Molecular and functional observations on the donor intestinal muscularis during human small bowel transplantation

doi:10.1053/gast.2002.33628

Gastroenterology

Volume 122, Issue 7, June 2002, Pages 1886-1897

https://doi.org/10.1053/gast.2002.33628 Get rights and content

Abstract

Background & Aims: Ischemia-reperfusion injury or intestinal manipulation evokes an inflammatory response within the intestinal muscularis that is associated with intestinal dysmotility. We hypothesize that human small intestinal transplantation induces an analogous response. Methods: Human intestinal graft specimens were obtained during transplantation and compared with specimens removed early during elective bowel resections. Inflammatory gene expression was quantified by real-time reverse-transcription polymerase chain reaction. Histochemistry and immunohistochemistry were used to characterize leukocyte infiltration and macrophage activation. In vitro circular muscle contractility and intracellular electric neuromuscular transmission in response to electric field stimulation (EFS) were measured. Results: Messenger RNA (mRNA) values were significantly elevated before reperfusion and further increased during reperfusion (4 hour reperfusion: interleukin [IL]-6, 311-fold; monocyte chemoattractant protein [MCP-1, 122-fold; IL-8, 338-fold; epithelial neutrophil-activating peptide-78 [ENA-78], 56-fold; intercellular adhesion molecule-1 [ICAM-1], 9-fold; and cyclooxygenase-2 [COX2], 37-fold) over elective specimens. Neutrophils and monocytes extravasated in increased numbers in whole mounts before and after reperfusion over the elective specimens. Activated resident macrophages were identified as a major source of inflammatory mediators. Muscle contractions and neuromuscular transmission were markedly attenuated in the grafts. Conclusions: The data suggest that manipulation during organ harvesting initiates a functionally relevant molecular and cellular inflammatory response within the graft muscularis that is potentiated during the reperfusion period. Significant mechanical and neuromuscular functional alterations occurred during the transplant process.

GASTROENTEROLOGY 2002;122:1886-1897

Section snippets

Patients and small bowel specimens

Adult patients undergoing small intestinal transplantation at the University of Pittsburgh were prospectively included in the study. Specimens were obtained during back table preparation of the donor graft and intraoperatively during graft implantation from patients undergoing small intestinal transplantation. Intestinal segments (1.5-cm length) were removed from the aboral graft border at the end of the cold preservation period after the cumulative insult of harvesting (2.8 ± 0.2 hours between

Molecular inflammatory events

Gene expression analysis focused on the multifunctional cytokine IL-6, on MCP-1, and ICAM-1 as regulators of cellular inflammation, and on COX-2 as a kinetically active mediator. Real-time reverse-transcription PCR analysis showed a significant successive up-regulation in the messenger RNA (mRNA) values of all these mediators as the transplant procedure progressed: preserved donor grafts, before reperfusion of the implanted graft, and after reperfusion (Figures 1 and 2).

. Real-time

Discussion

Our results show that the human donor intestinal muscularis itself is an immunologically active participant in the development of a complex inflammatory response during the transplantation process. Additionally, that these local molecular events functionally result in the recruitment of circulating host leukocytes, a suppression in intestinal smooth muscle contractility, and a dysfunction in neuromuscular transmission.

We have shown that the intestinal muscularis of mammals is populated with a

References (57)

RM Ghobrial et al.
Advances in pediatric liver and intestinal transplantation
Am J Surg
(2000)
V Garrido et al.
Late severe rejection of intestinal allografts: risks and survival outcome
Transplant Proc
(2001)
L Cicalese et al.
Segmental living related small bowel transplantation in adults
J Gastrointest Surg
(2001)
RG Lee et al.
Pathology of human intestinal transplantation
Gastroenterology
(1996)
CP Johnson et al.
Effects of intestinal transplantation on postprandial motility and regulation of intestinal transit
Surgery
(2001)
DW Vane et al.
Impaired bowel motility following small intestinal transplantation
J Surg Res
(1989)
H Pernthaler et al.
Altered pacemaker frequency gradient of a human small bowel allograft
Transplant Proc
(1997)
JC Kalff et al.
Surgically induced leukocytic infiltrates within the rat intestinal muscularis mediate postoperative ileus
Gastroenterology
(1999)
NT Schwarz et al.
Prostanoid production via COX-2 as a causative mechanism of rodent postoperative ileus
Gastroenterology
(2001)
JC Kalff et al.
Selective coinduction of cyclooxygenase-2 and inducible nitric oxide synthase in human intestinal smooth muscle during abdominal surgery (abstr)
Gastroenterology
(1998)

N Kruse et al.

Quantification of cytokine mRNA expression by RT PCR in samples of previously frozen blood

J Immunol Methods

(1997)

CA Power et al.

Cloning of a full-length cDNA encoding the neutrophil-activating peptide ENA-78 from human platelets

Gene

(1994)

DE Staunton et al.

Primary structure of ICAM-1 demonstrates interaction between members of the immunoglobulin and integrin supergene families

Cell

(1988)

TD Schmittgen et al.

Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: comparison of endpoint and real-time methods

Anal Biochem

(2000)

C Hierholzer et al.

Intraoperative upregulation of muscularis IL-6, Stat3 activation and an association between neutrophil infiltration and muscle dysfunction in human (abstr)

Gastroenterology

(1998)

KJ Walgenbach et al.

Increase of interleukin-6 in the intestinal mucosa and muscularis following chronic rejection after small bowel transplantation

Transplant Proc

(1998)

CE Eberhart et al.

Eicosanoids and the gastrointestinal tract

Gastroenterology

(1995)

MD Josephs et al.

Products of cyclooxygenase-2 catalysis regulate postoperative bowel motility

J Surg Res

(1999)

JC Kalff et al.

Biphasic response to gut manipulation and temporal correlation of cellular infiltrates and muscle dysfunction in rat

Surgery

(1999)

NT Schwarz et al.

Prostanoid production via COX-2 as a causative mechanism of rodent postoperative ileus

Gastroenterology

(2001)

GM Rovera et al.

Quality of life of patients after intestinal transplantation

Transplantation

(1998)

SS Kaufman et al.

Indications for pediatric intestinal transplantation: a position paper of the American Society of Transplantation

Pediatr Transplantation

(2001)

RA Veenendaal et al.

Clinical aspects of small-bowel transplantation

Scand J Gastroenterol Suppl

(2000)

O Goulet et al.

Intestinal transplantation: indications, results and strategy

Curr Opin Clin Nutr Met Care

(2000)

S Satoh et al.

Circulating ICAM-1, E-selectin, IL-2 receptor, and HLA class I in human small bowel, liver, and small bowel-plus-liver transplant recipients

Transplantation

(1995)

SS Kaufman et al.

Disaccharidase activities and fat assimilation in pediatric patients after intestinal transplantation

Transplantation

(2000)

H Ishii et al.

Changes of intestinal motility after small bowel transplantation in the rat

Transplantation

(1994)

M Kusunoki et al.

Long-term effects of small bowel transplantation on intestinal motility

Transplantation

(1995)

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Intestinal transplantation (ITx) remains challenged by frequent/severe rejections and immunosuppression-related complications (infections/malignancies/drug toxicity). We developed the Leuven Immunomodulatory Protocol (LIP) in the lab and translated it to the clinics. LIP consists of experimentally proven maneuvers, destined to promote T-regulatory (Tregs)-dependent graft-protective mechanisms: donor-specific blood transfusion (DSBT); avoiding high-dose steroids/calcineurin-inhibitors; and minimizing reperfusion injury and endotoxin translocation. LIP was tested in 13 consecutive ITx from deceased donors (2000–2014) (observational cohort study). Recipient age was 37 years (2.8–57 years). Five-year graft/patient survival was 92%. One patient died at 9 months due to aspergillosis, another at 12 years due to nonsteroidal anti-inflammatory drug–induced enteropathy. Early acute rejection (AR) developed in two (15%); late AR in three (23%); all were reversible. No chronic rejection (CR) occurred. No malignancies developed and estimated glomerular filtration rate remained stable post-Tx. At last follow-up (3.5 years [0.5–12.5 years]), no donor-specific antibodies were detected and 11 survivors were total parenteral nutrition free with a Karnofsky score >90% in 8 recipients (follow-up >1 years). A high frequency of circulating CD4⁺CD45RA⁻Foxp3^hi memory Tregs was found (1.8% [1.39–2.21]), comparable to tolerant kidney transplant (KTx) recipients and superior to stable immunosuppression (IS)-KTx, KTx with CR, and healthy volunteers. In this ITx cohort we show that DSBT in a low-inflammatory/pro-regulatory environment activates Tregs at levels similar to tolerant-KTx, without causing sensitization. LIP limits rejection under reduced IS and thereby prolongs long-term survival to an extent not previously attained after ITx.
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The body weight and food intake of the mice administered with distilled water decreased significantly compared with before RT. After the same dose of abdominal RT, tumor necrosis factor-α, diamine oxidase, intestinal fatty acid-binding protein in plasma and malonalhehyde and apoptosis of the intestine were significantly greater in the control group than in the mice administered berberine (p < .05–.01). In contrast, interleukin-10 in the mice with berberine treatment was significantly greater than in the control group (p < .01). A similar result was found in the fractionated RT experiment and at different points after 16 Gy abdominal RT (p < .05–.01). Berberine treatment significantly delayed the point of death after 20 Gy, but not 16 Gy, abdominal RT (p < .01).
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^☆: Address requests for reprints to: Anthony J. Bauer, Ph.D., Department of Medicine/Gastroenterology, University of Pittsburgh Medical School, S849 Scaife Hall, 3550 Terrace Street, Pittsburgh, Pennsylvania 15261.e-mail: [email protected]; fax: (412) 648-9731.

^☆☆: Supported by National Institute of Health grants R01-GM-58241 and P50-GM-53789 and a grant from the Deutsche Forschungsgemeinschaft (TU 116/2-1).

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Basic ResearchMolecular and functional observations on the donor intestinal muscularis during human small bowel transplantation☆,☆☆

Abstract

Section snippets

Patients and small bowel specimens

Molecular inflammatory events

Discussion

Am J Surg

Transplant Proc

J Gastrointest Surg

Gastroenterology

Surgery

J Surg Res

Transplant Proc

Gastroenterology

Gastroenterology

Gastroenterology

J Immunol Methods

Gene

Cell

Anal Biochem

Gastroenterology

Transplant Proc

Gastroenterology

J Surg Res

Surgery

Gastroenterology

Quality of life of patients after intestinal transplantation

Transplantation

Indications for pediatric intestinal transplantation: a position paper of the American Society of Transplantation

Pediatr Transplantation

Clinical aspects of small-bowel transplantation

Scand J Gastroenterol Suppl

Intestinal transplantation: indications, results and strategy

Curr Opin Clin Nutr Met Care

Circulating ICAM-1, E-selectin, IL-2 receptor, and HLA class I in human small bowel, liver, and small bowel-plus-liver transplant recipients

Transplantation

Disaccharidase activities and fat assimilation in pediatric patients after intestinal transplantation

Transplantation

Changes of intestinal motility after small bowel transplantation in the rat

Transplantation

Long-term effects of small bowel transplantation on intestinal motility

Transplantation

Basic Research
Molecular and functional observations on the donor intestinal muscularis during human small bowel transplantation☆,☆☆