Anti-Human CTLA4 Recombinant Antibody (Ipilimumab) (CAT#: TAB-067)

Figure 1 Intratumoral blood vessel changes from treatment with bevacizumab plus ipilimumab in subcutaneous melanoma deposits.

A, characterization of tumor-associated vasculature before and while on therapy. Endothelial cells lining small vessels within melanomas of ipilimumab plus bevacizumab–treated patients were rounded and columnar as assessed by hematoxylin and eosin (H&E) and CD31 staining, in contrast with pretreatment samples and on-treatment samples from patients receiving ipilimumab alone (column to extreme right; v, vessel). Scale bar, 50 microns. Endothelial cells in tumor deposits of patients receiving ipilimumab plus bevacizumab were also associated with increased expression of E-selectin, and adhesion and diapedesis of CD8+ T cells. Enlarged central panels highlight the focally occlusive appearance of this endothelial activation [top, H&E; bottom, CD31; inset, E-selectin)]. Base membrane of vessels approximated by dotted lines.

Hodi, F. S., Lawrence, D., Lezcano, C., Wu, X., Zhou, J., Sasada, T.,... & Friedlander, P. (2014). Bevacizumab plus ipilimumab in patients with metastatic melanoma. Cancer immunology research, 2(7), 632-642.

Figure 2 Histologic changes in tumor deposits resulting from treatment with bevacizumab plus ipilimumab.

A, phenotypic characterization of immune-cell infiltrates in biopsies from responders before and after initiation of therapy. Tumors after initiation (ON) of ipilimumab–bevacizumab therapy were characterized as compared with pretreatment samples (PRE). Significant infiltration by CD3+CD8+ T cells and CD163+ macrophages with minimal change in Foxp3+ component was observed. The enlarged panels (center) emphasize the tumor-infiltrating architecture of the immune response (top left, skeletal muscle). In contrast, patients treated only with ipilimumab showed a lesser degree of immune-cell infiltration while on therapy. The two ipilimumab–bevacizumab specimens are subcutaneous tissues, and the ipilimumab-alone specimen was from the oropharyngeal submucosa.

Hodi, F. S., Lawrence, D., Lezcano, C., Wu, X., Zhou, J., Sasada, T.,... & Friedlander, P. (2014). Bevacizumab plus ipilimumab in patients with metastatic melanoma. Cancer immunology research, 2(7), 632-642.

Figure 3 Cellular and humoral immune responses in the peripheral blood are altered by the addition of bevacizumab to ipilimumab.

All samples were obtained pretreatment and at week 12 at the completion of ipilimumab or ipilimumab–bevacizumab induction. A, example of changes as a function of treatment in CD4+CCR7+CD45RO+ and CD4+CCR7−CD45RO+ T-cell populations to ipilimumab plus bevacizumab treatment, compared with changes with ipilimumab treatment alone. B, example of changes as a function of treatment in CD8+CCR7+CD45RO+ and CD8+CCR7−CD45RO+ T-cell populations to ipilimumab plus bevacizumab treatment, compared with the responses to ipilimumab treatment alone. C, number of patients with melanoma who have at least 50% enhancement in CD4+/CD8+ CCR7+CD45RO+ and CD4+/CD8+ CCR7−CD45RO+ T-cell populations following treatment with ipilimumab (3 mg/kg), or ipilimumab (3 mg/kg) plus bevacizumab, or ipilimumab (10 mg/kg) plus bevacizumab. *, P < 0.05 between ipilimumab and ipilimumab plus bevacizumab; **, P < 0.01. D, representative immunoblots of antibody responses in 4 bevacizumab plus ipilimumab–treated patients to a total of zero, one, two, or three galectins. Arrows, increased antibody levels in posttreatment sera samples. Galectin-1, -3, and -9 proteins were mixed together and equally loaded and separated by gel electrophoresis. Following transfer onto a membrane, strips were incubated with equally diluted pretreatment and posttreatment sera. Density analysis using NIH ImageJ software confirmed the increases in densities of the indicated bands after subtracting background taken from nearby areas of each band. As in most of the cases, density change was seen in only one or two of the three galectins; protein(s) without density change served as loading controls. E, antibody responses to galectins in patients treated with bevacizumab plus ipilimumab and ipilimumab alone. Anti-galectin-1, anti-galectin-3, and anti-galectin-9 antibodies were detected in pretreatment and posttreatment patient sera by immunoblot analyses. Percentages of patients with increased levels of antibodies to a total of zero, one, two, or three galectins in bevacizumab plus ipilimumab-treated patients (n = 45) and ipilimumab-alone patients (n = 18). Density of each band was measured using NIH ImageJ software and the antibody fold change was calculated using the formula: fold change = (densityPost − densityPre)/densityPre. Antibody levels were considered as increased when the fold change ≥ 0.5.

Hodi, F. S., Lawrence, D., Lezcano, C., Wu, X., Zhou, J., Sasada, T.,... & Friedlander, P. (2014). Bevacizumab plus ipilimumab in patients with metastatic melanoma. Cancer immunology research, 2(7), 632-642.

Figure 1 Anti-CTLA4 Antibody (TAB-067) in WB

Western blot analysis of TAB-067 was performed with CTLA-4 Protein, Human, Recombinant (His Tag).

TAB-067 incubation concentration: 2ng/μL.
The secondary antibody: HRP-Anti-Human IgG (H+L)
Lane 1: Reducing Antigen (0.1μg)
Lane 2: Reducing Antigen (0.3μg)
Lane 3: Reducing Antigen (0.6μg)

Figure 2 Anti-CTLA4 Antibody (TAB-067) in ELISA

ELISA analysis of TAB-067 was performed by coating with CTLA-4 Protein, Human, Recombinant (His Tag).

The secondary antibody: HRP-Anti-Human IgG (H+L)