Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: MCP in biopsies from six children at IBD onset and after treatment harvested from colon (in some patients, it was not possible to analyze all MCPs).

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques:

Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: Six cytokine profiles in mucosal biopsies according to mRNA levels measured by q-PCR at disease onset and after EEN treatment. ( A ). IFN-γ at onset and after EEN treatment. ( B ). CSF-2 at onset and after EEN treatment. ( C ). TNF-α at onset and after EEN treatment. ( D ). IL-10 at onset and after EEN treatment. ( E ). IL-12β at onset and after EEN treatment. ( F ). IL-23α at onset and after EEN treatment.

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques:

Journal: Nutrients

Article Title: Exclusive Enteral Nutrition: Clinical Effects and Changes in Mucosal Cytokine Profile in Pediatric New Inflammatory Bowel Disease

doi: 10.3390/nu11020414

Figure Lengend Snippet: Real-time PCR values per patient pre- and post-EEN and mean values and SD for group per cytokine.

Article Snippet: Probes were obtained from Applied Biosystems, TaqMan ® MGB probes, FAM™ dye-labeled: IL-1β (Hs00174097_m1), IL-4 (Hs00174122_m1), IL-6 (Hs00985639_m1), IL-10 (Hs00961622_m1), IL-12β (Hs001011518_m1), IL-22 (Hs01574154_m1), IL-23 (Hs00900828_g1), IL-36γ (Hs00219742_m1), TGF-β1 (Hs00998133_m1), TNF-α (Hs01113624_g1), IFN-γ (Hs00989291_m1), GM-CSF-2 (Hs00929873_m1) and ABL1 (Hs01104728_m1) according to the manufacturer protocol.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-2 is not required for the initiation of CD8 T cell cycling but sustains expansion.

doi: 10.4049/jimmunol.171.11.5727

Figure Lengend Snippet: FIGURE 2. Antiviral CD8 T cells undergo IL-2-independent cell cy- cling but are dependent on IL-2 for sustained expansion. LN cells isolated from OT-I-RAG/ (Ly5.1/5.2) or OT-I-CD25/-RAG/ (Ly5.1) mice were labeled with CFSE and adoptively transferred i.v. as a mixture (1 106 cells each) into naive (Ly5.2) B6 recipients. The presence of a 1:1 ratio of the two cell types was confirmed before transfer. Mice were immunized by i.v. injection with 1 106 PFU VSV-OVA 1 day post- transfer. Fifty-seven hours (top panels) and 104 h (bottom panels) later, cells from the PLN, spleen, and lung were isolated and analyzed for the presence of donor cells by fluorescence flow cytometry. Histograms are gated on the indicated donor cells (gray histograms, OT-I-RAG/ cells; black histograms, OT-I-CD25/-RAG/ cells) isolated from tissues of the same immunized animal. As a control, cells were also isolated from the LN of mice that were left unimmunized (PLN, 57 h, rear histogram). Num- bers indicated are the percentage of CD8 lymphocytes. Four mice were analyzed at 57 h and two mice were tested at 104 h. Similar results were obtained in a second experiment using OT-I-CD25/ and OT-I cells.

Article Snippet: C57BL/6J, C57BL/6-IL-2 / , and C57BL/6-CD25 / mice were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Isolation, Labeling, Injection, Cytometry, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-2 is not required for the initiation of CD8 T cell cycling but sustains expansion.

doi: 10.4049/jimmunol.171.11.5727

Figure Lengend Snippet: FIGURE 3. CD8 T cells are capable of IL-2-independent cell cycle ini- tiation in the absence of overt inflammation. LN cells isolated from OT-I (Ly5.1/5.2) or OT-I-CD25/ (Ly5.1) mice were labeled with CFSE and adoptively transferred i.v. (1 106 cells each) as a mixture into naive (Ly5.2) B6 recipients. Mice were immunized by i.p. injection with 5 mg soluble OVA 1 day posttransfer. At the later indicated times, cells from the PLN (A), spleen (B), and lung (C) were isolated and analyzed for the presence of donor cells by fluorescence flow cytometry. Histograms are gated on the indicated donor cells (gray front histograms, OT-I cells; black middle histograms, OT-I-CD25/ cells) isolated from tissues of the same animal that had been challenged. As a control, cells were also isolated from the LN of mice that were left unchallenged (rear histograms in B and C). Numbers indicated are the percentage of CD8 lymphocytes and the ratios indicated are the ratios of OT-I cells:OT-I-CD25/ cells in the respective plots. This experiment was repeated twice with two or three mice per group at each time point.

Article Snippet: C57BL/6J, C57BL/6-IL-2 / , and C57BL/6-CD25 / mice were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Isolation, Labeling, Injection, Cytometry, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-2 is not required for the initiation of CD8 T cell cycling but sustains expansion.

doi: 10.4049/jimmunol.171.11.5727

Figure Lengend Snippet: FIGURE 4. The optimal expansion of autoreactive CD8 T cells, but not their initial proliferation is de- pendent on IL-2. A, LN cells isolated from OT-I or OT-I-IL-2/ mice were adoptively transferred i.v., either separately or as a mixture (MIX; 1 106 cells each whether singly or in a mixture), into 232-4 mice. IEL were isolated and analyzed by flow cytometry 6 days posttransfer. The OT-I mice used were Ly5.2, while the 232-4 mice and the OT-I-IL-2/ mice were Ly5.1 and hence OT-I-IL-2/ cells were identified as those that were V2V5CD8 and Ly5.2. Data shown in the transfer of separate cell populations was derived from gating on CD8 cells and represents the entire V2V5CD8 population which will in- clude transferred and endogenous cells (the latter of which were always 0.4% of total lymphocytes in that compartment). Data shown in the transfer of mix- tures is derived from gating on V2CD8 cells. The number indicated is the percentage of total lympho- cytes. B, OT-I (Ly5.2) and OT-I-CD25/ (Ly5.1) LN cells were transferred either separately or as a mix- ture (MIX) into 232-4 (Ly5.1/5.2) mice. IEL were isolated and analyzed by flow cytometry at 6 days posttransfer. Data shown are derived from gating on V2CD8 cells and numbers indicate percentage of total lymphocytes. C, OT-I-RAG/ (Ly5.2) and OT-I-IL-2/-RAG/ (Ly5.1) LN cells were la- beled with CFSE and transferred separately into 232-4 (Ly5.1/5.2) mice or into B6 mice as controls. Mes- enteric LN and IEL were isolated and analyzed by flow cytometry 3 days posttransfer. Histograms are gated on the donor cells and the percentages indicated are percentages of total lymphocytes. Also indicated on the plots is the number of cell divisions. Each ex- periment was repeated twice with two mice per group.

Article Snippet: C57BL/6J, C57BL/6-IL-2 / , and C57BL/6-CD25 / mice were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Isolation, Cytometry, Derivative Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-2 is not required for the initiation of CD8 T cell cycling but sustains expansion.

doi: 10.4049/jimmunol.171.11.5727

Figure Lengend Snippet: FIGURE 5. Tumor-specific CD8 T cells undergo IL-2-independent pro- liferation. CFSE-labeled mixtures of OT-I (Ly5.1/5.2) and OT-I- CD25/ (Ly5.1) cells were adoptively transferred into naive Ly5.2 B6 mice. One day posttransfer, mice were challenged i.d. on the lower right flank with 5 106 E.G7 tumor cells or parental EL4 cells. Seven days later, cells from the draining or nondraining inguinal LN were isolated and analyzed for the presence of the donor cell types. Histograms are gated on the indicated donor cells isolated from the draining LN of mice that were challenged with the E.G7 cells or EL4 cells. Numbers indicated are the percentage of CD8 lymphocytes and the ratios of OT-I cells:OT-I- CD25/ cells are indicated in the respective plots. This experiment was repeated twice with three to four mice per group.

Article Snippet: C57BL/6J, C57BL/6-IL-2 / , and C57BL/6-CD25 / mice were obtained from The Jackson Laboratory (Bar Harbor, ME).

Techniques: Labeling, Isolation