2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a well-known immunotoxic environmental pollutant. However, most immunotoxicology studies of TCDD were based on the animal models and the inner mechanisms have just focused on a few genes/proteins. In this study, the immune functions of THP-1-derived macrophages was measured with in-vitro bioassays after 24-h exposure of TCDD including environmentally relevant concentrations. RNA-seq and Weighted Gene Co-expression Network Analysis were used to characterize the immunotoxicity molecular mechanisms. Our study is the first report on the TCDD-induced effects of cell adhesion, morphology, and multiple cytokines/chemokines production on THP-1 macrophages. After TCDD treatment, we observed an inhibited cell adherence, probably attributed to the suppressed mRNA levels of adhesion molecules ICAM-1, VCAM-1 and CD11b, and a decrease in cell pseudopodia and expression of F-actin. The inflammatory cytokines TNF-α, IL-10 and other 8 cytokines/chemokines regulating granulocytes/T cells and angiogenesis were disrupted by TCDD. Alternative splicing event was found to be a sensitive target for TCDD. Using WGCNA, we identified 10 hub genes (TNF, SRC, FGF2, PTGS2, CDH2, GNG11, BDNF, WNT5A, CXCR5 and RUNX2) highly relevant to these observed phenotypes, suggesting AhR less important in the effects TCDD have on THP-1 macrophages than in other cells. Our findings broaden the understanding of TCDD immunotoxicity on macrophages and provide new potential targets for clarifying the molecular mechanisms.

The aim of this study was to assess the impact of treated wastewaters on native wild Prussian carp inhabiting effluent-receiving waters (ERC) receiving municipal and sugar plant treated wastewaters, further downstream waters (DW), and a detached canal unaffected by the WWTP activities. To that end, general fish health status was determined, including plasma biochemical, haematological, oxidative stress and tissue histopathological indices, over three seasons. The greatest tissue alterations were in fall in ERC during sugar beet processing, as hypertrophy of gill epithelial and interlamellar cells, necrosis and lymphocytic infiltration, hyperplasia and hypertrophy of renal tubules, distention of hepatic sinusoids. In fall the lowest leukocytes, lymphocytes and granulocytes (2467 ± 565, 1333 ± 264, 1133 ± 488 cells/μL respectively), as well as highest plasma ALP (52.7 ± 19.39 U/L) were measured. ERC in fall had the highest ammonium (20 mg/L), nitrite (1.48 mg/L), nitrate (13.4 mg/L), and lowest dissolved O2 (1.23 mg/L). Gill, kidney and liver alterations, and the highest plasma cholesterol (9.1 ± 1.98 mmol/L) were noted in DW fish in fall. Tissue morphology during sugar cane processing seems a consequence of cellular and structural tissue integrity loss. Structural heterogeneity of gills and spleen was enhanced with increasing concentrations of heavy metals and correlated with oxidative stress (SOD 392.5 ± 77.28 U/L). Monogenean infestation was moderate in ERC fish in all seasons compared with DW fish. Prussian carp biological responses to multiple stressors, measured by the effects of WWTP on blood and tissue parameters, reached far downstream and were not of localized nature. This study demonstrated that in aquatic environments impacted with complex contaminants acting synergistically, causal relationships between biological responses and environmental stressors should be interpreted. Integrated histopathological, haematological and biochemical findings are valuable biomarkers for native fish adaptive patterns and monitoring of water quality/pollution of freshwater ecosystems.

Hemocytes are the main immune cells in bivalve mollusks and one of the sensitive targets for nanoparticle toxicity. Bivalve hemocytes consist of multiple functional heterogeneous cell types, but their different roles in immune system against foreign particles remain largely unknown. In order to clarify the different immune responses of hemocyte subpopulations to silver nanoparticles (AgNPs) and Ag ions, in this study, the Hong Kong oyster (Crassostrea hongkongensis) hemocytes were employed and separated into three subpopulations based on their cell size and granularity, including agranulocytes (R1), semigranulocytes (R2), and granulocytes (R3). We first demonstrated that AgNPs could rapidly enter into the oyster hemocytes within 3 h by phagocytosis process and resulted in different immune responses in hemocyte subpopulations. The most affected cell subtype by AgNPs was the granulocytes, followed by semigranulocytes, whereas agranulocytes were not affected following exposure to AgNPs. Interestingly, AgNPs induced the granule formation in semigranulocytes and further increased the proportion of granulocytes, whereas their ionic counterparts had no such effects on hemocyte composition, indicating the different detoxification mechanisms for nanoparticulate and ionic form. Following AgNP exposure, the dissolved Ag ions were accumulated in lysosomes and caused lysosomal dysfunction, indicating that lysosomes were the main targets for AgNP toxicity and the dissolved Ag ions were the main contributor of AgNP toxicity. Furthermore, AgNP exposure induced reactive oxygen production and impeded the lysosome function and phagocytosis in granulocytes, with impaired immunity system in oysters. Our study identified the different immune responses of oyster hemocyte subpopulations to AgNPs based on the in vitro short-term exposure assays, which may be applied to rapidly evaluate the ecotoxicological risks of different nanoparticles in aquatic systems.

Hemocyte parameters have been used as a proxy to characterize the health condition of marine bivalves, as the effects of external and internal stresses are reflected well in these parameters. Marine bivalve hemocyte functions are often depressed during the post-spawning period due to physiological stress and energy depletion. In this study, we analyzed temporal changes in hemocyte parameters of the oyster Saccostrea kegaki on Jeju Island, off the south coast of Korea, using flow cytometry. Total hemocyte count (THC), hemocyte types, hemocyte mortality, and phagocytosis capacity were analyzed. S. kegaki spawned during June and August, when the sea surface temperature increased from 18 to 23 °C. Most of the oysters were in the spent and resting phases from September to January. THC dropped dramatically from September to October, when most oysters completed spawning. Histology revealed that the residual eggs or sperm were actively resorbed through phagocytosis by hemocytes during the spent stage. Hemocyte mortality also showed its annual peak in October, possibly due to increased resorbing activities. The phagocytosis capacities of the granulocytes decreased dramatically from September to February. The level of energy reserves (glycogen) in post-spawning tissues was significantly lower than that prior to spawning. The low energy level reserve during the post-spawning period likely reduced the THC and immune capacities, as oysters may have been unable to acquire sufficient food from the ambient environment.

Marine bivalves are often used as a sentinel species in coastal environmental monitoring since changes in the environmental quality are often well preserved in their cells and tissues. Anadara and Tegillarca species of Arcidae, the blood cockles, are considered to be good sentinel species in monitoring coastal pollution and ecosystem health because they are distributed widely in the subsurface of intertidal mudflats. Internal cellular defense of the blood cockles to physical and biological stresses is mediated by the circulating hemocytes, while their hemocyte types and functions are poorly studied. In this study, we first characterized morphology and immune-related activities of hemocytes of three common blood cockles Anadara broughtonii, A. kagoshimensis, and Tegillarca granosa using flow cytometry. Based on cell morphology and immunological functions, we described five types of hemocytes identically in the three blood cockles: erythrocytes type-I (erythrocytes-I), erythrocytes type-II (erythrocytes-II), granulocytes, hyalinocytes, and blast-like cells. Erythrocytes were round cells containing hemoglobin with numerous granules in the cytoplasm and these cells consist of two central populations. Erythrocytes-I were the most abundant cells accounting for 80–89% of the total circulating hemocytes and exhibited a certain level of lysosome and oxidative capacity. Erythrocytes-II were the largest cells and displayed high lysosome content and the most active oxidative capacity. Both erythrocytes-I and erythrocytes-II did not show phagocytosis capacity. Granulocytes were intermediated-sized hemocytes characterized by granules in the cytoplasm and long pseudopodia on the cell surface, and these cells were mainly engaged in the cellular defense exhibiting the largest lysosome content, the most active phagocytosis, and high oxidative capacity. Contrary to granulocytes, hyalinocytes were comparatively small and round cells and exhibited no granules in the cytoplasm. Hyalinocytes displayed a certain level of lysosome and phagocytosis and oxidative capacities. Blast-like cells characterized by the smallest size and small quantity of cytoplasm and exhibited an absence of phagocytosis and extremely low oxidative capacity, suggesting that this population is not directly involved in the cell-mediated immune activities. In conclusion, flow cytometry indicated that three blood cockles had five types of hemocytes, and the erythrocytes and granulocytes were mainly involved in the immunological activities.

It now is widely recognised that the global temperature is rising, a phenomenon which could alter the effects of pollution on wildlife. In order to assess the role of temperature and exposure to chlorine due to cooling water discharges, a battery of metabolic, oxidative stress and histological parameters were evaluated in Mytilus galloprovincialis after 15 and 30 days at 15 °C and at two increased temperatures (+5 and +10 °C). Diverse gill pathologies such as haemolymphatic sinus dilatation, an increased number of mucocytes and granulocytes as well as a lower number of cilia were observed after 30 days exposure at higher temperatures. Protein, amino acid, triglyceride and fatty acid levels decreased when the temperature increased, as a consequence of higher energetic demand. Similarly, acetylcholinesterase, catalase and glutathione S-transferase activities showed an inhibition at higher temperatures, although gill lipid peroxidation levels remained unaffected. Our results suggest that increased temperatures induce deterioration in the health status of the mussels and in their defensive capacity against a polluted environment.

Although the toxicity of the pesticide sulfentrazone in some aquatic organisms is known, its effects on edaphic organisms such as earthworms remain completely unknown. Thus, we aimed at evaluating the behavior and immune response of Eisenia fetida exposed to sulfentrazone at environmentally relevant concentrations (EC). E. fetida representatives exposed to this contaminant (for 48 h) were divided in the following groups: environmental concentration (EC1x: 318 ng sulfentrazone/g of dry weight soil) and EC100x (concentration 100 times higher than in EC1x). Based on the avoidance test results, earthworms responded to this pesticide and proved the toxicity of sulfentrazone. The observed immune response induction was expressed by increased granulocytes presenting phagocytic vacuoles and agglomerations/encapsulations, mainly in animals belonging to groups EC1x and EC100x. However, the reduced frequency of plasmocytes in these animals’ hemolymphs suggested that the phagocytic immune response was not efficient to assure 100% survival. Our study is the first to report sulfentrazone toxicity in an edaphic organism, at environmental concentration.

The impact of chronic low-level groundwater arsenic (As) exposure [in the range above the WHO-recommended limit of 10 μg/L but ≤50 μg/L (permissible limit of As for many Asian countries)] was investigated for cross talk of inflammatory changes and expression of phagocytic receptors of exposed rural women (N, 45) from districts of 24 Parganas (south) and in matched control groups (N, 43) [As ≤10 μg/L] from the same district. Systemic inflammation was evident from the upregulated levels of pro-inflammatory mediators like tumor necrosis factor-α (TNF-α); interleukins (ILs) like IL-6, IL-8, and IL-12; and C-reactive protein (CRP) in the sera and upregulated expression of protein kinase B phosphorylated at ser473 (pAKTser473)/nuclear factor-κB (NF-κB)/TNF-α axis in the leukocytes of exposed women with respect to control. We found that low-dose As exposure apart from inflicting inflammation altered the expression of phagocytic receptors—Fcγ receptors (FcγRs) and complement receptors (CRs). The leukocytes of the low-As-exposed women exhibited suppression of CD64, CD35, and CD11b and increased expression of CD16 with respect to control. Groundwater As showed a negative correlation with CD64 expression on monocytes [Pearson’s r, −0.8205; 95% confidence interval (CI), −0.8789 to −0.7379] and granulocytes [r, −0.7635; 95% CI, −0.8388 to −0.6595] and a positive correlation with CD16 on granulocytes [r, 0.8363; 95% CI, 0.7599 to 0.8899]. A negative correlation of groundwater As was also observed with expression of CD35 on granulocytes [r, −0.8780; 95% CI, −0.9185 to −0.8192] and monocytes [r, −0.7778; 95% CI, −0.8490 to −0.6790] and CD11b on monocytes [r, −0.6035; 95% CI, −0.7218 to −0.4511]. Therefore, it may be indicated that chronic low-level As exposure (11–50 μg/L) not only evoked chronic inflammatory changes but also suppressed the expression of FcγRs and CRs in the exposed women. This, in turn, may lead to susceptibility towards pathogenic infections or in long run may even contribute towards chronic inflammatory diseases including cancer.

It has been shown that static magnetic field (SMF) of moderate intensity produces considerable impact on biological systems. SMF can be homogeneous or inhomogeneous. In many studies, inhomogeneous SMF was employed. Aware that inhomogeneous SMF could result in experimental variability, we investigated the influence of a vertical homogeneous SMF of different orientation. Male Swiss-Webster 9- to 10-week-old mice were subacutely exposed to upward- and downward-oriented SMF of 128 mT generated by a cyclotron for 1 h/day during a 5-day period. We found that SMF affected various organs and that these effects were, to some degree, dependent on SMF orientation. Both upward- and downward-oriented SMF caused a reduction in the amount of total white blood cells (WBC) and lymphocytes in serum, a decrease of granulocytes in the spleen, kidney inflammation, and an increase in the amount of high-density lipoprotein (HDL). In addition, upward-oriented SMF caused brain edema and increased spleen cellularity. In contrast, downward-oriented SMF induced liver inflammation and a decrease in the amount of serum granulocytes. These effects might represent a specific redistribution of pro-inflammatory cells in blood and among various organs. It appears that homogeneous SMF of 128 mT affected specific organs in the body, rather than simultaneously and equally influencing the entire body system.

PCBs are persistent organic pollutants that are carcinogenic and immunotoxic and have developmental toxicity. This suggests that they may interfere with normal cell maturation. Cancer and stem/progenitor cells have telomerase activity to maintain and protect the chromosome ends, but lose this activity during differentiation. We hypothesized that PCBs interfere with telomerase activity and the telomere complex, thereby disturbing cell differentiation and stem/progenitor cell function. HL-60 cells are cancer cells that can differentiated into granulocytes and monocytes. We exposed HL-60 cells to PCB126 (dioxin-like) and PCB153 (nondioxin-like) 6 days before and during 3 days of differentiation. The differentiated cells showed G0/G1 phase arrest and very low telomerase activity. hTERT and hTR, two telomerase-related genes, were downregulated. The telomere shelterins TRF1, TRF2, and POT1 were upregulated in granulocytes, and TRF2 was upregulated and POT1 downregulated in monocytes. Both PCBs further reduced telomerase activity in differentiated cells, but had only small effects on the differentiation and telomere-related genes. Treatment of undifferentiated HL-60 cells for 30 days with PCB126 produced a downregulation of telomerase activity and a decrease of hTERT, hTR, TRF1, and POT1 gene expression. With PCB153, the effects were less pronounced and some shelterin genes were increased after 30 days of exposure. With each PCB, no differentiation of cells was observed and cells continued to proliferate despite reduced telomerase activity, resulting in shortened telomeres after 30 days of exposure. These results indicate cell-type and PCB congener-specific effects on telomere/telomerase-related genes. Although PCBs do not seem to strongly affect differentiation, they may influence stem or progenitor cells through telomere attrition with potential long-term consequences for health.