Human CCR6+ Memory T-helpers Cells form a Heterogenous Population Including Th17/Th22 and Th17.1 Cells, which Differ in Transcription Factor and Cytokine Expression but all Activate Synovial Fibroblast in an IFNγ-independent Manner

BackgroundChronic synovial inflammation is an important hallmark of inflammatory arthritis, but the cells and mechanisms involved are incompletely understood. Previously, we have shown that CCR6+ memory T-helper (memTh) cells and synovial fibroblasts (SF) activate each other in a pro-inflammatory feedforward loop, which potentially drives persistent synovial inflammation in inflammatory arthritis. However, the CCR6+ memTh cells are a heterogeneous population, containing Th17/Th22 and Th17.1 cells. Currently it is unclear which of these subpopulations drive SF activation and how they should be targeted. In this study, we examined the individual contribution of these CCR6+ memTh subpopulations to SF activation and examined ways to regulate their function.MethodsTh17/Th22 (CXCR3-CCR4+), Th17.1 (CXCR3+CCR4-), DP (CXCR3+CCR4+) and DN (CXCR3-CCR4-) CCR6+ memTh, cells sorted from PBMC of healthy donors or treatment-naïve early rheumatoid arthritis (RA) patients, were cocultured with SF from RA patients with or without anti-IL17A, anti-IFNγ or 1,25(OH)2D3. Cultures were analyzed by RT-PCR, ELISA or flow cytometry.ResultsTh17/Th22, Th17.1, DP and DN cells equally express RORC, but differ in production of T-bet and cytokines like IL-17A and IFNγ. Despite these differences, all the individual CCR6+ memTh subpopulations, both from healthy individuals and RA patients, were more potent in activating SF than the classical Th1 cells. SF activation was partially inhibited by blocking IL-17A, but not by inhibiting IFNγ or T-bet. However, active vitamin D inhibited the pathogenicity of all subpopulations leading to suppression of SF activation.ConclusionsHuman CCR6+ memTh cells contain several subpopulations that equally express RORC but differ in T-bet, IFNγ and IL-17A expression. All individual Th17 subpopulations are more potent in activating SF than classical Th1 cells in an IFNγ-independent manner. Furthermore, our data suggest that IL-17A is not dominant in this T cell-SF activation loop but that a multiple T cell cytokine inhibitor, such as 1,25(OH)2D3, is able to suppress CCR6+ memTh subpopulation-driven SF activation.

Neem (Azadiracta indica) a possible silver bullet in Covid 19

In past few years, multiple new viral diseases like SARS,MARS and Ebola are emerging all over the world. Due to global warming and pollution, people are becoming more susceptible to all infectious diseases. Recurrent mutations in these viruses make it difficult to create vaccine. So again there is need of safer and natural remedies which can help to sustain health of the present world and which can be used without potential hazards. The recent pandemic of Covid 19 is one of the example of a global crisis emerged by SARS-CoV-2 virus. It is causing acute as well as silent chronic damage to various systems in human body. Neem (Azadiracta indica) is found to have following properties as per various studies- It has Broad spectrum antiviral , antibacterial, antifungal , antiretroviral and antimalerial effect. It has proinflammatory cytokine inhibitor and immunomodulator effect. It has hepatoprotective and antioxidant effect. It has thrombolytic properties. It has ACE inhibitor action. As per its docking study, it has high inhibition against COVID-19 Main Protease. As Covid 19 is responsible for severe cytokine storm induced complications and coagulopathies, the Neem can be useful as a single Silver bullet in Covid 19 in both prophylactic and curative aspect. It can even be helpful in post Covid complications.

Effect of treating glioblastoma with a cytokine inhibitor, ibudilast, in combination with temozolomide on survival in a patient-derived xenograft model.

2062 Background: Recurrence in patients with glioblastoma (GBM) is inevitable, even in patients with O-6-Methylguanine-DNA Methyl Transferase ( MGMT) methylation. We identified increased expression of the inflammatory cytokine, Macrophage Inhibitory Factor (MIF) and its receptor CD74 in patients with recurrent tumors. High levels of MIF and CD74 were associated with poor overall survival in GBM patients. This study aims to determine efficacy of Ibudliast (MN-166; 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine) to block MIF expression and decrease tumor burden. Ibudilast is an anti-inflammatory drug that was developed for the treatment of bronchial asthma. Methods: The patient derived cell lines (PDCLs) RN1 ( MGMT unmethylated), BAH1 ( MGMT methylated), and HW1 ( MGMT methylated) were treated in vitrowith different concentrations of ibudilast in combination with temozolomide (TMZ). Patient derived xenograft (PDX) models of GBM were developed and treated with the combination of ibudilast and TMZ. Overall survival was calculated. Results: Regardless of MGMT status, significant synergism between ibudilast and TMZ was observed in the PDCLs. Efficacy was associated with significantly decreased expression of its targets, MIF and CD74. Downstream proteins such as Src and Akt were also significantly inhibited. The combination induced apoptosis. RN1 tumors were established intracranially in Balb/c nude mice. Significant increases in survival times of the mice were recorded when treated with the combination. Conclusions: Ibudilast in combination with TMZ resulted in significant blockage of MIF expression, increased apoptosis and longer survival in vivo. A human pilot study for recurrent GBM patients is underway.