Understanding immune-modulatory efficacy in vitro

Abstract

Boosting or suppressing our immune system represents an attractive adjunct in the treatment of infections including SARS-CoV-2, cancer, AIDS, malnutrition, age related problems and some inflammatory disorders. Thus, there has been a growing interest in exploring and developing novel drugs, natural or synthetic, that can manipulate our defence mechanism. Many of such studies, reported till date, have been designed to explore effect of the therapeutic on function of macrophages, being a key component in innate immune system. Indeed, RAW264.7, J774A.1, THP-1 and U937 cell lines act as ideal model systems for preliminary investigation and selection of dose for in vivo studies. Several bioassays have been standardized so far where many techniques require high throughput instruments, cost effective reagents and technical assistance that may hinder many scholars to perform a method demanding compilation of available protocols. In this review, we have taken an attempt for the first time to congregate commonly used in vitro immune-modulating techniques explaining their principles. The study detected that among about 40 different assays and more than 150 sets of primers, the methods of cell proliferation by MTT, phagocytosis by neutral red, NO detection by Griess reaction and estimation of expression of TLRs, COX-2, iNOS, TNF-α, IL-6 and IL-1β by PCR have been the most widely used to screen the therapeutics under investigation.

Immunomodulatory effects of Abelmoschus esculentus water extract through MAPK and NF-κB signaling in RAW 264.7 cells

“…Immune-enhancing compounds predominantly activate macrophage function after being specifically combined with cell surface acceptors. 5 Toll-like receptor 4 (TLR4) is a type 1 transmembrane glycoprotein with a molecular weight of 100 kDa which is known to be expressed in macrophages and other cells. 6 A number of downstream signaling pathways, including the nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways, are triggered by various adapter complexes that interact with TLR4, resulting in the production of nitric oxide (NO) and inflammatory cytokines/chemokines.…”

PCSK9 Confers Inflammatory Properties to Extracellular Vesicles Released by Vascular Smooth Muscle Cells

“…Third, instead of using murine J774 macrophages, the human U937 could have been used. However, these two cell lines are readily exchangeable when evaluating phagocytosis, cytokine production and morphological analyses [ 84 ]. Fourth, to disentangle the role of CD36 in the uptake of oxLDL, an experiment in which CD36 was silenced could have been performed.…”

COX-2 is required to mediate crosstalk of ROS-dependent activation of MAPK/NF-κB signaling with pro-inflammatory response and defense-related NO enhancement during challenge of macrophage-like cell line with Giardia duodenalis

“…The macrophage J774A.1 and RAW264.7 cell lines [ 29 ] used to interact with Giardia trophozoites in this study were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). J774A.1 and RAW264.7 cells were cultured in high-glucose DMEM (Hyclone, Logan, USA) containing 10% heat-inactivated FBS (Cellmax, Beijing, China) and maintained in a 37°C humidified incubator with 5% CO 2 .…”