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Interactions between NCR+ILC3s and the Microbiome in the Airways Shape Asthma Severity

  • Jongho Ham;Jihyun Kim;Sungmi Choi;Jaehyun Park;Min-gyung Baek;Young-Chan Kim;Kyoung-Hee Sohn;Sang-Heon Cho;Siyoung Yang;Yong-Soo Bae;Doo Hyun Chung;Sungho Won;Hana Yi;Hye Ryun Kang;Hye Young Kim
    • IMMUNE NETWORK
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    • v.21 no.4
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    • pp.25.1-25.16
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    • 2021
  • Asthma is a heterogeneous disease whose development is shaped by a variety of environmental and genetic factors. While several recent studies suggest that microbial dysbiosis in the gut may promote asthma, little is known about the relationship between the recently discovered lung microbiome and asthma. Innate lymphoid cells (ILCs) have also been shown recently to participate in asthma. To investigate the relationship between the lung microbiome, ILCs, and asthma, we recruited 23 healthy controls (HC), 42 patients with non-severe asthma, and 32 patients with severe asthma. Flow cytometry analysis showed severe asthma associated with fewer natural cytotoxicity receptor (NCR)+ILC3s in the lung. Similar changes in other ILC subsets, macrophages, and monocytes were not observed. The asthma patients did not differ from the HC in terms of the alpha and beta-diversity of the lung and gut microbiomes. However, lung function correlated positively with both NCR+ILC3 frequencies and microbial diversity in the lung. Sputum NCR+ILC3 frequencies correlated positively with lung microbiome diversity in the HC, but this relationship was inversed in severe asthma. Together, these data suggest that airway NCR+ILC3s may contribute to a healthy commensal diversity and normal lung function.

Symbiotic effectiveness and intrinsic antibiotic resistance of Rhizobium meliloti populated in Korean pasture soils (국내(國內) 초지토양(草地土壤)에 분포(分布)한 Rhizobium meliloti의 질소고정력(窒素固定力)과 항균제반응(抗菌劑反應) 특성(特性))

  • Kang, Ui-Gum;Ha, Ho-Sung;Jung, Yeun-Tae
    • Applied Biological Chemistry
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    • v.35 no.3
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    • pp.179-185
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    • 1992
  • Rhizobium meliloti populated in five Korean pasture soils were characterized by symbiotic effectiveness and intrinsic antibiotic resistance using whole-soil inoculum and 11 antibiotics, respectively. Most probable number (MPN) of naturalized rhizobia counted with alfalfa Vernal[Medicago sativa (L.)] as a host ranged $1.7{\times}10^2\;cells/g$. soil(Chunghyo, Kyeongiu)-$1.0{\times}10^5\;cells/g$. soil(Gampo, Kyeongiu) and ended to be positively associated with soil pH. On the whole, the effectiveness of population as compared to TAL mix inoculum (TAL 380+TAL 1372+TAL 1373) was very low. Nevertheless, there were two highly effective strains, YCK 539 and YCK 542, which were not inferior to TAL 1372, from Ogpo, Dalseong among the total of 30 of 6 isolates per each soil. As long as mean $N_2$ fixing ability of each soil isolate, the isolates from Hyeongog, Kyeonju were outstanding and the rest were in order of Ogpo, Dalseong>Chunghyo, Kyeongju>Hwaweon, Dalseong>Gampo, Kyeongiu. Isolates as a whole were resistant to erythromycin(67崙), nalidixic acid(77%), and streptomycin sulfate(8051), which had the concentration of $100\;{\mu}g/ml$, $160\;{\mu}g/ml$, and $10\;{\mu}g/ml$, respectively and divided into 14 patterns of resistance. Association between resistances in each soil was not clear. And there was no relationship of resistance pattern to effectiveness. The best effective strain YCKa 542 exclusively fell into No. X pattern having resistance to erythromycin, nalidixic acid, and neomycin sulfate.

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QUANTITATIVE ANALYSIS OF TRANSFORMING GROWTH $FACTOR-{\beta}_1$ IN HUMAN FIBROBLASTS INDUCED WITH STAPHYLOCOCCUS ENTEROTOXIN B AND LIPOPOLYSACCHARIDE (Staphylococcus enterotoxin B와 lipopolysaccharide를 작용시킨 사람 섬유아 세포에서 생성된 Transforming Growth $Factor-{\beta}_1$의 정량적 분석)

  • Lee, Seong-Geun;Kim, Kwang-Hyuk;Kim, Uk-Kyu;Kim, Jong-Ryoul;Chung, In-Kyo;Yang, Dong-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.123-132
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    • 2000
  • $TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

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Hepatitis B Virus-Induced TNF-a Expression in Hepa-lc1c7 Mouse Hepatoma Cell Line (마우스 Hepa-1c1c7 세포주에서 B형 간염 바이러스에 의한 tumor necrosis factor-a의 발현 유도)

  • Yea Sung Su;Jang Won Hee;Yang Young-Il;Lee Youn Jae;Kim Mi Seong;Seog Dae-Hyun;Park Yeong-Hong;Paik Kye-Hyung
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.38-44
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    • 2005
  • Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model (천식 마우스 모델의 알러지 반응에서 Enterococcus faecalis 전체 추출물의 억제 효과)

  • Chang, Jeong Hyun;Yang, EunJu;Yu, Sun Nyoung;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1168-1175
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    • 2017
  • Probiotics are usually defined as intestinal bacteria that provide healthy benefit to the host and may offer new therapeutic materials for the treatment of inflammatory diseases. Lactobacillus, Bifidobacterium and Enterococcus are known as typical probiotics. But, these bacteria have mostly a weak viability and thus decreased probiotics-mediated effects in the intestinal tract. Asthma is an inflammatory airway disease, which is characterized by the releases of inflammatory mediators including cytokine and IgE. They are mainly associated with the recruitment, activation and disregulation of specific inflammatory cells, especially mast cells, monocytes, T cells, eosinophils and neutrophils in asthma. We performed these studies as in vitro and in vivo test the human inflammatory cell lines and ovalbumin (OVA)-induced asthma mouse model. And then the inhibitory effects of Enterococcus faecalis whole extract on inflammatory responses were examined. For our examinations, the E. faecalis whole extract (Ef extract) was acquired from whole bacteria of E. faecalis using freeze/thawing after ultrasonication method. As results, OVA-mediated THP-1 cell viability was decreased by the treatment of Ef extract. In the asthmatic mouse model, Ef extract inhibited the infiltration of inflammatory cells into the inflammatory sites and blood. This whole extract may have anti-asthmatic effects associated with the regulation of IL-5 and IgE expression. It may also be a promising candidate in anti-allergic medicine for the treatment of asthma.

Antitumor and Antimetastatic Effects of Toxoplasma Gondii in Mice with Lewis Lung Carcinoma (Lewis 폐암 마우스 모델에서 톡소포자충에 의한 항암 및 항전이 효과)

  • Kim, Ju-Ock;Jung, Sung-Soo;Park, Hee-Sun;Kim, Myoung-Hoon;Kim, Sun-Young;Lee, Young-Ha
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.4
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    • pp.317-329
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    • 2002
  • Background : Immunotherapy is another treatment modality for various cancers. There is little information on the antitumor effects of immunotherapy on implanted lung cancer mouse models. Toxoplasma gondii is able to potently induce a nonspecific stimulation of the host immune system. Therefore, this study evaluated the antitumor and antimetastatic effect of nonspecific immune stimulation by T. gondii in a Lewis lung cancer mouse model. Methods : Female C57BL/6 mice were injected with either Lewis lung cancer cells ($1{\times}10^6$ per mouse) or 5 cysts from the T. gondii Me49 strain with various schedules. The number of survival days, the tumor size of the implanted muscle and the histopathological findings of each group were noted. In addition to these mice, the Toxoplasma antigen($50{\mu}g$ per mouse) or a lymphokine (0.5 ml per mouse) was added to boost the immunotherapy. Results : No mouse in the Toxoplasma-infected group had died, whereas the mice receiving only the cancer cells (cancer control) survived for $29.1{\pm}4.4$ days. Cancer cells were revealed from 1 week after cancer cell inceulation in the muscle and from 3 weeks in the lung of the cancer control, whereas cancer cells were found in both the preinfection control and coinfection control groups from 2 weeks and 4 weeks in the lung respectively. The in the number of survival days were $32.4{\pm}3.3$ in the mice receiving T. gondii 2 weeks prior to the cancer cells inoculation (preinfection control), $30.9{\pm}5.1$ in mice received both simultaneously (coinfection control), and $34.9{\pm}2.9$ in mice received T. gondii 2 weeks after cancer cells implantation (postinfection control). These 3 infection groups had significantly longer survival days and suppressed tumor growth than those of the cancer control. In addition to these mice, and injection with the Toxoplasma antigen alone or in combination with lymphokine resulted in a significant increase in the number of survival days. Conclusion : These findings suggest that an injection with T. gondii can induce the antitumor and antimetastatic effects in Lewis lung cancer mouse models. Moreover, these effects were increased with an injection of the Toxoplasma antigen alone or in combination with lymphokine. However, this therapy can not prevent the development of cancer.

Physiological and Ecological Characteristics of Indigenous Soybean Rhizobia Distributed in Korea -III. Symbiotic Effectiveness and Nitrate Reductase Characteristics of Indigenous Soybean Rhizobia (우리나라 토착대두근류균(土着大豆根瘤菌)의 분포상태(分布狀態)와 생리(生理) 및 생태학적(生態學的) 특성(特性) -제(第)III보(報) 토착대두근류균(土着大豆根瘤菌)의 질소고정효율 및 Nitrate reductase 특성(特性))

  • Ryu, Jin-Chang;Suh, Jang-Sun;Lee, Sang-Kyu;Park, Jun-Kyu;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.1
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    • pp.43-53
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    • 1987
  • In order to improve effectiveness of rhizobia- legume symbiotic nitrogen fixation, ecological and physiological characteristics of indigenous rhizobia distributed in Korea, that is, symbiotic effectiveness of indigenous soybean rhizobia, nitrate reductase activities of the soybean bacteroid from five different soils, and differences of host-infection abilities among the soybean cultivars under population densities of the same indigenous soybean rhizobia, were investigated. The results were summarized as follows: 1. The number of indigenous soybean rhizobia was ranged from $9.2{\times}10^2$ cells per gram of soil in calcareous soil II to $42.4{\times}10^3$ cells per gram of soil in calcareous soil I in Danyang. 2. The symbiotic effectiveness of indigenous soybean rhizobia from five different soils was high in the case of soybean continuously cultivated, and calcareous soil I that population densities of indigenous soybean rhizobia were observed highly. 3. Inverse relationship was observed between total nitrogenase activity (TNA) and nitrate reductase activity (NRA) from the soybean bacteroids ($r=-0.502^*$), but the correlation between nitrate reductase and specific nitrogenase activities (SNA) could be devided into two groups. It was classified into group I which is high in SNA and low in NRA, and group II which is low in SNA and high in NRA. 4. The infection ability of the indigenous soybean rhizobia in the same soil conditions showed the reciprocal difference among each soybean cultivars. In Kwangkyo and Jangyeup, the symbiotic effectiveness appeared by infection of indigenous soybean rhizobia was higher than it of the other soybean cultivars.

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Spatial Variation in the Reproductive Effort of Mania Clam Ruditapes philippinarum during Spawning and Effects of the Protozoan Parasite Perkinsus olseni Infection on the Reproductive Effort (여름철 산란기에 있어 바지락 번식량의 공간적 변이와 기생 원생생물 Perkinsus olseni 감염이 바지락 번식에 미치는 영향)

  • Kang, Hyun-Sil;Hong, Hyun-Ki;Yang, Hyun-Sung;Park, Kyung-Il;Lee, Taek-Kyun;Kim, Young-Ok;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.37 no.1
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    • pp.49-59
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    • 2015
  • Spatial variation in the reproductive effort of Manila clam Ruditapes philippinarum is often closely associated with variation in the seawater temperature and food availability, which determines gonad maturity and the quantity of gamates produced during spawning. Previous studies also have reported that severe infection by the protozoan parasite Perkinsus olseni exerts a negative impact on clam reproduction, retarding gonad maturation or decreasing the reproductive effort. In the present study, we investigated impacts of P. olseni infection on the reproductive condition of Manila clam during a spawning season. Histology revealed that 54% of female clams in Wando off the south coast were in spawning, while only 10% of the female from Gomso and 0% of the female from Seonjaedo in Gyeonggi bay off the west coast were engaged in spawning at the end of May in 2004. Ray's fluid thioglycollate media (RFTM) assay was applied to assess P. olseni infection and indicated that the infection intensity in Wando ($3,608,000{\pm}258,000cells/g$ wet tissue) was significantly higher than the levels in Gomso ($1,305,000{\pm}106,000cells/g$ wet tissue) and Seonjaedo ($1,083,000{\pm}137,000cells/g$ wet tissue, p < 0.001). The size of the ripe female follicle determined from histology was significantly smaller in Wando ($0.032mm^2$) compared to the sizes in Gomso ($0.059mm^2$) and Seonjaedo ($0.052mm^2$, p < 0.05). Accordingly, the number of ripe eggs in the follicle was significantly fewer among clams in Wando (14) compared to the numbers determined in Gomso (23) and Seonjaedo (22). The absolute quantity of egg in ripe clams from Wando (31.01 mg) was also significantly smaller than Seonjaedo (61.79 mg) and Gomso (133.3 mg). Quantity of total protein, carbohydrate, and lipid in the tissue in the Wando samples was significantly smaller than the quantities determined in Gomso and Seonjaedo (p < 0.001). The observed poor reproductive condition and proximate tissue composition of the females in Wando were, in part, explained by the extremely high level of the parasites, sapping the ability to store energy in the host tissues, which is used in tissue growth and the egg production.

Rhizobium meliloti Populations and Alfalfa Yields Due to Nitrogen Fertilization and Inoculation Methods at Cultivated Upland Soil (숙전(熟田)에서 질소시용(窒素施用)과 Rhizobium meliloti의 접종방법(接種方法)이 근류균(根瘤菌) 밀도(密度) 및 알팔파 수량(收量)에 미치는 영향(影響))

  • Kang, Ui-Gum;Jung, Yeun-Tae;Lee, Soo-Kwan;Ha, Ho-Sung
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.3
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    • pp.275-280
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    • 1992
  • Inoculation responses of alfalfa[Medicago sativa(L.) Vernal] to Rhizobium meliloti with nitrogen fertilization were characterized by soil populations and plant yields from 1989 to 1991 at cultivated upland soil, which contained $3.1{\times}10$cells of R. meliloti/g.soil. The results obtained accordingly to fertilizing with 8Kg N/10a and differently inoculating the host such as initial and annual inoculation methods were as follows : 1. Soil populations of R. meliloti were increased more at no nitrogen fertilized condition compared to nitrogen fertilized condition to 2nd year experiment from 1st year that came up $1.6{\times}10^3$cells/g.soil, but at 3rd year the trend was reversed. Between inoculation methods, on the whole, the annual inoculation caused more populations, which were $2.0{\times}10^4$cells/g.soil in maximum number. And the populations declined after winter but recovered passing through summer season. 2. Alfalfa yields were mainly influenced by rhizobial populations rather than by fertilizing nitrogen showing a significant correlation(Y=0.36+0.287X, $r^2=0.58^{**}$) with the former. The increased extents of yields obtained by inoculation at no nitrogen and nitrogen fertilized conditions, respectively, were 66 and 10% in 1st : 13 and 20% in 2nd : 19 and 13% in 3rd year experiment with the initial inoculation, and were 66 and 10% in 1st ; 30 and 20% in 2nd : 35 and 36% in 3rd year experiment with the annual inoculation. 3. The results demonstrated the importance of inoculating, if possible, annual inoculating alfalfa to get much yields even at cultivated upland soil.

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Regulation of Tumor Neceosis Factor-${\alpha}$ Receptors and Signal Transduction Pathways

  • Han, Hyung-Mee
    • Toxicological Research
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    • v.8 no.2
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    • pp.343-357
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    • 1992
  • Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

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