Limits...
Virological and preclinical characterization of a dendritic cell targeting, integration-deficient lentiviral vector for cancer immunotherapy.

Odegard JM, Kelley-Clarke B, Tareen SU, Campbell DJ, Flynn PA, Nicolai CJ, Slough MM, Vin CD, McGowan PJ, Nelson LT, Ter Meulen J, Dubensky TW, Robbins SH - J. Immunother. (2015 Feb-Mar)

Bottom Line: Specificity of ID-VP02 for antigen-presenting cells in the mouse was confirmed through biodistribution studies showing that following subcutaneous administration, transgene expression was only detectable at the injection site and the draining lymph node.Upon homologous readministration, ID-VP02 induced a level of high-quality secondary effector and memory cells characterized by stable polyfunctionality and expression of IL-7Rα.Importantly, a single injection of ID-VP02 also induced robust cytotoxic responses against an endogenous rejection antigen of CT26 colon carcinoma cells and conferred both prophylactic and therapeutic antitumor efficacy.

View Article: PubMed Central - PubMed

Affiliation: *Immune Design Corp. †TRIA Bioscience Corp., Seattle, WA.

ABSTRACT
Dendritic cells (DCs) are essential antigen-presenting cells for the initiation of cytotoxic T-cell responses and therefore attractive targets for cancer immunotherapy. We have developed an integration-deficient lentiviral vector termed ID-VP02 that is designed to deliver antigen-encoding nucleic acids selectively to human DCs in vivo. ID-VP02 utilizes a genetically and glycobiologically engineered Sindbis virus glycoprotein to target human DCs through the C-type lectin DC-SIGN (CD209) and also binds to the homologue murine receptor SIGNR1. Specificity of ID-VP02 for antigen-presenting cells in the mouse was confirmed through biodistribution studies showing that following subcutaneous administration, transgene expression was only detectable at the injection site and the draining lymph node. A single immunization with ID-VP02 induced a high level of antigen-specific, polyfunctional effector and memory CD8 T-cell responses that fully protected against vaccinia virus challenge. Upon homologous readministration, ID-VP02 induced a level of high-quality secondary effector and memory cells characterized by stable polyfunctionality and expression of IL-7Rα. Importantly, a single injection of ID-VP02 also induced robust cytotoxic responses against an endogenous rejection antigen of CT26 colon carcinoma cells and conferred both prophylactic and therapeutic antitumor efficacy. ID-VP02 is the first lentiviral vector which combines integration deficiency with DC targeting and is currently being investigated in a phase I trial in cancer patients.

Show MeSH

Related in: MedlinePlus

Antibody against SIGNR1 blocks transduction with ID-VP02. HT1080 cells expressing SIGNR1 (HT1080-SIGNR1) or the parental cells (HT1080) were preincubated with anti-SIGNR1 antibody (10 µg/mL) for 1 hour, after which increasing doses of (A) ID-VP02 or (B) VSV-G vectors encoding GFP were added. Transduction was assessed 72 hours later. Nevirapine was included on highest vector dose as a control for real transduction (data not shown). Vector titers of ID-VP02 and VSV-G were comparable (115 µg/mL p24, and 111 µg/mL p24, respectively).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4323576&req=5

Figure 1: Antibody against SIGNR1 blocks transduction with ID-VP02. HT1080 cells expressing SIGNR1 (HT1080-SIGNR1) or the parental cells (HT1080) were preincubated with anti-SIGNR1 antibody (10 µg/mL) for 1 hour, after which increasing doses of (A) ID-VP02 or (B) VSV-G vectors encoding GFP were added. Transduction was assessed 72 hours later. Nevirapine was included on highest vector dose as a control for real transduction (data not shown). Vector titers of ID-VP02 and VSV-G were comparable (115 µg/mL p24, and 111 µg/mL p24, respectively).

Mentions: HT1080 cells stably expressing either mouse SIGNR1, SIGNR3, or SIGNR5 were generated (see Materials and methods section). Expression of the each receptor was confirmed by flow cytometry or RT-PCR (Supplementary Fig. 1, Supplemental Digital Content 1, http://links.lww.com/JIT/A370). These cells were incubated with varying concentrations of integration-deficient GFP-encoding vector that was pseudotyped with Sindbis virus envelope E1001, kifunensine-modified high mannose E1001, or with pantropic VSV-G. In previous studies, we have established that the E1001 envelope produced in the presence of the mannosidase I inhibitor kifunensine is required for DC-SIGN binding and human DC transduction.6 In this experiment, therefore, HT1080 cells expressing human DC-SIGN were used as a positive control. Of the 3 mouse DC-SIGN orthologs tested, E1001-pseudotyped vector-transduced only mouse SIGNR1-expressing cells and did so in a kifunensine-dependent manner (Supplementary Fig. 2, Supplemental Digital Content 2, http://links.lww.com/JIT/A371) and the ability of E1001-pseudotyped vector to transduce SIGNR1-expressing cells was completely blocked in the presence of an anti-SIGNR1 antibody (Fig. 1A). The transduction efficiency of kifunensine-modified E1001 vector on human DC-SIGN-expressing and mouse SIGNR1-expressing cells was comparable, indicating that SIGNR1 is a functionally orthologous receptor for ID-VP02 in the mouse.


Virological and preclinical characterization of a dendritic cell targeting, integration-deficient lentiviral vector for cancer immunotherapy.

Odegard JM, Kelley-Clarke B, Tareen SU, Campbell DJ, Flynn PA, Nicolai CJ, Slough MM, Vin CD, McGowan PJ, Nelson LT, Ter Meulen J, Dubensky TW, Robbins SH - J. Immunother. (2015 Feb-Mar)

Antibody against SIGNR1 blocks transduction with ID-VP02. HT1080 cells expressing SIGNR1 (HT1080-SIGNR1) or the parental cells (HT1080) were preincubated with anti-SIGNR1 antibody (10 µg/mL) for 1 hour, after which increasing doses of (A) ID-VP02 or (B) VSV-G vectors encoding GFP were added. Transduction was assessed 72 hours later. Nevirapine was included on highest vector dose as a control for real transduction (data not shown). Vector titers of ID-VP02 and VSV-G were comparable (115 µg/mL p24, and 111 µg/mL p24, respectively).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4323576&req=5

Figure 1: Antibody against SIGNR1 blocks transduction with ID-VP02. HT1080 cells expressing SIGNR1 (HT1080-SIGNR1) or the parental cells (HT1080) were preincubated with anti-SIGNR1 antibody (10 µg/mL) for 1 hour, after which increasing doses of (A) ID-VP02 or (B) VSV-G vectors encoding GFP were added. Transduction was assessed 72 hours later. Nevirapine was included on highest vector dose as a control for real transduction (data not shown). Vector titers of ID-VP02 and VSV-G were comparable (115 µg/mL p24, and 111 µg/mL p24, respectively).
Mentions: HT1080 cells stably expressing either mouse SIGNR1, SIGNR3, or SIGNR5 were generated (see Materials and methods section). Expression of the each receptor was confirmed by flow cytometry or RT-PCR (Supplementary Fig. 1, Supplemental Digital Content 1, http://links.lww.com/JIT/A370). These cells were incubated with varying concentrations of integration-deficient GFP-encoding vector that was pseudotyped with Sindbis virus envelope E1001, kifunensine-modified high mannose E1001, or with pantropic VSV-G. In previous studies, we have established that the E1001 envelope produced in the presence of the mannosidase I inhibitor kifunensine is required for DC-SIGN binding and human DC transduction.6 In this experiment, therefore, HT1080 cells expressing human DC-SIGN were used as a positive control. Of the 3 mouse DC-SIGN orthologs tested, E1001-pseudotyped vector-transduced only mouse SIGNR1-expressing cells and did so in a kifunensine-dependent manner (Supplementary Fig. 2, Supplemental Digital Content 2, http://links.lww.com/JIT/A371) and the ability of E1001-pseudotyped vector to transduce SIGNR1-expressing cells was completely blocked in the presence of an anti-SIGNR1 antibody (Fig. 1A). The transduction efficiency of kifunensine-modified E1001 vector on human DC-SIGN-expressing and mouse SIGNR1-expressing cells was comparable, indicating that SIGNR1 is a functionally orthologous receptor for ID-VP02 in the mouse.

Bottom Line: Specificity of ID-VP02 for antigen-presenting cells in the mouse was confirmed through biodistribution studies showing that following subcutaneous administration, transgene expression was only detectable at the injection site and the draining lymph node.Upon homologous readministration, ID-VP02 induced a level of high-quality secondary effector and memory cells characterized by stable polyfunctionality and expression of IL-7Rα.Importantly, a single injection of ID-VP02 also induced robust cytotoxic responses against an endogenous rejection antigen of CT26 colon carcinoma cells and conferred both prophylactic and therapeutic antitumor efficacy.

View Article: PubMed Central - PubMed

Affiliation: *Immune Design Corp. †TRIA Bioscience Corp., Seattle, WA.

ABSTRACT
Dendritic cells (DCs) are essential antigen-presenting cells for the initiation of cytotoxic T-cell responses and therefore attractive targets for cancer immunotherapy. We have developed an integration-deficient lentiviral vector termed ID-VP02 that is designed to deliver antigen-encoding nucleic acids selectively to human DCs in vivo. ID-VP02 utilizes a genetically and glycobiologically engineered Sindbis virus glycoprotein to target human DCs through the C-type lectin DC-SIGN (CD209) and also binds to the homologue murine receptor SIGNR1. Specificity of ID-VP02 for antigen-presenting cells in the mouse was confirmed through biodistribution studies showing that following subcutaneous administration, transgene expression was only detectable at the injection site and the draining lymph node. A single immunization with ID-VP02 induced a high level of antigen-specific, polyfunctional effector and memory CD8 T-cell responses that fully protected against vaccinia virus challenge. Upon homologous readministration, ID-VP02 induced a level of high-quality secondary effector and memory cells characterized by stable polyfunctionality and expression of IL-7Rα. Importantly, a single injection of ID-VP02 also induced robust cytotoxic responses against an endogenous rejection antigen of CT26 colon carcinoma cells and conferred both prophylactic and therapeutic antitumor efficacy. ID-VP02 is the first lentiviral vector which combines integration deficiency with DC targeting and is currently being investigated in a phase I trial in cancer patients.

Show MeSH
Related in: MedlinePlus