Deteksi populasi Cd3+Cd4+Cd25+Foxp3+ T-regulator pada Limpa Mencit Galur Balb/c dan Swiss-webster yang Mudah dan Cepat dengan Metode Flow Cytometry

Sel T-regulator (Treg) memainkan peran kunci dalam mengendalikan autoimunitas, respon alergi, peradangan, dan respons terhadap infeksi. Sel T Cd4+ yang secara konstitutif mengekspresikan Cd25 dan protein forkhead 3 (Foxp3; yaitu, Cd3+Cd4+Cd25+Foxp3+) atau disebut juga sebagai Treg fungsional, telah terbukti memainkan peran utama dalam pemeliharaan toleransi dan homeostasis imun, sehingga deteksi Treg fungsional sangat penting dilakukan sebagai salah satu parameter imunomodulasi. Penelitian ini bertujuan untuk mendeteksi populasi sel Treg (Cd3+Cd4+Cd25+Foxp3+) pada organ limpa mencit galur Balb/c dan Swiss-webster dengan menggunakan metode yang mudah, cepat, serta terjangkau. Metode deteksi sel Treg dilakukan dengan cara isolasi organ limpa mencit Balb/c dan Swiss-webster kemudian dilakukan preparasi dan deteksi sel menggunakan instrumen flow cytometry. Hasil populasi sel Treg yang didapatkan berturut-turut pada mencit Balb/c dan Swiss-webster adalah 2,02 ± 0,36% dan 3,14 ± 1,64% dari total populasi yang dideteksi dengan hanya membutuhkan waktu 145 menit sejak pembedahan. Berdasarkan hasil tersebut dapat disimpulkan bahwa pengamatan populasi sel Treg pada splenosit mencit galur Balb/c dan Swiss-webster dengan flow cytometry dapat dilakukan dengan mudah dan praktis dengan pewarnaan kombinasi antibodi yang tertera pada manuskrip ini. Selain itu, metode pewarnaan yang tertera dapat digunakan sebagai salah satu acuan untuk mendeteksi imunomodulator pada berbagai model hewan coba yang menggunakan mencit galur Balb/c dan Swiss-webster, khususnya yang berhubungan dengan populasi sel Treg

Pengaruh asam 2-(3-(klorometil)benzoiloksi)benzoat terhadap rasio limfosit-T CD3CD4 dan CD3CD8 pada limpa mencit Swiss webster metode fluorescence-activated cell sorting (FACS)

Inflamasi merupakan sistem imunitas tubuh ketika mengalami induksi yang berbahaya seperti patogen, sel-sel rusak, ataupun senyawa beracun. Inflamasi terjadi karena terbentuknya mediator yaitu COX-2 yang dilepaskan ke matriks ekstraseluler sehingga sel-sel imunitas akan terstimulasi. OAINS memiliki mekanisme kerja yaitu dengan menghambat sintesis prostaglandin pada jalur asam arakidonat. Sel imunitas adaptif tubuh mengalami maturasi salah satunya pada organ limpa. Sel limfosit-T akan berikatan dengan reseptor αβ lalu berdiferensiasi menjadi CD4 dan CD8. Asam Asetilsalisilat (AAS) diketahui dapat meningkatkan ekspresi Foxp3 pada sel CD3posCD4posCD25pos dan CD3posCD8posCD25pos limfosit-T sehingga dapat menjadi sel T regulator (Treg) dan memiliki kemampuan dalam menekan proliferasi sel T CD4 sehingga memiliki keuntungan sebagai imunosupresan. Penelitian ini akan menggunakan hewan coba mencit Swiss Webster usia 12 minggu dan terbagi menjadi 5 kelompok yaitu kelompok kontrol negatif, kontrol positif, AAS, 3-CH2Cl, dan Celecoxib. Hewan coba akan diinduksi inflamasi dengan senyawa Lipopolisakarida (LPS) lalu diikuti dengan pemberian senyawa uji selama 2 hari. Hasil yang didapatkan adalah pemberian asam 2-(3-(klorometil)benzoiloksi)benzoat (3-CH2Cl) dosis 60 mg/KgBB dapat meningkatkan rasio sel limfosit-T CD3posCD4pos dan menurunkan CD3posCD8pos sehingga menunjukkan kemampuan serupa dengan senyawa AAS sebagai imunosupresan dengan metode Fluorescence-Activated Cell Sorting (FACS) setelah diinduksi lipopolisakarida

2-(3-(chloromethyl)benzoyloxy)benzoic Acid Increases CD4+ Regulatory T-Cell Population and FoxP3 Expression in Lipopolysaccharide-induced Mice

BACKGROUND: Lipopolysaccharide (LPS) has been reported to increase CD4+ regulatory T-cell (CD4+ Treg) populations. Acetylsalicylic acid (ASA) has been reported to have immunomodulatory activity, but it may induce chronic gastric ulceration. Another salicylic acid-bearing compound, 2-(3-(chloromethyl)benzoyloxy)benzoic acid (3-CH2Cl), has been reported to have less gastric mucosal damage. However, the effect of 3-CH2Cl on CD4+ Tregs in LPS-induced mice is still unknown. Therefore, the present study was conducted to investigate the immunomodulatory effect of 3-CH2Cl on CD4+ T-cell and CD4+ Treg populations as well as FoxP3 expression in LPS-induced mice. METHODS: Synthesis of 3-CH2Cl was performed by mixing salicylic acid and chloromethylbenzoylchloride with the catalyzation of pyridine, acetone and heat. The 3-CH2Cl tablets were prepared using direct compression method. After intraperitoneal injection of 1 mg/kg BW LPS to mice, 60 mg/kg BW ASA or 60 mg/kg BW 3-CH2Cl was given orally for 3 days. The splenocyte was obtained through splenectomy and collagenase digestion. The population of CD4+ T-cells and CD4+ Tregs, as well as the splenic FoxP3 expression were determined using flow cytometry technique. RESULTS: CD4+ T-cell populations in mice treated with LPS and 3-CH2Cl or ASA were lower than those treated with LPS merely. Meanwhile, CD4+ Treg populations and FoxP3 expression levels in mice treated with LPS and 3-CH2Cl or ASA were higher than those treated with LPS merely. CONCLUSION: Since 3-CH2Cl could decrease CD4+ T-cell population and increase CD4+ Treg population mediated by the increase of FoxP3 expression in LPS-induced inflammation, it may act as a potential therapeutic drug to reduce inflammatory conditions. KEYWORDS: 2-(3-(chloromethyl)benzoyloxy)benzoic acid, acetylsalicylic acid, CD4, T-regulatory cells, FoxP3, LP

Data formats and standards for opportunistic rainfall sensors [version 2; peer review: 2 approved]

Opportunistic sensors are increasingly used for rainfall measurement. However, their raw data are collected by a variety of systems that are often not primarily intended for rainfall monitoring, resulting in a plethora of different data formats and a lack of common standards. This hinders the sharing of opportunistic sensing (OS) data, their automated processing, and, at the end, their practical usage and integration into standard observation systems. This paper summarises the experiences of the more than 100 members of the OpenSense Cost Action involved in the OS of rainfall. We review the current practice of collecting and storing precipitation OS data and corresponding metadata, and propose new common guidelines describing the requirements on data and metadata collection, harmonising naming conventions, and defining human-readable and machine readable file formats for data and metadata storage. We focus on three sensors identified by the OpenSense community as prominent representatives of the OS of precipitation: Commercial microwave links (CML): fixed point-to-point radio links mainly used as backhauling connections in telecommunication networks Satellite microwave links (SML): radio links between geostationary Earth orbit (GEO) satellites and ground user terminals. Personal weather stations (PWS): non-professional meteorological sensors owned by citizens. The conventions presented in this paper are primarily designed for storing, handling, and sharing historical time series and do not consider specific requirements for using OS data in real time for operational purposes. The conventions are already now accepted by the ever growing OpenSense community and represent an important step towards automated processing of OS raw data and community development of joint OS software packages