| [Objectives]1. To observe preservation injury of liver graft using light microscopy and electron microscopy.2. To study changes of humoral and cellular immune markers before and after reperfusion in liver transplantation.3. To study the difference between HBV positive graft and HBV negative graft in preservation injury, humoral and cellular immune markers.4. To study whether humoral immune participate in two cases with acute central lobular rejection.[Methods]1. 68 liver specimens were obtained from the donors and recipients at the time of transplantation from May 2007 to January 2009. The median age at transplantation was 26 years (range 26～71) including 30 male and 4 female. All recipients were hepatitis B positive, 18 cases with liver cancer, one with acute liver failure, one with sub-acute liver failure, and the rest with chronic hepatitis B and decompensated liver cirrhosis.All donors were dead with traumatic brain injury. The median age of all donors was 29 years (range 25～30 years). All donors were divided into HBV positive (HB-D, n=13) and HBV negative (NHB-D, n=21). The control group were 20 specimens with moderate hepatitis B.Two liver specimens were obtained from two patients with central lobular rejection.2. All biopsy specimens were 10% phosphate-buffered formalin fixed and paraffin embedded for light microscopy. Immunohistochemical staining using the peroxidase method was carried out on each specimen for IgG, IgM, IgA, the components of complement C3, C4d, C1q, HBsAg and HBcAg. Three samples of NHB liver grafts including before and after reperfusion were prepared for Electron Microscopy.3. Optical microscope was used to observe the deposition of immunoglobulin and complement fragments in the liver. Scoring of staining is based on the percentage of stained tissue on IHC that has a linear, circumferential staining pattern in both vascular walls of portal area(PA) and hepatic sinusoidal endothelial cell (SEC): negative for 0, <25% for 1, 26 ~ 50% for 2, >50% for 3. All recipients were followed up for six months for post-transplantation rejection.[Results]1. All donors'liver had different preservation injury (PI) after reperfusion. NHB-D had 9.5% moderate PI, while HB-D had more 38%(p<0.05). The rest were mild PI.2. Under EM: Mitochondria were round, swelling, and glycogen granules were reduced, apoptotic bodies were forming.3. The markers of humoral immuneThere were various degrees of injury in all donors'livers.(1)The deposition of SEC-IgM, SEC-IgA, PA-IgG,PA-IgM,PA-IgA and PA-C1q statistically significant increased in NHB(all p<0.05);(2)The HB group IgG,IgM,IgA,C3c,C1q statistically significant increased in both SEC and PA(all p<0.05);(3)There were no difference between NHB and HB before reperfusion (p>0.05). HB-D had a higher SEC-C1q, SEC-C4d than NHB-D after reperfusion (p<0.05).4. The markers of cellular immuneThe main infiltrated cells in all specimens were CD45ro + T cells, followed by CD8+T cells, CD20+B cells and CD138+ plasma cells and CD4+T cells. Perforin were negative in all specimens. There were no difference between NHB-D and HB-D(p>0.05).5. Various spherical bodies were cattered in hepatic sinusoid and cells. It were eosinophilic with a diameter of 1–3μM, expressing different levels of IgG, IgM, IgA, C3c. 6. Immunoglobulin deposition and complement activation were seen in the specimens in the two cases with central lobular rejection.[Conclusions]1. Immunoglobulin and complement expression increased in all donors'liver after reperfusion with little changes in the immune cells.2. Donors'livers with HBV can more active the humoral immune syterm after reperfusion.3. Spherical bodies cattered in hepatic sinusoid and cells may be associated with plasma protein aggregation due to preservation. It has not been described in the literature.4. Humoral immune participate in the acute central lobular rejection.
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