- CD0329 - Invitrogen Antibodies CD247 antibody

Submitted references Tumor organoid-T-cell coculture systems.

Cattaneo CM, Dijkstra KK, Fanchi LF, Kelderman S, Kaing S, van Rooij N, van den Brink S, Schumacher TN, Voest EE
Nature protocols 2020 Jan;15(1):15-39

Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids.

Dijkstra KK, Cattaneo CM, Weeber F, Chalabi M, van de Haar J, Fanchi LF, Slagter M, van der Velden DL, Kaing S, Kelderman S, van Rooij N, van Leerdam ME, Depla A, Smit EF, Hartemink KJ, de Groot R, Wolkers MC, Sachs N, Snaebjornsson P, Monkhorst K, Haanen J, Clevers H, Schumacher TN, Voest EE
Cell 2018 Sep 6;174(6):1586-1598.e12

Tumor specific regulatory T cells in the bone marrow of breast cancer patients selectively upregulate the emigration receptor S1P1.

Rathinasamy A, Domschke C, Ge Y, Böhm HH, Dettling S, Jansen D, Lasitschka F, Umansky L, Gräler MH, Hartmann J, Herold-Mende C, Schuetz F, Beckhove P
Cancer immunology, immunotherapy : CII 2017 May;66(5):593-603

HLA Class II tetramers reveal tissue-specific regulatory T cells that suppress T-cell responses in breast carcinoma patients.

Schmidt HH, Ge Y, Hartmann FJ, Conrad H, Klug F, Nittel S, Bernhard H, Domschke C, Schuetz F, Sohn C, Beckhove P
Oncoimmunology 2013 Jun 1;2(6):e24962

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Human PBMCs stained with: CD45 Pacific Orange™ (Product # MHCD4530TR), CD4 PerCP-Cy®5.5 (Product # A15858), CD20 APC, CD14 APC-Cy®7 (Product # A15453), CD19 Pacific Green™ (Product # C11210), CD3 Alexa Fluor® 700 (CD0329), HLA-DR PE-Cy®7 (Product # A18558), CD8 Pacific Blue™ (Product # MHCD0828), CD33 FITC (Product # A16185), and CD11c PE (Product # A18674) using the Attune® NxT Acoustic Focusing Cytometer with 405 nm excitation and 440/50 emission filter (Pacific Blue™), 512/25 emission filter (Pacific Green™), and 603/48 emission filter (Pacific Orange™); 488 nm excitation and 530/30 emission filter (FITC) and 695/50 emission filter (PerCP-Cy®5.5); 561 nm excitation and 585/16 emission filter (PE) and 780/60 emission filter (PE-Cy®7); 637 nm excitation and 660/20 emission filter (APC), 720/30 emission filter (Alexa Fluor®700), and 780/60 emission filter (APC-Cy®7). Within the CD3 negative CD45 positive gate, B cells can be identified based on expression of CD19 and CD20. Conventional dendritic cells in peripheral blood are generally negative for T and B cell lineage markers and co-express the integrin CD11c and HLA-DR. Values noted are a percent of the parent CD19 negative CD20 negative gate (top value, no parenthesis) or percent of FSC/SSC lymphocyte gate (bottom value, in parentheses).
Conjugate: Near infrared dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Human PBMCs stained with: CD45 Pacific Orange™ (Product # MHCD4530TR), CD4 PerCP-Cy®5.5 (Product # A15858), CD20 APC, CD14 APC-Cy®7 (Product # A15453), CD19 Pacific Green™ (Product # C11210), CD3 Alexa Fluor® 700 (CD0329), HLA-DR PE-Cy®7 (Product # A18558), CD8 Pacific Blue™ (Product # MHCD0828), CD33 FITC (Product # A16185), and CD11c PE (Product # A18674) using the Attune® NxT Acoustic Focusing Cytometer with 405 nm excitation and 440/50 emission filter (Pacific Blue™), 512/25 emission filter (Pacific Green™), and 603/48 emission filter (Pacific Orange™); 488 nm excitation and 530/30 emission filter (FITC) and 695/50 emission filter (PerCP-Cy®5.5); 561 nm excitation and 585/16 emission filter (PE) and 780/60 emission filter (PE-Cy®7); 637 nm excitation and 660/20 emission filter (APC), 720/30 emission filter (Alexa Fluor®700), and 780/60 emission filter (APC-Cy®7). Within the CD3 negative CD45 positive gate, B cells can be identified based on expression of CD19 and CD20. Conventional dendritic cells in peripheral blood are generally negative for T and B cell lineage markers and co-express the integrin CD11c and HLA-DR. Values noted are a percent of the parent CD19 negative CD20 negative gate (top value, no parenthesis) or percent of FSC/SSC lymphocyte gate (bottom value, in parentheses).
Conjugate: Near infrared dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Decreased frequencies of Treg subpopulations in the BM of breast cancer patients. a Representative plot of a healthy donor showing the CD4+ T-cell gating strategy. b-c CD4+ cells were analyzed for FoxP3 and CD25 expression. CD25+ FoxP3+ cells were gated as Treg, and CD25- FoxP3- cells were gated as Tcon. Representative plots illustrating Treg frequencies in BM and PB of a healthy donor ( b ) and a breast cancer patient ( c ). Red circle and square represents the CD4 gate and CD25+ FoxP3+ Treg gate, respectively. d Cumulative data of CD25+ FoxP3+ Treg frequencies in PB and BM of all patients and healthy donors analyzed--Healthy donor PB ( n = 7) and BM ( n = 8), patients with matched PB and BM samples ( n = 50). Data distribution in 1 d is represented by mean with SEM. For healthy donors unpaired t test and for patient samples, paired t test was used for statistical analysis. Epigenetic PCR was performed on DNA isolated from tumor areas from FFPE sections obtained from 42 patients. Samples that passed quality control [Treg ( n = 33) and CD3 ( n = 40)] were taken into analysis. e, f Treg and CD3 T-cell percentage ( e ) and counts per mm 3 volume of tumor ( f ). Data distribution is represented by median with interquartile range in e and f. g Graph bridging data of Treg frequencies in BM, PB, and tumor--ratio of Treg in BM to PB correlated to ratio of Treg in tumor to PB [for all patients with Treg frequencies >4.6% in PB ( n = 18)]. Non-parametric Spearman Correlation wa
Conjugate: Near infrared dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. Tetramer staining of conventional and regulatory T cells from breast cancer patients. ( A-G ) Peripheral blood mononuclear cells (PBMCs) of a breast cancer patient were analyzed, upon gating on living CD3 + CD4 + T cells. The patient sample was stained with tetramers presenting either mam 34-48 ( B ) or the CLIP peptide ( C ). Numbers indicate the percentage of cells in the respective gate, referring to lymphocytes ( A ) or CD3 + CD4 + T cells ( B ) and ( C ). Within CD3 + CD4 + T cells, regulatory T cells (Tregs) were identified as CD25 high CD127 low ( D ) and ( G ) and CD25 high CD127 low FOXP3 + ( E ), ( F ) and ( H ). ( G ) reports the percentage of Tregs within tetramer-positive cells in a representative patient. ( I ) Frequency of mam 34-48 - and CLIP-presenting tetramer-positive cells among CD3 + CD4 + T cells of breast cancer patients and healthy donors (HD) (p values as per Mann-Whitney U tests are indicated). ( J and K ) Frequency of mam 34-48 -specific Tcon ( J ) cells and Tregs ( K ) of breast cancer patients and HDs.
Conjugate: Near infrared dye

CD0329

Antibody data