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Lymph node transfer improves immune function and decreases swelling in a transgenic mouse model of lymphedema
Gabriela D. Garcia Nores, MD, Jung-Ju Huang, MD, Walter J. Joseph, MD, Jason C. Gardenier, MD, Seth Z. Aschen, MD, Daniel A. Cuzzone, MD, Geoffrey E. Hespe, BS, Raghu P. Kataru, PhD, Babak J. Mehrara, MD.
Memorial Sloan Kettering Cancer Center, New York, NY, USA.

BACKGROUND: Lymphedema is a morbid complication commonly encountered after lymphadenectomy for cancer treatment and affects almost 6 million Americans. Complications of lymphedema include recurrent infections, poor wound healing and decreased antibody responses in the affected limb. Lymph node transfer (LNT) has emerged as a promising therapy for this disease but little is known of its effectiveness in restoring immune responses or improving lymphedema outcomes. The purpose of this study was therefore to analyze lymphatic and immune function after LNT. METHODS: Selective lymphatic ablation was achieved in a transgenic mouse model though diphtheria toxin injection in the hindlimb or forepaw. All animals underwent ablation followed by lymph node dissection 2 weeks later (axillary or popliteal lymph nodes). Experimental animals were then transplanted with a healthy lymph node while control animals underwent only lymph node dissection. Limb swelling, lymphatic fluid transport function, immune cell trafficking, bacterial clearance, and humoral and cellular immune responses were then analyzed 3-8 weeks later. RESULTS: LNT resulted in decreased hindlimb diameter (85%) when compared to baseline (i.e. 2 weeks after lymphatic ablation) and diminished adipose deposition (90%) when compared to control animals which showed no improvement in swelling over this period of time. Furthermore, animals treated with LNT had decreased T cell infiltration and marked increase in lymphangiogenesis and lymphatic regeneration as assessed by histology and NIR imaging after ICG injection. LNT animals demonstrated better B cells responses than controls when stimulated with OVA antigen (P=0.01) as well as improved T cell responses, bacterial particle clearance and dendritic cell trafficking (426% increase). Lymphatic fluid transport was also improved in these animals as assessed by lymphoscintigraphy (P=0.01). CONCLUSIONS: LNT markedly improves B and T cell responses and immune cell trafficking in a mouse model of lymphatic injury and lymphedema. LNT also induces lymphangiogenesis and improves lymphatic function.

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