Laparoscopic Technique for Serial Collection of Liver and Mesenteric Lymph Nodes in Macaques

J Vis Exp. 2017 May 2;(123):55617. doi: 10.3791/55617.

Abstract

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them immunologically unique. The GI tract and associated MLN are sites of early viral replication in human immunodeficiency virus (HIV) infection and the MLN are likely important reservoir sites that harbor latently-infected cells even after prolonged antiretroviral therapy (ART). The liver has been shown to play a significant role in immune responses to lentiviruses and appears to play a significant role in clearance of virus from circulation. Nonhuman primate (NHP) models for HIV and Acquired Immunodeficiency Syndrome (AIDS) closely mimic these aspects of HIV infection and serial longitudinal sampling of primary sites of viral replication and the associated immune responses in this model will help to elucidate critical events in infection, pathogenesis, and the impact of various intervention strategies on these events. Current published techniques to sample liver and MLN together involve major surgery and/or necropsy, which limits the ability to investigate these important sites in a serial fashion in the same animal. We have previously described a laparoscopic technique for collection of MLN. Here, we describe a minimally invasive laparoscopic technique for serial longitudinal sampling of liver and MLN through the same two port locations required for the collection of MLN. The use of the same two ports minimizes the impact to the animals as no additional incisions are required. This technique can be used with increased sampling frequency compared to major abdominal surgery and reduces the potential for surgical complications and associated local and systemic inflammatory responses that could complicate interpretation of results. This procedure has potential to facilitate studies involving NHP models while improving animal welfare.

Publication types

  • Video-Audio Media

MeSH terms

  • Anesthesia
  • Animals
  • Biopsy / methods*
  • Female
  • Flow Cytometry
  • Laparoscopy / methods*
  • Leukocyte Common Antigens
  • Liver / pathology*
  • Lymph Nodes / pathology*
  • Lymphocytes / metabolism
  • Macaca mulatta
  • Sentinel Lymph Node Biopsy / methods*
  • Specimen Handling / methods*

Substances

  • Leukocyte Common Antigens