3. Hematopoeisis is not completely understood. For example, dendritic cells (DC)

Question

3. Hematopoeisis is not completely understood. For example, dendritic cells (DC) found in peripheral tissues are thought to originate in the bone marrow, partly differentiate into monocyte/DC precursors that can leave the BM and enter the circulation, and further differentiate into either monoblasts monocytes macrophages, or common DC precursors pDC and cDC (see class notes for details). However, in the study described (Crawford, 1999), blood PBMC were analyzed by FACS (A) and immunoblotting (B) to directly detect the surface marker proteins CD14, TCR, and CD2. (GAPDH is a control protein in all cells). A. B. 6% 71% T cells CD2tMo CD2 TCR CD14 GAPDH ACD2 Mo 12% 10 10' CD14 FITC - (1) what kinds of cells and fraction (%) of the PBMC are: a. CD2+CD14- b. CD2- CD14+ c. CD2+ CD14+ d. CD2- CD14- cells, cells, cells,- (2) Study the immune cells' protein expression in the immunoblot (B). a. b. c. Do CD2+ monocytes express TCR? Do T cells express CD14? Do B cells express CD2?

Explanation / Answer

1. CD2+ CD14-       T cell                           71%

2.CD2- CD14+        Monocytes                   12%

3.CD2+CD14+        Monocytes                   6%

4. CD2-CD14-        T cell                            11%

(2) a. According to the immunoblot there is no band observed when CD2+ expressing cells are immunoblotted with TCR antibody. Thus CD2+ monocytes do not express TCR.

b. T cell do not express CD14, as no band observed for CD14 antibody.

c. B cells do not express CD2 as there isno band observed for CD2.

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