Sentences with word «splenocytes»

(C) In vitro expanded human Lin - CD127 + cells or (D) mouse Lin - CD127 + c - Kit + Thy1.2 + cells expanded from splenocytes of Rag1 - / - mice were treated with lethal toxin for 2 hr followed by IL - 23 stimulation for 30 mins.

Once optimal detector gains were determined, an extensive brightness index of dyes was established on this instrument using sequential single staining of mouse splenocyte with 43 different antibody conjugates against mouse CD8a (53 - 5.8).

Positive controls were CD4 + T cells isolated from animals prior sensitized with two immunizations of 3 × 107 donor splenocytes over 10 days.

T cell lines were maintained by biweekly stimulations in complete DMEM supplemented with 10 % prescreened FCS and 5 % conditioned medium that was prepared from culture supernatant of rat splenocytes stimulated with Con A for 48 h.

Since our flow cytometric data had shown that the HPCs poorly expressed MHC antigens [17], we wondered whether they stimulate allogenic MRL splenocytes (H2k).

All three groups of animals responded equally to third - party Balb / c splenocytes.

The ILCs present in Rag1 - / - splenocytes bound protective antigen in a dose - dependent manner (S1C Fig), suggesting the cells were direct targets for lethal toxin.

100,000 — 500,000 splenocytes per well were incubated in a round bottom 96 well plate in IMDM media supplemented with 10 % FBS, 100 U / ml pencillin, 100 U / ml streptomycin, IL - 2 (20 ng / ml) and IL - 7 (10 ng / ml).

For initial lethal toxin experiments splenocytes / tonsillar lymphocytes were cultured in media containing in 0.1 % serum to minimize the effect of serum on lethal factor enzyme activity.

As we found in the heterogeneous splenocyte cultures, lethal toxin down - regulated secreted IL - 22 (Fig 1H).

Boerner et al. «Production of Antigen - Specific Human Monoclonal Antibodies From In Vitro - Primed Human Splenocytes», The Journal of Immunology, 147 (1): 86 - 95, 1991.

In addition, mixed lymphocyte cultures using the cells of the chimeric animals as responder cells to donor splenocytes failed to demonstrate any sensitization or clonal deletion of alloreactive T cells; i.e. T cell responses were identical to those of control animals (n = 4)(data not shown).

An equal number of MRL responder splenocytes were added and the cells cultured for 48 h before pulsing.

For comparison, we used splenocytes and bone marrow cells, separately, as stimulator cells.

(A) CD45.1 + OTII splenocytes were labeled with cell trace violet and adoptively transferred into Ebi2 + / − and Ebi2 − / − mice.

Additional controls were splenocytes stimulated with ConA.

One of the assays widely used in the literature is the application of splenocytes from tolerant animals as responder cells in a 4 - day mixed lymphocyte culture.

Controls were animals pre-sensitized with two separate IP injections of donor splenocytes and non-transplanted control animals.

Balb / c splenocytes were used as third - party responder cells assuming that these would induce the same degree of T cell proliferation in tolerant and control animals.

Briefly, single - cell suspensions of splenocytes were prepared from either experimental or control host strain mice, as described earlier.

In total, 106 splenocytes were cultured without peptide or with 0.2 μM LCMV gp33 peptide, NP396 peptide for 5 h. IFN - γ positivity by flow cytometry was used to identify LCMV - specific cells (13).

Mice were sacrificed on day 7 postinfection, serum samples were obtained, and splenocytes were stimulated in vitro and analyzed.

Splenocytes were obtained following single - cell suspension and RBC lysis.

(C) Sorted Lin - CD127 + cells from Rag1 - / - splenocytes were pretreated with DMSO or indicated concentrations of PD98059 or SB203580 as in A and analyzed for IL - 22 secretion by ELISA.

Splenocytes were isolated from Rag1 - / - mice and cultured in IMDM (Corning) containing 0.1 % FBS, 100 U / ml penicillin, 100 U / ml streptomycin and 2 mM glutamine supplemented with IL - 2 (20 ng / ml) and IL - 7 (10 ng / ml).

(A-C) Rag1 - / - splenocytes were pretreated with (A) either lethal toxin (LeTx, lethal factor + protective antigen) or lethal factor (LF)(1 μg / ml) or (B) increasing doses of lethal toxin or (C) with enzymatic mutant toxin (E687C) for 3 hr followed by IL - 23 (50 ng / ml) stimulation for 18 hr.

We used Rag1 - / - mouse splenocytes, a source of ILC3s, because the absence of T cells in this model makes ILC3s the dominant IL -22-expressing cell type [40].

Rag1 - / - splenocytes were pretreated with DMSO or indicated concentration of (A) PD98059 (MEK1 inhibitor) or (B) SB203580 (p38 MAPK inhibitor) for 1 hr followed by IL - 23 stimulation for 18 hr.

Reduced levels of secreted IL - 22 in heterogeneous lymphocyte mixtures of Rag1 - / - splenocytes could be due to another cell affecting ILC3 production of IL - 22 and / or modulation in IL - 22 from other innate immune cells that have been reported to produce low levels of IL - 22, such as DCs or neutrophils [22, 42, 43].

For mouse ILC3s, splenocytes from Rag1 - / - mice were sorted (Lin -(CD3, CD45R, CD11c, GR - 1, NK1.1) c - kit + Thy1.2 +) and cultured as described for human cells using recombinant mouse cytokines.

Both inhibitors significantly reduced IL -23-mediated IL - 22 production from Rag1 - / - splenocytes (Fig 5A and 5B).

For isolating mouse primary ILCs, single cell preparations of Rag1 - / - splenocytes were surface stained for NK1.1 and CD127.

(D - F) Rag1 - / - splenocytes were treated with lethal toxin followed by IL - 23 stimulation for 6 hr.

The addition of this pendant catalyst created an MnP - PVPON / TA capsule with the following characteristics: 1) the microcapsules synergistically remove reactive oxygen species, including superoxide and hydrogen peroxide, at dramatically increased rates compared to unmodified TA / PVPON microcapsules; 2) the microcapsule does not degrade with long exposure to reactive oxygen species; and 3) the microcapsules are nontoxic to mouse splenocytes.

For measuring responses of chimeric or transplanted animals, splenocytes from these animals were used as responder animals similarly.

The sensitized animals showed a higher number of IL - 2 spots against the donor splenocytes, as expected.

Additionally, splenocytes of recipient animals were used as responder cells in mixed lymphocyte reactions and ELISPOT to determine responses to donor alloantigen.