Determination and comparison of freely dissolved PAHs using different types of passive samplers in freshwater.

Polycyclic aromatic hydrocarbons (PAHs) are well-known for their recalcitrant properties and biotoxicity in organisms, leading to serious environmental and health problems. Despite various analytical methods available, accurate determination of the bioavailable fraction is warranted in order to evaluate the precise toxic potentials of these compounds. Currently, the passive sampler is used worldwide to measure the bioavailable PAHs in the environment using the equilibrium partitioning principle. In this study, we co-deployed different types of passive samplers, which are linear low-density polyethylene (LLDPE) and low-density polyethylene (LDPE), to determine freely dissolved concentrations (Cfree) of PAHs using the performance reference compounds (PRCs) in Kentucky Lake (KL), Ohio River (OH), and Mississippi River (MS). The fractional equilibrium (feq) of BeP-d12 was observed to be high in LLDPE compared with LDPE in OH and MS. In contrast, the feq of all PRCs was similar in both passive samplers in KL due to slow flow velocity. The sum of average freely dissolved PAH concentrations in LLDPE and LDPE during the exposure period were 2.89 and 1.27 ng/L in KL, 8.13 and 3.31 ng/L in OH, and 5.19 and 3.82 ng/L in MS, respectively. The results revealed that LLDPE is a suitable alternative tool to LDPE for both short-term and long-term monitoring of PAHs.

Phthalate Metabolites, Hydroxy-Polycyclic Aromatic Hydrocarbons, and Bisphenol Analogues in Bovine Urine Collected from China, India, and the United States.

Human exposure to endocrine-disrupting chemicals (EDCs) has aroused considerable public concern over the last three decades. Nevertheless, little is known with regard to the exposure of EDCs in farm animals. In this study, concentrations of 22 phthalate metabolites (PhMs), 15 hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs), and 8 bisphenols (BPs) were determined in 183 bovine urine samples collected from China, India, and the United States. The median concentrations of urinary PhMs, OH-PAHs, and BPs in bovines, collectively, were 66, 4.6, and 16 ng/mL, respectively. Mono-n-butyl phthalate (mBP; median, 14 ng/mL) and ∑4DEHP (four secondary metabolites of di(2-ethylhexyl) phthalate; 13 ng/mL) were the dominant PhMs; hydroxy-fluorene (OH-Fluo; 1.2 ng/mL) and -phenanthrene (OH-Phen; 1 ng/mL) were the dominant OH-PAHs; and 4,4'-di-hydroxydiphenylmethane (BPF; 10 ng/mL) and 2,2-bis(4-hydroxyphenyl) propane (BPA; 6.7 ng/mL) were the dominant BPs. Bovine urine samples from India and China contained the highest concentrations of PhMs and OH-PAHs, whereas those from India and the United States contained the highest concentrations of BPs. PhM and OH-PAH concentrations were significantly higher in the urine of bulls than those of cows; no such difference was found for BPs. Our findings establish baseline exposure information about three classes of EDCs in domestic farm animals.

Occurrence and Profiles of Melamine and Cyanuric Acid in Bovine Feed and Urine from China, India, and the United States.

Melamine and cyanuric acid have been reported to occur in animal products. Nevertheless, information that pertains to the occurrence of melamine and cyanuric acid in cattle feed and urine is lacking. In this study, the occurrence of melamine and its three derivatives (i.e., cyanuric acid, ammeline, and ammelide) was determined in 183 bovine urine and 29 matched feed samples collected from China, India, and the United States. ∑Melamine (sum of four target compounds) was found in all urine samples at concentrations that ranged from 4.2 to 5280 ng/mL (median: 370 ng/mL); cyanuric acid was the major derivative, accounting for 97% of the total concentrations, followed by melamine (2.2%). The ubiquitous occurrence of ∑Melamine in feed (21-6230 ng/g) suggests that it is the major source of melamine and its derivatives in bovines. Urinary concentrations of melamine and cyanuric acid varied significantly among the three countries, with samples from China as having the highest concentrations, followed by the United States and India. The calculated cumulative daily intakes of melamine and cyanuric acid were at least 10-fold below the current tolerable daily intake recommended for humans. Our study provides evidence-based data on exposure patterns and sources of melamine and cyanuric acid in cattle.

Source identification and health risk of polycyclic aromatic hydrocarbons associated with electronic dismantling in Guiyu town, South China.

In this study the concentrations and distribution of sixteen polycyclic aromatic hydrocarbons (PAHs) were investigated in gas and total suspended particle (TSP) samples collected during daytime and night time. The sampling locations included an electronic waste dismantling workshop (EW), a plastic recycling workshop (PW) and a waste incineration field (WF) in Guiyu, China. A large residential area (RA) in this region was used as a control site. In the daytime, the highest concentration was found at WF (1041 ng m(-3)); while in the night time the highest concentration was found outside of EW (744 ng m(-3)). Comparison between work hours (daytime) and rest hours (night time) displayed that the total PAHs (gas+particulate phase) concentrations and the percentages of PAHs associated with TSP were higher at night than those in the daytime in all sampling workshops except WF. Source diagnostic-ratio analysis revealed that unwanted materials and smoldering honeycomb coals were the main sources of PAHs in EW, WF and PW. Benzo[a]pyrene equivalent [BaPeq] concentrations calculated by using the toxic equivalent factors [TEFs] suggested that the occupational exposure levels were not significantly high when compared with other occupational exposure. Additionally, our study suggested that the smoldering of unwanted materials could produce much more toxic PAHs compounds.

Deposition of organochlorine pesticides, PCBs (Aroclor 1268), and PBDEs in selected plant species from a Superfund Site at Brunswick, Georgia, USA.

Deposition of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) were measured in Loblolly pine needles (Pinus taeda) collected in and around a Linden Chemicals and Plastics (LCP) Superfund Site at Brunswick, Georgia, USA. For the comparison, foliage of eastern red cedar (Juniperus virginiana) was also collected to monitor contaminant levels. This study revealed that concentrations of OCPs, PCBs and PBDEs ranged from 0.75-10, 3.4-15 to 0.05-3, ng/g wet wt, respectively in both plant species. Total OCPs concentrations in pine needles decreased from 10 to 2.3 ng/g; and total PCBs decreased from 28 to 9.3 ng/g between 1997 and 2006. To our knowledge, this is the first report on PBDEs concentrations in pine needles and red cedar foliage samples from the Superfund Site at Brunswick, Georgia, USA.

Distribution of persistent organohalogen compounds in pine needles from selected locations in Kentucky and Georgia, USA.

Epicuticular wax of pine needles accumulates organic pollutants from the atmosphere, and the pine needle samples have been used for monitoring both local and regional distributions of semivolatile organic air pollutants. One-year-old pine needles collected from residential and industrial locations in western Kentucky and the vicinity of Linden Chemicals and Plastics, a Superfund Site at Brunswick, Georgia, were analyzed for polychlorinated biphenyls (PCBs), major chlorinated pesticides, and polychlorinated naphthalenes (PCNs). Total PCB concentrations in pine needles from Kentucky ranged from 5.2 to 12 ng/g dry weight (dw). These sites were comparatively less polluted than those from the Superfund Site, which had total PCB concentrations in pine needles in the range of 15-34 ng/g dw. Total chlorinated pesticides concentrations in pine needles ranged from 3.5 to 10 ng/g dw from Kentucky. A similar range of concentrations of chlorinated pesticides (7.3-12 ng/g dw) was also found in pine needle samples from the Superfund site. Total PCN concentrations in pine needles ranged from 76 to 150 pg/g dw in Kentucky. At the Superfund Site, total PCN concentrations ranged from 610 pg/g dw to 38,000 pg/g dw. When the toxic equivalencies (TEQs) of PCBs in pine needles were compared, Kentucky was relatively lower (0.03-0.11 pg/g dry wt) than the TEQs at the Superfund Site (0.24-0.48 pg/g dry wt). The TEQs of PCNs from Kentucky (0.004-0.067 pg/g dw) were much lower than the TEQs from locations near the Superfund Site (0.30-19 pg/g dry wt). The results revealed that pine needles are excellent, passive, nondestructive bioindicators for monitoring and evaluating PCBs, chlorinated pesticides, and PCNs.

Polychlorinated dibenzo-p-dioxins, dibenzofurans, and dioxin-like polychlorinated biphenyls in sediment and mussel samples from Kentucky Lake, USA.

Sediment and mussel tissues from the Kentucky Dam Tailwater (KDTW) and Ledbetter Embayment (LE) of Kentucky Lake, Kentucky, USA, were analyzed to examine the presence of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and non-, mono-, and di-ortho-chlorine-substituted polychlorinated biphenyls. Concentrations of target compounds varied with locations and sample matrices. In general, KDTW sediment samples contained slightly higher amounts of PCDD/DFs (average: 1100, range: 120-2400) than the LE sediments (average: 920, range: 580-1300) on a pg/g dry wt (dw) basis. Dioxin-like PCBs in KDTW were (average: 550, range: 70-2,000) higher than in LE (average: 320, range: 44-1000) on a ng/g dw basis. In contrast, mussel tissues had greater concentrations of PCDD/DFs in LE (average: 6500, range: 2200-13,000) than in KDTW (average: 3500, range: 2500-4800). Dioxin-like PCBs were slightly higher in KDTW (average: 76, range: 18-100) than in LE (average: 49, range: 24-96) on a ng/g fat wt basis. Biota sediment accumulation factors (BSAFs) were calculated using tissue concentrations and sediment concentrations based on dry weight. PCDD/DFs BSAF was in the range of 0.21-25 in LE and 0.093-13 in KDTW. 1,2,3,7,8,9-HxCDF in LE and 2,3,7,8-TCDF in KDTW had a greater BSAF, while BSAF for dioxin-like PCBs ranged from 0.84 to 13 in LE and from 2.3 to 12 in KDTW in which PCB-169 had the greatest BSAF in LE and PCB-167 in KDTW. Toxic equivalency (TEQ) was greatest in mussel from LE (mean: 193 pgTEQ/g fat wt) followed by mussel from KDTW (32 pgTEQ/g fat wt), sediment in KDTW (13 pgTEQ/g dry wt), and sediment in LE (7.6 pgTEQ/g dry wt). In general, PCDD/DF had a greater contribution to toxicity in mussels, while dioxin-like PCBs had a greater contribution to toxicity in sediment at both locations.

Perfluoroalkyl sulfonates and perfluorocarboxylates in two wastewater treatment facilities in Kentucky and Georgia.

Discharge of effluents from municipal wastewater treatment plants (WWTPs) is a route for the introduction of certain organic contaminants into aquatic environments. Earlier studies have reported the occurrence of perfluorochemicals in effluents from WWTPs. In this study, contamination profiles of perfluorinated compounds (PFCs), including perfluoroalkyl sulfonates (PFASs; PFOS, PFOSA, PFHxS) and perfluoroalkyl carboxylates (PFACs; PFOA, PFNA, PFDA, PFDoDA, PFUnDA), were determined in samples collected at various stages of wastewater treatment during different seasons. The two WWTPs selected for this study represent rural (Plant A, Kentucky) and urban (Plant B, Georgia) areas. PFOS was a major contaminant in samples from Plant A (8.2-990 ng/g dry wt in solid samples and 7.0-149 ng/L in aqueous samples), followed by PFOA (8.3-219 ng/g dry wt in solid samples and 22-334 ng/L in aqueous samples). PFOA was the predominant contaminant in samples from Plant B (7.0-130 ng/g dry wt in solid samples and 1-227 ng/L in aqueous samples), followed by PFOS (<2.5-77 ng/g dry wt in solid samples and 1.8-22 ng/L in aqueous samples). PFHxS, PFNA, PFDA, and PFOSA were detected in most of the samples, whereas PFUnDA and PFDoDA were detected in very few samples. Concentrations of some PFCs, particularly PFOA, were slightly higher in effluent than in influent, suggesting that biodegradation of some precursors contributes to the increase in PFOA concentrations in wastewater treatment processes. No large-magnitude seasonal variations in concentrations were found, although mass flow of PFCs was higher in winter than in summer. In general, samples from the rural plant in Kentucky contained greater concentrations of PFCs than did those from the urban plant in Georgia. Incineration of sludge reduced the PFC levels significantly. The mass flows of PFCs in these two plants were several hundreds of mg/day, comparable to flow values reported earlier.

Occurrence and fate of polycyclic musks in wastewater treatment plants in Kentucky and Georgia, USA.

Wastewater treatment plants (WWTPs) are a potential of source of polycyclic musks in the aquatic environment. In this study, contamination profiles and mass flow of polycyclic musks, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[gamma]-2-benzopyran (HHCB), 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphthalene (AHTN), and HHCB-lactone (oxidation product of HHCB), in two WWTPs, one located in Kentucky (Plant A, rural area) and the other in Georgia (Plant B, urban), USA, were determined. HHCB, AHTN and HHCB-lactone were detected in the influent, effluent, and sludge samples analyzed. The concentrations in wastewater samples varied widely, from 10 to 7,030 ng/l, 13 to 5,400 ng/l, and 66 to 790 ng/l, for HHCB, AHTN, and HHCB-lactone, respectively. Sludge samples contained HHCB at <0.02-36 microg/g dry weight, AHTN at <0.02-7.2 microg/g dry weight, and HHCB-lactone at <0.05-17 microg/g dry weight. Based on the daily flow rates and mean concentrations of polycyclic musks, the estimated discharge of total polycyclic musks to the rivers was 21 g/day from Plant A and 31 g/day from Plant B. Mass balance analysis suggested that only 30% of HHCB and AHTN entering the plants was accounted for in the effluent and the sludge. Removal efficiencies of HHCB and AHTN in the two WWTPs ranged from 72% to 98%. In contrast, HHCB-lactone concentrations increased following the treatment. Concentrations of polycyclic musks in sludge were on the order of several parts per million. Incineration of sludge at one plant reduced the concentration of polycyclic musks.

Immunomodulation of human natural killer cell cytotoxic function by organochlorine pesticides.

Organochlorine pesticides are used worldwide. To our knowledge there have been no studies dealing with the effects of these agents under in vitro conditions on human natural killer (NK) cell cytotoxic function. NK cells play a central role in immune defense against tumor development and viral infections. Thus, any agent that interferes with the ability of NK cells to lyse their targets could increase the risk of tumor incidence and/or viral infections. In this study, we examined the effects of organochlorine pesticides and some of their breakdown products on the ability of human NK cells to lyse tumor cells. A total of 11 compounds were tested. The compounds were tested in both purified NK cells as well as a cell preparation that contained other mononuclear cells (predominantly T cells) and NK lymphocytes (referred to as T/NK cells). Lymphocytes were exposed to the compounds for periods of time ranging from 1 hour to 6 days. Exposure of highly purified NK cells to 5 microM alpha-chlordane, gamma-chlordane, 4,4'-DDT, heptachlor, oxychlordane, or pentachlorophenol (PCP) inhibited their ability to destroy K562 tumor-cells by 88+/-5, 92+/-8, 61+/-13%, 64+/-10%, 69+/-11%, 76+/-12%, respectively, after a 24 h exposure. The loss of cytotoxic function seen with alpha-and gamma-chlordane remained essentially constant out to 6 days, while that seen with 4,4'-DDT, oxychordane and PCP increased with longer exposures (6 d). PCP was the most effective of the compounds tested at decreasing NK function. Of the compounds that caused decreased lytic function when tested in purified NK cells, only PCP and oxychordane decreased the lytic function of the T/NK cell preparation after any exposure. The results provide evidence of relative toxic potential for the 11 compounds and their immunomodulatory effects on other mononuclear cells (such as T-cells, B-cells, and monocytes) as well as NK lymphocyte function.

Perfluorooctanesulfonate and related fluorochemicals in human blood from several countries.

Perfluorooctanesulfonyl fluoride based compounds have been used in a wide variety of consumer products, such as carpets, upholstery, and textiles. These compounds degrade to perfluorooctanesulfonate (PFOS), a persistent metabolite that accumulates in tissues of humans and wildlife. Previous studies have reported the occurrence of PFOS, perfluorohexanesulfonate (PFHxS), perfluorooctanoate (PFOA), and perfluorooctanesulfonamide (PFOSA) in human sera collected from the United States. In this study, concentrations of PFOS, PFHxS, PFOA, and PFOSA were measured in 473 human blood/serum/plasma samples collected from the United States, Colombia, Brazil, Belgium, Italy, Poland, India, Malaysia, and Korea. Among the four perfluorochemicals measured, PFOS was the predominant compound found in blood. Concentrations of PFOS were the highest in the samples collected from the United States and Poland (>30 ng/mL); moderate in Korea, Belgium, Malaysia, Brazil, Italy, and Colombia (3 to 29 ng/mL); and lowest in India (<3 ng/mL). PFOA was the next most abundant perfluorochemical in blood samples, although the frequency of occurrence of this compound was relatively low. No age- or gender-related differences in the concentrations of PFOS and PFOA were found in serum samples. The degree of association between the concentrations of four perfluorochemicals varied, depending on the origin of the samples. These results suggested the existence of sources with varying levels and compositions of perfluorochemicals, and differences in exposure patterns to these chemicals, in various countries. In addition to the four target fluorochemicals measured, qualitative analysis of selected blood samples showed the presence of other perfluorochemicals such as perfluorodecanesulfonate (PFDS), perfluoroheptanoic acid (PFHpA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorododecanoic acid (PFDoA), and perfluoroundecanoic acid (PFUnDA) in serum samples, at concentrations approximately 5- to 10-fold lower than the concentration of PFOS. Further studies should focus on identifying sources and pathways of human exposure to perfluorochemicals.

Brief exposure to triphenyltin produces irreversible inhibition of the cytotoxic function of human natural killer cells.

Phenyltin (PT) compounds (mono-, di-, and triphenyltins) are used in agricultural and consumer products. They contaminate the environment and have toxic effects on aquatic and terrestrial animals including humans. In an earlier study we demonstrated that PTs (1 micro M, for 1h in vitro exposure) could cause considerable inhibition of the tumor-killing function of human natural killer (NK) cells (as much as 85%). In this study we examined whether cytotoxic function can be recovered after a brief exposure (1h) to PTs. Freshly isolated lymphocytes were exposed to triphenyltin (TPT) or diphenyltin (DPT) for 1h. The compound was then removed and the cells were incubated in PT-free medium for as long as 6 days. The results indicated that exposure to 750nM TPT for 1h caused an approximately 63+/-10% decrease in NK-cytotoxic function. However, if the cells were exposed to 750nM TPT for 1h and then allowed to incubate in TPT-free medium for 24h, there was a 91+/-12% loss of cytotoxic function. NK-cytotoxic function remained inhibited for as long as 6 days after removal of the TPT. A 1-h exposure to as much as 5 micro M DPT caused no loss of NK-cytotoxic function when the cells were tested immediately after the exposure. However, if the cells were allowed to incubate in DPT-free medium for 24h after the 1-h exposure to 5 micro M DPT, cytotoxicity was inhibited by 68+/-29% and this inhibition persisted for at least 6 days. These results indicated that short-term exposure to PTs caused persistent negative effects on human NK-cell function. The persistent effects of PTs are compared to those of the butyltins (BTs).

Immunomodulation of human natural killer cell cytotoxic function by triazine and carbamate pesticides.

Triazine (atrazine) and carbamates (maneb, metiram, and ziram) are used as pesticides on a variety of crops around the world. To our knowledge, there have been no studies dealing with the effects of these compounds on human natural killer (NK) cells cytotoxic function. NK cells play a central role in immune defense against tumor development and viral infections. Thus, any agent that interferes with the ability of NK cells to lyse their targets could increase the risk of tumor incidence and/or viral infections. In this study, we examined the effects of atrazine, maneb, metiram, zineb, and ziram on the ability of human NK cells to lyse tumor cells. The compounds were tested in both purified NK cells as well as a cell preparation that contained both T and NK lymphocytes (T/NK cells). Lymphocytes were exposed to the compounds for periods of time ranging from 1 h to 6 days. Exposure of highly purified NK cells to 10 microM atrazine, maneb, or metiram inhibited K562 tumor cell lysis by 63+/-25, 95+/-4, and 50+/-6%, respectively, after a 24 h exposure and by 83+/-21, 70+/-39, and 48+/-41% after a 6-day exposure. Exposure to 2.5 microM ziram for 24 h caused a 99+/-2% decrease in lytic function and at 1 microM for 6 days caused a 96+/-4% decrease. However, when T/NK cells were exposed to atrazine, maneb, or metiram for 24 h only 10 microM atrazine and maneb caused a significant decreases in lytic function (61+/-13 and 38+/-18%) and after 6 days only atrazine was inhibitory (54+/-12%). A 24-h exposure to 2.5-microM ziram caused a 41+/-51% decrease in function, but a 6-day exposure to 1 microM ziram caused no inhibition of lytic function. The results provide evidence of relative toxic potential for the five compounds and the immunomodulatory effects on both T and NK lymphocyte function.

Interleukins 2 and 12 produce recovery of cytotoxic function in tributyltin-exposed human natural killer cells.

Cytotoxic function of human natural killer (NK) cells is modulated by a variety of cytokines. Interleukins (IL) 2 and 12 are both potent stimulators of NK cell cytotoxic function. Tributyltin (TBT) is used in a variety of consumer products and industrial applications. TBT is found in dairy products, meat, and fish. We and others have shown that there are measurable levels of TBT in human blood. Butyltins appear to increase the risk of cancer and viral infections in exposed individuals. We have demonstrated that the ability of NK cells to kill tumor cells is greatly diminished after a l-h exposure to TBT and that this inhibition persists even after removal of the compound. In the current study we examine the effects of the NK-stimulatory ILs, IL2 and IL12, on the ability of NK cells to recover from the persistent inhibitory effects of a 1-h TBT treatment. Highly purified NK cells (> 95% CD16(+)) or a lymphocyte preparation containing both T lymphocytes and NK cells were treated with 300 nM TBT and then allowed to recover for 24 h, 48 h, 4 days, and 6 days in TBT-free media containing no interleukin, 1000 U/mL IL2, 20 ng/mL IL l2, or a combination of IL2 plus IL12. Tumor killing function was then tested using a radioactive chromium release assay. As seen in our previous studies there is no recovery of NK cell cytotoxic function even after a 6-day recovery period when no interleukin is present in the medium. However, there is significant recovery of NK cytotoxic function when IL2, IL12, or the combination of IL2 plus IL12 is present in the medium during the recovery period.

Brief butyltin exposure induces irreversible inhibition of the cytotoxic function on human natural killer cells, in vitro.

Despite mounting evidence on butyltin (BT) contamination and related immunotoxic effects on wildlife, very little is known about BT-associated immunotoxic effects on humans, particularly the effects on human natural killer (NK) lymphocyte function. Our earlier studies demonstrated that in vitro exposure to environmentally relevant concentrations of BTs negatively affect human NK cells and that there are measurable levels of BTs in human blood. In this study we examined whether the inhibition of NK cell cytotoxic function induced by a brief exposure (1 h) to BTs is reversible when the cells are allowed to recover in BT-free media for up to 6 days. Standard methods were used in chemical preparation, blood sampling, NK cell isolation, and 51-Chromium release assay. The results revealed that exposure to 300 nM TBT for 1 h caused an approximately 65- decrease in NK cytotoxic function, whether the lymphocytes were given as long as a 6-day recovery period or no recovery period. There was no recovery (nor any further loss) of NK cytotoxic function following removal of the compound. Exposure to 5 microM DBT for 1 h showed a 41% decrease in cytotoxic function with 0-h recovery and an 83% decrease after a 24-h recovery period. Thus, not only is there no significant recovery of NK cytotoxic function when the lymphocytes are allowed to incubate in BT-free medium for up to 6 days but there is additional loss of cytotoxic function. The results indicated that short-term exposure to BTs causes persistent negative effects on NK cell ability to kill cancer cells.

Interleukins 2 and 12 produce significant recovery of cytotoxic function in dibutyltin-exposed human natural killer cells.

Cytotoxic function of human natural killer (NK) cells is modulated by a variety of cytokines. Interleukins (IL) 2, 12, 15, and 18 and Interferon gamma (IFNgamma) are potent stimulators of NK cell cytotoxicity. Butyltins (BTs) are used in a variety of consumer products and industrial applications. Dibutyltin (DBT) is found in plastic products, beverages stored in PVC pipes during manufacturing, and poultry products. BTs appear to increase the risk of cancer and viral infections in exposed individuals. Recently, we have demonstrated that the ability of NK cells to kill tumor cells is greatly diminished after a 1-h exposure to dibutyltin. This inhibition of tumor killing function continues even after removal of the compound. There is no significant recovery of NK cytotoxic function even when the cells are allowed to recover for 6 days. In the current study we examine the effects of NK-stimulatory cytokines on the ability of NK cells to recover from the inhibitory effects of a 1-h DBT treatment. Highly purified NK cells (>95% CD16(+)) or a lymphocyte preparation containing both T lymphocytes and NK cells were treated with 5 microM DBT and then allowed to recover for 24 h, 48 h, 4 days, and 6 days in DBT-free medium containing either no cytokine or a maximally stimulatory dose of several NK-stimulatory cytokines. Tumor killing function was tested using a radioactive chromium release assay. As seen in our previous studies there is no recovery of NK cell cytotoxic function even after a 6-day recovery period when no cytokine is present in the medium. However, there is significant recovery of NK cytotoxic function when IL2, IL12, or the combination of IL2 plus IL12 is present in the medium during the recovery period. The other cytokines tested (IL15, IL18, and IFNgamma) were unable to increase the cytotoxicity of DBT-exposed NK cells.

Expression of CD16, CD18 and CD56 in tributyltin-exposed human natural killer cells.

Tributyltin (TBT) was produced in large quantities for use in wood preservation, marine antifouling paints, disinfection of circulating industrial cooling waters and slime control in paper mills. TBT is found in dairy products, meat and fish. We and others have shown that there are measurable levels of TBT in human blood. BTs appear to increase the risk of cancer and viral infections in exposed individuals. In previous studies, we demonstrated that the NK-cytotoxic function of lymphocytes was greatly diminished after a 1-h exposure to 300 nM TBT or a 24-h exposure to 200 nM TBT. Inhibition induced by a 1-h exposure to 300 nM TBT continues even after removal of the compound. There is also decreased ability of NK cells to bind to tumor target cells when they have been exposed to 200 nM TBT for 24 h. This loss of binding function is not seen when NK cells are exposed to 300 nM TBT for 1 h. However, NK cells exposed to 300 nM TBT for 1 h and then incubated in TBT-free media for 24, 48 or 96 h, show a significant loss of tumor-binding function by 96 h. The effects of TBT on cell surface molecules that are crucial to NK cell function is investigated. The data indicate there is a loss of expression of CD16 and CD56 on NK cells exposed to 200 nM TBT for 24 h. There is no decrease in expression of any of the markers studied when NK cells are exposed to 300 nM TBT for 1 h, consistent with the fact that a 1-h exposure has no effect on the ability of NK cells to bind to tumor targets. However, when NK cells are exposed to 300 nM TBT followed by 24, 48 or 96 h incubations in TBT-free media, there is a significant loss of CD16 and CD18 expression after 24 h and of CD16 and CD56 expression after 48 and 96 h.