Conditions affecting the immunohistochemical detection of HIV in fixed and embedded renal and nonrenal tissues.
A number of studies have suggested that HIV infection can be detected in a variety of routinely fixed archival tissues using antibodies to various viral proteins. In order to study this immunocytochemical approach, paraffin sections were examined with a large panel of commercially available monoclonal antibodies against the various HIV proteins (5 antibodies to p24, 1 to p17, 1 to gp41, and 1 to gp120) using a streptavidin-biotin method. A polyclonal antibody against p24 was also tested. Formalin-fixed, paraffin-embedded HIV infected CEM E5 T cells were used as positive controls. Tissues from AIDS patients included 31 kidneys, 8 lymph nodes, 2 spleens and 3 brains. Non-AIDS tissues examined were 6 renal biopsies with focal segmental glomerulosclerosis, 5 with interstitial nephritis, 6 reactive lymph nodes, and a brain with encephalitis, all from patients not known to be at high risk for HIV infection. Additional negative controls included: 1) replacement of primary antibody with a hybridoma derived mouse monoclonal IgG1 standard, 2) omission of the primary antibody, and 3) sections of formalin-fixed paraffin-embedded CEM E5 T cells cultures not infected with HIV. Competition experiments with excess recombinant p24 protein were also performed. False positive staining with the IgG1 standard or with the antibodies to HIV proteins was frequently seen in tissues with pathologic findings (inflammation, hyalin degeneration), particularly following protein digestion. Protein digestion also had a major impact on specific staining. Digestion with proteinase K abolished specific staining for the core proteins of the virus (p17, p24) on the positive control sections.(ABSTRACT TRUNCATED AT 250 WORDS)
Nadasdy
T
T
Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City.
Hanson-Painton
O
O
Davis
L D
LD
Miller
K W
KW
DeBault
L E
LE
Burns
D K
DK
Silva
F G
FG
D016263:AIDS-Associated Nephropathy / Q000382:microbiology*; D000368:Aged; D000911:Antibodies, Monoclonal; D002648:Child; D002675:Child, Preschool; D006678:HIV / Q000302:isolation & purification*; D016655:HIV Core Protein p24 / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007668:Kidney / Q000382:microbiology* / Q000473:pathology; D008875:Middle Aged; D013194:Staining and Labeling; D016610:Tissue Embedding; D016707:Tissue Fixation
D000911:Antibodies, Monoclonal; D016655:HIV Core Protein p24