- Email: [email protected]
2110 ADIPOSE TISSUE-DERIVED STEM CELLS PREVENT ACUTE CELLULAR REJECTION AND PROLONG GRAFT SURVIVAL IN RAT KIDNEY TRANSPLANTATION
Vol. 189, No. 4S, Supplement, Wednesday, May 8, 2013
RESULTS: On the basis of antibody responses against the strain-specific gH epitopes, the D⫹/R⫹ pairs were classified into two groups, “matched gH (gH-m⫹)” and “mismatched gH (gH-m-)”, on the basis of combinations of antibody responses between the recipients and their donors. A total of 77 D⫹/R⫹ pairs were classified into 4 subgroups based on the combinations of antibody responses against both gH and gB (gH-m⫹/gB⫹, gH-m⫹/gB-, gH-m-/gB⫹ and gH-m-/ gB-). Among the 77 transplant recipients, 37 (48.1%) were found to be positive for CMV infection using the pp65 antigenemia assay during the 6-month follow up. The maximum numbers of pp65-positive cells in the patients with CMV infection were statistically different among the subgroups. CMV disease was significantly more prevalent in the subgroups lacking antibodies against gH and/or gB than in the matched gH-m⫹/gB⫹ subgroup (p⬍0.008). The acute rejection rates were 67%, 27%, 29% and 7% for the gH-m-/gB⫹, gH-m-/gB-, gH-m⫹/gB⫹ and gH-m⫹/gB- subgroups, respectively. Acute rejection was more frequent in the recipients with antibodies against gB (gB⫹ 44% vs. gB16%, p⫽0.015). CONCLUSIONS: The combination of presence or absence of antibodies against gB and gH in D⫹/R⫹ transplantation recipients can be a good indicator of acute rejection and CMV disease. Source of Funding: Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (No.16591609).
2110 ADIPOSE TISSUE-DERIVED STEM CELLS PREVENT ACUTE CELLULAR REJECTION AND PROLONG GRAFT SURVIVAL IN RAT KIDNEY TRANSPLANTATION Taigo Kato*, Koji Yazawa, Osaka, Japan; Masahiro Tanemura, Hiroshima, Japan; Masayoshi Okumi, Koichi Tsutahara, Yoichi Kakuta, Shiro Takahara, Norio Nonomura, Osaka, Japan INTRODUCTION AND OBJECTIVES: Mesenchymal stem cells (MSCs) have exhibited immunomodulatory effects in vitro. Especially, in addition to its abundance and easy accessibility, adipose tissue yields far more stem cells (adipose tissue-derived stem cells, ADSCs) than bone marrow per gram. However, the beneficial effects of ADSCs on alloreactivity in solid organ transplant models are scarcely known. In this study, we evaluated the beneficial effects of ADSCs in a rat kidney transplantation model. METHODS: ADSCs obtained from male Lewis rats used in the experiments. The immunomodulatory capacity of ADSCs was tested in vitro using co-cultures with rat lymphnode cells. The immunomodulatory capacity of ADSCs was tested in vitro using co-cultures with rat lymphnode cells. For in vivo experiments, fully MHC-disparate left kidneys from Dark Agouti rats were orthotopically transplanted into bilateral nephrectomized Lewis rats without immunosuppressant. ADSCs (2⫻106) were injected via the left renal artery of the donors prior to the nephrectomy (ADSCs group), while PBS was injected into the graft kidney as the control group. Histological and immunohistochemical (CD4⫹, CD8⫹ T cells and ED-1⫹ macrophages) analyses were performed on post-transplant day 5. Quantitative real-time PCR was also performed to assess the mRNA expressions of several cytokines in the graft. Finally, we investigated whether ADSCs prolonged graft survival. RESULTS: Similar to previous reports, ADSCs showed a dosedependent immunomodulatory capacity in vitro. However, in experiments in the transwell setting, their inhibitory effect on T cell proliferation was absent. Histological findings of the ADSCs group revealed a reduced rejection grade, while the mean number of infiltrated CD4⫹, CD8⫹ T cells was also significantly decreased as compared to the control group (p⫽0.028 and p⫽0.016, respectively). Moreover, injected ADSCs suppressed macrophages infiltration. mRNA expressions of pro-inflammatory cytokines, IL-1, TNF␣, and IFN in the graft, were suppressed on day 5 in the ADSCs group relative to the suppression of acute cellular rejection. Finally, injection of ADSCs led to significantly prolonged mean graft survival as compared to the control group (7.6⫾1.3 vs 6.6⫾0.6 days, p⫽0.013).
THE JOURNAL OF UROLOGY姞
e865
CONCLUSIONS: Our findings clearly show that intra-arterial injection of ADSCs attenuated acute rejection by down-regulation of pro-inflammatory cytokines and the cell-cell contact mechanism. Further studies are required to clarify the immunomodulative mechanism in greater detail. Source of Funding: None
2111 SUB-FASCIAL CONTINUOUS LOCAL ANESTHESIA DRAMATICALLY REDUCES NARCOTIC MEDICATION REQUIREMENTS FOLLOWING LAPARO-ENDOSCOPIC SINGLE SITE DONOR NEPHRECTOMY Miguel A. Mercado*, Lambros Stamatakis, Shawn Mathur, Judy M. Choi, Edward M. Sanchez, Wesley A. Mayer, Richard E. Link, Houston, TX INTRODUCTION AND OBJECTIVES: Improving pain control after donor nephrectomy may shorten convalescence and reduce barriers to kidney donation. In thoracic and abdominal surgery, sub-fascial continuous local anesthesia (SFCLA) has proven beneficial in reducing the need for narcotic pain medications in the postoperative setting. Herein, we present our experience with SFCLA with laparo-endoscopic single site donor nephrectomy (LESS-DN). METHODS: We retrospectively compared all cases of LESS-DN with or without SFCLA performed at our institution by two surgeons (WAM, REL) from October 2011 to August 2012. The On-Q Pain Buster 姞 with Silver Soaker ™ (I-Flow, Lake Forest CA) catheter system was used for SCFLA. Bilateral catheters were tunneled into the abdominal wall under direct laparoscopic visualization just superficial to the anterior peritoneum. A bolus of 20 mL of 0.5% ropivicaine was administered per catheter, followed by infusion of the same drug at 5 mL/hour from a 725 mL reservoir. The quantity of narcotic pain medicine used ⫹/⫺ SFCLA during the postoperative course was converted to morphine equivalents and compared. Visual analog pain scores at discharge were also recorded. Statistical analysis was performed using the student’s t-test. RESULTS: Fifty cases of LESS-DN with SCFLA and 77 cases without SCFLA were performed. SCFLA significantly reduced the administered postoperative morphine equivalents by 51% (32.7 mg with SCFLA and 63.2 without SCFLA, p⬍0.001). No significant differences were noted in hospital length-of-stay or discharge visual analog pain scores between the groups.SCFLA placement increased operative time by 13.6 minutes (p⫽ 0.04). No SCFLA related complications were noted. CONCLUSIONS: SFCLA significantly reduces postoperative pain following LESS-DN surgery, as assessed by the amount of narcotic needed to achieve adequate pain control. Although operative time was increased slightly, there were no complications noted. This technique may be valuable in decreasing the perioperative morbidity of laparoscopic donor nephrectomy. Source of Funding: None
2112 LONG TERM RENAL FUNCTION AFTER DONOR NEPHRECTOMY: A DOUBLE-BLINDED RANDOMIZED CONTROLLED TRIAL OF KETOROLAC VERSUS PLACEBO Gwen Grimsby*, Erik Castle, Paul Andrews, Laurie Mihalik, Yu-Hui Chang, Mitchell Humphreys, Phoenix, AZ INTRODUCTION AND OBJECTIVES: Ketorolac is a powerful non-opiod analgesic commonly used for post-surgical pain control but may be associated with renal impairment. We previously reported the short term outcomes of a novel continuous infusion of ketorolac versus placebo with no evidence of renal impairment. The goal of this study was to evaluate the long term safety of ketorolac versus placebo after donor nephrectomy.
RESULTS: On the basis of antibody responses against the strain-specific gH epitopes, the D⫹/R⫹ pairs were classified into two groups, “matched gH (gH-m⫹)” and “mismatched gH (gH-m-)”, on the basis of combinations of antibody responses between the recipients and their donors. A total of 77 D⫹/R⫹ pairs were classified into 4 subgroups based on the combinations of antibody responses against both gH and gB (gH-m⫹/gB⫹, gH-m⫹/gB-, gH-m-/gB⫹ and gH-m-/ gB-). Among the 77 transplant recipients, 37 (48.1%) were found to be positive for CMV infection using the pp65 antigenemia assay during the 6-month follow up. The maximum numbers of pp65-positive cells in the patients with CMV infection were statistically different among the subgroups. CMV disease was significantly more prevalent in the subgroups lacking antibodies against gH and/or gB than in the matched gH-m⫹/gB⫹ subgroup (p⬍0.008). The acute rejection rates were 67%, 27%, 29% and 7% for the gH-m-/gB⫹, gH-m-/gB-, gH-m⫹/gB⫹ and gH-m⫹/gB- subgroups, respectively. Acute rejection was more frequent in the recipients with antibodies against gB (gB⫹ 44% vs. gB16%, p⫽0.015). CONCLUSIONS: The combination of presence or absence of antibodies against gB and gH in D⫹/R⫹ transplantation recipients can be a good indicator of acute rejection and CMV disease. Source of Funding: Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (No.16591609).
2110 ADIPOSE TISSUE-DERIVED STEM CELLS PREVENT ACUTE CELLULAR REJECTION AND PROLONG GRAFT SURVIVAL IN RAT KIDNEY TRANSPLANTATION Taigo Kato*, Koji Yazawa, Osaka, Japan; Masahiro Tanemura, Hiroshima, Japan; Masayoshi Okumi, Koichi Tsutahara, Yoichi Kakuta, Shiro Takahara, Norio Nonomura, Osaka, Japan INTRODUCTION AND OBJECTIVES: Mesenchymal stem cells (MSCs) have exhibited immunomodulatory effects in vitro. Especially, in addition to its abundance and easy accessibility, adipose tissue yields far more stem cells (adipose tissue-derived stem cells, ADSCs) than bone marrow per gram. However, the beneficial effects of ADSCs on alloreactivity in solid organ transplant models are scarcely known. In this study, we evaluated the beneficial effects of ADSCs in a rat kidney transplantation model. METHODS: ADSCs obtained from male Lewis rats used in the experiments. The immunomodulatory capacity of ADSCs was tested in vitro using co-cultures with rat lymphnode cells. The immunomodulatory capacity of ADSCs was tested in vitro using co-cultures with rat lymphnode cells. For in vivo experiments, fully MHC-disparate left kidneys from Dark Agouti rats were orthotopically transplanted into bilateral nephrectomized Lewis rats without immunosuppressant. ADSCs (2⫻106) were injected via the left renal artery of the donors prior to the nephrectomy (ADSCs group), while PBS was injected into the graft kidney as the control group. Histological and immunohistochemical (CD4⫹, CD8⫹ T cells and ED-1⫹ macrophages) analyses were performed on post-transplant day 5. Quantitative real-time PCR was also performed to assess the mRNA expressions of several cytokines in the graft. Finally, we investigated whether ADSCs prolonged graft survival. RESULTS: Similar to previous reports, ADSCs showed a dosedependent immunomodulatory capacity in vitro. However, in experiments in the transwell setting, their inhibitory effect on T cell proliferation was absent. Histological findings of the ADSCs group revealed a reduced rejection grade, while the mean number of infiltrated CD4⫹, CD8⫹ T cells was also significantly decreased as compared to the control group (p⫽0.028 and p⫽0.016, respectively). Moreover, injected ADSCs suppressed macrophages infiltration. mRNA expressions of pro-inflammatory cytokines, IL-1, TNF␣, and IFN in the graft, were suppressed on day 5 in the ADSCs group relative to the suppression of acute cellular rejection. Finally, injection of ADSCs led to significantly prolonged mean graft survival as compared to the control group (7.6⫾1.3 vs 6.6⫾0.6 days, p⫽0.013).
THE JOURNAL OF UROLOGY姞
e865
CONCLUSIONS: Our findings clearly show that intra-arterial injection of ADSCs attenuated acute rejection by down-regulation of pro-inflammatory cytokines and the cell-cell contact mechanism. Further studies are required to clarify the immunomodulative mechanism in greater detail. Source of Funding: None
2111 SUB-FASCIAL CONTINUOUS LOCAL ANESTHESIA DRAMATICALLY REDUCES NARCOTIC MEDICATION REQUIREMENTS FOLLOWING LAPARO-ENDOSCOPIC SINGLE SITE DONOR NEPHRECTOMY Miguel A. Mercado*, Lambros Stamatakis, Shawn Mathur, Judy M. Choi, Edward M. Sanchez, Wesley A. Mayer, Richard E. Link, Houston, TX INTRODUCTION AND OBJECTIVES: Improving pain control after donor nephrectomy may shorten convalescence and reduce barriers to kidney donation. In thoracic and abdominal surgery, sub-fascial continuous local anesthesia (SFCLA) has proven beneficial in reducing the need for narcotic pain medications in the postoperative setting. Herein, we present our experience with SFCLA with laparo-endoscopic single site donor nephrectomy (LESS-DN). METHODS: We retrospectively compared all cases of LESS-DN with or without SFCLA performed at our institution by two surgeons (WAM, REL) from October 2011 to August 2012. The On-Q Pain Buster 姞 with Silver Soaker ™ (I-Flow, Lake Forest CA) catheter system was used for SCFLA. Bilateral catheters were tunneled into the abdominal wall under direct laparoscopic visualization just superficial to the anterior peritoneum. A bolus of 20 mL of 0.5% ropivicaine was administered per catheter, followed by infusion of the same drug at 5 mL/hour from a 725 mL reservoir. The quantity of narcotic pain medicine used ⫹/⫺ SFCLA during the postoperative course was converted to morphine equivalents and compared. Visual analog pain scores at discharge were also recorded. Statistical analysis was performed using the student’s t-test. RESULTS: Fifty cases of LESS-DN with SCFLA and 77 cases without SCFLA were performed. SCFLA significantly reduced the administered postoperative morphine equivalents by 51% (32.7 mg with SCFLA and 63.2 without SCFLA, p⬍0.001). No significant differences were noted in hospital length-of-stay or discharge visual analog pain scores between the groups.SCFLA placement increased operative time by 13.6 minutes (p⫽ 0.04). No SCFLA related complications were noted. CONCLUSIONS: SFCLA significantly reduces postoperative pain following LESS-DN surgery, as assessed by the amount of narcotic needed to achieve adequate pain control. Although operative time was increased slightly, there were no complications noted. This technique may be valuable in decreasing the perioperative morbidity of laparoscopic donor nephrectomy. Source of Funding: None
2112 LONG TERM RENAL FUNCTION AFTER DONOR NEPHRECTOMY: A DOUBLE-BLINDED RANDOMIZED CONTROLLED TRIAL OF KETOROLAC VERSUS PLACEBO Gwen Grimsby*, Erik Castle, Paul Andrews, Laurie Mihalik, Yu-Hui Chang, Mitchell Humphreys, Phoenix, AZ INTRODUCTION AND OBJECTIVES: Ketorolac is a powerful non-opiod analgesic commonly used for post-surgical pain control but may be associated with renal impairment. We previously reported the short term outcomes of a novel continuous infusion of ketorolac versus placebo with no evidence of renal impairment. The goal of this study was to evaluate the long term safety of ketorolac versus placebo after donor nephrectomy.