Intra-arterial Versus Intravenous Adoptive Cell Therapy in a Mouse Tumor Model

Abstract

Adoptive cell transfer therapy for cancer has existed for decades and is experiencing a resurgence in popularity that has been facilitated by improved methods of production, techniques for genetic modification, and host preconditioning. The trafficking of adoptively transferred lymphocytes and infiltration into the tumor microenvironment is sine qua non for successful tumor eradication; however, the paradox of extremely poor trafficking of lymphocytes into the tumor microenvironment raises the issue of how best to deliver these cells to optimize entry into tumor tissue. We examined the route of administration as a potential modifier of both trafficking and antitumor efficacy. Femoral artery cannulation and tail vein injection for the intra-arterial (IA) and IV delivery, respectively, were utilized in the B16-OVA/OT-I mouse model system. Both IV and IA infusions showed decreased tumor growth and prolonged survival. However, although significantly increased T-cell tumor infiltration was observed in IA mice, tumor growth and survival were not improved as compared with IV mice. These studies suggest that IA administration produces increased early lymphocyte trafficking, but a discernable survival benefit was not seen in the murine model examined.

FIGURE 1

A, In patients adoptive transfer via the regional arterial supply of the tumor (right) was hypothesized to increase exposure of adoptively transferred cells (green) to the tumor vasculature and avoid trapping in the lungs as seen with intravenous delivery (left). Adoptively transferred lymphocyte retention in the pulmonary capillaries is multifactorial (inset). B, Experimental design to examine this hypothesis in a murine model of cancer.

FIGURE 2

Retention of adoptively transferred lymphocytes 3 hours after IV injection. 1×10⁷ cells were injected into recipient mice. Blocking integrin interactions before transfer partially abrogates lung retention suggesting both active and passive mechanisms. Transferred cells recovered from subcutaneous tumor only account for 0.4% of total cells recovered (*P<0.05). Ab indicates antibody.

FIGURE 3

A–C, Representative photomicrographs of immunofluorescence images of tumor tissue 24 hours after adoptive transfer with 1×10⁷ green CFSE-labeled cells IV (arrows) and 1×10⁷ orange CTO-labeled cells intra-arterial (arrow heads). DAPI-stained nuclei of endogenous cells within the tumor appear as blue background. CFSE indicates carboxyfluorescein succinimidyl ester; CTO, Cell Tracker Orang; DAPI, 4’,6-diamidino-2-phenylindole. *P<0.05.

FIGURE 4

Percentage of total adoptively transferred cells (A) and total number of adoptively transferred cell/10⁴ total cells (B) identified by flow cytometry in various tissues after ACT. Intra-arterial injection resulted in increased tumor infiltration of T cells and lymph node accumulation at 24 hours (*P<0.05). ACT indicates adoptive cell transfer; LN, lymph node.

FIGURE 5

Treatment of established B16 melanoma tumor with 5×10⁶ T cells given IA (given as a bolus) or IV. IA compared with IV ACT shows no difference in tumor growth no treatment control. However, both forms of ACT show improvement as compared with no treatment control (day 0, tumor inoculation; day 7, T-cell adopted transfer; ***P<0.0001). ACT indicates adoptive cell transfer; IA, intra-arterial.

FIGURE 6

Treatment of established B16 melanoma tumor with various doses of T cells given IA (infusion over 10 min) or IV. IA compared with IV ACT shows no improvement in tumor growth curves at given doses (day 0, tumor inoculation; day 7, T-cell adoptive transfer; n=5). IA indicates intra-arterial.

FIGURE 7

Treatment of established B16 melanoma tumor with 10×106 T cells given IA or IV. IA and IV ACT both significantly improve survival over no treatment control (**P<0.001). However, No difference was seen in IA ACT versus IV ACT (P=0.3). ACT indicates adoptive cell transfer; IA, intra-arterial; ns, not significant.