The tissue specimens in patients who underwent revision were fixed in 10 % neutral buffered formalin prior to processing and embedding in paraffin wax. Sample sections 5-μm-thick were stained with hematoxylin and eosin and examined by light microscopy [13 (link)], and ALVAL scoring was performed. Sections were also analyzed with immunohistochemistry using antibodies to T cells (CD3; DAKO, Glostrup, Denmark) and B cells (CD20; DAKO) to characterize the immunophenotype. Sections 5-μm-thick were treated with superblock solution (Scytek Laboratories, Logan, UT) before incubation with antibodies to T cells and B cells overnight at 48 °C. After washing, sections were incubated in 0.3 % H2O2 in methanol for 15 min to block endogenous peroxidase activity. After being washed, sections were treated with peroxidase-conjugated antimouse IgG Fab0 (1:500 MBL, Nagoya, Japan) for 1 h, followed by color development with diaminobenzidine/H2O2 solutions. Light counterstaining with hematoxylin was performed to aid orientation.
The predominant lymphocyte was determined based on whether T cells or B cells were dominant. T-cell dominant hips were further analyzed using antibodies to T helper (Th) cells (CD4; DAKO) and T cytotoxic (Tc) cells (CD8; DAKO) to determine whether Th cells or Tc cells were dominant.
The predominant lymphocyte was determined based on whether T cells or B cells were dominant. T-cell dominant hips were further analyzed using antibodies to T helper (Th) cells (CD4; DAKO) and T cytotoxic (Tc) cells (CD8; DAKO) to determine whether Th cells or Tc cells were dominant.