Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: List of antibodies used in this study

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Blocking Assay

Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: List of flow cytometry antibodies used in this study

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Flow Cytometry, Control

Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: ILP and ISP promotes a phenotypic switch in helper T cells toward a T reg phenotype. a , b Flow cytometric assessment revealed no apparent difference in the overall infiltration of helper T cells (CD45 + /CD3 + /CD4 + ) in the injured spinal cord at 3 and 7 days post-injury between vehicle and ILP/ISP-treated animals. c , d However, ILP/ISP-treated animals exhibited a significant decrease in the number of effector T cells (CD45 + /CD3 + /CD4 + /IFNγ + ) at 7 days post-injury. e , f A significant increase in the total number of regulatory T cells (CD45 + /CD3 + /CD4 + /IL10 + ) was observed at 3 days post-injury in ILP/ISP-treated animals. g , h Representative flow cytometry gates are shown. i Western blot analysis showed upregulation of CD4 protein expression, at 7 and 14 days post-SCI compared to uninjured control group confirming infiltration of helper T cells in the injured spinal cord. Confirming our flow cytometry, ILP and ISP had no apparent effect on the overall protein expression of CD4. j However, ILP and ISP significantly increased FOXP3 protein expression, a marker of regulatory T cells, at both 7 and 14 days post-SCI compared to SCI vehicle control. Western blot results have been normalized to the actin loading control prior to subsequent normalization to the control values. The data show mean ± SEM, * p < 0.05, Student t test ( a – f ), one-way ANOVA ( i , j ), N = 4-6/group

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Flow Cytometry, Western Blot, Expressing, Control, Marker

Journal: The Journal of Neuroscience

Article Title: Neuroprotection by Encephalomyelitis: Rescue of Mechanically Injured Neurons and Neurotrophin Production by CNS-Infiltrating T and Natural Killer Cells

doi: 10.1523/JNEUROSCI.20-14-05283.2000

Figure Lengend Snippet: Immunohistochemical colocalization of neurotrophins and leukocyte markers (C, F, I, L, yellow) in spinal cord sections from immunized animals (14 d after immunization) subjected to ventral root avulsion. The depicted cells are located in the ventral horn of the lesioned side. Examples of double-labeled cells are indicated by arrowheads inA–L. The NT-3 immunolabeling displays a high degree of colocalization with W3/25 (CD4+) (A–C), OX8 (CD8+) (D–F), and NKR-P1 (NK cells) (G–I)-labeled cells. The immunolabeling for BDNF is more diffuse and does not colocalize to cells to the same degree as NT-3. However, a moderate number of W3/25-labeled cells also display positive BDNF staining (J–L). Only very weak immunofluorescence is seen in control sections where the primary antibody was pre-absorbed with the corresponding peptide for NT-3 and BDNF, respectively (M and N display double labeling with W3/25). Scale bar, 0.05 mm.

Article Snippet: The following antisera were used: anti-rat NKR-P1 (a marker for NK cells; clone 3.2.3; mouse IgG1; Harlan Sera-Lab, Loughborough, UK), anti-rat CD25 (clone OX-39; mouse IgG1; Serotec, Oxford, UK), anti-rat CD4 (clone W3/25; mouse IgG1; Serotec), anti-rat CD8 (clone OX-8; mouse IgG1; Serotec), anti-human glial fibrillary acidic protein (goat polyclonal IgG; Santa Cruz Biotechnology, Santa Cruz, CA), anti-rat CD11b (clone OX-42; mouse IgG2a; Serotec), anti-rat macrophage antigen (clone ED1; mouse IgG1; Serotec), anti-human NT-3, and BDNF (cross-reacting with rat; rabbit polyclonal IgG; Santa Cruz Biotechnology).

Techniques: Immunohistochemical staining, Labeling, Immunolabeling, Staining, Immunofluorescence, Control

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

Article Title: A novel antibody-dependent cellular cytotoxicity mechanism involved in defense against malaria requires costimulation of monocytes FcgammaRII and FcgammaRIII.

doi: 10.4049/jimmunol.178.5.3099

Figure Lengend Snippet: FIGURE 6. A, Identification of the FcRs involved in the antiplasmo- dial ADCI effect. The role of FcRI, FcRII, and FcRIII was determined in a two-step ADCI assay by preincubation of MN with F(ab)2 derived from the mAbs 10.1, AT10, and 3G8 respectively, to block selectively each FcR family expressed on MN. After a 30-min incubation, MSP-3CT- coated PMz and PIAG were added to the ADCI wells and incubated for 2 h. Supernatants were then transferred to asynchronous culture of P. falcipa- rum (0.5% parasitemia) without MN for 72 h and the SGI was determined. Results are the mean SD of three independent experiments. B, Identi- fication of FcRs involved in phagocytosis of parasites. The role in phago- cytosis of each of the three FcR expressed on MN was determined by blocking FcRI, FcRII, and FcRIII by either mAb 10.1, AT10, or 3G8, respectively. To MN were added combinations of blocking mAb 30 min before the addition of MSP-3-CT PMz opsonized by either PIAG or N-IgG. After 45 min, the percentage of phagocytosis by MN was determined by light microscopy of over 100 MN. An MN was considered as having phagocytosed particles when at least three beads were ingested. Data are mean SD of two distinct experiments.

Article Snippet: Mouse mAbs of either the IgG1 (clone W3/25; Serotec) or the IgG2a class (catalog number 16-4724; eBioscience) were used as negative controls.

Techniques: Derivative Assay, Blocking Assay, Incubation, Light Microscopy