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Chicken macrophages and thrombocytes share a common cell surface antigen defined by a monoclonal antibody

1993; Elsevier BV; Volume: 36; Issue: 4 Linguagem: Inglês

10.1016/0165-2427(93)90029-4

1873-2534

Bernd Kaspers, Hyun S. Lillehoj, Erik P. Lillehoj,

Protein purification and stability

A monoclonal antibody (mAb), designated K1, reacted with a cell surface antigen shared on chicken macrophages and thrombocytes. By immunofluorescence staining, the mAb K1 was reactive with 31.8% of peripheral blood lymphocytes (PBL) separated on Histopaque. In contrast, only 3.2% of PBL separated by slow-speed centrifugation were K1 positive. This antibody did not react with B- or T-lymphocytes, as demonstrated by the very small percentage of positive cells in thymus, bursa and spleen. Furthermore, no staining was observed with avian T-cell (MDCC-RP1 and SK3) or B-cell (LSCC-RP9) lines. Adherent cells derived from PBL separated on Histopaque and cultured for 48 h in plastic cell culture dishes were 81.5% positive with K1. These cells were also 82.6% positive with an antibody detecting the major histocompatibility complex (MHC) Class II antigen in chickens, indicating that they were monocyte-derived macrophages. Sheep red blood cell phagocytosis by these cells could be demonstrated, further supporting their macrophage lineage. In addition, K1 stained virtually 100% of HD11 cells, a chicken macrophage cell line, as well as 86.7% of peritoneal exudate cells. Eighty five percent of plastic adherent cells from PBL collected after 2 h of adherence reacted with the mAb K1, but only 8% of these cells were MHC Class II positive. These cells were morphologically identified as thrombocytes. Immunoprecipitation analysis demonstrated that the K1-reactive antigen consisted of a heterodimer with constituent polypeptide chains of 135 kDa and 61-68 kDa.