Journal: mSphere

Article Title: Differential innate immune responses of human macrophages and bronchial epithelial cells against Talaromyces marneffei

doi: 10.1128/msphere.00258-22

Figure Lengend Snippet: Cytokine responses of human and murine macrophages cultured in the presence or absence of plasma/serum upon Talaromyces marneffei infection. Upon T. marneffei infection, the relative expression of ( A ) TNF and ( B ) CXCL8 mRNAs in hPBDMs could only be stimulated in the presence of autologous plasma; whereas no significant change in the mRNA expression of these two cytokines was observed when the culture medium was not supplemented with autologous plasma. The infection experiment was conducted in triplicate using blood cells isolated from three independent donors. The inability of T. marneffei to induce ( C ) TNF and ( D ) CXCL8 mRNAs upon infection was also observed in the murine macrophage cell lines J774A.1 and RAW264.7. Data are presented as mean ± SD and comparisons were analyzed using unpaired t -test (two-tailed). **, P < 0.01; ***, P < 0.001.

Article Snippet: Expression of cytokines were measured semi-quantitatively using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction; and all samples were tested in duplicates.

Techniques: Cell Culture, Clinical Proteomics, Infection, Expressing, Isolation, Two Tailed Test

Journal: mSphere

Article Title: Differential innate immune responses of human macrophages and bronchial epithelial cells against Talaromyces marneffei

doi: 10.1128/msphere.00258-22

Figure Lengend Snippet: Changes of cytokine mRNA expression in hPBDMs upon Aspergillus fumigatus and Talaromyces marneffei infections. Relative mRNA expressions of CXCL8 and TNF in hPBDMs infected with both fungi were significantly induced mainly after 4 h post-infection and the expression of CXCL10 mRNA in hPBDMs was only significantly induced by T. marneffei infection, in which the expression level reached the peak at 8 h post-infection. The infection experiment was conducted in triplicate using blood cells isolated from three independent donors. The relative mRNA expression levels of CXCL8 , CXCL10 , and TNF were detected by qRT-PCR. Data are presented as mean ± SD and comparisons were analyzed using unpaired t -test (two-tailed). *, P < 0.05; †, P < 0.01; ‡, P < 0.001; ×, undetectable.

Article Snippet: Expression of cytokines were measured semi-quantitatively using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction; and all samples were tested in duplicates.

Techniques: Expressing, Infection, Isolation, Quantitative RT-PCR, Two Tailed Test

Journal: Virus research

Article Title: Paracrinal regulation of neutrophil functions by coronaviral infection in iPSC-derived alveolar type II epithelial cells.

doi: 10.1016/j.virusres.2024.199391

Figure Lengend Snippet: Fig. 5. Identification of cytokines secreted by infected iAECIIs and their effect on HL-60 cells. (A) Cytokine array analysis showing the levels of the indicated cy tokines in the medium conditioned by iAECIIs infected with HCoV-229E for 24 or 48 h (24 hpi and 48 hpi). Ctrl – uninfected control. Right panel: cytokine label legend. (B) Western blot analysis of the expression of IL-8 and ICAM-1 in HCoV-229E-infected iAECIIs. (C) Transwell migration assay showing the migration capacity of HL-60 neutrophils in response to a medium conditioned by iAECIIs infected with HCoV-229E for 48 h (48 hpi) in the absence or presence of an IL-8-neutralizing antibody (IL-8 nAb). Ctrl – uninfected control conditioned medium. Data shown as means with SD error bars, n = 3, *p < 0.005, (ANOVA). (D) Representative fluorescence microscopy images of adherent HL-60 neutrophils after incubation with infected iAECIIs in the absence or presence of an ICAM-1-neutralizing antibody (ICAM-1 nAb). Ctrl – uninfected control. (E) Cell count of adherent HL-60 neutrophils. Mean values are shown with SD error bars, n = 3, *p < 0.005.

Article Snippet: Cytokines in the conditioned medium were detected using the Human Cytokine Array Kit (#ARY005B; R&D Systems), and the test strips were moistened and activated according to the manufacturer’s user manual.

Techniques: Infection, Control, Western Blot, Expressing, Transwell Migration Assay, Migration, Fluorescence, Microscopy, Incubation, Cell Counting

Journal: Virus research

Article Title: Paracrinal regulation of neutrophil functions by coronaviral infection in iPSC-derived alveolar type II epithelial cells.

doi: 10.1016/j.virusres.2024.199391

Figure Lengend Snippet: Fig. 6. RNA-seq analysis identifies upstream cytokine pathways in the immune responses by neutrophils. (A and B) Cytoscape ClueGO networks of upregulated genes by infection of iAECIIs with HCoV-229E (A) or triggered by infected HCoV-229 conditioned medium in HL-60 cells (B). (C and D) Bubble plots showing the most enriched GO-BP terms among the genes upregulated by the infection of iAECIIs with HCoV-229E (C) or the genes upregulated in HL-60 cells by cultivation in the infected HCoV-229-conditioned medium (D). (E and F) Hierarchical clustering heatmaps showing the signatures of genes differentially regulated in infected iAECIIs (E) and conditioned medium-stimulated HL-60 cells (F). (G and H) X2K network analysis showing the kinases and transcription factors predicted to regulate differentially expressed genes in infected iAECIIs (G) and conditioned medium-stimulated HL-60 cells (H).

Article Snippet: Cytokines in the conditioned medium were detected using the Human Cytokine Array Kit (#ARY005B; R&D Systems), and the test strips were moistened and activated according to the manufacturer’s user manual.

Techniques: RNA Sequencing, Infection

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 3. Cytokine production from autologous exosome stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio-Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). The exosomes appeared to contain significant amounts of CCL5 (RANTES) immediately after the addition of exosomes (at 0 h) since the supernatants showed relatively large amounts of RANTES. These levels were decreased at day five. doi:10.1371/journal.pone.0049723.g003

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 5. Cytokine production from IL-2 stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio- Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). Cytokines IL-5, MIF, and GM-CSF (CSF2) were present at a high level in the supernatant after five days. doi:10.1371/journal.pone.0049723.g005

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 6. Cytokine production from exosome+IL-2 stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio-Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). The cytokines IL-5, IL-13 and GM-CSF as well as the chemokines CCL3 and CCL4 were present at higher levels at day five. doi:10.1371/journal.pone.0049723.g006

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software