Biased expression of T cell receptor genes characterizes activated T cells in multiple sclerosis cerebrospinal fluid.

To better characterize the inflammatory response that occurs in the nervous system in multiple sclerosis (MS), T-cell receptor (TCR) gene expression was quantified from cerebrospinal fluid (CSF) cells of 21 patients with active disease. Unstimulated CSF cells expressed each of 22 different TCR beta chain variable region (V beta) gene families in proportion to their expression in simultaneously sampled peripheral blood. When CSF cells from individuals with MS were expanded by in vitro culture in T-cell growth factor/interleukin 2 and 4-containing medium (TCGF/IL2/IL4), restricted numbers of V beta genes were expressed. In many subjects, expanded CSF cells expressed predominantly V beta 2. In contrast to CSF, expansion of corresponding peripheral blood mononuclear cells (PBMC) in TCGF/IL2/IL4 resulted in persistent expression of all V beta gene families. Within individuals, different V beta genes were overexpressed by PBMC compared with CSF cells. No effect of the HLA haplotype of the individual on CSF V beta gene expression was observed. Expanded CSF cells retained their capacity to respond to mitogen stimulation, but the proliferative response to myelin basic protein (MBP) was not enhanced. Finally, freshly obtained CSF cells stimulated directly with MBP also expressed a limited number of V beta genes, although these were generally different from patterns observed following stimulation with TCGF/IL2/IL4. Thus, restricted populations of T cells capable of responding to TCGF/IL2/IL4, presumably reflecting in vivo activated cells, are compartmentalized in the nervous system in MS.

Unusual organization of the human T-cell receptor beta-chain gene complex is linked to recombination hotspots.

Two rare Sfi I polymorphisms of 360 kb and 280 kb present within the human T-cell antigen receptor beta-chain gene complex were revealed by pulsed-field gel electrophoresis. They represent allelic variants of the polymorphic 330- and 300-kb Sfi I fragments previously described. The 360-kb polymorphism results from duplication of the 30-kb DNA fragment responsible for the 330/300-kb insertion/deletion-related polymorphism. The 280-kb polymorphism results from a 20-kb deletion from the 300-kb SfiI allele. The rare polymorphisms also map on either side of a Sal I site located near a recombination hotspot, suggesting that germline duplications and deletions arose from nonhomologous crossover events.

The human T-cell receptor beta-chain repertoire: longitudinal fluctuations and assessment in MHC matched populations.

The influence of the environment and of the major histocompatibility complex (MHC) in shaping the human T-cell receptor beta-chain variable region (TCRBV) repertoire has not been systematically studied. Here, expression of TCRBV gene families was estimated by a sensitive polymerase chain reaction (PCR)-based method. Serial studies of peripheral blood, performed at 2-week intervals over a 3-month period, revealed that fluctuation in the expression of many TCRBV genes occurred in healthy individuals and in the absence of clinically evident infections. Fluctuation of TCRBV4, TCRBV5.2, TCRBV9, and TCRBV13.1 genes were present in all subjects. Additional TCRBV genes fluctuated in some but not in other individuals. Comparison of the TCRBV repertoire between these unrelated individuals indicated differences in the mean expression of TCRBV5.1, TCRBV9, TCRBV11, TCRBV15, TCRBV17, and TCRBV20 genes. For any TCRBV gene, intersubject differences were generally of a magnitude of twofold or less. Larger differences characterized the TCRBV repertoire of CD4 compared to CD8 cells. Some differences, for example over-representation of TCRBV2 and TCRBV5.1 on CD4, and TCRBV10, TCRBV14, and TCRBV16 on CD8 cells, were present in most subjects. Individuals homozygous for DR2- or DR3-bearing extended MHC haplotypes displayed similar individual variability of TCRBV expression. These data indicate that the circulating TCRBV repertoire in humans is both dynamic and diverse. Both environment and MHC effects contribute to the diversity of TCRBV expression.

Comparing ambulatory care practices of primary care and traditional medicine residents.

Although special residency programs preparing internists for primary care have been in existence for a decade, little is known about whether these tracks have achieved their goals. As part of a multicenter evaluation of ambulatory care at four university hospitals, 1,040 patient care encounters were reviewed for 16 primary-care and 41 traditional medicine residents. Using a chart-based audit, the authors examined 16 discrete items of patient care to assess resident management in the following areas: screening for colorectal carcinoma, management of hypertension, benzodiazepine drug prescribing, and management of chronic lung disease. Their hypothesis that primary care residents would score higher than traditional medicine residents in the areas of screening, prevention, and prescribing of drugs was not supported. There was no association between type of training and performance of a task with the following exception: second-year primary care residents screened for colorectal carcinoma in 86% (126) of patients whose charts were audited, while second-year traditional medicine residents did so in 77% (160) (P less than 0.025). This difference was not maintained when the residents were reaudited 1 year later. Both groups of residents scored high in all areas with the following exceptions: documentation of the amount of sedative dispensed and immunization of susceptible patients against pneumococcus and influenza. The ambulatory practices of both groups of residents exceeded expectations, probably because of the wider influence of primary care training.