Biomedical subjects

E H Leonard

Publications and source records attributed to E H Leonard.

3 recordsLinked to original sources

PubMed1998

Consistent, persistent expression from modified retroviral vectors in murine hematopoietic stem cells.

Retroviral vectors based on the Moloney murine leukemia virus (MoMuLV) have shown inconsistent levels and duration of expression as well as a propensity for the acquisition of de novo methylation in vivo. MoMuLV-based vectors are known to contain sequences that are capable of suppressing or preventing expression from the long terminal repeat. Previously, we constructed a series of modified retroviral vectors and showed that they function significantly better than MoMuLV-based vectors in vitro. To test the efficacy of the modified vectors in hematopoietic stem cells in vivo, we examined gene expression and proviral methylation in differentiated hematopoietic colonies formed in the spleens of mice after serial transplantation with transduced bone marrow (2 degreesCFU-S). We found a significant increase in the frequency of expression with our modified vectors (>90% expression in vector DNA containing 2 degreesCFU-S) over the frequency observed with the standard MoMuLV-based vector (28% expression in vector containing 2 degreesCFU-S). Expression from the modified vectors was highly consistent, with expression in >50% of the vector-containing 2 degreesCFU-S from all 20 transplant recipients analyzed, whereas expression from the standard MoMuLV-based vector was inconsistent, with expression in 0-10% of the vector containing 2 degreesCFU-S from 8 recipients and expression in >50% of the vector-containing 2 degreesCFU-S from 4 other recipients. In addition, we established that the modified vectors had a lower level of DNA methylation than the control vector. These findings represent significant advances in the development and evaluation of effective retroviral vectors for application in vivo.

Animals↗

PubMed1998

Effectiveness of physicians-in-training counseling for smoking cessation in African Americans.

This study examined the effectiveness of smoking cessation counseling by physicians-in-training (residents) with African-American patients. One hundred fifty-eight family and internal medicine residents at a large urban public general hospital participated in the study; two thirds of the residents underwent a 2-hour smoking cessation training program. Ninety-two of the trained physicians counseled from 1 to 18 patients. The majority of physicians were male, with 8% being current smokers. Over a 26-month period, 1086 patients were randomly assigned to intervention and control (usual care) groups. Mean patient age was 44 years, mean years smoking was 25, and mean number of cigarettes smoked per day was 14. There were no differences in biochemically validated smoking cessation rates between the intervention and control groups at 3 or 12 months postenrollment (2% versus 1.8% and 2.2% versus 2.8%, respectively). Losses to follow-up were high at both 3 and 12 months (38% and 40% respectively). Implications for future trials in minority populations are discussed. A brief physician-based smoking cessation message does not appear to be an effective strategy for use with African-American smokers in a large urban public general hospital.

Black or African American↗

PubMed1997

Infection of human marrow stroma by human immunodeficiency virus-1 (HIV-1) is both required and sufficient for HIV-1-induced hematopoietic suppression in vitro: demonstration by gene modification of primary human stroma.

Patients with human immunodeficiency virus-1 (HIV-1) infection often present with bone marrow (BM) failure that may affect all hematopoietic lineages. It is presently unclear whether this failure reflects a direct viral impairment of the CD34+ hematopoietic progenitor cells or whether the virus affects the BM microenvironment. To study the effects of HIV-1 on the BM microenvironment, we examined the stromal cell monolayers in long-term BM culture (LTBMC), which are the in vitro equivalent of the hematopoietic microenvironment. We assessed the hematopoietic support function (HSF) of human stromal layers by determining the cellular proliferation and colony-forming ability of hematopoietic progenitors from BM cells grown on the stromal layers. We show that the HSF is reduced by in vitro infection of the human stromal cell layer by a monocytotropic isolate of HIV-1 (JR-FL). There is no loss of HSF when the stromal cell layer is resistant to HIV-1 replication, either using murine stromal cell layers that are innately resistant to HIV-1 infection or using human stromal cells genetically modified to express a gene that inhibits HIV-1 replication (an RRE decoy). Decreased HSF was seen using either human or murine hematopoietic cells, if the stromal cells were human cells that were susceptible to HIV-1 infection. These in vitro studies implicate HIV-1 replication in the stroma as the essential component causing decreased hematopoietic cell production in HIV-1 infection.

Bone Marrow↗