polyclonal biotinylated anti-mouse ifn-γ antibody  (PeproTech)

Journal:

Article Title: Gamma Interferon Can Prevent Herpes Simplex Virus Type 1 Reactivation from Latency in Sensory Neurons

doi: 10.1128/JVI.75.22.11178-11184.2001

Figure Lengend Snippet: IFN-γ is produced in day 35 TG cultures. TG were excised 35 days after HSV-1 corneal infection, and single-cell suspensions were prepared and incubated in culture medium. Culture supernatant fluids were sampled every other day and assayed for IFN-γ content by a standard ELISA and for infectious HSV-1 (indicating HSV-1 reactivation from latency) in a plaque assay. The data (n = 8) are expressed as means ± standard errors of the mean in picograms of IFN-γ per milliliter (●) and percent reactivation from latency (○).

Article Snippet: At various times after culture initiation, 150 μl of medium was removed from each culture and tested for IFN-γ content using a standard enzyme-linked immunosorbent assay (ELISA) incorporating anti-IFN-γ MAb (clone R4-6A2) as the capture antibody and biotinylated polyclonal anti-mouse IFN-γ antibody (R & D Systems, Inc.) as the detection antibody.

Techniques: Produced, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Plaque Assay

Journal:

Article Title: Gamma Interferon Can Prevent Herpes Simplex Virus Type 1 Reactivation from Latency in Sensory Neurons

doi: 10.1128/JVI.75.22.11178-11184.2001

Figure Lengend Snippet: rIFN-γ alone cannot block HSV-1 reactivation in day 35 TG cultures. TG were excised 35 days after HSV-1 corneal infection. Single-cell suspensions were prepared, and replicate cultures (n = 8) were incubated in culture medium alone (diamonds) or medium containing rIFN-γ (1,000 U/ml) (circles). The supernatant fluids were sampled every other day and assayed for infectious virus titer in a plaque assay. The data are expressed as means ± standard errors of the mean in PFU per culture (open symbols) and percentage of cultures with detectable virus (percent reactivation) (solid symbols). Observed differences in the reactivation frequency and the viral titer between IFN-γ-treated and control cultures were not significant (P > 0.05).

Article Snippet: At various times after culture initiation, 150 μl of medium was removed from each culture and tested for IFN-γ content using a standard enzyme-linked immunosorbent assay (ELISA) incorporating anti-IFN-γ MAb (clone R4-6A2) as the capture antibody and biotinylated polyclonal anti-mouse IFN-γ antibody (R & D Systems, Inc.) as the detection antibody.

Techniques: Blocking Assay, Infection, Incubation, Virus, Plaque Assay

Journal:

Article Title: Gamma Interferon Can Prevent Herpes Simplex Virus Type 1 Reactivation from Latency in Sensory Neurons

doi: 10.1128/JVI.75.22.11178-11184.2001

Figure Lengend Snippet: Transient ACV treatment cannot block HSV-1 reactivation from latency in day 35 TG cultures. Day 35 TG single-cell suspensions were prepared and pooled; half of the TG cells were depleted of CD8+ T cells by immunomagnetic separation. CD8+ T-cell-depleted (triangles) and nondepleted (circles) TG cells were incubated in culture medium with ACV for 4 days. ACV was removed, and the cultures were incubated with medium alone for an additional 10 days. Culture supernatant fluids were sampled every other day and assayed for IFN-γ content by a standard ELISA and for infectious HSV-1 (indicating HSV-1 reactivation from latency) in a plaque assay. The data are expressed as means ± standard errors of the mean in picograms of IFN-γ per milliliter (panel A, closed symbols), percent reactivation from latency (panel A, open symbols), and PFU per culture (panel B). For CD8+ T-cell-depleted cultures, n = 23, and for nondepleted cultures, n = 26, except in IFN-γ determination, where n = 5. Depletion of CD8+ T cells significantly accelerated the reactivation rate (P < 0.05) and increased viral titer on day 10 (P < 0.05).

Article Snippet: At various times after culture initiation, 150 μl of medium was removed from each culture and tested for IFN-γ content using a standard enzyme-linked immunosorbent assay (ELISA) incorporating anti-IFN-γ MAb (clone R4-6A2) as the capture antibody and biotinylated polyclonal anti-mouse IFN-γ antibody (R & D Systems, Inc.) as the detection antibody.

Techniques: Blocking Assay, Immunomagnetic Separation, Incubation, Enzyme-linked Immunosorbent Assay, Plaque Assay

Journal:

Article Title: Gamma Interferon Can Prevent Herpes Simplex Virus Type 1 Reactivation from Latency in Sensory Neurons

doi: 10.1128/JVI.75.22.11178-11184.2001

Figure Lengend Snippet: IFN-γ can block HSV-1 reactivation from latency. Day 35 TG cell cultures were treated with ACV for 4 days, rinsed, and then incubated for an additional 10 days in medium alone (○, n = 26) or culture medium that was supplemented with rIFN-γ at 0 h (●, n = 25), 24 h (▴, n = 10), 48 h (▾, n = 10), or 72 h (♦, n = 10) after removal of ACV. Culture supernatant fluids were sampled every other day and assayed for infectious HSV-1 (indicating HSV-1 reactivation from latency) in a plaque assay. Reactivation frequency was significantly reduced (P < 0.001) (relative to medium-only controls) by IFN-γ treatment at 0 and 24 h only.

Article Snippet: At various times after culture initiation, 150 μl of medium was removed from each culture and tested for IFN-γ content using a standard enzyme-linked immunosorbent assay (ELISA) incorporating anti-IFN-γ MAb (clone R4-6A2) as the capture antibody and biotinylated polyclonal anti-mouse IFN-γ antibody (R & D Systems, Inc.) as the detection antibody.

Techniques: Blocking Assay, Incubation, Plaque Assay

Journal:

Article Title: Gamma Interferon Can Prevent Herpes Simplex Virus Type 1 Reactivation from Latency in Sensory Neurons

doi: 10.1128/JVI.75.22.11178-11184.2001

Figure Lengend Snippet: IFN-γ can directly and indirectly inhibit HSV-1 reactivation from latency in day 35 TG cultures. TG were excised 35 days after HSV-1 corneal infection and single cell suspensions were pooled, depleted of either CD8+ T cells (n = 23) (A) or CD45+ cells (n = 12) (B), and incubated with culture medium containing ACV for 4 days. After removal of ACV, cultures were incubated with medium alone (○) or medium supplemented with rIFN-γ (●) for an additional 10 days. Culture supernatant fluids were sampled every other day and assayed for infectious HSV-1 (indicating HSV-1 reactivation from latency) in a plaque assay. IFN-γ significantly reduced the reactivation frequency (P < 0.001) in both CD8+ T-cell-depleted and CD45+ cell-depleted TG cultures.

Article Snippet: At various times after culture initiation, 150 μl of medium was removed from each culture and tested for IFN-γ content using a standard enzyme-linked immunosorbent assay (ELISA) incorporating anti-IFN-γ MAb (clone R4-6A2) as the capture antibody and biotinylated polyclonal anti-mouse IFN-γ antibody (R & D Systems, Inc.) as the detection antibody.

Techniques: Infection, Incubation, Plaque Assay

Journal:

Article Title: Regulation of secretion of IL-4 and IgG1 isotype by melatonin-stimulated ovalbumin-specific T cells

doi: 10.1046/j.1365-2249.1998.00493.x

Figure Lengend Snippet: (a) Melatonin enhances IL-4 and down-regulates the production of IL-2. The levels of IL-4 (▪) and IL-2 (□) were monitored in the serum of animals immunized with ovalbumin (OVA) and followed by melatonin and control, PBS and PBS–ethanol. Sera were collected 24 h after the last injection of melatonin and assayed for IL-4 and IL-2 using HT-2 cells. IL-2 activity was assayed in the presence of 11B11 MoAb (600 ng/ml) and IL-4 secretion was measured in the presence of anti-IL-2 MoAb (cocktail of 1:12 dilution of culture SN of CRL 1698, HB 8794 and TIB222). The specificity of IL-4 and IL-2 was established by blocking the proliferation of HT-2 cells by their respective MoAbs (for details see Materials and Methods). All data calculated as U/ml are from the mean ± s.d. of triplicate determinations. (b) Melatonin decreases the secretion of IFN-γ. The level of IFN-γ was measured in the serum by ELISA. Sera were collected from the immunized animals as mentioned in the legend to (a). All data calculated from the mean ± s.d. of triplicate determinations as pg/ml were computed by comparison with the standard curve using recombinant IFN-γ (Genzyme).

Article Snippet: Recombinant murine IL-2, IFN-γ, IL-4, monoclonal anti-mouse IFN-γ and biotinylated polyclonal goat anti-mouse IFN-γ antibodies were purchased from Genzyme (Cambridge, MA).

Techniques: Injection, Activity Assay, Blocking Assay, Enzyme-linked Immunosorbent Assay, Recombinant