Combination of Id2 Knockdown Whole Tumor Cells and Checkpoint Blockade: A Potent Vaccine Strategy in a Mouse Neuroblastoma Model
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{"title"=>"Combination of Id2 knockdown whole tumor cells and checkpoint blockade: A potent vaccine strategy in a mouse neuroblastoma model", "type"=>"journal", "authors"=>[{"first_name"=>"Lina", "last_name"=>"Chakrabarti", "scopus_author_id"=>"15058814400"}, {"first_name"=>"Clifford", "last_name"=>"Morgan", "scopus_author_id"=>"56735363500"}, {"first_name"=>"Anthony D.", "last_name"=>"Sandler", "scopus_author_id"=>"35553413600"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"605265705", "pmid"=>"26079374", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0129237", "scopus"=>"2-s2.0-84937908293", "sgr"=>"84937908293"}, "id"=>"c0795627-708d-3109-9936-3663c362e8a8", "abstract"=>"Tumor vaccines have held much promise, but to date have demonstrated little clinical success. This lack of success is conceivably due to poor tumor antigen presentation combined with immuno-suppressive mechanisms exploited by the tumor itself. Knock down of Inhibitor of differentiation protein 2 (Id2-kd) in mouse neuroblastoma whole tumor cells rendered these cells immunogenic. Id2-kd neuroblastoma (Neuro2a) cells (Id2-kd N2a) failed to grow in most immune competent mice and these mice subsequently developed immunity against further wild-type Neuro2a tumor cell challenge. Id2-kd N2a cells grew aggressively in immune-compromised hosts, thereby establishing the immunogenicity of these cells. Therapeutic vaccination with Id2-kd N2a cells alone suppressed tumor growth even in established neuroblastoma tumors and when used in combination with immune checkpoint blockade eradicated large established tumors. Mechanistically, immune cell depletion studies demonstrated that while CD8+ T cells are critical for antitumor immunity, CD4+ T cells are also required to induce a sustained long-lasting helper effect. An increase in number of CD8+ T-cells and enhanced production of interferon gamma (IFNγ) was observed in tumor antigen stimulated splenocytes of vaccinated mice. More importantly, a massive influx of cytotoxic CD8+ T-cells infiltrated the shrinking tumor following combined immunotherapy. These findings show that down regulation of Id2 induced tumor cell immunity and in combination with checkpoint blockade produced a novel, potent, T-cell mediated tumor vaccine strategy.", "link"=>"http://www.mendeley.com/research/combination-id2-knockdown-whole-tumor-cells-checkpoint-blockade-potent-vaccine-strategy-mouse-neurob", "reader_count"=>18, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Other"=>4, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Other"=>4, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>3, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>5, "Neuroscience"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>3}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2115868"], "description"=>"<p>(<b>A</b>) Representative flow cytometry plots showingCD4+ and CD8+ T-cells in CD45+ splenocytes of naïve (n = 5), tumor bearing (n = 3) and tumor free (n = 5) mice. (<b>B</b>) Graphical representation of (A) indicating significant (** p = 0.013) increase in CD8+ cells in the spleens of mice cleared of tumor. (<b>C</b>) Splenocytes of mice that cleared tumor had enhanced IFNγ secretion following stimulation with CD3 (* p<0.02),wtN2a or Id2kdN2a cells (*** p<0.0001). (<b>D</b>) Remarkable increase in CD45+ cells detected in the shrinking tumor.(<b>E</b>, <b>F</b>) Tumor infiltrating lymphocytes (TIL) following vaccination were quantified and a massive infiltration of CD8+ T-cells was detected in the shrinking tumors (n = 5) as opposed to the growing tumors (* p<0.02). (<b>G</b>) Chromium<sup>51</sup> release assay exhibited potent cytotoxic activity of CD8+ TIL from shrinking tumor (n = 3); whereas the TIL isolated from growing tumors (n = 4) show no activity at all. Data presented as mean ± S.D.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451061, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g006", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Enhanced_in_vivo_immune_response_mediates_tumor_clearance_/1451061", "title"=>"Enhanced <i>in vivo</i> immune response mediates tumor clearance.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115853"], "description"=>"<p>(<b>A</b>) Schematic diagram of the therapeutic vaccine strategy. Two established tumor models, namely Neuro2a (wtN2a) and AgN2a were tested, where mice challenged with either wtN2a or AgN2a cells were subjected to a combination immunotherapy with Id2kdN2a and α-CTLA4 antibody starting at day 6 after inoculation. Neuroblastoma tumors are normally visible (5mm in diameter) in AJ mice by day 6. Tumor growth curves in individual mice of wtN2a (<b>B</b>) and AgN2a (<b>C</b>) cells show that the Id2kd tumor cell vaccination combined with immune-modulation cures mice with established tumor. Parentheses indicate the number of mice that survived tumor free.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451057, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g003", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Combination_of_Id2kd_N2a_and_945_CTLA4_antibody_as_a_therapeutic_vaccine_/1451057", "title"=>"Combination of Id2kd N2a and α-CTLA4 antibody as a therapeutic vaccine.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115852"], "description"=>"<p>(<b>A</b>) Schematic diagram of the prophylactic Neuro2a tumor models. Two models (0-day and 5-day) were tested, in which three doses of α-CTLA4 antibody were administered either days 0,3,6 or 5,8,11 following Id2kdN2a cell inoculation into the right leg of the mice (n = 5 for each model). (<b>B, C</b>) Tumor growth curves show that 60% of mice challenged with only Id2-kd N2a cells (no α-CTLA4) survived tumor free (B) and in the 0-day model (C) 40% of mice never grew tumor and another 40% cleared the tumor slowly and became tumor free. Graph depicts tumor growth in individual mice. The parenthesis indicates number of mice that survived tumor free. (<b>D</b>) All mice in the 5-day model survived tumor free, in contrast to 80% in the 0-day model or 60% in the Id2kd cells model. (<b>E</b>) Six weeks after tumor clearance, the tumor-free mice from (B) (n = 9) were re-challenged with wtN2a cells into their left leg and 100% were completely immunized against wild type tumor growth. Only 1 of 5 mice (20%) treated previously with anti-CTLA4 antibody alone survived tumor free after wtN2a challenge at 6 weeks.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451056, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CTLA_4antibody_enhances_anti_tumor_effect_of_Id2_kd_N2a_cell_vaccine_in_prophylactic_tumor_models_/1451056", "title"=>"CTLA-4antibody enhances anti-tumor effect of Id2-kd N2a cell vaccine in prophylactic tumor models.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115850"], "description"=>"<p>(<b>A</b>) Sixty percent of mice (n = 9/15) challenged (right leg) with Id2 knock down Neuro2a (Id2kdN2a) cells rejected tumor and survived tumor-free, whereas all mice challenged with either wild type Neuro2a (wtN2a, n = 20) or scrambled shRNA lentivirus transfected Neuro2a (sc-shRNA-N2a, n = 5)) cells died from tumor burden. (<b>B</b>) Tumor free survivors from (A) were re-challenged with wtN2a cells into their left leg 6 weeks after they cleared the tumor and 8 out of 9 mice were protected from tumor growth. (<b>C</b>) SCID and nude mice grew tumors aggressively following inoculation with Id2kdN2a cells. (<b>D</b>) Following inoculation of wtN2a cells in the right leg, Id2kdN2a (n = 10) and wtN2a (n = 5) cells were vaccinated into the left leg of mice, 3 and 5 days later respectively. The wild type tumor growth on the right leg was delayed in Id2kdN2a vaccinated mice when compared to control unvaccinated mice or wtN2a vaccinated mice.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451054, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g001", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Id2_knock_down_attenuates_tumorigenicity_and_induces_host_immunity_/1451054", "title"=>"Id2 knock down attenuates tumorigenicity and induces host immunity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115869"], "description"=>"<p>Five freshly harvested human neuroblastoma specimens were analyzed for TIL; two of the tumors were from patients with Opsoclonus/myoclonus syndrome (OMS). (<b>A</b>) Representative flow cytometry plots showing CD4+ and CD8+ T-cell population in the neuroblastoma tumor lymphocytes. (<b>B</b>) Graphical representation of (A) indicating a remarkable infiltration of CD8+ T-cells in the two tumors associated with OMS (NB-OMS) as opposed to the other three neuroblastoma (NB) tumors that had minimal TIL.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451062, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g007", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TIL_in_human_neuroblastoma_samples_/1451062", "title"=>"TIL in human neuroblastoma samples.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115855"], "description"=>"<p>Mice were depleted of specific T cell subsets by systemic administration of antibodies against CD4, CD8 and NK cells and subjected to combined Id2kdN2a cells and α-CTLA-4 antibody treatment strategy (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129237#pone.0129237.g003\" target=\"_blank\">Fig 3A</a>) (<b>A</b>) <i>In vivo</i> depletion of CD4+, CD8+ and NK+ cells (n = 5 for all groups) shows that the therapeutic effect of the combined treatment was completely abrogated by CD8+ cell depletion. Mice lacking NK cells were for the most part able to reject their tumors following therapy. CD4+ T cell depletion initially appeared to have minimal adverse effects on the therapeutic vaccine strategy, but after 4 weeks all mice in the CD4 depletion group developed delayed tumors. (<b>B</b>) All CD8+ T cell depleted mice developed tumors at the site of Id2kdN2a cell vaccination as well. (<b>C</b>) Cell infiltrates from the tumors of CD4 depleted mice were stained with anti-CD45-FITC, CD4-APC and FoxP3-PerCPCy5.5 antibodies to identify Treg cells; there was no evidence of Treg cell infiltrate in the late developing tumors from CD4 depleted mice. Purified Treg cells were used as positive control.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451059, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_T_cell_immunity_required_for_tumor_eradication_following_combination_therapy_/1451059", "title"=>"T cell immunity required for tumor eradication following combination therapy.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/2115854"], "description"=>"<p>(<b>A</b>, <b>B</b>) Representative mouse from untreated and combination therapy group showing growing and shrinking tumor respectively. All bio-luminescent images were analyzed under the same scale.(<b>C</b>, <b>D</b>) Irradiated Id2 knock down Neuro2a (IR-Id2kdN2a, n = 5) and wild type Neuro2a (IR-wtN2a, n = 5) were compared as whole tumor cell vaccine antigen source in combination with α-CTLA-4 antibody against AgN2a (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129237#pone.0129237.g003\" target=\"_blank\">Fig 3A</a>). IR-Id2kdN2a vaccine was able to eradicate 60% of tumors in comparison to IR-wtN2a cells which had no effect on growth of the AgN2a tumors. All mice in (C) were sacrificed on day 20 due to large tumor burden. The parenthesis indicates number of mice that survived tumor free.</p>", "links"=>[], "tags"=>["differentiation protein 2", "Id 2 Knockdown", "ifn", "tumor antigen presentation", "Neuro 2A cells", "cell depletion studies", "tumor cell immunity", "checkpoint blockade", "tumor vaccine strategy", "Mouse Neuroblastoma Model Tumor vaccines", "Potent Vaccine Strategy"], "article_id"=>1451058, "categories"=>["Biological Sciences"], "users"=>["Lina Chakrabarti", "Clifford Morgan", "Anthony D. Sandler"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0129237.g004", "stats"=>{"downloads"=>3, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Tumor_clearance_and_effect_of_irradiation_on_tumor_cell_immunogenicity_/1451058", "title"=>"Tumor clearance and effect of irradiation on tumor cell immunogenicity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-16 02:58:10"}

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{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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