title: Differential antigen preservation during tissue autolysis.
abstract: Immediate fixation or snap freezing of tissue is ordinarily done to maximize antigen preservation for immunocytochemistry; however, delay in tissue allocation or spontaneous lymph node infarction can render tissue suboptimal for immunostaining. To test the effects of tissue autolysis/necrosis on the preservation of various lymphoid, epithelial, and mesenchymal markers, two lymph nodes (one with reactive lymphoid hyperplasia and one with metastatic ductal breast carcinoma) were evaluated for immunocytochemically demonstrated antigen preservation at 0-, 4-, 8-, 12-, 24-, 48-, and 72-hour intervals of autolysis at 37 degrees C. All specimens were stained by frozen section and formalin-fixed paraffin section immunocytochemical reactions with antibodies against CLA (CD45), UCHL-1 (CD45RO), L-26, kappa, lambda, anti-epithelial keratins (AE-1 and AE-3), epithelial membrane antigen, and vimentin. Frozen sections were additionally stained for Leu-1 (CD5), Leu-2a (CD8), Leu-3a+b (CD4), Leu-4 (CD3), and Leu-14 (CD22). The most resilient lymphoid antigen preservation was observed with CLA and UCHL-1, both exhibiting immunoreactivity at 72 hours in both frozen and fixed preparations. L-26 showed similar reactivity in frozen sections, but detectable antigen was observed only up to 24 hours in formalin-fixed tissue. Leu-2a proved to be the most labile antigen, persisting for only 12 hours in frozen sections. The epithelial markers epithelial membrane antigen and AE-1 exhibited excellent antigenic preservation in both frozen and fixed preparations; AE-3 persisted well in frozen section but was not demonstrated in fixed tissue. Vimentin immunoreactivity was vastly superior in frozen, as compared with fixed, tissue sections. Most antigens showed remarkable preservation despite morphologic degradation; however, differential antigenic resilience was demonstrated. Knowledge of this variation in antigen decay is critical for evaluation of immunoperoxidase phenotypic studies of autolyzed or necrotic tissue.
authors.affiliation: Department of Pathology, University of Texas Health Science Center, San Antonio 78284-7750.
authors.forename: R J
authors.initials: RJ
authors.lastname: Pelstring
authors.forename: D C
authors.initials: DC
authors.lastname: Allred
authors.forename: R J
authors.initials: RJ
authors.lastname: Esther
authors.forename: S R
authors.initials: SR
authors.lastname: Lampkin
authors.forename: P M
authors.initials: PM
authors.lastname: Banks
mesh_terms: D000941:Antigens / Q000032:analysis*; D015703:Antigens, CD / Q000276:immunology; D001329:Autolysis; D001706:Biopsy; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007633:Keratins / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D008562:Membrane Glycoproteins / Q000276:immunology; D018396:Mucin-1; D009336:Necrosis / Q000276:immunology / Q000473:pathology; D014021:Tissue Preservation; D014746:Vimentin / Q000276:immunology
chemical_list: D000941:Antigens; D015703:Antigens, CD; D008562:Membrane Glycoproteins; D018396:Mucin-1; D014746:Vimentin; D007633:Keratins