canine il12  (GenScript corporation)

Journal: Frontiers in veterinary science

Article Title: Establishing an experimental model for canine atopic dermatitis through epicutaneous application of Dermatophagoides farinae .

doi: 10.3389/fvets.2022.1015915

Figure Lengend Snippet: FIGURE 7 Serum cytokine analysis. Change in the serum level of (A) IFN-gamma, (B) IL-2, (C) IL-4, (D) IL-10, (E) IL-12, (F) IL-13, (G) IL-31, and (H) TGF-beta. IL-4, IL-13 and IL-31, which belong to the Th2 cytokine group, were increased significantly after provocation. IFN-gamma, IL-2, and IL-12, which belong to the Th1 cytokine group, did not show a common pattern over the time course. IL-10 and TGF-beta, which belong to the Treg cytokine group, were decreased significantly after provocation. Zero week, pre-sensitization; 12 week, post-sensitization; 16 week, post-provocation. All values are expressed as the mean ± standard error. *P < 0.05, **P < 0.01.

Article Snippet: Commercially available ELISA kits, which are labeled to detect canine IFN-gamma (DY781B, R&D Systems, Minneapolis, MN, USA), IL-2 (DY1815, R&D Systems, Minneapolis, MN, USA), IL-4 (DY754, R&D Systems, Minneapolis, MN, USA), IL-10 (DY735, R&D Systems, Minneapolis, MN, USA), IL12 (DY1969, R&D Systems, Minneapolis, MN, USA), IL13 (SEA060Ca, Cloud-Clone Corp., Katy, TX, USA), IL31 (ECI0041, ABclonal, Woburn, MA, USA), and TGF-beta (DB100B, R&D Systems, Minneapolis, MN, USA), were used.

Techniques:

Journal: EBioMedicine

Article Title: In situ administration of cytokine combinations induces tumor regression in mice

doi: 10.1016/j.ebiom.2018.09.050

Figure Lengend Snippet: A lentivector-based inducible system to evaluate the antitumor effects of cytokines. (a) Schematic representation of the two lentivectors of the inducible system. (b) Dox-induced iRFP expression in B16F10-rtTA(TRE-iRFP) tumors. (c) Effects of dox-induced single cytokine expression, including IL12, GMCSF or IL2, on the growth of established tumors. The tumor growth curves are data from one of two replicate experiments (five mice in each replicate). (d) Effects of dox-induced two-cytokine combinations expression, including IL12 + GMCSF, IL12 + IL2 or GMCSF+IL2, on the growth of established tumors. The tumor growth curves are data from two of three replicate experiments (five mice in each replicate). (e) Effects of dox-induced expression of the IL12 + GMCSF+IL2 combination on the growth of established tumors. The tumor growth curves are data from two of three replicate experiments (five mice in each replicate). (f) Representative photographs of tumor regression after induced IL12 + GMCSF+IL2 expression. (g) Effects of dox-induced expression of the IL12 + GMCSF+IL2 + IFNγ combination on the growth of established tumors. The tumor growth curves are data from two of three replicate experiments (five mice in each replicate). Each curve represented the tumor growth of an individual mouse after dox administration. The numbers in the graphs indicate tumor-cleared mice/total dox-treated mice.

Article Snippet: Canine IL12, GMCSF and IL2 (Novas Biologicals) were reconstituted to 150 ng/μl.

Techniques: Expressing

Journal: EBioMedicine

Article Title: In situ administration of cytokine combinations induces tumor regression in mice

doi: 10.1016/j.ebiom.2018.09.050

Figure Lengend Snippet: Induced expression of the IL12 + GMCSF + IL2 combination elicites systemic antitumor immunity. (a) Effects of dox-induced expression of the IL12 + GMCSF+IL2 combination on the growth of large established tumors (10 mm < diameter < 15 mm or diameter > 15 mm). Each curve represented the tumor growth of an individual mouse after dox administration. The numbers in the graphs indicate tumor-cleared mice/total dox-treated mice. X indicates a death case within four days after induction. The tumor growth curves are data from two of three replicate experiments (five mice in each replicate). (b) Cured mice rejected rechallenged B16F10 tumor cells, either subcutaneously or intravenously injected. The numbers indicated death case/injected mice. (c) At different times post subcutaneous inoculation of inducible tumor cells into the flank of C57BL/6 mice, parental tumor cells were subcutaneously injected into the contralateral side. Dox induction was initiated when the size of bilateral tumors was at different ratios (inducible tumor larger, parental tumor larger, or size of both tumors were comparable). Mice bearing only parental tumors served as the control. The survival of tumor bearing mice was recorded. n = 5 for the control group and n = 10 for the other groups. ***, p < .001. (d) Parental tumor cells were intravenously injected into mice bearing subcutaneously established inducible tumors. Dox induction was initiated just after injection or two days later. Mice without inducible tumors served as the control. The survival of tumor-bearing mice was recorded. n = 5 for the control group and n = 10 for the other groups. ***, p < .001. (e) Vitiligo at the tumor site after tumor regression caused by the induced expression of IL12 + GMCSF+IL2.

Article Snippet: Canine IL12, GMCSF and IL2 (Novas Biologicals) were reconstituted to 150 ng/μl.

Techniques: Expressing, Injection, Control

Journal: EBioMedicine

Article Title: In situ administration of cytokine combinations induces tumor regression in mice

doi: 10.1016/j.ebiom.2018.09.050

Figure Lengend Snippet: Induced expression of cytokine combinations eliminates established tumors. Mixtures of different cytokine inducible B16F10 cells, including IL12 + GMCSF+IL15 (a), IL12 + GMCSF+IL21 (b), IL12 + FLT3L + IL15 (c), IL12 + FLT3L + IL21 (d), IL12 + FLT3L + IL2 (e), were subcutaneously inoculated at the flank of C57BL/6 mice. Dox induction was initiated after tumor nodule formation. Tumor growth was recorded after induction. Each curve represented the tumor growth of an individual mouse after dox administration. The numbers in the graphs indicate tumor-cleared mice/total dox treated mice. The tumor growth curves are data from two of three replicate experiments (five mice in each replicate). The photographs indicate a representative tumor regression progress.

Article Snippet: Canine IL12, GMCSF and IL2 (Novas Biologicals) were reconstituted to 150 ng/μl.

Techniques: Expressing

Journal: EBioMedicine

Article Title: In situ administration of cytokine combinations induces tumor regression in mice

doi: 10.1016/j.ebiom.2018.09.050

Figure Lengend Snippet: Treatment of syngeneic tumors by intratumoral injection of chitosan/IL12 + GMCSF + IL2. B16F10 (a), EL4 (b), LLC (c), CT26 (d) and 4 T1 (e) tumors were established by subcutaneous injection of tumor cells into the flank of syngeneic mice. Intratumoral injection of chitosan/IL12 + GMCSF+IL2 was performed when the tumor diameter reached ~6 mm. Tumor growth was monitored and additional injections were carried out at the signs of tumor size increase or relapse after regression, indicated by arrows in the graphs. X indicates a death case. The numbers in the graphs indicated tumor-cleared mice/total treated mice. The tumor growth curves are data from one of two replicate experiments (five mice in each replicate). The photographs indicate a representative tumor regression progress.

Article Snippet: Canine IL12, GMCSF and IL2 (Novas Biologicals) were reconstituted to 150 ng/μl.

Techniques: Injection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Safe and effective treatment of spontaneous neoplasms with interleukin 12 electro-chemo-gene therapy

doi: 10.1111/jcmm.12382

Figure Lengend Snippet: Details of the enrolled subjects

Article Snippet: To produce the canine IL12 used in this study, canine IL12 subunits P35 and P40 DNA were synthesized based on Genebank sequences NM_001003293 and NM_001003292 (GenScript USA) and subcloned to control vector pVC1157 under CMV promoter , .

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Safe and effective treatment of spontaneous neoplasms with interleukin 12 electro-chemo-gene therapy

doi: 10.1111/jcmm.12382

Figure Lengend Snippet: ECGT with IL12 pDNA is equally effective with gemcitabine or bleomycin. Analysing all treatment cycles with IL12 pDNA and either bleomycin (IL12 + blm) or gemcitabine (IL12 + gem) in squamous cell carcinoma (A, SCC, n + 9, 10), plasmacytoma (B, PC, n + 2, 3) and sarcoma (C, Sarc, n + 5, 4) reveals that ECGT is equally effective with blm or gem. (D) Comparison of all tumour histotypes shows that sarcomas do not respond to ECGT while PC and SCC are very responsive. (E) ECGT treatments extends survival compared to non-surgical standard treatments. Survival curve comparing ECGT ( n + 16) and non-surgical standard treatments in subjects with complete records at the clinical trial host site. Error bars represent SEM.

Article Snippet: To produce the canine IL12 used in this study, canine IL12 subunits P35 and P40 DNA were synthesized based on Genebank sequences NM_001003293 and NM_001003292 (GenScript USA) and subcloned to control vector pVC1157 under CMV promoter , .

Techniques: Comparison

Journal: Journal of Cellular and Molecular Medicine

Article Title: Safe and effective treatment of spontaneous neoplasms with interleukin 12 electro-chemo-gene therapy

doi: 10.1111/jcmm.12382

Figure Lengend Snippet: IL12 pDNA can confer an antitumour immune response. (A) In squamous cell carcinoma, ECGT with IL12 pDNA and a chemotherapeutic induces tumour regression equally compared to IL12 pDNA alone plus electroporation, but the tumour growth resumes by day 21 after treatment ( n + 5 for IL12 pDNA alone and n + 20 for ECGT). (B) In acanthomatous ameloblastoma, IL12 pDNA alone plus electroporation appears more effective than ECGT with IL12 pDNA and a chemotherapeutic, although the difference is not significant ( n + 4). (C) In Sarcomas, IL12 pDNA alone plus electroporation ( n + 2) appears to stabilize the tumour volume while ECGT with IL12 pDNA and a chemotherapeutic ( n + 9) cannot inhibit tumour growth; however, the low sample size of IL12 pDNA prohibits statistical analysis. (D) SCC tumours located anywhere except the nasal planum (non-NP SCC, n + 20) respond to ECGT with an average 30% tumour reduction in only 21 days, but nasal planum SCC tumours (NP SCC, n + 8) do not respond to ECGT. (E) A non-treated metastatic lymph node SCC tumour steadily regressed after two treatments in the initial dorsal carpus tumours on days 0 and 7. At 96 days after the treatments in the dorsal carpus tumours, progression of the metastatic nodule was noted. X denotes surgical removal of the metastatic tumour. Error bars represent SEM.

Article Snippet: To produce the canine IL12 used in this study, canine IL12 subunits P35 and P40 DNA were synthesized based on Genebank sequences NM_001003293 and NM_001003292 (GenScript USA) and subcloned to control vector pVC1157 under CMV promoter , .

Techniques: Electroporation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Safe and effective treatment of spontaneous neoplasms with interleukin 12 electro-chemo-gene therapy

doi: 10.1111/jcmm.12382

Figure Lengend Snippet: IL12 pDNA alone or with chemotherapy can equally affect large and small tumours. (A) EP-mediated EGT (IL12 pDNA alone) equally regresses small (<3 cm, n + 4) and large (>3 cm, n + 5) tumours for 14 days, but tumour progression returns by day 21. (B) ECGT (IL12 pDNA plus either bleomycin or gemcitabine) equally regresses small (<3 cm, n + 11) and large (>3 cm, n + 19) tumours. Error bars represent SEM.

Article Snippet: To produce the canine IL12 used in this study, canine IL12 subunits P35 and P40 DNA were synthesized based on Genebank sequences NM_001003293 and NM_001003292 (GenScript USA) and subcloned to control vector pVC1157 under CMV promoter , .

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Safe and effective treatment of spontaneous neoplasms with interleukin 12 electro-chemo-gene therapy

doi: 10.1111/jcmm.12382

Figure Lengend Snippet: ECGT with IL12 pDNA and either bleomycin or gemcitabine can eradicate or debulk large squamous cell carcinoma tumours. (A) Squamous cell carcinoma tumours on Day 0 prior to ECGT with IL12 pDNA and bleomycin. The larger tumour is outlined in blue and the samller tumour is outlined in red. (B) Eradicated tumours on Day 70. (C) Tumour volume curves showing the eradication as per cent of tumour volume on Day 0. The coloured lines corresponds to the tumours outlined in A. (D) Maxillary Squamous cell carcinoma tumours on Day 0 prior to ECGT with IL12 pDNA and gemcitabine. (E) Healthy maxillary tissue with no evidence of residual SCC tumour cells as determined by histopathological analysis. (F) Tumour volume curve showing the eradication as per cent of tumour volume on Day 0. CT scan (G) and photograph (H) of an invasive squamous cell carcinoma tumour on Day 0 prior to ECGT with IL12 pDNA and bleomycin. (I) Photograph illustrating quick debulking of SCC on day 7 after only 1 ECGT treatment. (J) Tumour volume curve showing tumour reduction as per cent of tumour volume on Day 0. (K) Tumour volume curve showing tumour reduction as per cent of tumour volume on Day 0 of a subject with an SCC tumour in the retromolar trigone after only 1 ECGT treatment with IL12 pDNA and bleomycin. (L) Average tumour volume reduction in SCC tumours with ECGT with IL12 pDNA and either bleomycin or gemcitabine. Error bars represent SEM ( n + 8).

Article Snippet: To produce the canine IL12 used in this study, canine IL12 subunits P35 and P40 DNA were synthesized based on Genebank sequences NM_001003293 and NM_001003292 (GenScript USA) and subcloned to control vector pVC1157 under CMV promoter , .

Techniques: Computed Tomography

Journal:

Article Title: Angiogenesis blockade as a new therapeutic approach to experimental colitis

doi: 10.1136/gut.2006.114314

Figure Lengend Snippet: Figure 6 Reduction of cytokine production in the inflamed gut mucosa by angiogenesis blockade. Intraperitoneal injections of ATN‐161 (n = 12) or its scrambled peptide ATN‐163 (n = 10) were given on alternate days to 15‐week‐old mice with fully established colitis. After 6 weeks, animals were killed, their colons removed, maintained in organ cultures and supernatants collected for the analysis of interleukin (IL) 12 and IL6 content by ELISA. *p<0.05.

Article Snippet: After five washes with PBS/Tween, 40 μl of supernatant or murine recombinant IL12 standard (5000–78 pg/ml in twofold serial dilutions; R&D Systems, Minneapolis, Minnesota, USA) was added to the plate in duplicate at room temperature for 2 h. The plate was washed five times with PBS/Tween, followed by 1 μg/ml of biotinylated C17.8 (PharMingen, San Diego, California, USA) for 1 h at room temperature.

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Angiogenesis blockade as a new therapeutic approach to experimental colitis

doi: 10.1136/gut.2006.114314

Figure Lengend Snippet: Figure 7 Lack of effect of ATN‐161 on lamina propria mononuclear cell (LPMC) cytokine production and proliferation. (A) LPMCs isolated from the normal colon of interleukin (IL) 10‐deficient mice kept under the ultra‐barrier facility were cultured alone or stimulated with lipopolysaccharide (LPS), in the presence or absence of ATN‐161. After 3 days, supernatants were collected and the content of IL12 was measured by ELISA. (B) Isolated LPMCs were cultured alone or stimulated with anti‐CD3 in the presence or absence of ATN‐161, and proliferation was measured by thymidine incorporation.

Article Snippet: After five washes with PBS/Tween, 40 μl of supernatant or murine recombinant IL12 standard (5000–78 pg/ml in twofold serial dilutions; R&D Systems, Minneapolis, Minnesota, USA) was added to the plate in duplicate at room temperature for 2 h. The plate was washed five times with PBS/Tween, followed by 1 μg/ml of biotinylated C17.8 (PharMingen, San Diego, California, USA) for 1 h at room temperature.

Techniques: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay