Isolation and phenotypic characterization of CD16, CD8 and CD4 positive cells from LBC cell line

RUYBAL PAULA; MONGINI CLAUDIA; GRAVISACO MARÍA JOSÉ; SÁNCHEZ LOCKHART MARIANO; WALDNER CLAUDIA

Punta del Este, Uruguay

Congreso; V° Congreso de la Asociación Latinoamericana de Inmunología (ALAI); 1999

Asociación Latinoamericana de Inmunología

LBC is a murine cell line derived from LB tumor, which is a spontaneous T cell lymphoma originated in a Balb/c mouse. LBC cells were stained using specific, PE or Fitc labeled, monoclonal antibodies. Phenotypic analysis by FACS revealed that they are CD3+/-, C3+ (cytoplasmatic), CD8+, CD16+, CD25+, TCR ab+/-, CD2+/-, CD4-, TCR gd-. Immunomagnetic isolation of CD16+ cells was performed by positive selection using dynabeads. The resulting cell population contained 99.9% CD16 expressing cells. Morphologically, the cells displayed a lymphoblstoid appearance. By flow cytometry, a cell population with higher forward scatter pattern than normal mouse lymphocytes was found. The following phenotype was expressed CD2-, CD3+/-, CD3+ (cytoplasmatic), CD4+, CD8+, CD16+, CD25+, TCR ab+/- y TCR gd-. By using a CD4+ immunomagnetic isolation method, we demonstrated a subset of LBC cells expressing CD4 marker (1%). This characterization suggests that LBC cells bear large granular lymphocyte phenotype (LGL) from T cell origin and that LBC tumor could be an interesting model for large granular T cell lymphoproliferative disorders. By using an immunomagnetic separation we further demonstrated a CD4+ cell subpopulation which was no detected in the original LBC line by FACS.