Tandem rhodium catalysis: exploiting sulfoxides for asymmetric transition-metal catalysis.

Sulfoxides are uncommon substrates for transition-metal catalysis due to their propensity to inhibit catalyst turnover. In a collaborative effort with Ken Houk, we developed the first dynamic kinetic resolution (DKR) of allylic sulfoxides using asymmetric rhodium-catalyzed hydrogenation. A detailed mechanistic analysis of this transformation using both experimental and theoretical methods revealed rhodium to be a tandem catalyst that promoted both hydrogenation of the alkene and racemization of the allylic sulfoxide. Using a combination of deuterium labelling and DFT studies, a novel mode of allylic sulfoxide racemization via a Rh(III)-π-allyl intermediate was identified.

CD28-independent costimulation of T cells in alloimmune responses.

T cell costimulation by B7 molecules plays an important role in the regulation of alloimmune responses. Although both B7-1 and B7-2 bind CD28 and CTLA-4 on T cells, the role of B7-1 and B7-2 signaling through CTLA-4 in regulating alloimmune responses is incompletely understood. To address this question, we transplanted CD28-deficient mice with fully allogeneic vascularized cardiac allografts and studied the effect of selective blockade of B7-1 or B7-2. These mice reject their grafts by a mechanism that involves both CD4(+) and CD8(+) T cells. Blockade of CTLA-4 or B7-1 significantly accelerated graft rejection. In contrast, B7-2 blockade significantly prolonged allograft survival and, unexpectedly, reversed the acceleration of graft rejection caused by CTLA-4 blockade. Furthermore, B7-2 blockade prolonged graft survival in recipients that were both CD28 and CTLA-4 deficient. Our data indicate that B7-1 is the dominant ligand for CTLA-4-mediated down-regulation of alloimmune responses in vivo and suggest that B7-2 has an additional receptor other than CD28 and CTLA-4 to provide a positive costimulatory signal for T cells.

Bacterial pathogens induce abscess formation by CD4(+) T-cell activation via the CD28-B7-2 costimulatory pathway.

Abscesses are a classic host response to infection by many pathogenic bacteria. The immunopathogenesis of this tissue response to infection has not been fully elucidated. Previous studies have suggested that T cells are involved in the pathologic process, but the role of these cells remains unclear. To delineate the mechanism by which T cells mediate abscess formation associated with intra-abdominal sepsis, the role of T-cell activation and the contribution of antigen-presenting cells via CD28-B7 costimulation were investigated. T cells activated in vitro by zwitterionic bacterial polysaccharides (Zps) known to induce abscess formation required CD28-B7 costimulation and, when adoptively transferred to the peritoneal cavity of naïve rats, promoted abscess formation. Blockade of T-cell activation via the CD28-B7 pathway in animals with CTLA4Ig prevented abscess formation following challenge with different bacterial pathogens, including Staphylococcus aureus, Bacteroides fragilis, and a combination of Enterococcus faecium and Bacteroides distasonis. In contrast, these animals had an increased abscess rate following in vivo T-cell activation via CD28 signaling. Abscess formation in vivo and T-cell activation in vitro required costimulation by B7-2 but not B7-1. These results demonstrate that abscess formation by pathogenic bacteria is under the control of a common effector mechanism that requires T-cell activation via the CD28-B7-2 pathway.

The role of CD154-CD40 versus CD28-B7 costimulatory pathways in regulating allogeneic Th1 and Th2 responses in vivo.

We used signal transducer and activator of transcription 4 (STAT4) and STAT6 gene knockout (-/-) mice as recipients of fully mismatched cardiac allografts to study the role of T-cell costimulatory pathways in regulating allogeneic T-helper 1 (Th1) versus Th2 responses in vivo. STAT4(-/-) mice have impaired Th1 responses, whereas STAT6(-/-) mice do not generate normal Th2 responses. Cardiac allografts from C57BL/6 mice were transplanted into normal wild-type (WT), STAT4(-/-), and STAT6(-/-) BALB/c recipients. STAT4(-/-) and STAT6(-/-) mice rejected their grafts with the same tempo as untreated WT recipients. CD28-B7 blockade by a single injection of CTLA4Ig induced long-term engraftment and donor-specific tolerance in all three groups of recipients. CD154 blockade by a single injection of MR1 was effective in prolonging allograft survival and inducing tolerance in STAT4(-/-) mice but was only marginally effective in STAT6(-/-) recipients and WT controls. In addition, a similar protocol of MR1 was ineffective in prolonging graft survival in CD28(-/-) BALB/c recipients, suggesting that the lack of efficacy seen in WT and STAT6(-/-) mice is not due to the presence of a functional CD28-B7 pathway. Furthermore, there was a similar differential effect of CD28-B7 versus CD154-CD40 blockade in inhibiting immune responses in animals immunized with ovalbumin and complete Freund's adjuvant. These novel data indicate that Th1 and Th2 cells are differentially regulated by CD28-B7 versus CD154-CD40 costimulation pathways in vivo and may have potential implications for the development of therapeutic strategies such as T-cell costimulatory blockade in humans.

The role of costimulatory molecules as targets for new immunosuppressives in transplantation.

T-cell costimulation is critical for all T-cell-mediated immune responses that are responsible for endpoints such as allograft rejection or autoimmune disease. Recent experimental data elucidate specific pathways for T-cell activation and negative regulatory mechanisms via unique costimulatory molecules. These data have implications for the development of novel therapeutic strategies to prevent graft rejection and improve long-term graft survival in transplant patients, and to treat autoimmune diseases in humans.

Transplantation tolerance: the concept and its applicability.

Recent advances have enabled researchers to induce tolerance in animal transplant models. Although it has been relatively easy to do so in rodents, it has been much more difficult to translate such strategies into primates. Understanding the cellular and molecular mechanisms of the alloimmune response has prompted the development of novel strategies that may obviate the need for immunosuppression in humans. Mechanisms of tolerance and promising new therapies, as well as the inherent difficulties in bringing them into clinical practice, are reviewed.

Coronary angiographic rates after stress single-photon emission computed tomographic scintigraphy.

BACKGROUND: There is speculation that coronary angiography may be overused for the assessment of coronary artery disease. Because of its proved ability to differentiate high- and low-risk subsets of patients with coronary artery disease, myocardial perfusion scintigraphy should be an effective strategy in patient selection for angiography. This retrospective clinical study analyzed the relation between scintigraphic findings and subsequent angiography. METHODS AND RESULTS: Coronary angiographic rates were determined by following up on all stress single-photon emission computed tomographic (SPECT) myocardial perfusion studies performed in a cardiology practice nuclear laboratory during a 26-month interval. All patients were followed up for at least 3 months; mean follow-up was 8.9 months. Subsequent angiography was determined from catheterization laboratory logs, medical records, and telephone contact. Scintigraphic studies were graded according to presence or absence of reversible perfusion defects, affected coronary territories, and lung uptake of 201Tl. Scans were categorized high risk if more than two of the following three criteria were met: reversibility of left anterior descending or multivessel distributions or abnormal lung uptake of thallium. Of 4162 studies, 60% had reversible perfusion defects. Of such studies, 32% were followed up by angiography versus 3.5% without reversible defects. Among studies with reversible defects, the subsequent angiography rate was 60% for those showing high-risk reversibility, compared with 9% for all other studies demonstrating reversibility. Multivariate logistic regression identified high-risk reversibility (odds ratio 20.96) and any reversibility (odds ratio 8.22) as the strongest predictors of angiography. Other correlates of lesser statistical significance were angina and absence of prior infarction or coronary bypass. CONCLUSION: In this large retrospective study, the results of SPECT scintigraphy overpowered all other clinical and treadmill characteristics in determining the likelihood of subsequent coronary angiography. Only rarely were patients categorized as relatively low risk by scintigraphy referred for angiography. As such, SPECT scintigraphy as used in a private practice, self-referral environment appears to be effective in stratifying potential candidates for coronary angiography.