Background
Expression of the immunostimulatory xenoantigen alphaGal on malignant cells is being investigated as a means to formulate anticancer vaccines. Expression
Conclusion
Our method provides an effective, highly reproducible means to efficiently express alphaGal antigens on cells obtained from patients with a spectrum of hematological malignancies. This method can provide a simple, safe alternative to viral-mediated gene transfer or enzymatic alteration to express alphaGal antigens on human tumor cells. By virtue of its simplicity, our technique presents a novel approach to the preparation of polyvalent autologous or syngeneic anticancer vaccines.
Methods
Freshly isolated white blood cells (WBC) were obtained from patients with malignant hematological disease and combined with diluted PBC. Cell mixtures were labeled with human CD mAbs, followed by IB4 lectin or M86 mAb to detect alphaGal antigens and then co-incubated with PEG. Back-gated, dual-color flow cytometry was used to detect alphaGal on human cells.
Results
alphaGal antigens were detected on sizeable numbers of human WBC (approximately 45%) after incubation with PEG. Antigen expression was profuse as assessed by the strong fluorescent intensity demonstrated by IB4-FITC and M86 labeling. Human cells combined with PBC without PEG were not reactive with IB4-FITC or M86.