Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: A, B. Two million CEM cells were exposed to NL-EGFP preparations (see Table 1) at MOI = 0.05 infectious units (IU, P4R5 cell assay) for 4 hr in 1.5 ml total volume. Cells were washed once and and cultured for 7 days. Results from one representative experiment. C, D. Five million primary CD4 cells were infected overnight with MOI = 0.05 IU (P4R5) of NL-EGFP, with or without 1 hour preincubation with CD44 MicroBeads. Three independent experiments. A, C. GFP expression. B. Secreted p24, net values. D. Secreted p24, total values.

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Cell Culture, Infection, Expressing

Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: Comparison of NL4-3 infections (A, B) of primary CD4 T cells at MOI = 0.01 TCID50 with CD44 MicroBead pretreatment (right panel) and without (left panel). Primary CD4 cells were infected with the R5 clone, JR-CSF (C, E) or clade C, primary X4 isolate 98IN017 (D, F) with an MOI of 0.01 pg p24. Infections were monitored for % infected cells by ICp24 (A,C,D) and soluble p24 (B,E,F). Mean + SEM. n = 3, except for A, left panel (n=4) and B, left panel (n=7).

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Infection

Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: Primary CD4 cells were infected with the R5 clone, JR-CSF (left panels) or clade C, X4 isolate 98IN017 (right panels) with or without pretreatment with CD44 MicroBeads. At the end of 2 weeks of culture, genomic DNA was isolated for qPCR quantification of HIV integrants per 200 ng input DNA (A) and harvested cells were reactivated with αCD3/αCD28 to determine replication competent infectious units per million (IUPM) by limiting dilution assay (B). Results from three independent cell donors shown (x axis: A, B, C). nd = not done.

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Infection, Isolation, Limiting Dilution Assay

Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: Primary CD4 T cells were infected with NL4-3 produced from transfection in: 293T (CD44H−), HeLa P4R5 (CD44H+), and CEM (CD44H+) cells, with or without prior incubation with CD44 MicroBeads. The fraction of infected cells was measured by ICp24 with FACS (A) and secreted p24, assayed by ELISA (B). n = 3 except at one time point, as noted. Mean + SEM. Fold change in CD25 (C) and CD69 (D) expression with CD44 MicroBead treatment of virus produced by various cell lines (x axis), assessed one day after infection in unstimulated cells. Mean and range of three experiments.

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Infection, Produced, Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Expressing

Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: Treatments with CD44 MicroBeads, streptavidin-complexed biotinylated αCD44, free αCD44 and CD45 MicroBeads were compared in spreading CEM infections with NL-EGFP (A, B). The fraction of CEM cells expressing GFP was analyzed by FACS (A) and soluble secreted p24 was assayed by ELISA (B). Mean + SEM of three experiments. C. CD69 expression in unstimulated primary CD4 cells, measured one day after exposure to αCD44 or αCD45 reagents alone, or in conjunction with NL4-3 infection. Mean and range of three experiments.

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Infection

Journal:

Article Title: CD44 MicroBeads Accelerate HIV-1 Infection in T Cells

doi: 10.1016/j.virol.2009.03.022

Figure Lengend Snippet: Binding of CD44 MicroBeads to virus particles may accelerate sedimentation onto target cells (1). Simultaneous binding of MicroBeads to virions and cell membrane CD44 may stabilize virion-CD4 interactions and promote coreceptor binding. CD44 MicroBead attachment to cellular CD44 may initiate intracellular signaling that promotes infection (2). Also, virus may spread more efficiently due to cell clustering induced by activated adhesion molecules or by cross-linking of cells directly through MicroBead binding (3). Illustration not to scale.

Article Snippet: Prescreened, pooled human AB serum for primary CD4 T cell culture was purchased from Omega Scientific (Tarzana, CA).

Techniques: Binding Assay, Sedimentation, Infection

Journal: bioRxiv

Article Title: Extracellular vesicle bioactivity and potential clinical utility is determined by mesenchymal stromal cell clonal subtype

doi: 10.1101/2024.09.05.609844

Figure Lengend Snippet: In vitro treatment of activated CD4+ T cells with Y201 and Y202 EVs (n=2, mean events >18,000 counted) A) Proliferative cycles. B) Proliferative index C) Polarisation of activated T cells in the absence and presence of Y201 EVs and Y202 EVs (n=2, mean events >32,000 counted). D/E) Total peritoneal exudate cell (PEC) counts following zymosan (D) or schistosome egg (E) induced inflammation in the absence and presence of Y201 EVs and Y202 EVs. F/G) Examination of TCR+CD4+, naïve and central memory T cells in zymosan or schistosome egg induced inflammation in the absence and presence of Y201 EVs and Y202 EVs (n=3). One-Way ANOVA with Bonferroni post hoc testing,*p<0.05, **p<0.01, ***p<0.001.

Article Snippet: To determine MSC-derived EV immunomodulation for deactivation and suppression of T cell proliferation, suspension cultures of 1.0×10 5 primary human peripheral blood-derived CD4+ T cells (Stem Cell Technologies) were grown in RPMI-1640 with 10% FBS 1% P/S (Gibco, Cat: 10363083) and were pre-treated for 6hrs with 20X EVs isolated from serum-free conditioned medium collected over 24hrs culture from 2.0×10 6 of Y201 or Y202 MSCs.

Techniques: In Vitro