Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD99 , REA1174 , 50 , 130-121-078 , PE , Miltenyi Biotec.

Techniques: Imaging

Journal: Hypertension

Article Title: Mechanisms of Enhanced Vascular Reactivity in Preeclampsia

doi: 10.1161/hypertensionaha.111.176602

Figure Lengend Snippet: Figure 2. Representative sections of omental fat vessels immunostained for CD66b, a neutrophil antigen; CD99, a lymphocyte antigen; and CD14, a monocyte/macrophage antigen. A, Isotype negative control for CD66b (preeclamptic). B, Normal pregnancy for CD66b. Vessels of normal pregnant patients had some brown staining for neutrophils primarily in the lumen. C and D, Preeclamptic pregnancy for CD66b. Vessels of preeclamptic patients showed extensive brown staining of neutrophils in the lumen, adhered and flattened along the endothelium, and infiltrated to the vascular smooth muscle. E and F, Lower magnification of CD66b staining showing little to no staining of vessels in normal pregnancy (E) as compared with almost all of the vessels showing extensive staining in preeclampsia (F). G, CD99 staining and (H) CD14 staining in preeclamptic vessels. Few vessels showed staining for CD99 or CD14, and, when they did, there were only 1 or 2 lymphocytes or monocytes/macrophages per vessel (arrows). A, adipocyte; VL, vessel lumen; NP, normal preg- nancy; PE, preeclamptic pregnancy. All of the images were taken with a 40 lens except E and F taken with a 10 lens.

Article Snippet: Tissues were stained with the following antibodies: 1) mouse monoclonal IgM anti-human CD66b (1:50, BD BioSciences, San Diego, CA), 2) rabbit polyclonal IgG anti-human CD14 (titer 1:100, ProteinTech, Chicago, IL), 3) mouse monoclonal IgG1 anti-human CD99 (titer 1:400, Serotec, Oxford, UK).

Techniques: Negative Control, Staining

Journal: Blood Cells, Molecules & Diseases

Article Title: In-depth phenotypic characterization of reticulocyte maturation using mass cytometry

doi: 10.1016/j.bcmd.2018.06.004

Figure Lengend Snippet: Antibodies, clones, metal isotopes tagged antibodies and companies used for flow cytometry and mass cytometry analysis. Commercially available metal isotopes tagged antibodies were purchased from Fluidigm. Other antibodies were purchased from other companies and custom-conjugated with heavy metal-preloaded maleimide-coupled MAXPAR chelating polymers by Fluidigm. Targets are termed by their cluster of differentiation (CD) and alternative names. Custom-conjugated antibodies went through an extra validation process by Fluidigm (*) (Supplemental Fig. 1).

Article Snippet: CD99 , MIC2/single-chain type-1 glycoprotein , HCD99 , Fluidigm , 148Nd , .

Techniques: Clone Assay, Flow Cytometry, Mass Cytometry, Biomarker Discovery, Conjugation Assay

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. DMG and normal cells were transduced with K27M mutant at the H3.3 locus. Western blot showing the changes in histone H3.3K27M and histone H3K27Me3 proteins in the transduced cells of (i) HSJD-GBM01, (ii) VUMC-DIPG10, and (iii) NHA, respectively. B. In the above-transduced cells, the changes in the expression of CD99 in (i) HSJD-GBM01, (ii) VUMC-DIPG10, and (iii) NHA, respectively are shown by western blotting. C. Flow cytometry plots for CD99 (i) HSJD-GMB01 transduced with H3.3K27M, and (ii) VUMC-DIPG10 transduced with H3.3K27M. The quantification of the mean fluorescence intensity is shown adjacent to the flow plots. (*p<0.05) D. H3.3K27M mutant in BT245 cells was deleted by Crispr-CAS9 gene editing. (i) Expression of histone H3.3K27M and histone H3K27Me3 in the paired samples by western blotting. (ii) H3.3K27M mRNA expression by RT-qPCR. (iii) Expression of CD99 in the paired cell lines by western blotting. (iv) Expression of the cell surface marker, GD2, by flow cytometry. (***p<0.001) E. HSJD-DIPG007 cells were transduced with H3.3 Wildtype using shRNA. (i) Expression of histone H3.3K27M and histone H3K27Me3 in the paired samples by western blotting. (ii) H3.3K27M mRNA expression by RT-qPCR. (iii) Expression of CD99 in the paired cell lines by western blotting. (**p<0.01, ****p<0.0001) The expression of total histone H3 or β-Actin was used as protein loading control in all western blots. IgG, respective to each antibody, was used for gating cells in flow cytometry. DAPI was used to gate for live cells. GAPDH was used as a control for qRT-PCR. Data represented as Mean ± SEM.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Transduction, Mutagenesis, Western Blot, Expressing, Flow Cytometry, Fluorescence, CRISPR, Quantitative RT-PCR, Marker, shRNA, Control

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. Volcano plot showing the statistical significance as -log 10 (p-value) plotted against the log 2 (fold change) between the genes expressed by HSJD-GBM01 transduced with K27M mutant at the H3.3 locus and the unmodified HSJD-GBM01 (control) cells. HSJD-GBM01 transduced with K27M mutant demonstrated a several-fold increase in CD99 (indicated with blue arrow) compared to the control cells. B. CD99 mRNA expression in the following paired cell lines transduced with H3.3K27M: (i) HSJD-GBM01, (ii) VUMC-DIPG10, and (iii) normal human astrocytes (NHA). (ns = not significant, **p<0.01, ****p<0.0001) C. Flow plot showing a significant decrease in the expression of CD99 in K27M mutant deleted BT245 cells as compared to the parental BT245 cells. D. ChIP-seq signal peaks of H3K27Me3 and H3K27ac in human embryonic stem cells (hES), mid-radial glia (MRG), and DMG (SU-DIPG04) cells suggesting an increase in the activating H3K27ac at the CD99 promoter region in the DMG cells but not in the hES or MRG cells. E. ChIP-seq signal peaks of the paired samples of HSJD-DIPG007 and cells transduced with the H3.3 wildtype transgene reveal a decrease in the acetylation (H3K27ac) activating mark and an increase in trimethylation (H3K27me3) repressive marker at the CD99 promoter region in compared to K27M Mutant parental HSJD-DIPG007 cells. F. Representative immunohistochemical (IHC) staining showing expression of CD99 in K27M-DMG tumor and its absence in normal pons. K27M staining was used to confirm the mutation status of the tumor. G. CD99 is highly expressed in DMG patient tumors as compared to normal pons by western blotting. The molecular weight of the long (32kDa) and short (28kDa) isoforms are denoted next to the western blot image. H. Flow plot showing the higher expression of CD99 in DMG cell lines in comparison to NHA. Data shown as the mean ± SEM. (See also and and Table S4 )

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Transduction, Mutagenesis, Control, Expressing, ChIP-sequencing, Marker, Immunohistochemical staining, Immunohistochemistry, Staining, Western Blot, Molecular Weight, Comparison

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. CD99 expression in a large cohort of DIPG patient samples (n=37) compared to its expression in normal brain regions (n=172) from the dataset using the R2 genomics. (Data represented as mean ± SD.*p<0.0001) B. Representative IHC staining showing expression of CD99 in H3.3 wildtype and H3.3K27M mutant DMG tumors. The mutation status was verified using H3K27M staining. C. CD99 protein expression in cultured DMG cell lines is shown by western blotting. D. The sonic hedgehog-activated medulloblastoma line, ONS76, has a relatively lower expression of CD99 compared to the K27M mutant DMG cell line, SU-DIPG04 as seen by flow cytometry. E. ONS76 demonstrates low expression of CD99 compared to the K27M mutant DMG cell lines, SF8628 and SU-DIPG04 as seen by western blotting. F. The overexpression of the K27M transgene in ONS76 was verified using RT-qPCR. (****p<0.0001) G. The overexpression of the K27M transgene in ONS76 significantly upregulated the long isoform of CD99 but not the short isoform by using the respective primers. (ns = not significant, ***p<0.001) H. Overexpression of the long isoform of CD99 transgene significantly enhanced the proliferation of ONS76 cells. (**p<0.01) I. Overexpression of the short isoform of CD99 transgene did not alter the proliferation rate of ONS76 cells. (ns = not significant) J. Expression of CD99 by flow cytometry in cultured DMG cell lines including (i) DMG H3 wild type lines, VUMC-DIPG10 and SU-DIPG48, (ii) H3K27M mutant lines, SU-DIPGXIII, SU-DIPG04, HSJD-DIPG007, (iii) a primary cell line, MAF-001, and (iv) NHA. K. (i) Violin-plot showing the expression of CD99 transcripts at single cell level in each of the four DMG tumor cohorts collected at the Children’s Hospital, Colorado. (ii) Corresponding Uniform Manifold Approximation and Projection (UMAP) plot showing the expression of CD99 transcript at single cell level. L. IHC for Normal human Tissue Array (FDA999-1) stained for the expression of CD99 and the adjacent table gives a brief description of the tissues in the array collected from respective organs (Refer to https://usbiolab.com/tissue-array/product/multiple-organ/FDA999-1 for the list of tissues on this array and absence of the H&E staining) Data represented as Mean ± SEM.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Expressing, Immunohistochemistry, Mutagenesis, Staining, Cell Culture, Western Blot, Flow Cytometry, Over Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. CD99 mRNA expression in shRNA-mediated knockdown of CD99 in (i) SU-DIPG04 and (ii) BT245 cells. B. CD99 protein expression in the above cells by flow cytometry. C. shRNA-mediated knockdown of CD99 in BT245 cells, with different constructs, slows cell proliferation as measured as phase confluency using incucyte. (*p<0.05) D. shRNA-mediated knockdown of CD99 enhances the differentiation in SU-DIPG04 cells. (i) IF images of (a) control shNull and (b) shCD99 cells. The nuclei are stained with DAPI (blue) and Nestin (green). Scale bar = 50μm. (ii) mRNA expression of Nestin and Olig2 in SU-DIPG04 after shRNA-mediated knockdown of CD99 by RT-qPCR. (*p<0.05, ****p<0.0001) E. Self-renewal and differentiation of SU-DIPG04 increased after the shRNA-mediated knockdown of CD99 as measured by the Aldefluor assay. (i) Representative flow plots. (ii) Quantification of the mean fluorescence intensity of the percent aldehyde dehydrogenase-bright cells (n=3). (**p<0.01) F. CD99 mRNA expression in Crispr-CAS9 deletion of CD99 in SU-DIPGXIII cells by RT-qPCR. (***p<0.001) G. CD99 expression in Crispr-CAS9 deletion of CD99 in SU-DIPGXIII cells by flow cytometry. H. Establishment of the CD99-sufficient (shNull) and CD99-deficient (shCD99) tumors in mouse xenograft models. (i) Bioluminescence as log 10 (Total Flux) values (photons/second/cm 2 /steradian) showing changes in tumor burden in the CD99-sufficient and CD99-deficient tumor groups. (ii) Cleaved caspase3 and (iii) GFAP quantified from n>5 different high-power fields from the stained IHC regions obtained in (v) from n=2 mice for each group. (***p<0.001) Data represented as Mean ± SEM.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Expressing, shRNA, Knockdown, Flow Cytometry, Construct, Control, Staining, Quantitative RT-PCR, Fluorescence, CRISPR

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. shRNA-mediated depletion of CD99 in SU-DIPG04 cells, using different constructs, showed a decrease in their proliferation as measured by xCELLigence. (**p<0.01) B. Gene ontology analysis found that several genes that were downregulated in HSJD-GBM01+K27M were also upregulated in SU-DIPG04 shCD99 #1. C. Metascape analysis of upregulated genes in SU-DIPG04 shCD99 #1 shows that these genes are involved in critical neuronal pathways including neuron differentiation, synaptic assembly and development, and cytokine production. D. Neurosphere growth was diminished by shRNA-mediated knockdown of CD99 (a) SU-DIPG04 and (b) BT245. (i) Representative images of neurosphere from incucyte. The adjacent graph shows the quantification of neurosphere size over time. (ii) shRNA-mediated knockdown of CD99 in DMG cells increased the mRNA expression of the differentiation marker, GFAP. (**p<0.01, ***p<0.001) E. (i) Representative IF images show that shRNA-mediated knockdown of CD99 in SU-DIPG04 enhances their differentiation as measured by an increase in GFAP. The nuclei are stained with DAPI (blue), CD99 (green), and GFAP (red). (ii) The quantification of CD99 and GFAP from at least five high-power fields of images. F. Crispr-CAS9 deletion of CD99 in SU-DIPGXIII cells slowed neurosphere growth and increased differentiation. (i) Representative neurosphere images from incucyte. The adjacent graph shows the quantification of neurosphere size over time. (ii) Increase in the mRNA expression of the differentiation marker, GFAP. (*p<0.05) G. shRNA-mediated depletion of CD99 in BT245-Luc2-GFP delayed the tumor initiation and establishment in vivo . (i) Representative in vivo bioluminescence images (BLI) of the Luc2-expressing DMG tumor cells after injection of the DMG tumor cells in the pons. The scale bar adjacent to the image displays bioluminescence counts (photons/second/cm 2 /steradian). (ii) Kaplan-Meier survival analysis of mice engrafted with CD99-sufficient (shNull, n = 8) and CD99-deficient (shCD99, n = 8) DMG cells. Log-rank (Mantel–Cox) test was used to calculate statistical significance (*p=0.0011). Expression of CD99 mRNA (iii) and GFAP mRNA (iv) in mice tumors from (ii) collected at end-point by RT-qPCR. (v) Representative IHC slides staining H&E and cleaved caspase3 and GFAP in tumors collected at the endpoint. Data shown as the mean ± SEM. (See also and Tables S4 and S5 )

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: shRNA, Construct, Knockdown, Expressing, Marker, Staining, CRISPR, In Vivo, Injection, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. Binding affinity of clinically relevant antibodies and 10D1. B. 10D1 selectively binds to the long isoform (32kDa) of CD99 in DMG and normal cells. C. Blocking of CD99 in SF8628 using the 10D1 antibody followed by the detection with CD99-0662 antibody shows that 10D1 effectively blocks CD99 even at low concentrations. D. CD99 protein expression in NHA and induced pluripotent stem cells (iC7-2) by flow cytometry. E. 10D1, at different concentrations, did not inhibit the growth of (i) NHA and (ii) iC7-2 cells measured using xCELLigence. F. CD99 protein expression in NHA and iC7-2 by western blotting. G. Dosing curve of cpd22 representing the percent viable cells against the log concentration of cpd22 for (i) SU-DIPG04 and (ii) HSJD-DIPG007 cell lines.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Binding Assay, Blocking Assay, Expressing, Flow Cytometry, Western Blot, Concentration Assay

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. Volcano plot showing the statistical significance as -log 10 (p-value) plotted against the log 2 (fold change) between the genes expressed by HSJD-GBM01 transduced with K27M mutant at the H3.3 locus and the unmodified HSJD-GBM01 (control) cells. HSJD-GBM01 transduced with K27M mutant demonstrated an increase in Integrin-linked kinase 1 (ILK1, indicated with blue arrow) in comparison to the control cells. B. ILK1 mRNA expression in (i) HSJD-GBM01, (ii) VUMC-DIPG10, and (iii) NHA, transduced with H3.3K27M. (ns = not significant, ***p<0.001, ****p<0.0001) C. ILK1 protein, similar to CD99, is highly expressed in DMG patient tumors as compared to normal pons ( C ) and cultured DMG cell lines as compared to normal pons ( D ) by western blotting. E. Western blot showing shRNA-mediated knockdown of CD99 in BT245 cells decreased ILK1. F. Western blot showing Crispr-CAS9-mediated deletion of H3.3K27M in BT245 cells decreased ILK1. G. Coimmunoprecipitation assay showed consistent and modest binding of ILK1 and CD99 proteins in (i) SU-DIPG04, (ii) HSJD-DIPG007, and (iii) SU-DIPGXIII using either pulldown antibodies for CD99 and ILK1. H. The changes in IC 50 values of the ILK1 inhibitor, cpd22, in CD99-modulated DMG cell lines show that CD99 is an upstream regulator of ILK1 as measured by the MTS assay. Data shown as the mean ± SEM. (See also and Table S4 )

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Transduction, Mutagenesis, Control, Comparison, Expressing, Cell Culture, Western Blot, shRNA, Knockdown, CRISPR, Co-Immunoprecipitation Assay, Binding Assay, MTS Assay

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. Dosing curve for BT245 H3.3K27M parental cell line. B. Dosing curve for SU-DIPG04 shNull (non-target control) cell line. C. Dosing curve for HSJD-DIPG007 shNull (non-target control) cell line. D. Dosing curve for BT245 H3.3K27M with Crispr-CAS9 deletion of K27M mutation cell line. E. Dosing curve for SU-DIPG04 shCD99 cell line. F. Dosing curve for HSJD-DIPG007 shCD99 cell line. G. Dosing curve for BT245 shNull (non-target control) cell line. H. Dosing curve for SU-DIPGXIII parental (control) cell line. I. Dosing curve for BT245 shCD99 cell line. J. Dosing curve for SU-DIPGXIII with Crispr-CAS9 deletion of CD99 cell line.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Control, CRISPR, Mutagenesis

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A. Schematic showing the human CD99 protein and the approximate location of the 10D1 antibody epitope compared to some of the commercially available CD99 antibody binding epitopes. B. Kinetics of peptide antigen interacting with 10D1 human IgG4/light chain1 using SA biosensor. OctetRED 384 monitored the association and dissociation of this peptide antigen. C. Flow plot showing the detection of CD99 in cultured DMG cells using the 10D1. D. Representative IHC image for staining of CD99 in SU-DIPGXIII cells using the 10D1. E. Detection of CD99 (long isoform) by the 10D1 antibody in DMG patient tumors and normal pons. F. 10D1 significantly blocked CD99 in DMG cells as detected by CD99-0662 antibody even at its low concentrations. G. 10D1 significantly inhibits the proliferation of (i) a DMG tumor line, SU-DIPG04, and (ii) a primary cultured cell line, MAF-001 as measured by xCELLigence. (**p<0.01, ***p<0.001) H. 10D1 significantly induces apoptosis in (i) SU-DIPG04, (ii) BT245, and (iii) SU-DIPGXIII, as measured by the green cleaved caspase3/7 activity on an incucyte. The number of green cells increased significantly 24hrs and 48hrs after treatment. (ns = not significant. *p<0.05, **p<0.01) Data shown as the mean ± SEM. (See also )

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Binding Assay, Cell Culture, Staining, Activity Assay

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A: Representative flow cytometry plots showing the expression of CD99 at 24hrs after (a) BT245, and (b) SU-DIPGXIII cells were exposed to a single dose of RT of 2Gy or 4Gy or 6Gy. B: Representative flow cytometry plots showing no apparent changes in the expression of CD99 in NHA at 24hrs after they were exposed to FFRT of 2Gy or 4Gy dose for 3 consecutive days. C: Quantification of mean fluorescence intensity representing the expression of CD99 after cells were exposed to FFRT with 2Gy or 4Gy dose for 3 consecutive days; (a) NHA from , (b) BT245 from (ii) and (c) SU-DIPGXIII from (ii). (ns = not significant, **p<0.01, ***p<0.001) D : BT245 and G: MAF-001 DMG models and their individual BLI of mice in each treatment cohort for different days. E & H : Analysis of BLI for the 10D1 and IgG treatment groups. F : Relative body weight of BT245 model in both treatment groups. I : CBC analysis from the blood collected immediately after animals were euthanized upon reaching their endpoint. Samples were analyzed for WBC (leukocytes), hemoglobin, neutrophils, lymphocytes, and platelets count. Data represented as the mean ± SEM.

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: Flow Cytometry, Expressing, Fluorescence

Journal: bioRxiv

Article Title: Targeting Diffuse Midline Glioma with a novel anti-CD99 Antibody

doi: 10.1101/2024.03.19.585814

Figure Lengend Snippet: A-B: RT increases CD99 in DMG cells in vitro . A. Representative flow cytometry plots showing the expression of CD99 at 24hrs after they were exposed to RT of 2Gy or 4Gy dose for three consecutive days in (i) SF8628, (ii) BT245, and (iii) SU-DIPGXIII, (quantitation of CD99 expression is given in ). B. A representative western blot shows the increased CD99 protein (only the long form) after SF8628 cells were exposed to RT with a 4Gy dose for three consecutive days. C-E: IT delivered 10D1, followed by focal fractionated RT (FFRT) increased the anti-tumor latency of RT in the BT245-Luc2 in vivo model . C. Schematics representing the treatment protocol followed in the BT245 model. D. Representative BLI images of mice after tumor implantation, antibody treatment, and RT. The adjacent plot shows the flux values comparison on day 27 between the control and treatment groups (**p<0.01). E. Kaplan Meier survival curve shows the increase in survival in BT245 tumor-bearing mice treated with a single IT dose of 10D1 followed by RT in comparison to that of control-treated mice. Log-rank (Mantel–Cox) test was used to compare groups (***p<0.0005). n = 5 to 6 mice per group. The median survival of the mice is given in the table below. Dotted lines in the survival curves indicate intrathecal alone treatment. F-H In vivo effect of intrathecally delivered 10D1, followed by FFRT in MAF-001-Luc2 model . F. Schematics representing the protocol followed in the MAF-001 PDX model. G. Representative BLI of mice xenografted with luciferase-expressing MAF-001 before and after 10D1 treatment, followed by RT. The adjacent plot shows the flux values comparison on day 47 between the control and treatment groups (**p<0.05). H. Kaplan Meier survival curve shows the increase in survival in MAF-001 tumor-bearing mice treated with a single IT dose of 10D1 followed by RT in comparison to that of control-treated mice. Log-rank (Mantel–Cox) test was used to compare groups (**p=0.0018). n = 5 mice per group. The median survival of the mice is given in the table below. Dotted lines in the survival curves indicate intrathecal alone treatment. Data represented as the mean ± SEM. (Also see )

Article Snippet: The novel chimeric anti-CD99 antibody was generated as human IgG4 molecules under contract with GenScript (Piscataway, MJ).

Techniques: In Vitro, Flow Cytometry, Expressing, Quantitation Assay, Western Blot, In Vivo, Tumor Implantation, Comparison, Control, Luciferase