αcd4 gk1, by BioXCell - Product details

1

Conditioning for Hematopoietic Cell Transplantation

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A graphical timeline of conditioning is shown in Figure 1B. Mice were given 500μg Diphenhydramine HCl i.p. approximately 10–15 min prior to αCD117. 500μg αCD117 was injected retro-orbitally into mice under isoflurane anesthesia on day −6 prior to HCT. Mice were irradiated on day −3 with 200 cGy (Figures 1 and 2) or 300 cGy (Figures 3 and 4) TBI. 300μg each of αCD4 and αCD8 was administered i.p. on days −2, −1, and 0. αCD117 mAb, ACK2 was purchased from Bio X Cell (Lebanon, NH) or BioLegend (San Diego, CA). αCD4 (GK1.5) and αCD8 (YTS169.4) were purchased from Bio X Cell. Diphenhydramine HCl was purchased from MedChem Express (Monmouth Junction, NJ).
Animal irradiation (XRT) was performed using a Kimtron Polaris IC-250 Biological Irradiator (Oxford, CT) with a 225 kV X-ray tube filtered by 0.5mm Cu source set at 225kV, 13.3mA. Mice were divided in irradiation pie cages from Braintree Scientific (Braintree, MA) when irradiated. Dosimetry calibration for our setup was performed using published methods on radiochromic film (Ma et al., 2001 (link)).

Chang C.A., Bhagchandani P., Poyser J., Velasco B.J., Zhao W., Kwon H.S., Meyer E., Shizuru J.A, & Kim S.K. (2022). Curative islet and hematopoietic cell transplantation in diabetic mice without toxic bone marrow conditioning. Cell reports, 41(6), 111615.

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2

Blocking Antibodies Enhance Influenza Immunity

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For blocking antibodies, mice were injected i.p. with 250µg of α−CD4 (GK1.5, BioXcell) or 200µg of α-IFNAR (MAR1.5A3, BioXCell) one day before receiving 100 PFU of BrightFlu followed by a second dose at day 4 post infection. For depleting antibodies, mice were injected i.p. with 250µg of α-Ly-6G (1A8, Biolegend) daily for 5 days starting from 1 day prior to BrightFlu. Poly(I:C) was injected intratumorally with 10µg of Poly(I:C)-Rhodamine in 10µl PBS. For intracellular IL-12 staining, mice were injected i.v. with 250µg of Brefeldin A (Invivogen) and tissues harvested 6hr post injection.

Pirillo C., Al Khalidi S., Sims A., Devlin R., Zhao H., Pinto R., Jasim S., Shearer P., Shergold A., Donnelly H., Bravo-Blas A., Loney C., Perona-Wright G., Hutchinson E, & Roberts E. (2023). Co-transfer of antigen and contextual information harmonises peripheral and lymph node cDC activation. Science immunology, 8(85), eadg8249.

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3

Immunotherapy Regimen for Preclinical Cancer

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All antibody dosing was performed via intraperitoneal injection in PBS. αCD4 (GK1.5, BioXCell) and αCD8 (2.43, BioXCell) depleting antibodies were administered at 200 ug every 4 days. αPD-1 (29F.1A12, BioXCell) was administered at 200 μg three times a week. αCTLA (9H10, BioXCell) was administered at an initial dose of 200 μg, with all subsequent doses at 100 μg, three times a week. αCD40 (FGK4.5, BioXCell) was administered once at the beginning of treatment at 100 μg.
The adjuvant amphiphile-CpG (amph-CpG) and antigen amphiphile (amph-peptide) were produced as previously described^{53 (link)}. Briefly, class B CpG 1826 oligonucleotide with a G₂ spacer (5’-diacyl lipid-GGTCCATGACGTTCCTGACGTT- 3’) was conjugated via the 5’ end to an 18 carbon diacyl tail. Antigen peptide OVA_250–270 (CGLEQLESIINFEKLTEWTSS) and non-specific mutant gp100_20–39 (optimized S27P, EGP long^{52 (link)}, CAVGALEGPRNQDWLGVPRQL) were conjugated via N’ cysteine residue to 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethyleneglycol-2000] (Avanti Polar Lipids). Mice were vaccinated subcutaneously at the base of the tail with 1.24 nmol amph-CpG and 25 μg of amph-peptide, with half dose given to each side. Vaccination was performed once weekly starting 14 days post-transplant of lo^SIIN organoids.

Westcott P.M., Sacks N.J., Schenkel J.M., Ely Z.A., Smith O., Hauck H., Jaeger A.M., Zhang D., Backlund C.M., Beytagh M.C., Patten J., Elbashir R., Eng G., Irvine D.J., Yilmaz O.H, & Jacks T. (2021). Low neoantigen expression and poor T-cell priming underlie early immune escape in colorectal cancer. Nature cancer, 2(10), 1071-1085.

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4

T-cell Depletion Alters TRAMP-C2 Tumor Growth

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To determine T-cell mediated growth of TRAMP-C2 tumors, T-cell depletion experiment was performed. WT and Ack1 KO male mice (6–8 weeks) were injected intraperitoneally with 250 μg/mouse αCD4 (GK1.5, BioXcell) or 250 μg/mouse αCD8β (53-5.8, BioXcell) or IgG (control). Three days post the antibody injection, mice were subcutaneously implanted with 1.5 × 10⁶ TRAMP-C2 cells that were suspended in 200 µl of PBS with 50% matrigel (BD Biosciences). Following tumor implantation, mice were injected with the above antibodies intraperitoneally, once a week, for 5 weeks. Tumor growth was monitored and measured with calipers. At the end of the treatment period, all mice were humanely euthanized by carbon dioxide inhalation, followed by cervical dislocation. Tumors were extracted and weighed.

Sridaran D., Chouhan S., Mahajan K., Renganathan A., Weimholt C., Bhagwat S., Reimers M., Kim E.H., Thakur M.K., Saeed M.A., Pachynski R.K., Seeliger M.A., Miller W.T., Feng F.Y, & Mahajan N.P. (2022). Inhibiting ACK1-mediated phosphorylation of C-terminal Src kinase counteracts prostate cancer immune checkpoint blockade resistance. Nature Communications, 13, 6929.

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5

LCMV infection and T cell modulation

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Mice were infected with 4×10⁶ PFU LCMV Clone 13 i.v., monitored for weight loss, and bled or sacrificed at day 8, 15, 22 or 30 post infection for flow cytometry analyses. To deplete CD4⁺ T cells, mice were injected i.p. with 200 μg αCD4 (GK1.5, BioXCell Cat# BE0003–1) on days −1 and 1 (relative to LCMV Clone 13 injection on day 0). To block IFNAR1, mice were injected i.p. with 1 mg αIFNAR (MAR1–5A3, BioXCell Cat# BE0241) or isotype (MOPC-21, BioXCell Cat# BE0083) on days −1 and 1 (relative to LCMV Clone 13 injection on day 0). To block IFN-γ, mice were injected i.p. daily with 250 μg αIFN-γ (XMG1.2 BioXCell Cat# BE0055) or isotype (HRPN BioXCell Cat# BE0088) on days 0–3 (relative to LCMV Clone 13 injection on day 0).

LaFleur M.W., Nguyen T.H., Coxe M.A., Miller B.C., Yates K.B., Gillis J.E., Sen D.R., Gaudiano E.F., Abosy R.A., Freeman G.J., Haining W.N, & Sharpe A.H. (2019). PTPN2 regulates the generation of exhausted CD8+ T cell subpopulations and restrains tumor immunity. Nature immunology, 20(10), 1335-1347.

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6

Immunomodulation in Orthotopic Tumors

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To deplete CD4⁺ or CD8⁺ T cells, Batf3^–/– mice bearing CB⁺ orthotopic tumors were treated with 200 μg of αCD4 (GK1.5, BioXcell) i.p. on days –1, 1, 4, and/or 200 μg of αCD8 (2.43, BioXcell) i.p. on days –1, 5, and 10. A total of 400 μg of αCD20 (SA271G2, BioLegend) on day –1 relative to tumor implantation was used to deplete B cells. A total of 400 μg of αCSF1R (AFS98, BioXcell), 200 μg of αGr-1 (RB6-8C5, BioXcell), or 200–250 μL of control liposomes or clodronate-loaded liposomes (Encapsula NanoSciences) were administered i.p. on days –1, 1, 3, 5, 7, 9, and 11 relative to orthotopic tumor implantation.

Burrack A.L., Schmiechen Z.C., Patterson M.T., Miller E.A., Spartz E.J., Rollins M.R., Raynor J.F., Mitchell J.S., Kaisho T., Fife B.T, & Stromnes I.M. (2022). Distinct myeloid antigen-presenting cells dictate differential fates of tumor-specific CD8+ T cells in pancreatic cancer. JCI Insight, 7(7), e151593.

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7

Conditioning for Hematopoietic Cell Transplantation

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A graphical timeline of conditioning is shown in Figure 1B. Mice were given 500μg Diphenhydramine HCl i.p. approximately 10–15 min prior to αCD117. 500μg αCD117 was injected retro-orbitally into mice under isoflurane anesthesia on day −6 prior to HCT. Mice were irradiated on day −3 with 200 cGy (Figures 1 and 2) or 300 cGy (Figures 3 and 4) TBI. 300μg each of αCD4 and αCD8 was administered i.p. on days −2, −1, and 0. αCD117 mAb, ACK2 was purchased from Bio X Cell (Lebanon, NH) or BioLegend (San Diego, CA). αCD4 (GK1.5) and αCD8 (YTS169.4) were purchased from Bio X Cell. Diphenhydramine HCl was purchased from MedChem Express (Monmouth Junction, NJ).
Animal irradiation (XRT) was performed using a Kimtron Polaris IC-250 Biological Irradiator (Oxford, CT) with a 225 kV X-ray tube filtered by 0.5mm Cu source set at 225kV, 13.3mA. Mice were divided in irradiation pie cages from Braintree Scientific (Braintree, MA) when irradiated. Dosimetry calibration for our setup was performed using published methods on radiochromic film (Ma et al., 2001 (link)).

Chang C.A., Bhagchandani P., Poyser J., Velasco B.J., Zhao W., Kwon H.S., Meyer E., Shizuru J.A, & Kim S.K. (2022). Curative islet and hematopoietic cell transplantation in diabetic mice without toxic bone marrow conditioning. Cell reports, 41(6), 111615.

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8

LCMV infection and T cell modulation

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Mice were infected with 4×10⁶ PFU LCMV Clone 13 i.v., monitored for weight loss, and bled or sacrificed at day 8, 15, 22 or 30 post infection for flow cytometry analyses. To deplete CD4⁺ T cells, mice were injected i.p. with 200 μg αCD4 (GK1.5, BioXCell Cat# BE0003–1) on days −1 and 1 (relative to LCMV Clone 13 injection on day 0). To block IFNAR1, mice were injected i.p. with 1 mg αIFNAR (MAR1–5A3, BioXCell Cat# BE0241) or isotype (MOPC-21, BioXCell Cat# BE0083) on days −1 and 1 (relative to LCMV Clone 13 injection on day 0). To block IFN-γ, mice were injected i.p. daily with 250 μg αIFN-γ (XMG1.2 BioXCell Cat# BE0055) or isotype (HRPN BioXCell Cat# BE0088) on days 0–3 (relative to LCMV Clone 13 injection on day 0).

LaFleur M.W., Nguyen T.H., Coxe M.A., Miller B.C., Yates K.B., Gillis J.E., Sen D.R., Gaudiano E.F., Abosy R.A., Freeman G.J., Haining W.N, & Sharpe A.H. (2019). PTPN2 regulates the generation of exhausted CD8+ T cell subpopulations and restrains tumor immunity. Nature immunology, 20(10), 1335-1347.

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9

Immunotherapy Regimen for Preclinical Cancer

Cited 1 time

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All antibody dosing was performed via intraperitoneal injection in PBS. αCD4 (GK1.5, BioXCell) and αCD8 (2.43, BioXCell) depleting antibodies were administered at 200 ug every 4 days. αPD-1 (29F.1A12, BioXCell) was administered at 200 μg three times a week. αCTLA (9H10, BioXCell) was administered at an initial dose of 200 μg, with all subsequent doses at 100 μg, three times a week. αCD40 (FGK4.5, BioXCell) was administered once at the beginning of treatment at 100 μg.
The adjuvant amphiphile-CpG (amph-CpG) and antigen amphiphile (amph-peptide) were produced as previously described^{53 (link)}. Briefly, class B CpG 1826 oligonucleotide with a G₂ spacer (5’-diacyl lipid-GGTCCATGACGTTCCTGACGTT- 3’) was conjugated via the 5’ end to an 18 carbon diacyl tail. Antigen peptide OVA_250–270 (CGLEQLESIINFEKLTEWTSS) and non-specific mutant gp100_20–39 (optimized S27P, EGP long^{52 (link)}, CAVGALEGPRNQDWLGVPRQL) were conjugated via N’ cysteine residue to 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethyleneglycol-2000] (Avanti Polar Lipids). Mice were vaccinated subcutaneously at the base of the tail with 1.24 nmol amph-CpG and 25 μg of amph-peptide, with half dose given to each side. Vaccination was performed once weekly starting 14 days post-transplant of lo^SIIN organoids.

Westcott P.M., Sacks N.J., Schenkel J.M., Ely Z.A., Smith O., Hauck H., Jaeger A.M., Zhang D., Backlund C.M., Beytagh M.C., Patten J., Elbashir R., Eng G., Irvine D.J., Yilmaz O.H, & Jacks T. (2021). Low neoantigen expression and poor T-cell priming underlie early immune escape in colorectal cancer. Nature cancer, 2(10), 1071-1085.

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αcd4 gk1

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Close spelling variants of this product

9 protocols using αcd4 gk1

Conditioning for Hematopoietic Cell Transplantation

Blocking Antibodies Enhance Influenza Immunity

Immunotherapy Regimen for Preclinical Cancer

T-cell Depletion Alters TRAMP-C2 Tumor Growth

LCMV infection and T cell modulation

Immunomodulation in Orthotopic Tumors

Conditioning for Hematopoietic Cell Transplantation

LCMV infection and T cell modulation

Immunotherapy Regimen for Preclinical Cancer

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