We are interested in studying the effects of ischemia-reperfusion injury on outcomes following lung transplantation with the goal of understanding underlying cellular and molecular mechanisms. Neutrophils that enter the lung contribute to ischemia-reperfusion injury, or IRI. We've recently discovered that B cells proceed neutrophils in entering the lung, and they play a role in the recruitment of neutrophils following IRI.
Left lung transplantation in the mouse is technically challenging, and requires full-time, dedicated training over several months to achieve consistent results. Additionally, it introduces allergenicity and surgical anastomotic trauma. Thus, we need a more accessible technique that can isolate the study of ischemia-reperfusion injury.
Using our hilar clamp model, we have found that B-cells enter the lung early after ischemia-reperfusion injury, and they recruit classical monocytes in a B-cell receptor into TLR4 dependent fashion. This ultimately results in increased neutrophil extravasation into the lung. Our protocol involves reversible suture ligation of the left pulmonary hilum with a slip knot.
The pliable and unobtrusive nature of the suture allows for chest closure, which is useful for studying longer periods of warm ischemia, as chest closure minimizes heat and insensible fluid losses in the mouse.
