Quantitative determination of the human MRP1 and MRP2 mRNA expression in FACS‐sorted peripheral blood CD4⁺, CD8⁺, CD19⁺, and CD56⁺cells

Abstract

On an mRNA level both MRP1 and MRP2 were expressed in peripheral blood cells, with more than sevenfold higher MRP1 expression in all cell populations investigated. The impact of the MRP1 and MRP2 transcription in these cells remains to study. The use of beta2-microglobulin as a housekeeping gene could have a critical impact on the interpretation of RT-PCR data.

Enhanced Separation and Counting of CD4+ T Lymphocytes from Whole Blood Using a Quartz Microfluidic Magnetic Immunoassay

“…The demonstrated device efficiency is similar or better than those reported elsewhere for microfluidic-based systems [10,23]. We did not segregate the T lymphocytes collected from whole blood by sub-types to get a ratio metric measurement of T-lymphocyte sub-types, as reported by other groups and would likely be more challenging but could be done using a multiplexed magnetic bead assay [4,15]. Another recent report by Briggs et al explored the linearity and reproducibility of the measurements [10].…”

“…Due to the lack of access to modern laboratory facilities having FACS machines, physicians in developing nations sometimes make their diagnosis and treatment evaluations without knowledge of the CD4+ T lymphocyte cell count in the patient's blood. The cost and maintenance of FACS machines make the technology expensive for medical centers and hospitals in developing nations [13][14][15][16]. For this reason, treatment options are often not customized to the patients even though it is known that the CD4+ T lymphocytes are highly variable between patients infected with HIV.…”

“…The guidelines from the Center of Disease Control and Prevention reports that T cell counts of 200 cells µl -1 , which is approximately 15% of the total CD4+ cells in the body, serve as one of the important reference points that is used to determine when an HIV infected person is considered immune deficient [11,12]. In developed nations, the standard method for performing CD4+ T lymphocyte counts is done using a flow activated cell-sorting (FACS) machine [13][14][15]. In resource-limited settings, current state of the art equipment like FACS may be too expensive and time-consuming to be used routinely.…”

“…Enzyme linked immunosorbent assays (ELISA) have been developed and used; however, the results from these methods are inconsistent [15][16][17][18][19][20][21][22]. Additionally, other immune-magnetic separation techniques have been explored but these techniques suffered from low throughput and were therefore relatively slow [18,19,22].…”

Expression of MRP1 gene in acute leukemia

“…We used β2M because it has no pseudogene and the use of β2M as a housekeeping gene has a critical impact on the interpretation of PCR data. 32 In our assay, the dynamic quantification range (from 6 x 10 5 copies to 6 x 10 10 copies) was satisfactory for clinical use. When this system was applied to clinical samples, we found that there was a statistically significant increase (p < 0.01) in the average MRP1 expression level at the time of nonresponse or early relapse, compared with patients in remission.…”