• International Journal of Industrial Entomology and Biomaterials
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• v.45 no.1
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• pp.17-21
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• 2022

The present study aimed to determine whether a hemolymph prepared from Antheraea yamamai larvae had the same biological activities using a Bombyx mori hemolymph prior to exposure to lipopolysaccharide (LPS) in order to induce an inflammatory response. The effects of the hemolymph were determined using a reverse transcription-quantitative polymerase chain reaction to assess the expression of pro-inflammatory molecules. The A. yamamai hemolymph exerted anti-inflammatory effects on LPS-activated human monocytic leukemia cells via Toll-like receptor (TLR) 4-mediated suppression, similar to the B. mori hemocyte extract. Treatment with the A. yamamai hemolymph significantly suppressed LPS-induced upregulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA expression at all tested concentrations compared with the control, similar to the B. mori immune-challenged hemolymph. Finally, the A. yamamai hemolymph, like the B. mori immune-challenged hemolymph, suppressed all of these concentrations in a dose-independent manner. These results demonstrate that the hemolymph of A. yamamai exhibited important biologically active substances. Further in-depth functional studies are required to fully understand the mechanisms underlying the biological activities of wild-type silkworm hemolymphs.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1539-1545
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• 2007

An experiment was conducted using 20 male buffalo calves to study the effect of vitamin E and selenium supplementation on their immune response and plasma ${\alpha}$-tocopherol and selenium status. These buffalo calves (10-12 months old, average body weight $75.30{\pm}2.20 $ kg) were randomly allotted to four treatments on the basis of their body weights and were fed on wheat straw and concentrate mixture to meet their nutrient requirements of 500 g/d body weight gain. The buffalo calves were fed either a control diet (neither supplemented with Se nor VE) or diets supplemented with Se at 0.3 ppm (+Se), DL-alpha tocopheryl acetate at 300 IU (+VE), and both DL-alpha tocopheryl acetate at 300 IU and Se at 0.3 ppm (+Se+VE). These experimental diets were fed for 180 days. Blood samples were collected at day 0 and subsequently at 45 day intervals up to 180 days of experimental feeding to monitor plasma ${\alpha}$-tocopherol and Se concentrations. To assess humoral immune response, all calves were sensitized with formalin inactivated Pasteurella multocida antigen at 135 days of experimental feeding and blood was collected on 0, 7, 14, 21 and 28 days post vaccination (DPV) to measure antibody production using indirect ELISA. Cell mediated immune response of calves was assessed after 180 days of experimental feeding by in vivo delayed type hypersensitivity (DTH) reaction using phytohaemaglutinin-P (PHA-P) as a mitogen. Results revealed that feeding of VE and Se improved the plasma levels of these nutrients. Plasma levels of Se were affected by supplementation of both VE (p<0.001) and Se (p<0.001); however, no interaction ($Se{\times}VE$) was observed. Supplementation of Se improved the humoral immune response (p<0.008), whereas, VE showed a tendency towards improvement in cell mediated immune response (p<0.064). It was concluded that vitamin E and Se supplementation improved the status of these micronutrients and humoral immune response in buffalo calves.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.3
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• pp.483-489
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• 2002

APA-01, which is an aqueous extract of five Chinese herbs, is a modified formula of Huoxiang-Zhengqi-San. The effect of new herb extract on immune response was investigated. The parameter examined to assess apparent immune response of APA-01 in mice included changes of body weight, relative weight of immune organs, cell proliferation and cytokine gene expression. The body weight and relative weight of immune organs were not significantly changed among the tested groups. In the spleen cell prolijeration assay, APA-01 increased the cell proliferation in a dose-dependent manner. Methotrexate (MTX), an agent of immune suppression, inhibited the spleen cell proliferation (IC/sub 50/: 800㎍/㎖). However, APA-01 significantly inhibited the suppression of mouse spleen cell proliferation. Therefore, it seems that APA-01 has a reducing effect of immune suppression. Immunomodulatory effect of APA-01 was further investigated using reverse transcription polymerase chain reaction (RT-PCR) in mouse spleen cells. In RT-PCR test, APA-01 enhanced the expression of cyclooxygenase-2 (COX-2) mRNA in a dose-dependent manner. In spite of immune suppression by MTX, COX-2 mRNA was induced by co-treatment with APA-01. These results suggest that APA-01 stimulates the proliferation of spleen cells, regulates the expression of COX-2 mRNA, and accelerates the recovery of inhibition of spleen cell proliferation induced by MTX, thus providing the immunological basis for clinical benefit of APA-01.

• Parasites, Hosts and Diseases
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• v.37 no.2
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• pp.109-115
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• 1999

The present study observed the variation of antigenicity of Pneumocystis carinii and serum IgG antibody reastion to the antigens from different localtities in Korea. Antigens of rat P,carinii and sera of inhabitants were collected at Chunchon, Chungju, Kwangju and Seoul during 1995-1996. Enzyme-Linked Immunosorbent Assay and immunoblot were used for immune reaction. Absorbance of 1,294 human sera ranged between 0.01and 0.93. Sera from Chunchon showed higher absorbances than those from other areas. Immunoblotting revealed IgG antibody reactions to 116,100, and 45-55 kDa antigenic bands of rat P.carinii, but the frequencies of positive reaction to individual bands were variable by localities. Total 62.6% reacted to 100kDa band and 32.0% to 45-55 kDa bands. For the reaction to 116kda, the reaction rate was 60.0% of the sera showed the reaction to 116kda band while 37.7% reacted to 100kda, the reaction rate was 60.0% to 82.6% by localities. It is found that the reaction rates of the human sera to rat P.carinii antigen are variable according to the localities. Also, the high molecular antigen of 116kDa of rat P.carinii is the most frequent antigenic band reaction to human sera.

• Journal of Chest Surgery
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• v.12 no.1
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• pp.16-22
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• 1979

Clinical evaluation of contact sensitization to 2, 4-dinitro-chlorobenzene [DNCB] was performed in 2 groups: group A [30 patients with non-malignant disease] and group B [30 patients with bronchogenic carcinoma]. Initial sensitization was elicited out by applying 2, 000 ug of DNCB to skin surface of the both group A and B. Subsequently a relatively weak challenge dose, 200 ug of DNCB, was applied 14 days later, showing the satisfactory results of sensitization with minimizing non-specific irritative inflammatory skin response. Delayed cutaneous hypersensitivity reactions shown by spontaneous flare phenomena appeared at the challenge site, and they were assessed 48 hours later. The reaction were graded from +1 to +4 according to the degree of flare or vesicular reaction. The results were as follows: 1. 28 cases [93%] of group A, however, only 18 cases [67%] of group B exhibited delayed cutaneous hypersensitivity reaction to DNCB contact sensitization [P<0.02]. 2. Of group A, the delayed cutaneous hypersensitivity reactions above +2 of DNCB score were 25 cases [83%], meanwhile 11 cases [37%] in group B [P<0.001]. 3. Undifferentiated carcinomas showed highest incidence of anergy to DNCB contact sensitization in the all histologic types of group B. 4. In group B, 8 [42%] of 19 cases who react to DNCB were resectable, whereas only 2 [18 %] of 11 cases who failed to react to DNCB were resectable for curative cancer surgery. These study suggests that cellular immune reaction of group B was depressed remarkably comparing with that of group A.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.425-441
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• 1997

The neuropeptide substance P(SP) has been implicated in the mediation of inflammation and immune-mediated disease such as arthritis. Recently, it was reported that SP was markedly increased around the blood vessels in inflamed gingiva as well as in close association with the inflammatory cell infiltrate. These results support that SP may contribute to the pathophysiology of neuronal inflammation in human periodontal tissues. SP may regulate inflammatory/immune responses by stimulating the proliferation of human T cells, differentiation and antibody-secreting potential of B cells, macrophage respiratory burst, connective tissue proliferation, and the secretion of cytokines from monocytes and T cells. Here, I studied potential role of SP as a costimulatory chemical signal in inflammatory/immune responses, by determining the released proinflammatory cytokines such as $MIP-1{\alpha}$, $IL-1{\beta}$, and IL-6 from culture supernatants of homogeneous immune cell lines. Serum free cell supernatants were concentrated with TCA precipitation, fractionated with SDS-PAGE, and subjected into western blot analysis. Among 15 cell lines tested, macrophage/monocyte cell line RAW264.7 and WRl9m.1 showed the highest level of induction of $MIP-1{\alpha}$ when stimulated with LPS. Discrete IL-6 bands with multiple forms of molecular mass were detected from supernatants of B cell lines A20(32kDa), Daudi(32, 35kDa), and SKW6.4(29kDa), which were expressed constitutively. $IL-1{\beta}$ could not be detected by the method of western blot analysis from supernatants of all cell lines tested except RAW264.7, WRl9m.1, and erythroid cell line K562 which showed the least amount of $IL-{\beta}$ secretion. SP $10^{-9}M$ with suboptimal dose of LPS treatment showed synergistic induction of $MIP-1{\alpha}$ release from RAW264.7 or WR19m.1, and also IL-6 release from A20, but this synergism is not the case in costimulation of RAW264.7 or WRl9m.1 with SP $10^{-9}M$ and TPA. Although treatment of T cell line CTLL-R8 with SP $10^{-7}M$ or PHA+TPA induced modest level of $MIP-1{\alpha}$ secretion, synergism was not observed when they are applied together. These findings all together suggest the possibility of a regulatory role of SP in inflammatory/immune reaction through differential modulation of bioactivities of other chemical cosignals.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.2.1-2.18
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• 2022

Background: Co-infections of the porcine reproductive and respiratory syndrome virus (PRRSV) and the Haemophilus parasuis (HPS) are severe in Chinese pigs, but the immune response genes against co-infected with 2 pathogens in the lungs have not been reported. Objectives: To understand the effect of PRRSV and/or HPS infection on the genes expression associated with lung immune function. Methods: The expression of the immune-related genes was analyzed using RNA-sequencing and bioinformatics. Differentially expressed genes (DEGs) were detected and identified by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and western blotting assays. Results: All experimental pigs showed clinical symptoms and lung lesions. RNA-seq analysis showed that 922 DEGs in co-challenged pigs were more than in the HPS group (709 DEGs) and the PRRSV group (676 DEGs). Eleven DEGs validated by qRT-PCR were consistent with the RNA sequencing results. Eleven common Kyoto Encyclopedia of Genes and Genomes pathways related to infection and immune were found in single-infected and co-challenged pigs, including autophagy, cytokine-cytokine receptor interaction, and antigen processing and presentation, involving different DEGs. A model of immune response to infection with PRRSV and HPS was predicted among the DEGs in the co-challenged pigs. Dual oxidase 1 (DUOX1) and interleukin-21 (IL21) were detected by IHC and western blot and showed significant differences between the co-challenged pigs and the controls. Conclusions: These findings elucidated the transcriptome changes in the lungs after PRRSV and/or HPS infections, providing ideas for further study to inhibit ROS production and promote pulmonary fibrosis caused by co-challenging with PRRSV and HPS.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.53-59
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• 2004

Background: Minor histocompatibility HY antigen, as a transplantation antigen, has been known to cause graft rejection in MHC (major histocompatibility complex) matched donor-recipient. The aim of our study is to investigate the role of male antigen (HY) disparity on MHC matched pancreatic islet transplantation and to examine the mechanism of the immune reaction. Methods: Pancreatic islets were isolated and purified by collagen digestion followed by Ficoll gradient. The isolated islets of male C57BL6/J were transplanted underneath the kidney capsule of syngeneic female mice rendered diabetic with streptozotocine. Blood glucose was monitored for the rejection of engrafted islets. After certain period of time, tail to flank skin transplantation was performed either on mouse transplanted with HY mismatched islets or on sham treated mouse. The rejection was monitored by scoring gross pathology of the engrafted skin. Results: HY mismatched islets survived more than 300 days in 14 out of 15 mice. The acceptance of second party graft (male B6 islets) and the rejection of third party graft (male BALB/c islets) in these mice suggested the tolerance to islets with HY disparity. B6 Skin with HY disparity was rejected on day $25{\pm}7$. However, HY mismatched skin transplanted on the mice tolerated to HY mismatched islets survived more than 240 days. Tetramer staining in these mice indicated the CTL recognizing MHC Db/Uty was not deleted or anergized. Conclusion: The islet transplantation across HY disparity induced tolerance to HY antigen in C57BL6 mouse, which in turn induced tolerance to HY mismatched skin, which otherwise would be rejected within 25 days. The MHC tetramer staining suggested the underlying mechanisms would not be clonal deletion or anergy.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.8
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• pp.1189-1195
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• 2014

Natural resistance-associated macrophage protein 1 encoding gene (NRAMP1) plays an important role in immune response against intracellular pathogens. To evaluate the effects of NRAMP1 gene on immune capacity in pigs, tissue expression of NRAMP1 mRNA was observed by real time quantitative polymerase chain reaction (PCR), and the results revealed NRAMP1 expressed widely in nine tissues. One single nucleotide polymorphism (SNP) (ENSSSCG00000025058: g.130 C>T) in exon1 and one SNP (ENSSSCG00000025058: g.657 A>G) in intron1 region of porcine NRAMP1 gene were demonstrated by DNA sequencing and PCR-RFLP analysis. A further analysis of SNP genotypes associated with immune traits including contain of white blood cell (WBC), granulocyte, lymphocyte, monocyte (MO), rate of cytotoxin in monocyte (MC) and $CD4^-CD8^+$ T lymphocyte subpopulations in blood was carried out in four pig populations including Large White and three Chinese indigenous breeds (Wannan Black, Huai pig and Wei pig). The results showed that the SNP (ENSSSCG00000025058: g.130 C>T) was significantly associated with level of WBC % (p = 0.031), MO% (p = 0.024), MC% (p = 0.013) and $CD4^-CD8^+$ T lymphocyte (p = 0.023). The other SNP (ENSSSCG00000025058: g.657 A>G) was significantly associated with the level of MO% (p = 0.012), MC% (p = 0.019) and $CD4^-CD8^+$ T lymphocyte (p = 0.037). These results indicate that the NRAMP1 gene can be regarded as a molecular marker for genetic selection of disease susceptibility in pig breeding.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1613-1619
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• 2016

Francisella tularensis is a highly virulent pathogen of humans and other mammals. Moreover, F. tularensis has been designated a category A biothreat agent, and there is growing interest in the development of a protective vaccine. In the present study, we determine the in vitro and in vivo immune responses of a subunit vaccine composed of recombinant peptides Tul4 and FopA from epitopes of the F. tularensis outer membrane proteins. The recombinant peptides with adjuvant CpG induced robust immunophenotypic change of dendritic cell (DC) maturation and secretion of inflammatory cytokines (IL-6, IL-12). In addition, the matured DCs enabled ex vivo proliferation of naive splenocytes in a mixed lymphocyte reaction. Lastly, we determined the in vivo immune response by assessment of antibody production in C57BL/6 mice. Total IgG levels were produced after immunization and peaked in 6 weeks, and moreover, Tul4-specific IgG was confirmed in the mice receiving peptides with or without CpG. Based on these results, we concluded that the recombinant peptides Tul4 and FopA have immunogenicity and could be a safe subunit vaccine candidate approach against F. tularensis.