Bartonella henselae is the causative agent of cat scratch disease. Although cats are the main zoonotic reservoirs of Bartonella spp., unusual cases of cat scratch disease caused by a domestic dog scratch have been recently reported. For the in vivo B. henselae infection, eight dogs were inoculated intradermally with $2{\times}10^8CFU$ of B. henselae Houston-1 suspended in 1 ml of phosphate buffered saline on day 0 and subsequent injections of the same amount given intradermally on days 21, 28, 36, 58 and 64. After in vivo canine B. henselae infection was confirmed by nested PCR, the IFN-$\gamma$ levels of the culture supernatant of PBMC stimulated with B. henselae was significantly higher in the B. henselae-PCR positive group than the B. henselae-PCR negative group. Our results showed that the canine immune responses against B. henselae were different from those of cats. Th1 activation by B. henselae stimulation was characterized in dog peripheral blood mononuclear cells, whereas Th2 activation was reported in B. henselae-infected cats.
Apolipoprotein A-I (apoA-I) has anti-inflammatory and anti-oxidative properties. This study was designed to investigate whether apoA-I affects apoptosis and cytokine production of human blood neutrophils in an in vitro culture system. Spontaneous apoptosis of neutrophils was significantly delayed by apoA-I. In addition, high density lipoprotein containing apoA-I also delayed apoptosis of neutrophils. Apoptosis of neutrophils was inhibited by anti-scavenger receptor type B-I antibodies. The amounts of interleukin-8, interferon (IFN)-inducible protein 10 (IP-10), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) in the supernatants of cultured neutrophils treated with apoA-I were significantly increased. Combined treatment of neutrophils with IFN-$\gamma$ and apoA-I produced higher amounts of IP-10 and TNF-$\alpha$ than did treatment with IFN-$\gamma$ or apoA-I alone. The present study reveals that apoA-I activates neutrophils to produce cytokines and delays spontaneous apoptosis of neutrophils. These findings suggest that apoA-I, although a well-known negative acute-phase protein, has a pro-inflammatory effect in neutrophils.
Park, Jeong-Hae;Park, Jung-Ae;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
Journal of Life Science
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v.21
no.12
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pp.1778-1783
/
2011
Hematopoietic cytokines regulate production of blood cells by stimulating proliferation and differentiation of bone marrow cells. Among these hematopoietic cytokines, called hematopoitic growth factors, glranulocyte-colony stimulating Factor (G-CSF), which regulates growth of neutrophils, is one of important therapeutic factors because cancer patients suffer with neutropenia which is severe reduction of neutrophils after chemotherapy. Two groups of recombinant G-CSF have approved and used for therapeutic purposes and many researches are still on-going to produce recombinant G-CSF by different techniques. We engineered human G-CSF with Bombyx specific endoplasmic reticulum (ER) signal sequence, therefore, secretion of human G-CSF protein was improved in Bombyx mori-origined cell line, Bm5. The Bombyx ER signal sequence and human G-CSF matured protein region chimera was further remodeled at the N-terminus of matured G-CSF protein to understand roles of N-terminus on outer cellular secretion and/or production. Three different mutants were generated deleting three amino acids in non alpha-helical region in N-terminus in order to scan important amino acids for G-CSF secretion. One of 3 different N-terminal deletion mutants showed dramatically reduction of secreted amount of G-CSF indicating its important role on secretion. The data suggest that remodeling in non alpha-helical region of N-terminus is also important for recombinant G-CSF production.
We investigated the effect of the clay mineral fortified with complex trace elements by specific fermentation microbes on growth performance and a quality of pork in this study. For the declared experimental animal, 80 heads of crossbred with Large White and Yorkshire were randomly splitted into a control group and a test group. Each group were assigned with 4 replicates and 10 heads of each replicate. When 0.3% of the fermentation product of the clay mineral (FCM) was added into feed stuff and fed for 9 weeks of experimental feeding period, the results of the feed intake, weight gains, pH of carcass, lightness of meat color, and brightness of the muscle semimembranosus were similar to those of the control group. However, dressing percentage, reddishness of carcass, and water holding capacity were improved, respectively, as much as 2.7%, 12%, and 10%, and reduced by 6% in cooking loss. In order to investigate the immune modulatory effect of fermented clay mineral, pheripheral blood mononuclear cells (PBMC) were isolated and Immune modulatory parameters were measured. The proliferation activity of PBMC from pigs fed the fermented clay mineral were significantly increased compared to control group pigs, and also those results were more clearly observed as activated with lipopolysaccharide and concanavaline A. The secretion of TNF-${\alpha}$ of the FCM group pigs showed an increasing tendency. Therefore it was suggested that the feeding of FCM which was high in cation metathesis and the value of infrared ray, activated the immune responses, and thus the production of the environment-friendly high quality pork without the use of antibiotics would be possible.
Kim, Ji-Eun;Lee, Ho-Chan;Kang, Eun-Jeong;Choi, Jung-Wha;Kim, Jong-Han;Park, Soo-Yeon;Jung, Min-Yeong
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.32
no.3
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pp.58-76
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2019
Objectives : This study was designed to examine the effects of Daecheonglyong-tang(DCL) on atopic dermatitis induced by DNCB in mice Methods : The Nc/Nga mice were divided into 5 groups, and four groups excluding the normal group were applied by 2,4-dinitrochlorobenzene(DNCB), to cause AD and were orally administered with distilled water(negative control), dexamethasone(positive control), and DCL 200 or 400mg/kg once a day for 4 weeks respectively. The visual changes on skin, changes in skin tissue thickness and eosinophil infiltration were observed. IgE, Histamine, Cytokines, immune cells and the amount of gene expression of filaggrin, VEGF, $TGF-{\beta}1$, EGF were measured. Results : Dermatitis score showed a gross improvement on all DCL groups, similar to or better than positive control. All DCL groups showed no significant change in the basophils, while neutrophils and eosinophils decreased. In only DCL 400 mg/kg groups, white blood cells and mononuclear cells were decreased and lymphocytes were increased. In particular, neutrophils had similar or better effects than the positive control. In all DCL groups, IgE, Histamine, $IL-1{\beta}$, IL-4, IL-5, IL-6 and $TNF-{\alpha}$ were decreased and IL-2 was increased. In only DCL 400 mg/kg groups, IL-10 decreased and $IFN-{\gamma}$ increased. In particular, $IL-1{\beta}$ and $IFN-{\gamma}$ showed a similar rate of increase and decrease comparing positive control in DCL 400 mg/kg. $TGF-{\beta}$1 was increased in all DCL groups, filaggrin and VEGF were increased in only DCL 400 mg/kg groups. EGF did not make any changes. Epidermis, dermis thickness and eosinophil infiltration were also decreased in all DCL groups. Conclusions : By increasing Th1 cytokine and decreasing Th2 cytokine, DCL extracts appear to be effective in controlling immune response imbalances, anti-inflammatory and skin regeneration and are likely to be available as a treatment for AD.
Objective: The aims of this study were to analyze the deficiency-excess pattern identification (虛實辨證) and compare it to the sputum cytokines of asthma patients. Method: 50 asthma patients who met the inclusion and exclusion criteria were included in this study. They were divided into two groups: deficiency and excess syndrome groups. Sputum examinations were performed including $TNF-{\alpha}$, Interleukin (IL)-4, IL-5, IL-10, and IL-13. The Quality of Life Questionnaire for Adult Korean Asthmatics (QLQAKA), the Visual Analog Scale(VAS), and heart rate variability (HRV) were also measured. We also conducted laboratory tests, including the hematological indexes. Results: Based on the pattern identification, 50 asthma patients can be divided into two categories of groups: the deficiency syndrome group (N=24) and the excess syndrome group (N=26). In the analysis of sputum cytokines, although the $TNF-{\alpha}$, IL-4 and IL-13 were at a higher level in the deficient pattern group than in the excess pattern group, it was insignificantly different. There was a negative correlation in the analysis of QLQAKA and VAS. In the analysis of HRV, although the mean value of VLF, LF, and HF in the deficiency syndrome group was higher than in the excess syndrome group, it was insignificantly different. There was no significant difference in the hematological tests between the deficiency and the excess syndrome group. The mean value of the IgE in the blood tests was five times greater than the reference value. Conclusion: The cytokines of sputum including $TNF-{\alpha}$, IL-4, IL-5, IL-10, and IL-13 were indifferent statistically. Reinforcing the healthy and eliminating the pathogenic factors should be considered.
Choi, Eun Wha;Lee, Hee Woo;Lee, Jun Sik;Kim, Il Yong;Shin, Jae Hoon;Seong, Je Kyung
BMB Reports
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v.52
no.4
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pp.289-294
/
2019
The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.
Background: The existing research results on the combined toxicity of these pollutants using mammals, such as rodents, are insufficient, especially in relation to changes in the immune system. Objectives: This study aims at evaluating the cellular immune response to PE-MPs solely or when combined with Pb, which possess excellent adsorption capacity with PE-MPs and is commonly co-exposed in our daily lives. Methods: The study investigated the cellular immune function of 9-week ICR mice with 28 days exposure to PE-MPs (2 mg/mouse/day) and Pb (0.1 mM in distilled water) individually and in combination. PE-MPs were administered via gastric intubation while the lead intake was conducted via the oral drinking water route. Cellular immunity was evaluated by analyzing the production for TH1 cytokines namely, TNF-α and IFN-𝛾 and TH2 cytokines, IL-4 and IL-6 in culture supernatants from polyclonally activated splenic mononuclear cells ex vivo. Results: Both the PE-MPs only and the PE-MPs+Pb exposure group revealed an increased TH1 response with elevated TNF-α and IFN-𝛾 levels and downregulated TH2 response with low IL-4, and IL-6 production levels compared to the control group. Furthermore, an increased IFN-𝛾/IL-4 ratio was found in the PE-MPs only and PE-MPs+Pb exposure groups, which indicated the skewedness to TH1 response. Meanwhile, reduced blood hemoglobin levels and increased levels of IL-4, the dominant TH2 cytokine in the Pb-only exposure group, were observed. Conclusions: Our current findings on the predominance of TH1 immune response in the PE-MPs and PE-MPs+Pb groups suggest that PE-MPs could be responsible for the predominant induction of the cellular immune changes. This finding could be used as an important landmark in research related to TH1 predominance, such as autoimmune diseases. It suggests that additional research on immune modulation using longer exposure durations or the same exposure route is required to elucidate stronger findings.
Kim, Jae-Yeol;Choi, Hyung-Seok;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Min, Kyung-Up;Kim, Yoo-Young;Shim, Young-Soo;Yoo, Chul-Gyu
Tuberculosis and Respiratory Diseases
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v.49
no.2
/
pp.217-224
/
2000
Background : It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-$\alpha$ and IL-1$\beta$. However, there is an alteration in the macrophages' responsiveness when they are challenged with repeated bouts of endotoxin, termed "endotoxin tolerance" which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. Methods : Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE II score. Peripheral blood monocytes were isolated from the patients and diluted to $1{\times}10^5$ well. After stimulation with endotoxin (LPS of E. coli O114 : B4, 100 ng/ml), they were incubated at $37^{\circ}C$ in 5% $CO_2$ incubator for 24 hours. Supernatant was collected for the measurement of TNF-$\alpha$ and IL-1$\beta$ with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. Results : The APACHE II score (mean$\pm$SD) of the patients at the time of blood sampling was 12.2$\pm$5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods (10 cases), gram positive cocci (6 cases) with two cases of mixed infection. Serum TNF-$\alpha$ could be measured in 4 cases with 29.9$\pm$27.7 pg/ml. Serum IL-1$\beta$was measurable in only one patient. The TNF-$\alpha$ level of supernatant of cultured peripheral blood monocytes was 2,703$\pm$2,066 pg/ml in patients and 2,102$\pm$1914 pg/ml in controls. The IL-1$\beta$level of supernatant was 884$\pm$1,050 pg/ml in patients and 575$\pm$558 pg/ml in controls. There was no difference of TNF-$\alpha$ and IL-1$\beta$ level between patients and controls. Conclusion : We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.
Cytokines produced by immune cells infiltrating pancreatic islets have been incriminated as important mediators of $\beta$-cell destruction in insulin-dependent diabetes mellitus. In non insulin-dependent diabetes, cytokines are also associated with impaired $\beta$-cell function in high glucose condition. By the screening of various natural products blocking $\beta$-cell destruction, we have recently found that epigallocatechin gallate (EGCG) can prevent the in vitro destruction of RINm5F cell, an insulinoma cell line, that is induced by cytokines. In that study we suggested that EGCG could prevent cytokine-induced $\beta$-cell destruction by down-regulation of nitric oxide synthase (NOS) through inhibition of NF-kB activation. Here, to verify the in vivo antidiabetogenic effect of EGCG, we examined the possibility that EGCG could also prevent the experimental autoimmune diabetes induced by the treatment of multiple low doses of streptozotocin (MLD-STZ), which is recognized as an inducer of type I autoimmune diabetes. Administration of EGCG (100 mg/day/kg for 10 days) during the MLD-STZ induction of diabetes reduced the increase of blood glucose levels caused by MLD-STZ. Ex vivo analysis of $\beta$-islets showed that EGCG downregulates the MLD-STZ-induced expression of inducible NOS (iNOS). In addition, morphological examination showed that EGCG treatment ameliorated the decrease of islet mass induced by MLD-STZ. In combination these results suggest that EGCG could prevent the onset of MLD-STZ-induced diabetes by protecting pancreatic islets. Our results therefore revealed the possible therapeutic value of EGCG for the prevention of diabetes mellitus progression.
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