Journal: Journal of Inflammation Research

Article Title: Qing-Luo-Yin Alleviated Experimental Arthritis in Rats by Disrupting Immune Feedback Between Inflammatory T Cells and Monocytes: Key Evidences from Its Effects on Immune Cell Phenotypes

doi: 10.2147/JIR.S346365

Figure Lengend Snippet: General effects of QLY treatment on CIA in rats. ( A ) Morphological observation of hind paws and historical examination of ankle joints/subcutaneous tissues, black arrow: cartilage erosion, blue arrow, rheumatoid nodules; ( B ) local expression of Th17α in the thymus assessed by immunohistochemical method; ( C ) distribution of CD3 + CD4 + IL-17α + Th17 cells in peripheral blood evaluated by flow cytometry; ( D ) differentially expressed cytokines among different groups based on results from cytokine chip analysis; ( E ) protein expression of SCF, Flt-3L and c-kit in the marrow assessed by immunohistochemical method; ( F ) mRNA expression of SCF in marrow cells cultured in vitro evaluated by RT-qPCR; ( G ) distribution of inflammatory CD86 + CD11b + monocytes in peripheral blood evaluated by flow cytometry; ( H ) levels of IL-1β and TNF-α in the serum determined by ELISA. Statistical significance in image ( H ) *p < 0.05 and **p < 0.01 compared with CIA model rats; statistical significance in image F, *p < 0.05 compared with normal untreated cells.

Article Snippet: Rat enzyme-linked immunosorbent assay (ELISA) kits for the quantification of granulocyte-macrophage colony stimulating factor (GM-CSF, 70-EK363), interleukin 1 beta (IL-1β, 70-EK301B/3), interleukin 17 alpha (IL-17α, 70-EK317/3), TNF-α (70-EK382/3) and interleukin 6 (IL-6, 70-EK306/3) together with phorbol 12-myristate 13-acetate (PMA, CS1001) and brefeldin A (BFA, CS1002) were purchased from MultiScience Biotech (Hangzhou, Zhejiang, China).

Techniques: Expressing, Immunohistochemical staining, Flow Cytometry, Cell Culture, In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Qing-Luo-Yin Alleviated Experimental Arthritis in Rats by Disrupting Immune Feedback Between Inflammatory T Cells and Monocytes: Key Evidences from Its Effects on Immune Cell Phenotypes

doi: 10.2147/JIR.S346365

Figure Lengend Snippet: Impact of QLY treatment on immune milieu in AIA rats. ( A ) Flow cytometry analysis of CD3 + CD4 + IL-17α + Th17 cells in peripheral blood sampled on day 18; ( B ) flow cytometry analysis of inflammatory CD43 + CD86 + monocytes in peripheral blood sampled on day 18; ( C ) mRNA expression of iNOS, IL-1β, MCP-1, IL-10, Arg-1, SIRT1 in peripheral WBCs collected on day 18 and 28 assessed by RT-qPCR; ( D ) immunofluorescence method-based observation of co-localization of molecule CD43 and SIRT1 in peripheral monocytes sampled on day 18, yellow arrow: high expression of SIRT1, green arrow: high expression of CD43. Statistical significance: *p < 0.05 and **p < 0.01 compared with AIA model rats.

Article Snippet: Rat enzyme-linked immunosorbent assay (ELISA) kits for the quantification of granulocyte-macrophage colony stimulating factor (GM-CSF, 70-EK363), interleukin 1 beta (IL-1β, 70-EK301B/3), interleukin 17 alpha (IL-17α, 70-EK317/3), TNF-α (70-EK382/3) and interleukin 6 (IL-6, 70-EK306/3) together with phorbol 12-myristate 13-acetate (PMA, CS1001) and brefeldin A (BFA, CS1002) were purchased from MultiScience Biotech (Hangzhou, Zhejiang, China).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Immunofluorescence

Journal: Journal of Inflammation Research

Article Title: Qing-Luo-Yin Alleviated Experimental Arthritis in Rats by Disrupting Immune Feedback Between Inflammatory T Cells and Monocytes: Key Evidences from Its Effects on Immune Cell Phenotypes

doi: 10.2147/JIR.S346365

Figure Lengend Snippet: Interplay between lymphocytes and monocytes during QLY treatments in AIA. ( A ) Flow cytometry analysis of CD3 + CD4 + IL-17α + Th17 cells in normal splenocytes co-cultured from monocytes from different rats; ( B ) levels of IL-6 and IL-1β released by splenocytes co-cultured from monocytes derived from different rats, determined by ELISA; ( C ) mRNA expression of iNOS, IL-1β, IL-10, Arg-1, SIRT1 in normal monocytes co-cultured with either normal or AIA lymphocytes in the presence of different rat serums assessed using RT-qPCR; ( D ) levels of IL-6 and IL-1β released by normal monocytes co-cultured with either normal or AIA lymphocytes in the presence of different rat serums determined by ELISA; ( E ) Western blot analysis of p65, p-p65, JNK, p-JNK expression in normal monocytes co-cultured with either normal or AIA lymphocytes in the presence of different rat serums; ( F ) quantification of assay E. Statistical significance: *p < 0.05 and **p < 0.01 compared with the cells co-cultured with immune cells derived from AIA rats.

Article Snippet: Rat enzyme-linked immunosorbent assay (ELISA) kits for the quantification of granulocyte-macrophage colony stimulating factor (GM-CSF, 70-EK363), interleukin 1 beta (IL-1β, 70-EK301B/3), interleukin 17 alpha (IL-17α, 70-EK317/3), TNF-α (70-EK382/3) and interleukin 6 (IL-6, 70-EK306/3) together with phorbol 12-myristate 13-acetate (PMA, CS1001) and brefeldin A (BFA, CS1002) were purchased from MultiScience Biotech (Hangzhou, Zhejiang, China).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of Inflammation Research

Article Title: Qing-Luo-Yin Alleviated Experimental Arthritis in Rats by Disrupting Immune Feedback Between Inflammatory T Cells and Monocytes: Key Evidences from Its Effects on Immune Cell Phenotypes

doi: 10.2147/JIR.S346365

Figure Lengend Snippet: Effects of QLY-related anti-rheumatic compounds on AIA lymphocytes in vitro. ( A ) Levels of IFN-γ and IL-17α in the medium released by compounds-treated AIA lymphocytes, determined by ELISA; ( B ) expression of protein (p)-STAT3 in compounds-treated AIA lymphocytes, assessed by Western blot analysis; ( C ) quantification of assay B; ( D ) mRNA expression of IL-4, SIRT1, IFN-γ and IL-17α in compounds-treated AIA lymphocytes, evaluated by RT-qPCR. Statistical significance: *p < 0.05 and **p < 0.01 compared with untreated AIA lymphocytes; # p < 0.05 and ## p < 0.01 compared with AIA lymphocytes receiving combination stimulus (SIN + SCA + MT).

Article Snippet: Rat enzyme-linked immunosorbent assay (ELISA) kits for the quantification of granulocyte-macrophage colony stimulating factor (GM-CSF, 70-EK363), interleukin 1 beta (IL-1β, 70-EK301B/3), interleukin 17 alpha (IL-17α, 70-EK317/3), TNF-α (70-EK382/3) and interleukin 6 (IL-6, 70-EK306/3) together with phorbol 12-myristate 13-acetate (PMA, CS1001) and brefeldin A (BFA, CS1002) were purchased from MultiScience Biotech (Hangzhou, Zhejiang, China).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Quantitative RT-PCR