PCR with SYBR Green I technique, the expression level of CD3 zeta gene was analyzed in T-cells from 60 cases of UCB before and after T-cells culture at different time points (5–20days), β2-microglobulin gene was used as an endogenous reference. The relative mRNA expression level of CD3 zeta gene was used by the 2- ΔCt method. According to melting curve, polymorphism of nucleotide sequence was determined by PCR products direct sequencing. 60 cases healthy adults served as controls. The results showed that CD3 zeta gene was expressed in all cases from both UCB and healthy adults. The mean value 6.7%±5.56% of relative mRNA expression level of CD3 zeta gene was found in 60 UCB cases, the expression level under 1.0% was detected in 4 cases, over 10% in 14 cases and a high expression of 25.53% only in one case. In contrast, 3.1%±2.23% of relative mRNA expression level of CD3 zeta gene was detected in 60 cases healthy adults. The expression of the CD3 ζ gene from the healthy adults is more concentrated and the highest expression is only 9.34%. Compare with the healthy adults, a significant higher expression of CD3 zeta gene was found from UCB (P=0.000). Polymorphism and mutation of CD3 zeta gene were not identified by sequence analysis in both UCB and healthy samples. For the culture cells, CD3 zeta gene expression level in initial culture (5–10days) was increased after different stimulation. A higher expression lever was found in combined CD3 MCAb + CD28 McAb with or without PML-RARα peptide than in stimulation with PHA or IL-2 alone. The CD3 zeta gene expression lever in UCB T cells induced by combined PML-RARα peptide was 6.37 times as much as unstimulated cells at 10 days, whereas the CD3 zeta gene expression lever in UCB T cells stimulated by IL-2, CD3 McAb plus CD28 McAb was 5.83 times higher than that from un-stimulated cells. However, when the duration of T cells culture was prolonged, the expression of CD3 zeta was gradually reduced in all groups after 10 days. In conclusion, this is to our knowledge, the first description of CD3 zeta gene expression in UCB. Our data suggest that a higher expression of CD3 zeta gene could be found in UCB than in adult peripheral blood, and up-regulation of CD3 zeta gene can also be achieved after stimulation with PHA, IL-2, CD3 McAb + CD28 McAb + IL-2 with or without PML-RARα peptide, respectively. In addition, combined CD3 McAb + CD28 McAb with/without PML-RARα peptide showed a significantly higher capacity for promoting proliferation. The down-expression of CD3 zeta in cultured T cells after 10 days remains an open question.

WT-1 Expression Level In BM Is The Great Prognostic Marker In Three Of Classification IPSS, WPSS, and Latest Revised IPSS(IPSS-R)

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2795-2795
Author(s):  
Sumiko Kobayashi ◽  
Yasunori Ueda ◽  
Mineo Kurokawa ◽  
Kiyoyuki Ogata ◽  
Hirohiko Shibayama ◽  
...  
Keyword(s):  
Regression Analysis ◽  
Mrna Expression ◽  
Survival Time ◽  
Research Funding ◽  
Mrna Level ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Expression Levels ◽  
Wbc Count ◽  
Mrna Expression Levels

Abstract Objective/material/method It is known new classification named revised IPSS(IPSS-R) as a prognosis in MDS patients in 2012. To investigate the clinical utility of WT1 mRNA expression level including IPSS-R, we studied of MDS patients who provided consent at 17 medical institutions nationwide from Dec. 2008 to Sep. 2009 were enrolled in Japan. A total of 172 subjects including 115 MDS patients by IPSS (RA:69, RARS:9, RAEB:24, RAEB-t :13 ). 13 patients who developed AML from MDS, and 44 patients with non malignant hematological disorders who provided consent. This study was designed to follow up 82 patients, who gave secondary consent, among 115 MDS patients registered in study ODK-0801 for 5 years up to 2014 to analyze in detail the relationship between the WT1 mRNA expression level and prognosis. WT1 mRNA level in PB and BM were measured using the WT1 mRNA assay kit “OTSUKA” (OTSUKA PHARMACEUTICAL CO., LTD). WT1 mRNA expression levels in peripheral blood (and in bone marrow, if possible) were measured periodically to evaluate the usefulness of the WT1 mRNA expression level as a monitoring marker of MDS and analyze changes in pathology in individual patients by central review and changes in WT1 mRNA expression levels. In this study, value of 50 copies/μgRNA was set as the cut off value for WT-1mRNA expression. The differences in clinical and demographic data were assessed in the chi-square test and logistic regression analysis. The survival data was analyzed using Kaplan-Meier method and compared by the log-rank test. This study presents the results of interim analysis 3 years after the start of the investigation. Results The comparison study between the patients categorized into three groups by the WT1 mRNA level　in BM ,GroupI:less than 102 copies/μgRNA(n=35), Group II: 102 to 104 copies/μgRNA(n=30), and GroupIII: more than 104 copies/μgRNA(n=17) resulted that the survival rates decreased significantly as the WT1 mRNA level increased(GI vs GII P<0.01, GI vs GIII P<0.01, GII vs GIII p<0.067), respectively. In multivariate Cox proportional hazard regression analysis of IPSS, five out of fifteen parameters, which are WBC count (P=0.0001), IPSS score (P=0.0003), blast in PB(P=0.0011), WT1 mRNA level in BM(P=0.0055), sex(P=0.093) were independently associated with survival time, respectively. And we studied same analysis used WPSS. WBC count (P=0.0001), WPSS socre(P=0.0001), blast in PB(P=0.0037), WT1 mRNA level in BM(P=0.0029), sex(P=0.012) were independently associated with survival time, respectively. Also we analyzed using IPSS-R. In multivariate Cox proportional hazard regression analysis, five out of fifteen parameters, WBC count (P=0.0001), IPSS-R score(P=0.0001), blast in PB(P=0.0012), WT1 mRNA level in BM(P=0.01), sex(P=0.01) were independently associated with survival time, but not ANC(both score and absolute number), Hb, platelet, karyotype or other parameters. Using these selected five valiables, we provided the level of WT-1mRNA with 4 risk groups which classified :100, 50100, 100,100, 150,100copies/μRNA). According to increasing of WT-1mRNA level, survival duration shortened the increase with the increase in each risk group. Discussion The WT-1 mRNA level in BM was positively correlated with the prognosis of MDS stage and tended to be correlated with advanced risk of IPSS, WPSS, and IPSS-R. Therefore it was considered to be a useful prognostic marker for MDS. Conclusion We found that the survival time was shortened significantly with an increase in WT1 mRNA expression levels in both peripheral blood and bone marrow, demonstrating that WT1 mRNA expression levels are a highly useful prognostic factor in MDS even if we use any of classification of IPSS, WPSS, IPSS-R. This assay has great possibility to contribute to more appropriate therapeutic decisions in MDS patients as well as diagnosis and to evaluate the timing of allogeneic transplantation. Disclosures: Kurokawa: Novartis: Consultancy, Research Funding; Bristol-Myers Squibb: Research Funding; Celgene: Consultancy, Research Funding. Usuki:Alexion Pharmaceuticals, Inc.: Speakers Bureau. Ohyashiki:Novartis: Honoraria, Research Funding.

scholarly journals MARcKS and nAP-22 proteins mRnA expression in renal cortex and renal medulla of rats with spontaneous hypertension

2018 ◽  
Vol 24 (4) ◽  
pp. 435-440
Author(s):  
A. S. Aldekeeva ◽  
Y. S. Kraynova ◽  
E. D. Rudenko ◽  
N. Z. Klyueva
Keyword(s):  
Mrna Expression ◽  
Renal Cortex ◽  
Renal Medulla ◽  
Expression Level ◽  
Shr Rats ◽  
Mrna Expression Level ◽  
Interstrain Differences ◽  
Wky Rats ◽  
Mrna Expression Levels ◽  
Normotensive Control

Objective. To study the changes in mRNA expression level of two main protein kinase C substrates — MARCKS and NAP-22 — in rats with spontaneous hypertension (SHR rats) and in normotensive control rats (WKY rats) in renal cortex, renal medulla and total kidney. We also aimed at the identification of possible interstrain differences between the mRNA expression levels.Design and methods.We assessed the level of MARCKS and NAP-22 mRNA by real-time polymerase chain reaction in male SHR and WKY (as a normotensive control) rats.Results. In SHR rats, MARCKS mRNA expression level in renal cortex was 1,5 times higher than in renal medulla (p = 0,0001) and also higher than in total kidney (p = 0,001), in renal medulla it was lower than in total kidney (p = 0,002). In WKY rats, MARCKS mRNA expression level in renal cortex was higher than in renal medulla (p = 0,0005). There was no differences neither between renal cortex and total kidney (p = 0,011), nor between renal medulla and total kidney (p = 0,716). In SHR rats, NAP-22 mRNA expression level in renal cortex was twofold higher than in renal medulla (p = 0,001), in renal medulla it was lower than in total kidney (p = 0,005), the differences between renal cortex and total kidney were less significant (p = 0,011). In WKY rats, NAP-22 mRNA expression level in renal cortex was 1,5 times higher than in renal medulla (p = 0,001), while in renal medulla it was lower than in total kidney (p = 0,002). There was no significant difference in NAP-22 mRNA expression level between renal medulla and total kidney (p = 0,011). There were no significant interstrain differences in the animal groups either in the levels of MARCKS mRNA expression in renal cortex (p = 0,872), in renal medulla (p = 0,024) or in total kidney (p = 0,520). Neither there were differences in the levels of NAP-22 mRNA expression in cortex (p = 0,028), in medulla (p = 0,028) and in total kidney (p = 0,978).Conclusions. In both SHR and WKY rat strains, the level of MARCKS and NAP-22 mRNA expression in cortical and medullary kidney layers is different, in WKY rats these differences are less pronounced. At the same time, interstrain differences in NAP-22 and MARCKS mRNA expression levels in cortical, medullary layers and in total kidney of SHR and WKY rats were not found.

scholarly journals Th2 cytokine bias induced by silver nanoparticles in peripheral blood mononuclear cells of common bottlenose dolphins (Tursiops truncatus)

2018 ◽  
Author(s):  
Wen-Ta Li ◽  
Lei-Ya Wang ◽  
Hui-Wen Chang ◽  
Wei-Cheng Yang ◽  
Chieh Lo ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Mononuclear Cells ◽  
Bottlenose Dolphins ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Expression Levels ◽  
Th2 Cytokine ◽  
Tnf Α ◽  
Ifn Γ ◽  
Mrna Expression Levels

Background Silver nanoparticles (AgNPs) have been widely used in many commercial products due to their excellent antibacterial ability. The AgNPs are released into the environment, gradually accumulate in the ocean, and may affect animals at high trophic level, such as cetaceans and humans, via the food chain. Hence, the negative health impacts caused by AgNPs in cetaceans are of concern. Cytokines play a major role in the modulation of immune system and can be classified into two types, Th1 and Th2. Th1/Th2 balance can be evaluated by the ratios of their polarizing cytokines (i.e., interferon [IFN]-γ/ Interleukin [IL]-4), and animals with imbalanced Th1/Th2 response may become more susceptible to certain kinds of infection. Therefore, the present study evaluated the in vitro cytokine responses of cetacean peripheral blood mononuclear cells (cPBMCs) to 20 nm citrate-AgNPs (C-AgNP20) by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Methods Blood samples were collected from 6 captive common bottlenose dolphins (Tursiops truncatus). The cPBMCs were isolated and utilized for evaluating the in vitro cytokine responses. The cytokines evaluated included IL-2, IL-4, IL-10, IL-12, interferon (IFN)-γ, and tumor necrosis factor (TNF)-ɑ. The geometric means of two housekeeping genes (HKGs), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and β2-microglobulin (B2M), of each sample were determined and used to normalize the mRNA expression levels of target genes. Results The ratio of late apoptotic/necrotic cells of cPBMCs significantly increased with or without concanavalin A (ConA) stimulation after 24 h of 10 μg/ml C-AgNP20 treatment. At 4 h of culture, the mRNA expression level of IL-10 was significantly decreased with 1 μg/ml C-AgNP20 treatment. At 24 h of culture with 1 μg/ml C-AgNP20, the mRNA expression levels of all cytokines were significantly decreased, with the exceptions of IL-4 and IL-10. The IFN-γ/IL-4 ratio was significantly decreased at 24 h of culture with 1 μg/ml C-AgNP20 treatment, and the IL-12/IL-4 ratio was significantly decreased at 4 or 24 h of culture with 0.1 or 1 μg/ml C-AgNP20 treatment, respectively. Furthermore, the mRNA expression level of TNF-α was significantly decreased by 1 μg/ml C-AgNP20 after 24 h of culture. Discussion The present study demonstrated that the sublethal dose of C-AgNP20 (≤ 1 μg/ml) had an inhibitory effect on the cytokine mRNA expression levels of cPBMCs with the evidence of Th2 cytokine bias and significantly decreased the mRNA expression level of TNF-α. Th2 cytokine bias is associated with enhanced immunity against parasites but decreased immunity to intracellular microorganisms. TNF-α is a contributing factor for the inflammatory response against the infection of intracellular pathogens. In summary, our data indicate that C-AgNP20 suppresses the cellular immune response and thereby increases the susceptibility of cetaceans to infection by intracellular microorganisms.

scholarly journals Th2 cytokine bias induced by silver nanoparticles in peripheral blood mononuclear cells of common bottlenose dolphins (Tursiops truncatus)

2018 ◽  
Author(s):  
Wen-Ta Li ◽  
Lei-Ya Wang ◽  
Hui-Wen Chang ◽  
Wei-Cheng Yang ◽  
Chieh Lo ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Mononuclear Cells ◽  
Bottlenose Dolphins ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Expression Levels ◽  
Th2 Cytokine ◽  
Tnf Α ◽  
Ifn Γ ◽  
Mrna Expression Levels

Background Silver nanoparticles (AgNPs) have been widely used in many commercial products due to their excellent antibacterial ability. The AgNPs are released into the environment, gradually accumulate in the ocean, and may affect animals at high trophic level, such as cetaceans and humans, via the food chain. Hence, the negative health impacts caused by AgNPs in cetaceans are of concern. Cytokines play a major role in the modulation of immune system and can be classified into two types, Th1 and Th2. Th1/Th2 balance can be evaluated by the ratios of their polarizing cytokines (i.e., interferon [IFN]-γ/ Interleukin [IL]-4), and animals with imbalanced Th1/Th2 response may become more susceptible to certain kinds of infection. Therefore, the present study evaluated the in vitro cytokine responses of cetacean peripheral blood mononuclear cells (cPBMCs) to 20 nm citrate-AgNPs (C-AgNP20) by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Methods Blood samples were collected from 6 captive common bottlenose dolphins (Tursiops truncatus). The cPBMCs were isolated and utilized for evaluating the in vitro cytokine responses. The cytokines evaluated included IL-2, IL-4, IL-10, IL-12, interferon (IFN)-γ, and tumor necrosis factor (TNF)-ɑ. The geometric means of two housekeeping genes (HKGs), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and β2-microglobulin (B2M), of each sample were determined and used to normalize the mRNA expression levels of target genes. Results The ratio of late apoptotic/necrotic cells of cPBMCs significantly increased with or without concanavalin A (ConA) stimulation after 24 h of 10 μg/ml C-AgNP20 treatment. At 4 h of culture, the mRNA expression level of IL-10 was significantly decreased with 1 μg/ml C-AgNP20 treatment. At 24 h of culture with 1 μg/ml C-AgNP20, the mRNA expression levels of all cytokines were significantly decreased, with the exceptions of IL-4 and IL-10. The IFN-γ/IL-4 ratio was significantly decreased at 24 h of culture with 1 μg/ml C-AgNP20 treatment, and the IL-12/IL-4 ratio was significantly decreased at 4 or 24 h of culture with 0.1 or 1 μg/ml C-AgNP20 treatment, respectively. Furthermore, the mRNA expression level of TNF-α was significantly decreased by 1 μg/ml C-AgNP20 after 24 h of culture. Discussion The present study demonstrated that the sublethal dose of C-AgNP20 (≤ 1 μg/ml) had an inhibitory effect on the cytokine mRNA expression levels of cPBMCs with the evidence of Th2 cytokine bias and significantly decreased the mRNA expression level of TNF-α. Th2 cytokine bias is associated with enhanced immunity against parasites but decreased immunity to intracellular microorganisms. TNF-α is a contributing factor for the inflammatory response against the infection of intracellular pathogens. In summary, our data indicate that C-AgNP20 suppresses the cellular immune response and thereby increases the susceptibility of cetaceans to infection by intracellular microorganisms.

Hematopoietic Cell Expression of OPAL1, an Outcome Predictor in Pediatric Acute Lymphoblastic Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1457-1457
Author(s):  
Weihua Zeng ◽  
I.-Ming Chen ◽  
Yuexian Xu ◽  
Jeff Potter ◽  
Kerem Ar ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
B Cells ◽  
Mrna Expression ◽  
Mononuclear Cells ◽  
Lymphoblastic Leukemia ◽  
Expression Level ◽  
Healthy Individuals ◽  
Outcome Predictor ◽  
Expression Ratio ◽  
Mrna Expression Levels

Abstract Background: A novel sequence, NM_hypothetical protein FLJ20154, has been identified in our previous microarray study as one of the most powerful predictors of CCR in B precursor acute lymphoblastic leukemia (ALL). We have cloned, characterized, and named this gene as Outcome Predictor in Acute Leukemia 1(OPAL1). Our preliminary data indicates that OPAL1 distinguishes those ALL patients with t(12;21) who will achieve CCR, and predicts for CCR in children who have adverse prognostic features (normal karyotype, higher age, higher white blood cell count at presentation). To further investigate the role of OPAL1 in ALL, we first determined OPAL1 expression in cells from different hematopoietic lineages, compared the OPAL1 mRNA expression levels of normal hematopoietic cells with ALL patients with t(12;21) or t(1;19). Methods: Peripheral blood mononuclear cells from 46 healthy individuals (age range: 2–18 years) were collected and sorted into three groups: B enriched (CD19+), T enriched (CD3+) and others (mostly were nutrophil cells); along with pre-treatment ALL samples with high blast count (&gt;90%) from patient with t(12;21) or t(1;19) were used for study. mRNA expression levels of OPAL1 were determined by real-time quantitative PCR. The OPAL1 expression values were calculated by using the formula: Value=1/2^ΔCt, ΔCt=Cttarget−Ctcontrol.. The generated values were grouped into four multitude ranges categories: −10−2, 10−1, 100, 101. Results: Most of the relative expression ratio of OPAL1 (normalized to GAPDH) for the B enriched (29/45, 64.5%) or T enriched cells (6/12, 50%) in healthy individuals fell in the 100 level, while 75% (9/12) of the other residual mononuclear cells (mostly neutrophils cells), their OPAL1 relative expression ratio were in the 10−1 level which is ten times lower than those seen in normal T and B cells. OPAL1 expression ratio were significantly decreased in ALL patients with t(12;21) and t(1;19) compared with normal B enriched cells, p=0.0079 and p= 0.0006, respectively. Forty five percent (9/20) of t(12;21) patients their OPAL1expression ratio were at 10−1 level, and 40% (8/20) at 10−2 level. As for patients with t(1;19) or higher risk patients group (three patients in this group were bcr/abl positive), 60% (9/15) of the OPAL1 expression ratio were at 10−2 level. These results strongly suggested that the correlation of higher OPAL1 expression is associated with “good risk” genetic subtypes of ALL Summary: Although currently the biologic functions of OPAL1 remained to be determined, the high expression level of OPAL1 in normal T or B lymphocytes compares with non-T, non-B cells; low expression in ALL leukemia blast cells suggested that OPAL1 may play a role in lymphocyte differentiation, especially in the development and maturation of the normal B cell. The reverse relationship between OPAL1 expression level and the patient genetic risk factors indicates that OPAL1 might be another strong predictor for treatment outcome along with other well known clinical indicators. Hence, the levels of OPAL1 may be useful in identifying ALL patients with a potential good outcome, who may be able to benefit from less intensive treatment regimens and still achieve long term remission.

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