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"This volume compiles the latest advances in this rapidly evolving field as presented by eminent scientists at the 7th International Aegean Conference on Innate Immunity in Rhodes, Greece. It includes topics related to the biology and function of Toll-like and other pattern-recognition receptors, complement and its crosstalk with other physiological systems, inflammatory mechanisms and diseases, natural killer cells, and the cooperative interplay between innate and adaptive immune cells. This book is an excellent source of information for researchers and clinicians with interests in immunology, host-microbe interactions, and infectious and inflammatory diseases."

"This book will cover primary roles of NKT cells in immunity to cancer, in both mouse tumor models and cancer patients. There are several chapters describing general aspects of NKT cells."

"Acalypha indica (Ai) merupakan herbal yang diketahui memiliki efek anti-inflamasi dan anti-diabetik. Penelitian ini dilakukan dengan tujuan untuk mengetahui efek ekstrak Ai terhadap proses inflamasi sebagai bagian dari respons imun bawaan yang terjadi pada diabetes melitus tipe 2. Metode penelitian dilakukan dengan menggunakan dua puluh lima ekor tikus Sprague-Dawley yang dibagi menjadi lima kelompok yaitu kelompok normal, kelompok yang diinduksi diet tinggi fruktosa dan kolesterol  (DTFK), kelompok DTFK+Ai 250 mg/kgBB, kelompok DTFK+Ai 400 mg/kgBB dan DTFK+kelompok pioglitazone. Hasil penelitian menunjukkan bahwa ekstrak Ai dosis 400 mg/kgBB dapat menurunkan kadar glukosa darah sewaktu (GDS) (p=0.005) dengan mean±SEM berturut-turut (82±5.73), (199.9±27.55), (260.6±67.29), (147.2±12.18) dan (148.8±29.4). Ekstrak Ai dosis 400 mg/kgBB juga dapat menurunkan kadar glukosa darah puasa (GDP) (p=0.02) dengan mean ± SEM berturut-turut (76.65±2.612), (144.7±13.03), (193.0±46.17), (117.0±2.629) dan (130.5±8.890). Selain itu pemberian ekstrak Ai menunjukkan efek peningkatan pada kadar TLR4 (p=0.03) dengan mean±SEM berturut-turut (1.967±0.149), (2.632±0.146), (2.687±0.134), (2.902±0.136) dan (2.513±0.334). Pemberian ekstrak Ai juga menunjukkan peningkatan jumlah hitung sel radang di hati (p=0.0001) dengan mean ± SEM berturut-turut (25.92±2.03), (46.76±3.43), (44.28±3.69), (46.32±3.13) dan (30.72±2.94). Pemberian ekstrak Ai tidak menunjukkan perbedaan bermakna pada kadar TLR2 serta ekspresi gen Nos2 dan Arg1. Kesimpulannya, ekstrak Ai dapat menurunkan kadar GDS dan GDP serta meningkatkan kadar TLR4 dan jumlah sel radang di hati.

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Acalypha indica (Ai) is an herb that is known to have anti-inflammatory and anti-diabetic effects. This research was carried out with the aim of determining the effect of Ai extract on the inflammatory process as part of the innate immune response that occurs in type 2 diabetes mellitus. The research method was carried out using twenty-five Sprague-Dawley rats which were divided into five groups : the normal group, high fructose high cholesterol (DTFK) group, DTFK+Ai 250 mg/kgBW group, DTFK+Ai 400 mg/kgBW group and DTFK+pioglitazone group. The results showed that Ai extract at a dose of 400 mg/kgBW could reduce random blood glucose levels (GDS) (p=0.005) with mean ± SEM respectively (82 ± 5.73), (199.9 ± 27.55), (260.6 ± 67.29), (147.2±12.18) and (148.8±29.4). Ai extract at a dose of 400 mg/kgBW can also reduce fasting blood glucose (GDP) levels (p=0.02) with mean ± SEM respectively (76.65±2.612), (144.7±13.03), (193.0±46.17), (117.0± 2,629) and (130.5±8,890). In addition, administration of Ai extract showed an increasing effect on TLR4 levels (p=0.03) with mean ± SEM respectively (1.967 ± 0.149), (2.632 ± 0.146), (2.687 ± 0.134), (2.902 ± 0.136) and (2.513 ± 0.334). Administration of Ai extract also showed an increase in the number of inflammatory cells in the liver (p=0.0001) with mean ± SEM respectively (25.92±2.03), (46.76±3.43), (44.28±3.69), (46.32±3.13) and (30.72). ±2.94). Administration of Ai extract did not show significant differences in TLR2 levels and Nos2 and Arg1 gene expression. In conclusion, Ai extract can reduce GDS and GDP levels but increase TLR4 levels and the number of inflammatory cells."

"Innate and adaptive immunity play important roles in immunosurveillance and tumor destruction. This book discusses recent progress in innate immunity and Treg cells, and the regulation of innate immunity through Toll-like receptor (TLR) signalling."

"In this book, the author draws upon her own research, conducted over the past three decades, to provide a unique compilation of high-quality illustrations that offer illuminating insights in a readily accessible form. The ovarian follicles, the corpus luteum, and the interstitial cortex are presented as precisely controlled homeostatic compartments. Physiologic cell death forms and leukocyte subtypes are clearly depicted, and the various steps in the ovulatory process are demonstrated in detail. Further topics addressed include the microvascular bed, the plasticity of intraovarian nerves as part of the remodelling processes in the polycystic and the menopausal ovary, and the role of cytokeratin-positive cells as messengers of innate immunity. "

"This book will discuss several cellular pathways involved in controlling immune response in the context of infectious diseases, their biological consequences and potential "hijack" of these pathways for the benefit of pathogen leading towards pathogen persistence as opposed to clearance."

"Karsinoma hepatoseluler (KHS) adalah kanker primer liver dan penyebab kedua kematian akibat kanker. Eksosom pada lingkungan mikro KHS berfungsi untuk komunikasi antar sel dan bila endositosis ke sel NK dapat menyebabkan perubahan pada sel NK. Penelitian ini bertujuan menganalisis eksosom dari darah pasien KHS, perubahan fenotipe sel NK, dan uji pewarnaan histologi (peroksidase, toluidine blue) untuk mengamati perubahan granula azurofilik sel NK akibat endositosis eksosom. Metode penelitian meliputi isolasi sel NK dari donor sehat dan eksosom darah pasien KHS, karakterisasi eksosom dengan PSA, stimulasi sel NK dengan eksosom, flow cytometry reseptor pada sel NK dan CD81+ pada eksosom, imunofluoresens endositosis eksosom ke sel NK, pewarnaan toluidine blue dan peroksidase.Hasil menunjukkan eksosom berukuran 34,7 nm, bermuatan -4,33 mV dan positif CD81+. Perubahan reseptor sel NK sehat yang dipaparkan eksosom KHS tidak signifikan (P>0,05). Imunofluoresens memperlihatkan endositosis eksosom ke sel NK. Pewarnaan sel NK toluidine blue menunjukkan metakromasia dan peroksidase negatif. Sel NK+eksosom mengalami perubahan hasil pewarnaan. Peneliti menyimpulkan bahwa eksosom dari darah pasien KHS sesuai kriteria MISEV 2018.Tidak terjadi perubahan fenotipe sel NK sehat yang dipaparkan eksosom dari darah pasien KHS. Pewarnaan peroksidase dan toluidine blue dapat digunakan sebagai metode pengamatan endositosis eksosom ke sel NK.

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Hepatocellular carcinoma (HCC) is the primary liver cancer and the second leading cause of death from cancer. Exosomes in the HCC microenvironment function for communication between cells and when endocytosed to NK cells can cause changes in NK cells. This study aims to analyze exosomes from the blood of HCC patients, changes in NK cell phenotype, and histological staining tests (peroxidase, toluidine blue) to observe changes in NK cell azurophilic granules due to exosome endocytosis. NK cells from healthy donors and blood exosomes of KHS patients were isolated, exosomes characterized by PSA, stimulation of NK cells with exosomes, and flow cytometry of receptors on NK cells and CD81+ on exosomes were done. Endocytosis of exosomes onto NK cells were observed through immunofluorescence, then metacromasia and azurofilic granules of NK cells were observed after toluidine blue and peroxidase staining. Results showed The exosome is 34.7 nm in size, has a charge of -4.33 mV and is CD81+ positive. Changes in healthy NK cell receptors exposed to HCC exosomes were not significant (P>0.05). Immunofluorescence demonstrates exosome endocytosis in NK cells. Toluidine blue NK cell staining showed negative metachromasia and peroxidase. In NK cell+exosome there is a change in staining results. We concluded exosomes from the blood of HCC patients comply with MISEV 2018 criteria. There is no change in the phenotype of healthy NK cells exposed to exosomes from the blood of HCC patients. Peroxidase and toluidine blue staining can be used as a method of observing exosome endocytosis in NK cells."

"Latar BelakangKarsinoma Hepatoseluler (KSH) merupakan kanker yang umum terjadi dengan angka kejadian tinggi. Sel Natural Killer (NK) memiliki kemampuan sitotoksisitas terhadap sel kanker, namun efektivitasnya dapat dipengaruhi oleh eksosom. Eksosom dari serum sehat diduga mampu meningkatkan aktivitas sitotoksisitas sel NK. Penelitian ini bertujuan untuk menganalisis sitotoksisitas sel HepG2 oleh sel NK yang diinduksi eksosom serum sehat.MetodePenelitian ini merupakan studi in vitro menggunakan flow cytometry untuk menganalisis persentase sitotoksisitas HepG2 oleh sel NK yang diinduksi eksosom serum sehat. Tiga kelompok perlakuan diuji, yaitu NK tanpa induksi, NK diinduksi eksosom, dan NK diinduksi IL-2. Setiap kelompok perlakuan memiliki 1 kelompok data yang terdiri dari 8 well (n = 24). Analisis data menggunakan GraphPad Prism, menggunakan uji Normalitas dan homogenitas, uji One-Way Anova, dan uji t-test untuk melihat signifikansi setiap kelompok uji.HasilHasil persentase rata-rata sitotoksisitas HepG2 oleh sel NK yang diinduksi eksosom serum sehat adalah 14,6% ± 1,1%. Persentase sitotoksisitas kelompok kontrol induksi adalah 14% ± 1,7%. Tidak terdapat peningkatan persentase sitotoksisitas HepG2 oleh sel NK yang diinduksi eksosom serum sehat. Perbandingan rerata sitotoksisitas HepG2 antar kedua kelompok tidak signifikan secara statistik (P > 0,05)KesimpulanTidak terdapat peningkatan signifikan dalam sitotoksisitas sel NK terhadap sel HepG2 yang diinduksi oleh eksosom serum sehat dibandingkan dengan kontrol tanpa induksi.

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Introduction

Hepatocellular Carcinoma (HCC) is a commonly cancer with a high incidence rate. Natural killer cells have the ability to kill cancer cells, but their affectiveness may be affected by exosome. Exosomes from healthy serum have the potential to increase the cytotoxicity of NK cells. This study aims to analyze the cytotoxic effect of HepG2 cells by Natural Killer (NK) induced by healthy serum.

Method

This study is an in vitro investigation utilizing flow cytometry to analyze the percentage of cytotoxicity of HepG2 cells by Natural Killer (NK) cells induced by healthy serum exosomes. Three experimental groups were evaluated: NK cells without induction, NK cells induced by exosomes, and NK cells induced by IL-2. Each experimental group consisted of one dataset comprising 8 wells (n = 24). Data analysis was performed using GraphPad Prism, which included normatily and homogeneity tests, One-Way ANOVA, and t-tests to assess the significance of differences among the experimental groups. Results

The average percentage of cytotoxicity of HepG2 cells by NK cell induced by healthy serum exosome was 14,6% ± 1,1%. The percentage of cytotoxicity in the control group was 14% ± 1,7%. There was no increase in percentage of cytotoxicity HepG2 cells by NK cell induced by healthy serum exosome. Comparisone between mean of the two groups was not statistically significant (P > 0,05)

Conclusion

There was no significant increase in the cytotoxicity of NK cells agains HepG2 cells induced by healthy serum exosome compared to the non-induced control group."

"Latar Belakang: Anak-anak yang menderita stunting memiliki berbagai kekurangan jika dibandingkan anak-anak sebayanya yang memiliki HAZ normal, baik dari segi pertumbuhan fisik, emosional, maupun dalam sistem imun. Salah satu komponen sistem imun yang ada dalam tubuh adalah sitokin proinflamasi interleukin-18 yang berperan sebagai faktor kemotaksis sel T, basofil, serta neutrofil, penginduksi interleukin lainnya, serta menginduksi sel Th1 dan IFN- I³. Tujuan: Menganalisis ekspresi gen IL-18 pada anak stunting jika dibandingkan dengan anak dengan HAZ normal, menganalisis korelasi antara status stunting, ekspresi IL-18, status infeksi cacing, serta status OHI-S. Metode: Sampel diambil dari bahan biologis tersimpan berupa RNA cairan sulkus gingiva anak 6-8 tahun di Nusa Tenggara Timur (NTT) (n=8). Kemudian dilakukan ekstraksi RNA, sintesis cDNA, pre amplifikasi, dan kemudian dilakukan real-time PCR. Hasil: Tidak ditemukan perbedaan bermakna secara statistik pada ekspresi gen IL-18 anak stunting dibanding anak dengan HAZ normal (p ≥ 0,05) dan tidak pula ditemukan korelasi baik antara status stunting dan status infeksi cacing, ekspresi IL-18 dan status infeksi cacing, status stunting dan OHI-S, maupun ekspresi gen IL-18 dan status OHI-S (p ≥ 0,05). Kesimpulan: Meskipun ditemukan adanya downregulation pada ekspresi gen IL-18 anak stunting jika dibandingkan anak normal, perbedaan tersebut tidak bermakna secara statistik Tidak ditemukan korelasi pada ekspresi gen IL-18, status infeksi cacing, serta status OHI-S.

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Background: Stunted children have many handicaps compared to their normal age counterparts who have normal HAZ, either in physical growth, emotional growth, or in their immune system. Interleukin-18 is a part of the immune system, a proinflammatory cytokine that acts as a chemotaxis factor for T-cell, basophil, neutrophil, and inducts IFN- γ, Th1, and other cytokines.

Purpose: To analyze IL-18 expression in stunted children compared to their normal age counterpart, to analyze the correlation between stunting status, IL-18 expression, helminths infection status, and OHI-S.

Methods: Samples were stored biological material, taken from 6 to 7 years old’s gingival crevicular fluid from NTT (n=8). RNA was extracted from samples, then synthesized to cDNA, preamplified, and analyzed in RT-PCR. 

Results: The difference in IL-18 expression in stunted children compared to children with normal HAZ was not statistically significant.  There were no correlation between stunting status and helminths infection status, IL-18 expression and helminths infection status, stunting status, and OHI-S, nor IL-18 expression and OHI-S.

Conclusion: Even though a downregulation in IL-18 expression in stunted children compared to children with normal HAZ was found, the difference was not statistically significant. There was also no correlation between IL-18 expression, helminths infection status, and OHI-S status. "

"Sel Natural Killer (NK) adalah sel pelepas granul sitotoksik yang melisis patogen intracytoplasmic (yaitu infeksi virus atau bakteri) dan sel tumor/kanker sebagai bagian dari imunitas bawaan. Sel NK berasal dari diferensiasi sel punca hematopoietik (SPH) di jalur Common Lymphoid Progenitor (CLP). SPH diperoleh dari darah tali pusat, dengan jumlah SPH yang lebih tinggi daripada sumsum tulang. Berbagai protokol diferensiasi telah dilaporkan dengan jumlah sel NK dan fenotipe yang berbeda. Studi perbandingan efektivitas diferensiasi sel NK dengan sampel SPH yang dikultur dan SPH yang baru diisolasi masih minim. Penelitian ini bertujuan untuk membandingkan tahapan maturasi diferensiasi sel NK yang dihasilkan antara sampel SPH yang dikultur dan baru diisolasi menggunakan modifikasi protokol Dezell dkk. dengan menggunakan interleukin-2 (IL-2) tanpa keberadaan feeder cell. Kultur SPH dilakukan selama dua minggu sebelum diferensiasi untuk sampel SPH yang dikultur. Hasil kultur diferensiasi selama lima minggu dianalisis menggunakan flow cytometry untuk mengetahui keberadaan reseptor NKp46, pengamatan Giemsa untuk mengetahui tahapan maturasi sel NK, dan qRT-PCR untuk mengetahui ekspresi gen perforin dan granzyme B. Hasil penelitian menunjukkan bahwa sampel SPH yang dikultur menghasilkan jumlah sel NK di tahap dewasa (tahap 5) yang lebih tinggi dibandingkan sampel SPH yang diisolasi melalui pengamatan Giemsa. Hasil flow cytometry menunjukkan nilai MFI NKp46 yang berbeda signifikan pada kedua sampel, dengan keberadaan reseptor aktivasi NKp46 yang lebih tinggi dijumpai pada sampel isolasi di hari ke-35. Hal ini disebabkan oleh aktivitas ROS pada kultur SPH dan regulasi mikroRNA. Oleh karena itu, sampel SPH yang dikultur dan sampel SPH yang baru diisolasi mampu menghasilkan populasi sel NK yang dewasa.

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Natural Killer (NK) cells are cytotoxic-granule-releasing cells which lysis intracytoplasmic pathogens (ie. virus or bacteria infection) and tumor/ cancer cells as part of innate immunity. NK cells originate from differentiation of hematopoietic stem cells (HSCs) in the Common Lymphoid Progenitor (CLP) pathway. HSCs can be obtained from umbilical cord blood, with a higher number of HSCs than bone marrow. Various differentiation protocols have been reported with different NK cell yields and phenotypes obtained. Comparative studies on the effectiveness of NK cell differentiation with cultured HSC samples and freshly isolated HSC are still minimal. The aim of this study was to compare the different stages of NK cell differentiation maturation produced between cultured and newly isolated SPH samples using a modified protocol of Dezell et al. using interleukin-2 (IL-2) in the absence of feeder cells. For expanded HSC samples, cultures were carried out for two weeks before differentiation. The results of the differentiation culture for five weeks were then analyzed using flow cytometry to determine the presence of NKp46 receptors, Giemsa observations to determine the stages of NK cell maturation, and qRT-PCR to determine the expression of perforin and granzyme B genes. The results show that cultured HSC samples can produce a higher number of NK cells with a more mature stage than freshly isolated HSC samples by Giemsa's observations which showed the presence of NK cells at stages 5. The results of flow cytometry showed that the MFI NKp46 values ​​were significantly different in the two samples, with a higher NKp46 activation receptor found in the isolated samples on day 35. This is due to ROS activity on SPH culture and microRNA regulation. Therefore, the cultured HSC samples and freshly isolated HSC samples were able to produce mature NK cell populations."