anti human cd8a 146nd  (fluidigm)

Journal: Cell reports

Article Title: Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors

doi: 10.1016/j.celrep.2024.115202

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD8 , Standard BioTools , Cat3146001B; RRID:AB_3661846.

Techniques: Virus, Recombinant, Electron Microscopy, Clinical Proteomics, Modification, Antibody Labeling, Staining, Software

Journal: Cell reports

Article Title: Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors

doi: 10.1016/j.celrep.2024.115202

Figure Lengend Snippet: (A) Spleen, lymph node (LN), gut, male reproductive tract, kidney, liver, bone marrow (BM), heart, and lung were harvested postmortem from antiretroviral therapy (ART)-treated people with HIV (PWH; N = 7) and analyzed by VISOR-CyTOF. Sample sizes for individual tissues are indicated. Total numbers of immune, myeloid, B cells, CD4 + T, and CD8 + T cells analyzed are shown. Created with BioRender.com . (B and C) VISOR-CyTOF distinguishes myeloid, B, and naive and memory CD4 + and CD8 + T cells from tissues and blood. Immune cells from the tissues of PWH as described in panel (A) (B) and PBMCs from people without HIV (PWOH; N = 4 participants) (C) were phenotyped by VISOR-CyTOF and visualized by tSNE using all markers in the panel. (D and E) tSNE analysis using only VISOR markers of VISOR-CyTOF separates myeloid, B, and T cells from one another. Data from the tissues from PWH (D) and PBMCs from PWOH (E) are shown. In (B)–(E), overlaid tSNEs are shown on the left, while tSNEs separated out by each immune subset are shown on the right. Abbreviations: Tm, memory T cells; Tn, naive T cells.

Article Snippet: CD8 , Standard BioTools , Cat#3146001B; RRID:AB_3661846.

Techniques:

Journal: Cell reports

Article Title: Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors

doi: 10.1016/j.celrep.2024.115202

Figure Lengend Snippet: (A) Productive infection assays experimental design. PHA-stimulated PBMCs ( N = 4) were mock-treated or exposed to HIV-F4.HSA for 3 days and then analyzed by VISOR-CyTOF. Analyses were performed on events corresponding to bystander or productively infected cells, which were defined based on the expression of long terminal repeat-driven reporter gene HSA. Gating strategies are shown in and . Created with BioRender.com . (B) Identification of productively infected cells by VISOR-CyTOF. Events were pre-gated on live, singlet, CD19 – CD8 – cells. (C and D) tSNE plots of bystander and productively infected CD4 + T (C) or myeloid (D) cells analyzed by VISOR-CyTOF, highlighting dissimilarity of bystander cells to their productively infected counterparts. tSNE plots were generated using all markers in the CyTOF panel. (E and F) Productively infected cells express high levels of VISORs relative to their bystander counterparts. Shown are viral sensors and restriction factors that are significantly elevated in productively infected CD4 + T (E) and myeloid (F) cells relative to their bystander counterparts. Viral sensors significantly upregulated in productively infected CD4 + T and myeloid cells were capsid sensors (PQBP1 and MX2) and DNA sensors (TLR9, AIM2, IFI16, cGAS, and pIRF3). In productively infected CD4 + T but not myeloid cells, HIV RNA sensors RIGI and pSTING were upregulated. Restriction factors upregulated in both productively infected CD4 + T and myeloid cells were those blocking HIV reverse transcription (SAMHD1 and PAF1), nuclear import (MX2), integration (TRIM28), transcription (TRIM28, IFI16, and BRD4), and translation (IFITM1). Productively infected CD4 + T but not myeloid cells also upregulated inactive pSAMHD1. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, as assessed using Student’s two-sided paired t tests. Error bars correspond to SD.

Article Snippet: CD8 , Standard BioTools , Cat#3146001B; RRID:AB_3661846.

Techniques: Infection, Expressing, Generated, Blocking Assay, Reverse Transcription

Journal: Cell reports

Article Title: Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors

doi: 10.1016/j.celrep.2024.115202

Figure Lengend Snippet: (A) Experimental design for HIV infection of cGAMP-stimulated PBMCs. PHA-stimulated PBMCs ( N = 3) were incubated for 20 h with cGAMP-containing viral-like particles (VLPs), or control VLPs harboring catalytically inactive cGAS mutant, and then analyzed by VISOR-CyTOF. In parallel, VLP-treated cells from the same donor were mock-infected or exposed to HIV-F4.HSA, cultured for 3 days, and monitored for infection rates. Created with BioRender.com . (B) cGAMP-treated CD4 + T cells exhibited elevated expression of HIV DNA sensors (AIM2, cGAS, and pIRF3), as well as restriction factors targeting reverse transcription (SAMHD1) and translation (IFITM1). By contrast, inactive pSAMHD1 trended lower among cGAMP-treated cells. * p < 0.05, ** p < 0.01, as assessed using Student’s two-sided paired t tests. (C and D) cGAMP-treated CD4 + T cells restrict HIV infection. Representative CyTOF plots of uninfected and infected PBMC cultures treated with control or cGAMP VLPs (C) and cumulative results from 4 independent donors (D). Results were pre-gated on live, singlet, CD19 – CD3 + CD8 – cells. * p < 0.05, ** p < 0.01; n.s., non-significant, as assessed using one-way repeated measures ANOVA followed by Tukey’s multiple comparisons test.

Article Snippet: CD8 , Standard BioTools , Cat#3146001B; RRID:AB_3661846.

Techniques: Infection, Incubation, Control, Mutagenesis, Cell Culture, Expressing, Reverse Transcription

Journal: Cell reports

Article Title: Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors

doi: 10.1016/j.celrep.2024.115202

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD8 , Standard BioTools , Cat#3146001B; RRID:AB_3661846.

Techniques: Virus, Recombinant, Electron Microscopy, Clinical Proteomics, Modification, Antibody Labeling, Staining, Software