Effects of fixation and paraffin embedding on the immunohistological detection of cell-associated HIV-1 by different monoclonal antibodies.
This study evaluates a panel of monoclonal antibodies (MAbs) to HIV-1 antigens (DuPont anti-gp120, gp41, p24; Olympus anti-gp120/160, gp41, p24A, p24B, p55, p18A, p18B, reverse transcriptase) for their ability to detect the virus in tissues after exposure to various fixatives (100% acetone, 10% formaldehyde, 2.5% glutaraldehyde, 4% paraformaldehyde/1% glutaraldehyde, Bouin's fluid) and after paraffin embedding. Acetone, 10% formaldehyde, and Bouin's fluid all preserved a wide range of viral epitopes compared with other fixatives. The most robust MAbs were DuPont p24 and Olympus p55, which produced excellent staining regardless of the fixative used. Embedding in paraffin variability influenced the capacity of MAbs to detect HIV-1 epitopes on fixed cells. Certain antibodies (e.g., DuPont gp24, Olympus p24B) produced good staining, whereas other epitopes (e.g., DuPont gp120, formaldehyde) were destroyed. In some cases, paraffin embedding revealed antigenic sites that had been formerly masked (e.g., Olympus gp120 and p24A; formaldehyde and glutaraldehyde fixation). These results indicate that HIV-1 antigens can be detected by immunohistology on cells exposed to most common fixatives. Therefore, retrospective analysis of pathological material is possible, provided that the antibodies are matched to the fixative used to preserve the tissue.
Pudney
J
J
Fearing Research Laboratory, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Anderson
D
D
D000085:Acetates; D019342:Acetic Acid; D000096:Acetone; D000911:Antibodies, Monoclonal; D002460:Cell Line; D000939:Epitopes / Q000032:analysis; D005557:Formaldehyde; D015686:Gene Products, env / Q000032:analysis*; D005976:Glutaral; D015488:HIV Antigens / Q000032:analysis*; D016655:HIV Core Protein p24 / Q000032:analysis; D015699:HIV Envelope Protein gp120 / Q000032:analysis; D015700:HIV Envelope Protein gp41 / Q000032:analysis; D015497:HIV-1 / Q000302:isolation & purification* / Q000378:metabolism; D006652:Histological Techniques; D006801:Humans; D010232:Paraffin; D010851:Picrates; D011108:Polymers
D000085:Acetates; D000911:Antibodies, Monoclonal; D000939:Epitopes; D015686:Gene Products, env; D015488:HIV Antigens; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D010851:Picrates; D011108:Polymers; D000096:Acetone; D005557:Formaldehyde; C026239:Bouin's solution; D010232:Paraffin; D019342:Acetic Acid; D005976:Glutaral; C003043:paraform