Evaluation of SV40-transformed synovial fibroblasts in the study of rheumatoid arthritis pathogenesis

Abstract

The SV40 T antigen has been used to generate immortalized cells from rheumatoid arthritis (RA) synovial fibroblasts (RASFs) that are commonly used in lieu of primary RASFs. In the current study, we investigated the effect of stimulation by tissue necrosis factor alpha (TNF-α) and interleukin 17 (IL-17) on primary and immortalized RASFs in order to gauge the appropriateness of the use of immortalized RASFs, the MH7A cell line, in the study of RA pathogenesis. Changes in the levels of secretion and expression of 8 proteins associated with RA upon stimulation were assessed by multiplex immunoassay. IL-17 stimulation had a minimal impact on protein secretion and expression for primary and immortalized cells. Basic fibroblast growth factor (FGF-2) was not detectable for the primary cells but was detectable for the immortalized cells. In contrast, monocyte chemoattractant protein 1 (MCP-1) was detectable for primary cells but was undetectable for immortalized cells. In general, protein expression and secretion by cells stimulated with TNF-α were significantly increased. For primary cells, several proteins were below the limit of detection for unstimulated cells and cells stimulated with IL-17, while levels for TNF-α-stimulated cells were within the detectable range. For the same proteins, expression was observed for immortalized cells, regardless of stimulation, suggestive of constitutive activation of the NF-κB signaling pathway. The current study therefore provides strong evidence that immortalized and primary RASFs differ in regard to protein expression and secretion and therefore may not be appropriate for use in the study of RA pathogenesis.

Comparison of Stress and Suicide-Related Behaviors Among Korean Youths Before and During the COVID-19 Pandemic

“…The ORs for subjective stress, sadness or despair, suicidal thoughts, suicide planning, and suicide attempts of the 2020 participants were compared with those of the 2019 participants using multiple logistic regression analysis with complex sampling using the weighted value of each participant. 21 Both the male and female groups showed lower odds of experiencing severe stress, sadness or despair, and suicidality in 2020 than in 2019 (Table 3). In the male group, the 2020 participants had lower odds of very severe stress (aOR, 0.68 [95% CI, 0.61-0.77], P < .001 ), sadness or despair (aOR, (ie, the high-, middle-, and low-income groups) showed lower odds of severe stress, sadness or despair, suicidal thoughts, and suicide attempts in the 2020 participants than in the 2019 participants (Table 4).…”

“…The ORs for subjective stress, sadness or despair, suicidal thoughts, suicide planning, and suicide attempts of the 2020 participants were compared with those of the 2019 participants using multiple logistic regression analysis with complex sampling using the weighted value of each participant. 21 Crude and adjusted models (for age, BMI, physical exercise, sedentary time for study and leisure, sleep time, sex, economic level, educational levels of the father and mother, scholastic performance, and subjective health status) were designed. Subgroup analyses by sex, economic level, and scholastic performance were designed.…”

A comparison of strategies for immortalizing mouse embryonic fibroblasts

“…[10] found that SV40 immortalized MEFs have significant resistance toward D, L-sulforaphane (SFN) induced apoptosis compared with wild-type MEFs. While SV40 Ta has been effective for immortalizing different cell types, many have cited molecular and phenotypic differences between wild type and SV40 transformed cell lines [11-15]. Jeong et al .…”

Investigation of the cytokine response to NF-κB decoy oligonucleotide coated polysaccharide based nanoparticles in rheumatoid arthritis in vitro models

“…To further validate the ability of NP to serve as an effective treatment strategy for RA, in vitro experiments were also conducted with primary cells. Previous reports have noted discrepancies in cytokine production between cell line models and primary RASF cells [ 35 ]. Furthermore, a literature search revealed that immortalized cells, such as SW982, constitutively express the NF-κB pathway, indicating that they may be more susceptible to NF-κB interference than primary RASF cells [ 36 ].…”