Table 2.
Summary of TAPP expression profiles by glioma cell lines
| (A) Intracellular flow cytometric analysis of 12 TAPP expressed within the glioma cells* | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Antigen | Glioma |
|||||||||||
| A172 | D-54 | LN18 | LN229 | LNZ308 | SF767 | SNB19 | T98G | U-118 | U-251 | U-373 | U-87 | |
| B-cyclin | 1 | 1 | 2 | 2 | 2 | 2 | 3 | 1 | 1 | 1 | 2 | 1 |
| EphA2 | 1 | 1 | 1 | 2 | 2 | 1 | 1 | 0 | 1 | 2 | 3 | 1 |
| Her2/neu | 2 | 3 | 3 | 3 | 3 | 2 | 3 | 2 | 2 | 3 | 3 | 3 |
| IL13Rα2 | 3 | 4 | 2 | 2 | 3 | 2 | 3 | 2 | 3 | 3 | 3 | 3 |
| GnT-V | 2 | 1 | 2 | 2 | 1 | 2 | 2 | 3 | 1 | 3 | 2 | 2 |
| GP100 | 3 | 2 | 0 | 0 | 3 | 2 | 3 | 3 | 0 | 3 | 4 | 3 |
| Mage-1 | 4 | 3 | 2 | 3 | 3 | 3 | 3 | 2 | 2 | 3 | 2 | 3 |
| Mart-1 | 3 | 0 | 3 | 3 | 3 | 2 | 3 | 3 | 0 | 3 | 3 | 3 |
| Survivin | 3 | 2 | 2 | 2 | 3 | 2 | 2 | 3 | 1 | 3 | 2 | 3 |
| hTert | 4 | 4 | 3 | 3 | 4 | 4 | 2 | 4 | 4 | 4 | 3 | 4 |
| Trp-1 | 0 | 2 | 1 | 1 | 0 | 0 | 1 | 1 | 0 | 0 | 1 | 0 |
| Tyrosinase | 2 | 0 | 2 | 3 | 1 | 2 | 2 | 3 | 0 | 2 | 0 | 0 |
| Σ score | 28 | 23 | 23 | 26 | 28 | 24 | 28 | 27 | 15 | 30 | 28 | 26 |
| (B) Real-time PCR expression of Aim-2, Gage-1, Sart-1, and Trp-2 mRNA† | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Antigen | Glioma |
|||||||||||
| A172 | D-54 | LN18 | LN229 | LNZ308 | SF767 | SNB19 | T98G | U-118 | U-251 | U-373 | U-87 | |
| Aim-2 | 2 | 3 | 3 | 3 | 3 | 2 | 3 | 3 | 3 | 3 | 3 | 2 |
| Gage-1 | nd | nd | nd | nd | nd | nd | nd | nd | 2 | nd | nd | nd |
| Sart-1 | 2 | 2 | 3 | 2 | nd | 1 | 2 | 2 | 2 | 1 | 2 | nd |
| Trp-2 | nd | nd | nd | 2 | nd | 2 | nd | 2 | 2 | nd | 1 | nd |
| Σ score | 4 | 5 | 6 | 7 | 3 | 5 | 5 | 7 | 9 | 4 | 6 | 2 |
| Total‡ | 32 | 28 | 29 | 33 | 31 | 29 | 33 | 34 | 24 | 34 | 34 | 28 |
The cells were fixed, permeabilized, and stained with the antibodies directed toward the various tumor antigens. Cells were scored for the intensity of immunofluorescence staining obtained by flow cytometry. A score of 0 was given when the staining was not above that of the negative control cells. A score of 1+ to 4+ reflects the mean peak channel intensity of the cells. In the last row, we assign a cumulative score for each of the 12 cell lines (A score to summarize the overall strength of the 12 TAPP analyzed).
RNA was isolated from gliomas in exponential growth. The RNA was analyzed by real-time PCR after the RNA was converted into cDNA. The ΔCT value was calculated by taking the experimental values of the tumor antigen and subtracting the 18S value. nd indicates mRNA was not detected; the CT score was >35 cycles. A ΔCT value of 10 to 14 was a high degree of expression and assigned a 4; a value of 14 to 18 was assigned a 3; a value of 18 to 22 was assigned a 2; and a value of 22 to 26 was assigned a 1. In the next to last row, we assign a score for each of the 12 cell lines (A score) to summarize the overall strengths of the 4 TAPP analyzed.
The sum total of the Σ score from A + Σ score from B = total Σ score‡. The Σ score provides overall evaluation of the strength of expression of the 16 TAPP associated with each of the glioma cell lines.