14 research outputs found
Serum IL-10 levels increased following treatment with NHS-IL12.
<p>Each line represents a different dog, with individual colors representing different treatment groups. Serum IL-10 levels at 48 and 192 hours were significantly different from time points 0–8 hours (Kruskal-Wallis test followed by Dunn’s multiple comparison test) when data from all dogs was pooled. There was no difference in IL-10 levels at 48 hours between any of the treatment groups (Kruskal-Wallis test, p = .06, the 0.4 mg/m2 group was not included because it was a single dog).</p
No significant changes in circulating T- or B-cell populations were seen after treatment with NHS-IL12.
<p>Relative numbers of T- and B-cell populations were quantified in PBMCs by flow cytometric analysis on days 1, 2, 3, and 8 after treatment with NHS-IL12. Immune cell determination was based on labeling: B cells, CD21<sup>+</sup>MHCII<sup>+</sup> (A),T cells, CD4<sup>+</sup> (B) or CD8<sup>+</sup> (C); regulatory T cells, CD4<sup>+</sup>Foxp3<sup>+</sup> (D);. CD4<sup>+</sup> T cells trended downward at day 8 compared to their baseline measurements (B). Other differences were not evident over the time points evaluated.</p
Serial serum pharmacokinetics and IFN-γ/cytokine panel collections.
<p>IFN: interferon.</p><p>Serial serum pharmacokinetics and IFN-γ/cytokine panel collections.</p
Serum IFN-γ induction was associated with NHS-IL12 dose and the observation of adverse events.
<p>IFN-γ levels were measured using ELISA techniques. Induction of IFN-γ was detectable in dogs treated with NHS-IL12 at a dose of 1.6 mg/m<sup>2</sup> or higher. IFN-γ levels spiked sharply at 24 hours post-treatment and returned to undetectable levels by 48 hours. Elevated serum IFN-γ (> 100 pg/ml) was associated with increased risk for toxicity. However the highest level of IFN-γ induction was not directly linked to the most severe (Grade 4 or 5) adverse events.</p
RGD-A-<i>TNF</i> trafficking resulted in selective tumor endothelial cell localization and TNF <i>α</i> expression.
<p>(A, B) RGD-A-<i>TNF</i> selectively targeted tumor-associated vasculature (arrows) and was absent from normal tissues at 4–6 hours (A) and at 4 days (B). Magnification, 400-fold; scale bar, 100 µM. Pre-treatment tumor biopsies, post-treatment tumor biopsies, and post-treatment normal tissues in dogs that received a single-dose of RGD-A-<i>TNF</i> double-stained with an anti-CD31 specific antibody plus an anti-bacteriophage specific antibody. Detection was performed with Alexa Fluor 488 (green, blood vessels), Alexa Fluor 594 (red, AAVP), and DAPI (blue, cell nuclei). (C) Pre-treatment tumor biopsies (day 0), post-treatment normal biopsies (day 4) and post-treatment tumor biopsies (day 4) were used for extraction of total RNA. RT-PCR was performed to measure transcript levels of human TNFα in quadruplicate. The Y-axis represents the relative TNFα expression levels in post-treatment normal biopsies and post-treatment tumor biopsies compared to pre-treatment tumor biopsies after normalization to GAPDH expression (Kruskal-Wallis Test, p = 0.0107). All data are presented as means±standard deviations. (D–F) Presence of RGD-A-<i>TNF</i> was evaluated in post-treatment (day 28) necropsy samples of tumor (D) and normal tissues (E, F). Tissues were stained for RGD-A-<i>TNF</i> as described earlier. RGD-A-<i>TNF</i> selectively targeted tumor-associated vasculature in post-treatment tumor samples (arrows). In contrast, the vector was not apparent in pre-treatment tumor samples or in post-treatment normal control necropsy samples (such as lung, liver, spleen or intestine) after serial administrations of RGD-A-<i>TNF</i>.</p
Clinical activity of NHS-IL12 evident in dogs with malignant melanoma.
<p>Patient 0201 had a large malignant melanoma of the right maxilla and buccal surface. Prior to treatment on day 1, the tumor measured 4.4 cm in longest diameter (A). The dog received 0.8 mg/m<sup>2</sup> (1.04 mg) of NHS-IL12 administered subcutaneously. On day 29 RECIST response was assessed and a PR was noted. The tumor measured 2.4 cm, representing a 45% decrease (B). The dog continued monthly dosing of NHS-IL12 and had no evidence of disease after three cycles of therapy (C). On day 85, despite complete regression of the lesion, a formal CR could not be assigned to the observed clinical activity in this dog since canine malignant melanoma invades the underlying bone and a CT scan of the oral cavity was not performed at baseline. No gross disease was noted through nine cycles of therapy. PD eventually recurred in the neighboring nasal cavity.</p
RGD-A-<i>TNF</i> Dose Escalation Study: Patient Population and RGD-A-<i>TNF</i> Trafficking Data
<p>TU: transducing units; IF: immunoflourescent staining for presence of RGD-A-<i>TNF</i> in tumors; OCS: osteochondrosarcoma; PCT: plasma cell tumor; BCT: basal cell tumor; NST: nerve sheath tumor; STS: soft tissue sarcoma; OS: osteosarcoma; FS: fibrosarcoma; N.A.: not analyzed due to improper shipment.</p>*<p>Patient I.D.: dog is identified as participating in study 1 (dose escalation study), followed by patient number (i.e. patient 1.2 is dog 2 in study 1 and so on);</p>**<p>These two tumors originally diagnosed as soft tissue sarcomas were reclassified upon histopathologic review by a single pathologist (SN).</p
Pharmacokinetics of NHS-IL12 in tumor-bearing dogs after subcutaneous administration.
<p>AUC: area under the time-concentration curve; Cl: clearance; C<sub>max</sub>: maximum concentration; MRT: mean residence time; NA: not available, calculation not possible due to a small number of data points; ND, not detectable; T<sub>1/2</sub>: half-life.</p><p>Pharmacokinetics of NHS-IL12 in tumor-bearing dogs after subcutaneous administration.</p
Study schedule for dose escalation and cohort expansion of NHS-IL12 administered subcutaneously.
<p>CBC: complete blood count; IFN, interferon; PBMC, peripheral blood mononuclear cell; Tx: treatment; UA: urinalysis.</p><p>Study schedule for dose escalation and cohort expansion of NHS-IL12 administered subcutaneously.</p
RGD-A-<i>TNF</i> Multiple Dose Study: Patient Data and Tumor Responses
<p>FS: fibrosarcoma; OS: osteosarcoma; PNST: peripheral nerve sheath tumor (malignant); SCC: squamous cell carcinoma; S-C MS: subcutaneous myxosarcoma; MLT: multi-lobular tumor; Mel: LN mets: melanoma lymph node metastases; inc.: increase; dec.: decrease; PD: progressive disease; SD: stable disease (shown in yellow); PR: partial response (shown in green); Unevaluable: due to consent withdrawal by the owner, or death or euthanasia before completion of cycle 1</p>*<p>Patient I.D.: dog is identified as participating in study 2 (fixed multiple dose study), followed by patient number (i.e. patient 2.2 is dog 2 in study 2 and so on)</p><p>Cycle 1: dogs received 4 weekly doses of RGD-A-<i>TNF</i>; Cycle 2: dogs received additional 4 weekly doses of RGD-A-<i>TNF</i></p