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American Journal of Pathology, Vol 92, 619-635, Copyright © 1978 by American Society for Investigative Pathology

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Antiserums for immunofluorescent enumeration of human T lymphocytes utilizing fluoresceinated staphylococcal protein A

EW Ades, DS Gordon, DJ Phillips, M Hubbard, CM Black, LH Martin, MF LaVia and CB Reimer

Five lots (100 ml or more) of heterologous antiserums specific for human T lymphocytes were prepared using human or Rhesus monkey thymocytes as immunogens. After appropriate adsorptions, these antiserums reacted by immunofluorescence with 68% of human peripheral blood mononuclear cells and 98% of human thymocytes, with E-rosette-- positive cells but not with EAC-rosette--positive cells or five human B- lymphoblastoid-cell lines. Blocking experiments showed that Rhesus monkey thymocytes share thymic antigenic determinant(s) with humans. E- rosette receptors modulated independently from T-cell heteroantigens. Non-E--rosetting neoplastic T cells were identified in several patients with lymphoproliferative malignancies. Applying both the E-rosette assay and the anti-T-cell serum provides a better method of defining the biologic properties of normal and neoplastic T lymphocytes. Standardization of immunofluorescent conjugates for human T- or B-cell enumeration is simplified if large lots of well-characterized antiserums are available.