User:Joel Diaz

Contact Info



 * Joel Diaz
 * Loyola Marymount University
 * Address 1
 * Address 2
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I work in the Your Lab at XYZ University. I learned about OpenWetWare from Instructor, and I've joined because Required by instructor.

Education

 * 2009, BS in Biology, Loyola Marymount University

Research interests

 * 1) Interest 1
 * 2) Interest 2
 * 3) Interest 3

BIOL 598 - Advanced Topics in Gene Expression
Gene Therapy of Human Severe Combined Immunodeficiency (SCID)-X1 Disease

Severe Combined Immunodeficiency Disorder is characterized by a block in T and NK cells. A gene therapy trial for SCID was initiated by using complementary DNA containing a defective γc Moloney retrovirus. Two infants were used as patients and after a 10 month follow up period, both T and NK cells were detectable in the patients.

Figure 1 [[Media:Figure1.jpeg]] shows results from PCR and RT-PCR tests that were run to amplify a 904 base pair stretch of γc gene sequence.

Panel A shows the results of the PCR analysis. The top portion of panel A shows the Standard Curve of a SCID patient. The standard curve was set up with a cell from a γc gene that was not infected. The second portion of panel A shows the first patient, P1. The third portion of panel A is P2, is the second patient. The samples were taken at day 150 since the start of gene therapy (d. +150). The different cells represented in each portion are the following: T CELLS ARE CD3, B CELLS ARE CD19, MONOCYTES ARE CD14, GRANULOCYTES ARE CD15, AND NK CELLS ARE CD56. Actin DNA was amplified in parallel to the γc gene as a loading control. PBMC are used as negative controls.

Panel B shows the results of the RT-PCR analysis. The same blood sample that was taken from the patients for the PCR analysis was used for RT-PCR. Actin was also amplified as a loading control.

Figure 2 [[Media:figure2.jpeg]] shows a longitudinal study of lymphocyte subsets from both patients. This set of data shows that the longer the treatment went on, the larger the increase in number of T and NK cells.

Figure 3 [[Media:figure3.jpeg]] depicts γc protein expression and lymphocyte subsets. In panel A, γc expression on B cells from patient 2 after treatment was undetectable. Panel B depicts percentages of different cells in both patients.

Publications
// leave a comment about a paper here
 * 1) Paper1 pmid=6947258
 * 2) Paper2 pmid=13718526
 * 1) Book1 isbn=0879697164

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