Translation in action: Influence, collaboration, and evolution of COVID-19 research with Clinical and Translational Science Awards consortium support.

The National Institutes of Health (NIH)'s Clinical and Translational Science Awards (CTSA) consortium aims to accelerate translational processes that move discoveries from bench to bedside. The coronavirus disease 2019 (COVID-19) pandemic presented unmatched challenges and applications for CTSA hubs nationwide. Our study used bibliometrics to assess features of COVID-19 publications supported by the national CTSA program to characterize the consortium's response to the pandemic. Our goal was to understand relative scientific influence, collaboration across hubs, and trends in research emphasis over time. We identified publications from NIH's curated iSearch COVID-19 Publication Portfolio from February 2020 to February 2023; 3234 peer-reviewed articles relevant to COVID-19 cited a CTSA grant. All 66 CTSA hubs were represented, with large-size and longstanding hubs contributing more publications. Most publications cited UL1 grants, 457 cited KL2/TL1 training grants, and 164 cited multiple hub grants. Compared to a random sample of non-CTSA-supported COVID-19 publications, the CTSA portfolio exhibited greater clinical relevance, more human research, and higher altmetric and citation influence. Results were similar for multi-hub publications involving networked initiatives like multi-site clinical trials or the National COVID-19 Cohort Collaborative. Shifts from molecular/cellular-oriented research toward human-oriented research over time were evident, demonstrating translation in action. Results illuminate how the CTSA consortium confronted the pandemic through high-quality projects oriented toward human research, working across hubs on high-value collaborations, advancing along the translational spectrum over time. Findings validate CTSA hubs as critical support structures during health emergencies.

Translating Scientific Discovery Into Health Policy Impact: Innovative Bibliometrics Bridge Translational Research Publications to Policy Literature.

To understand how translational science efforts lead to outcomes, it is common to examine publications as a key step in the translational process. The National Institutes of Health's Clinical and Translational Science Awards (CTSA) program aims to accelerate that process by providing support to investigators. Although it is challenging to measure the impact of such support on translational outcomes, CTSA-supported research that arises in research publications can advance translation through use of these publications in public policy and guideline documents from government health agencies, intergovernmental organizations, and other outlets. Using cutting-edge bibliometric tools, the authors evaluated how CTSA-supported research has extended its impact beyond academic silos to influence public policy literature. The authors identified approximately 118,490 publications that acknowledged receiving support from a CTSA hub, from the inception of the program in 2006 through 2021. Articles were queried in the Overton policy database, which indexes references to publications in global policy literature. The search revealed 13% of CTSA-supported articles were referenced in policy documents, significantly more than the expected proportion (10%) calculated by Overton. References came from 576 policy source outlets across 87 countries, predominantly the United States and Europe. The most frequent sources included guidelines in PubMed Central, the World Health Organization, and the Centers for Disease Control and Prevention. The authors illustrate the bridge from translational research to public policy with case studies of 6 articles based on CTSA-supported research and having notable policy impact. They found articles with greater clinical relevance, altmetric attention (i.e., nonacademic community/public attention), and academic citation influence were more likely to be referenced in policy literature. Study findings help to characterize the kinds of research that have influenced and may be expected to influence health policy in the future.

Breaking Through Barriers: Factors That Influence Behavior Change Toward Leadership for Women in Academic Medicine.

Under-representation of women in leadership at Academic Medical Centers (AMCs) is a known challenge such that, in 2021, women made up only 28% of department chairs. AMCs are addressing the dearth of women leaders through targeted programming to create leadership pipelines of qualified women. The FLEX Leadership Development Program at the Case Western Reserve University (CWRU) School of Medicine prepares women faculty for increased leadership opportunities. FLEX includes the opportunity to leverage executive coaching to accomplish individual goals. The FLEX program has the explicit goal of increasing the number of women in visible leadership positions in academic medicine and health sciences. Semi-structured interviews were conducted with 25 graduates from seven FLEX cohorts (2012-2018). Participants reflected diversity in academic rank, terminal degree, racial/ethnic background, years of employment, and institutional affiliation. Interviews consisted of eight questions with additional probes to elicit lived experiences. Analysis consisted of two-stage open- and axial-coding of interview transcripts to understand: What factors facilitated behavior change following FLEX training? The analysis revealed five overarching themes: (1) Communication skills; (2) Self-Efficacy; (3) Networking; (4) Situational Awareness; and (5) Visioning. FLEX graduates reported achieving both personal and professional growth by drawing upon peer networks to proactively seek new leadership opportunities. These results suggest that the enduring benefits of the FLEX Program include improved communication skills, expanded situational awareness and relational capacity, greater self-efficacy and self-confidence, improved networking with an understanding of the value of networking. All these factors led FLEX graduates to have greater visibility and to engage with their colleagues more effectively. Similarly, FLEX graduates could better advocate for themselves and for others as well as paying it forward to mentor and train the next generation of faculty. Finally, participants learned to re-evaluate their goals and their career vision to be able to envision themselves in greater leadership roles. The five factors that strongly influenced behavior change provide valuable constructs for other programs to examine following leadership development training. Ongoing studies include examining successful leadership position attainment, personal goal attainment, and measuring changes in leadership self-efficacy.

Scholarly Productivity Evaluation of KL2 Scholars Using Bibliometrics and Federal Follow-on Funding: Cross-Institution Study.

BACKGROUND: Evaluating outcomes of the clinical and translational research (CTR) training of a Clinical and Translational Science Award (CTSA) hub (eg, the KL2 program) requires the selection of reliable, accessible, and standardized measures. As measures of scholarly success usually focus on publication output and extramural funding, CTSA hubs have started to use bibliometrics to evaluate the impact of their supported scholarly activities. However, the evaluation of KL2 programs across CTSAs is limited, and the use of bibliometrics and follow-on funding is minimal. OBJECTIVE: This study seeks to evaluate scholarly productivity, impact, and collaboration using bibliometrics and federal follow-on funding of KL2 scholars from 3 CTSA hubs and to define and assess CTR training success indicators. METHODS: The sample included KL2 scholars from 3 CTSA institutions (A-C). Bibliometric data for each scholar in the sample were collected from both SciVal and iCite, including scholarly productivity, citation impact, and research collaboration. Three federal follow-on funding measures (at the 5-year, 8-year, and overall time points) were collected internally and confirmed by examining a federal funding database. Both descriptive and inferential statistical analyses were computed using SPSS to assess the bibliometric and federal follow-on funding results. RESULTS: A total of 143 KL2 scholars were included in the sample with relatively equal groups across the 3 CTSA institutions. The included KL2 scholars produced more publications and citation counts per year on average at the 8-year time point (3.75 publications and 26.44 citation counts) than the 5-year time point (3.4 publications vs 26.16 citation counts). Overall, the KL2 publications from all 3 institutions were cited twice as much as others in their fields based on the relative citation ratio. KL2 scholars published work with researchers from other US institutions over 2 times (5-year time point) or 3.5 times (8-year time point) more than others in their research fields. Within 5 years and 8 years postmatriculation, 44.1% (63/143) and 51.7% (74/143) of KL2 scholars achieved federal funding, respectively. The KL2-scholars of Institution C had a significantly higher citation rate per publication than the other institutions (P<.001). Institution A had a significantly lower rate of nationally field-weighted collaboration than did the other institutions (P<.001). Institution B scholars were more likely to have received federal funding than scholars at Institution A or C (P<.001). CONCLUSIONS: Multi-institutional data showed a high level of scholarly productivity, impact, collaboration, and federal follow-on funding achieved by KL2 scholars. This study provides insights on the use of bibliometric and federal follow-on funding data to evaluate CTR training success across institutions. CTSA KL2 programs and other CTR career training programs can benefit from these findings in terms of understanding metrics of career success and using that knowledge to develop highly targeted strategies to support early-stage career development of CTR investigators.

What does team science look like across the CTSA consortium? A qualitative analysis of the Great CTSA Team Science Contest submissions.

INTRODUCTION: The Great CTSA Team Science Contest (GTSC) was developed to discover how Clinical and Translational Science Award (CTSA) hubs promote and support team science [1]. The purpose of this study was a secondary qualitative analysis of the GTSC submissions to better understand the diversity of team science initiatives across the CTSA consortium. METHODS: Secondary qualitative analysis of the GTSC data addressed the following research questions, which defined the top-level coding: (1) What CTSA component sponsored it? (2) Who was the team doing the work? (3) Who were the intended beneficiaries? (4) What was the intended outcome? (5) What strategies did they use? (6) What translational science (TS) stage was addressed? (7) How do they align with the NCATS team science strategic goals? (8) How do the CTSA's team science efforts align with the National Academies Research Council (NRC) recommendations for enhancing the effectiveness of team science? RESULTS: The GTSC received 170 submissions from 45 unique CTSA hubs. Qualitative analysis revealed a great variety of team science strategies for virtually all team science stakeholders. In addition to strategies to promote team science, results show successful examples that focus on outcomes and illustrate ways of measuring success. CONCLUSIONS: The GTSC shows that the CTSA consortium is involved in an extremely diverse array of team science activities, which align well with both the NRC recommendations for enhancing the effectiveness of team science and the NCATS strategic goals for team science. Future research should evaluate the efficacy of team science strategies.

A retrospective case study of successful translational research: Gazelle Hb variant point-of-care diagnostic device for sickle cell disease.

Evaluation researchers at Clinical and Translational Science Award (CTSA) hubs are conducting retrospective case studies to evaluate the translational research process. The objective of this study was to deepen knowledge of the translational process and identify contributors to successful translation. We investigated the successful translation of the HemeChip, a low-cost point-of-care diagnostic device for sickle cell disease, using a protocol for retrospective translational science case studies of health interventions developed by evaluators at the National Health Institutes (NIH) and CTSA hubs. Development of the HemeChip began in 2013 and evidence of device use and impact on public health is growing. Data collection methods included five interviews and a review of press, publications, patents, and grants. Barriers to translation included proving novelty, manufacturing costs, fundraising, and academic-industry relations. Facilitators to translation were CTSA pilot program funding, university resources, entrepreneurship training, due diligence, and collaborations. The barriers to translation, how they were overcome, and the key facilitators identified in this case study pinpoint areas for consideration in future funding mechanisms and the infrastructure required to enable successful translation.

A protocol for retrospective translational science case studies of health interventions.

The critical processes driving successful research translation remain understudied. We describe a mixed-method case study protocol for analyzing translational research that has led to the successful development and implementation of innovative health interventions. An overarching goal of these case studies is to describe systematically the chain of events between basic, fundamental scientific discoveries and the adoption of evidence-based health applications, including description of varied, long-term impacts. The case study approach isolates many of the key factors that enable the successful translation of research into practice and provides compelling evidence connecting the intervention to measurable changes in health and medical practice, public health outcomes, and other broader societal impacts. The goal of disseminating this protocol is to systematize a rigorous approach, which can enhance reproducibility, promote the development of a large collection of comparable studies, and enable cross-case analyses. This approach, an application of the "science of translational science," will lead to a better understanding of key research process markers, timelines, and potential points of leverage for intervention that may help facilitate decisions, processes, and policies to speed the sustainable translational process. Case studies are effective communication vehicles to demonstrate both accountability and the impacts of the public's investment in research.

Using bibliometrics to evaluate translational science training: evidence for early career success of KL2 scholars.

INTRODUCTION: Evaluating clinical and translational research (CTR) mentored training programs is challenging because no two programs are alike. Careful selection of appropriate metrics is required to make valid comparisons between individuals and between programs. The KL2 program provides mentored-training for early-stage CTR investigators. Clinical and Translational Awards across the country have unique KL2 programs. The evaluation of KL2 programs has begun to incorporate bibliometrics to measure KL2 scholar and program impact. METHODS: This study investigated demographic differences in bibliometric performance and post-K award funding of KL2 scholars and compared the bibliometric performance and post-K award federal funding of KL2 scholars and other mentored-K awardees at the same institution. Data for this study included SciVal and iCite bibliometrics and National Institutions of Health RePORTER grant information for mentored-K awardees (K08, K23, and KL2) at Case Western Reserve University between 2005 and 2013. RESULTS: Results showed no demographics differences within the KL2 program scholars. Bibliometric differences between KL2 and other mentored-K awardee indicated an initial KL2 advantage for the number of publications at 5 years' post-matriculation (i.e., the start of the K award). Regression analyses indicated the number of initial publications was a significant predictor of federal grant funding at the same time point. Analysis beyond the 5-year post-matriculation point did not result in a sustained, significant KL2 advantage. CONCLUSIONS: Factors that contributed to the grant funding advantage need to be determined. Additionally, differences between translational and clinical bibliometrics must be interpreted with caution, and appropriate metrics for translational science must be established.

Scope, Influence, and Interdisciplinary Collaboration: The Publication Portfolio of the NIH Clinical and Translational Science Awards (CTSA) Program From 2006 Through 2017.

The Clinical and Translational Science Awards (CTSA) program sponsors an array of innovative, collaborative research. This study uses complementary bibliometric approaches to assess the scope, influence, and interdisciplinary collaboration of publications supported by single CTSA hubs and those supported by multiple hubs. Authors identified articles acknowledging CTSA support and assessed the disciplinary scope of research areas represented in that publication portfolio, their citation influence, interdisciplinary overlap among research categories, and characteristics of publications supported by multihub collaborations. Since 2006, CTSA hubs supported 69,436 articles published in 4,927 journals and 189 research areas. The portfolio is well distributed across diverse research areas with above-average citation influence. Most supported publications involved clinical/health sciences, for example, neurology and pediatrics; life sciences, for example, neuroscience and immunology; or a combination of the two. Publications supported by multihub collaborations had distinct content emphasis, stronger citation influence, and greater interdisciplinary overlap. This study characterizes the CTSA consortium's contributions to clinical and translational science, identifies content areas of strength, and provides evidence for the success of multihub collaborations. These methods lay the foundation for future investigation of the best policies and priorities for fostering translational science and allow hubs to understand their progress benchmarked against the larger consortium.

A Consult Service to Support and Promote Community-Based Research: Tracking and Evaluating a Community-based Research Consult Service.

PURPOSE: This study describes the design, operation and evaluation of a community-based research (CBR) consult service within the setting of a Clinical and Translational Science Award (CTSA) institution. To our knowledge, there are no published evaluations of a CBR consult service at a CTSA hub. METHODS: A community-based research Consult Service was created to support faculty, health care providers/research coordinators, trainees, community-based organizations and community members. A framework was developed to assess the stages of client engagement and to foster clear articulation of client needs and challenges. A developmental evaluation system was integrated with the framework to track progress, store documents, continuously improve the consult service and assess research outcomes. RESULTS: This framework provides information on client numbers, types, services used and successful outreach methods. Tracking progress reveals reasons that prevent clients from completing projects and facilitates learning outcomes relevant to clients and funding agencies. Clients benefit from the expert knowledge, community connections and project guidance provided by the Consult Service team, increasing the likelihood of study completion and achieving research outcomes. CONCLUSION: This study offers a framework by which CTSA institutions can expand their capacity to conduct and evaluate community-based research while addressing challenges that inhibit community engagement.

RMS: a platform for managing cross-disciplinary and multi-institutional research project collaboration.

BACKGROUND: Cross-institutional cross-disciplinary collaboration has become a trend as researchers move toward building more productive and innovative teams for scientific research. Research collaboration is significantly changing the organizational structure and strategies used in the clinical and translational science domain. However, due to the obstacles of diverse administrative structures, differences in area of expertise, and communication barriers, establishing and managing a cross-institutional research project is still a challenging task. We address these challenges by creating an integrated informatics platform to reduce the barriers to biomedical research collaboration. RESULTS: The Request Management System (RMS) is an informatics infrastructure designed to transform a patchwork of expertise and resources into an integrated support network. The RMS facilitates investigators' initiation of new collaborative projects and supports the management of the collaboration process. In RMS, experts and their knowledge areas are categorized and managed structurally to provide consistent service. A role-based collaborative workflow is tightly integrated with domain experts and services to streamline and monitor the life-cycle of a research project. The RMS has so far tracked over 1,500 investigators with over 4,800 tasks. The research network based on the data collected in RMS illustrated that the investigators' collaborative projects increased close to 3 times from 2009 to 2012. Our experience with RMS indicates that the platform reduces barriers for cross-institutional collaboration of biomedical research projects. CONCLUSION: Building a new generation of infrastructure to enhance cross-disciplinary and multi-institutional collaboration has become an important yet challenging task. In this paper, we share the experience of developing and utilizing a collaborative project management system. The results of this study demonstrate that a web-based integrated informatics platform can facilitate and increase research interactions among investigators.

Visualizing and evaluating the growth of multi-institutional collaboration based on research network analysis.

Research collaboration plays an important role in scientific productivity and academic innovation. Multi-institutional collaboration has become a vital approach for integrating multidisciplinary resources and expertise to enhance biomedical research. There is an increasing need for analyzing the effect of multi-institutional research collaboration. In this paper, we present a collaboration analysis pipeline based on research networks constructed from publication co-authorship relationship. Such research networks can be effectively used to render and analyze large-scale institutional collaboration. The co-authorship networks of the Cleveland Clinical and Translational Science Collaborative (CTSC) were visualized and analyzed. SciVal Expert™ was used to extract publication data of the CTSC members. The network was presented in informative and aesthetically appealing diagrams using the open source visualization package Gephi. The analytic result demonstrates the effectiveness of our approach, and it also indicates the substantial growth of research collaboration among the CTSC members crossing its partner institutions.

Cellular proliferative response to cardiac troponin-I in patients with idiopathic dilated cardiomyopathy.

BACKGROUND: Approximately 20% of patients with idiopathic dilated cardiomyopathy (iDCM) have autoantibodies (AAbs) specific to cardiac troponin-I (cTnI). However, there has been no work evaluating active cellular autoimmunity. We aimed to identify a cTnI-stimulated cellular autoimmune response and to correlate our findings with cTnI AAb production. METHODS: Samples were obtained from stable ambulatory iDCM patients and healthy controls. Peripheral blood monocytes were incubated with cTnI, and cellular proliferation was measured using flow cytometry. AAbs against cTnI were detected by enzyme-linked immunosorbent assay. RESULTS: A positive cellular proliferative response to cTnI was identified in 20.5% (9/44) patients with iDCM and 5.7% (2/35) of healthy controls (p < 0.05). Positive cTnI AAbs were identified in 20% (7/35) of healthy controls and 13.6% (6/44) of patients with iDCM (p = NS). The presence of cTnI AAbs did not correlate with a positive cellular proliferative response. However, patients with iDCM who had an AAb response to cTnI were less likely to be taking a statin (p < 0.05). CONCLUSIONS: A cellular autoimmune response to cTnI is demonstrated in a subset of patients with iDCM. However, the presence of a cellular response did not correlate with the presence of AAbs to the same antigen.

Antiinflammatory autoimmune cellular responses to cardiac troponin I in idiopathic dilated cardiomyopathy.

BACKGROUND: Autoimmune mechanisms, particularly through generation of autoantibodies, may contribute to the pathophysiology of idiopathic dilated cardiomyopathy (iDCM). The precise role of cellular autoimmune responses to cardiac-specific antigens has not been well described in humans. The purpose of this study was to characterize the cellular autoimmune response to cardiac troponin I (cTnI), specifically, the release of cytokines by peripheral blood mononuclear cells (PBMCs), in subjects with iDCM and healthy control subjects. METHODS AND RESULTS: We performed enzyme-linked immunospot assays on PBMCs isolated from subjects with iDCM and healthy control subjects to examine the ex vivo interferon-gamma (IFN-γ) and interleukin-10 (IL-10) production in response to cTnI exposure. Thirty-five consecutive subjects with iDCM (mean age 53 ± 11 years, 60% male, left ventricular ejection fraction 23 ± 7%) and 26 control subjects (mean age 46 ± 13 years, 46% male) were prospectively enrolled. IFN-γ production in response to cTnI did not differ between the groups (number of secreting cells 26 ± 49 vs 38 ± 53, respectively; P = .1). In contrast, subjects with iDCM showed significantly higher IL-10 responses to cTnI compared with control subjects (number of secreting cells 386 ± 428 vs 152 ± 162, respectively; P < .05). Among iDCM subjects, heightened IL-10 response to cTnI was associated with reduced systemic inflammation and lower prevalence of advanced diastolic dysfunction compared with those with normal IL-10 response to cTnI. CONCLUSIONS: Our preliminary findings suggest that a heightened cellular autoimmune IL-10 response to cTnI is detectable in a subset of patients with iDCM, which may be associated with reduced systemic levels of high-sensitivity C-reactive protein and lower prevalence of advanced diastolic dysfunction.

Recent insights into the role of autoimmunity in idiopathic dilated cardiomyopathy.

Dilated cardiomyopathy is a devastating disease associated with poor outcomes. Although the etiology of this disease remains largely unknown, so-called "idiopathic" dilated cardiomyopathy (iDCM) is associated with evidence of an autoimmune process that may be contributing to the pathophysiology of this disease. Indeed, iDCM shares many characteristics with other autoimmune diseases, including an association with systemic and organ-specific inflammation, an association with viral infections, a genetic predisposition, and a correlation with specific human leukocyte antigen subtypes. Additionally, numerous pathologic cardiac-specific autoantibodies have been associated with iDCM, including those against alpha-myosin, the beta(1)-adrenoceptor, and cardiac troponin I. This review highlights the emerging evidence regarding autoimmune characteristics of iDCM, and summarizes the data of specific immunomodulatory therapies used to target autoimmune mechanisms in the treatment of patients with this devastating disease.

Interferon gamma allelic variants: sex-biased multiple sclerosis susceptibility and gene expression.

BACKGROUND: Interferon (IFN) gamma (IFNG) allelic variants are associated with susceptibility to multiple sclerosis (MS) in men but not in women. OBJECTIVES: To conduct a high-density linkage disequilibrium association study of IFNG and the surrounding region for sex-associated MS susceptibility bias and to evaluate whether IFNG allelic variants associated with MS susceptibility are associated with expression. DESIGN: Genotype case-control study, quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay expression analyses for IFN gamma. SETTING: Three independently ascertained populations from the Mayo Clinic, Rochester, Minnesota, Queen's University of Belfast, Belfast, Ireland, and University of Leuven, Leuven, Belgium. PATIENTS: For linkage disequilibrium, 861 patients with MS (293 men and 568 women) and 843 controls (340 men and 503 women) derived from the US (population-based) and the Northern Ireland and Belgium (clinic-based) cohorts were studied. For expression analyses, 50 US patients were selected to enrich for homozygotes and to achieve a balance between men and women. INTERVENTIONS: Twenty markers were genotyped over the 120-kilobase region harboring IFNG and the interleukin 26 gene (IL26). MAIN OUTCOME MEASURES: Expression of IFN gamma was evaluated by qPCR and enzyme-linked immunosorbent assay in stimulated peripheral blood mononuclear cells. RESULTS: Multiple markers were associated with MS susceptibility in men but not in women. The sex-specific susceptibility markers, of which rs2069727 was the strongest, were confined to IFNG. Carriers of rs2069727*G had higher expression than noncarriers. The effect of genotype in the qPCR experiments was also evident in men but not in women. CONCLUSIONS: IFNG is associated with sex bias in MS susceptibility and with expression of IFN gamma in MS. These observations add to a growing body of literature that implicates an interaction between sex and IFN gamma expression in a variety of disease states.

Multiple sclerosis patients show sexual dimorphism in cytokine responses to myelin antigens.

Multiple sclerosis affects more women than men. The reasons for this are unknown. Previously, we have shown significant differences in women versus men in inflammatory cytokine responses to the major protein component of myelin, proteolipid protein (PLP), which is thought to be a target in MS patients. Here, using the ELISPOT assay, we examined sex differences in single-cell secretion of Th1 and Th2 cytokines from freshly isolated PBMC between relapsing remitting (RR) MS patients and healthy individuals. Cells were stimulated with MS-associated antigens including proteolipid protein (PLP), myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), and non-disease related antigens. Our data show a sex bias in the cytokine responses to multiple MS-relevant myelin antigens: Women with MS show IFNgamma-skewed responses and men with MS show IL-5-skewed responses. These data extend our previous findings [Pelfrey, C.M., Cotleur, A.C., Lee, J.C., Rudick, R.A. 2002. Sex differences in cytokine responses to myelin peptides in multiple sclerosis. J. Neuroimmunol. 130, 211-223.]: (1) by demonstrating gender skewing in cytokine responses to an expanded myelin antigen repertoire, which includes MBP, MOG and PLP; (2) by showing TNFalpha and IL-10 do not display comparable gender skewing compared to IFNgamma and IL5; (3) by defining the patient population as early, untreated RRMS patients to avoid confounding factors, such as different disease stages/disability and immunomodulatory therapy; and (4) by showing HLA type does not appear to underlie the gender differences. These findings may explain increased susceptibility to MS in women and could contribute to the differences in disease severity between men and women.

Immunological studies of mitoxantrone in primary progressive MS.

Mitoxantrone (MX) has demonstrated efficacy in multiple sclerosis (MS), but its immunologic mechanisms of action are poorly understood. Furthermore, no study has examined the immunological effects of MX in primary progressive MS (PPMS). This study investigated the immunological effects of MX therapy in PPMS patients. Lymphocyte phenotypes and chemokine receptor (CCR) expression were evaluated by flow cytometry on fresh PBMC from 20 PPMS patients enrolled in a placebo (PLC)-controlled trial of MX. Longitudinal data were collected at weeks 0, 12, 24, 36 and short-term data (pre-/post-infusion) were collected at weeks 0 and 36. CXCR3, CCR1, CCR2 and CCR5 on CD4 and CD8 T cells and CD14 monocytes were evaluated. MX therapy induced a significant increase in the proportion of CD8 T cells (CD45RO(-)) over 9 months. Expression of several CCR increased following MX treatment. Two of the eight MX-treated patients demonstrated dramatic upregulation (70-76%) of CCR2 on monocytes. These two patients were the only MX-treated patients to demonstrate active inflammation by magnetic resonance imaging (MRI). PLC patients did not show significant changes in CCR expression. MX therapy was not associated with selective loss of CD4, CD8 or CD14 cells 1 month after treatment or over 9 months. These results suggest that MX therapy leads to a longitudinal increase in CD8 T cells and may increase CCR in patients with inflammation on MRI. Overall, MX did not show extensive immune changes in PPMS, although patients with active disease (gadolinium enhancing lesions) may identify a subset of PPMS subjects who respond immunologically to MX therapy. An improved understanding of MX may help identify markers of disease activity and response to therapy.

Effects of sex hormones on costimulatory molecule expression in multiple sclerosis.

Sex hormones play a central role as modulators of immune responses and autoimmune diseases. We hypothesized that suppression of MS disease during pregnancy may be mediated by sex steroid hormones via regulation of costimulatory molecules such as CD40L or CD80/CD86 (B7-1/B7-2). We tested two sex hormones that are implicated in immune suppression during pregnancy: estriol and progesterone. We also examined whether this regulation is gender-specific or disease-related. PBMC from untreated relapsing remitting multiple sclerosis (RR MS) patients and controls were examined for expression of T cell and monocyte costimulatory molecules following mitogen stimulation in the presence or absence of sex hormones. In the absence of hormones, we confirmed that mitogen stimulation induced significantly more CD40L on the surface of CD4(+)T cells in MS patients compared to controls, and we extend these findings by showing there were no gender differences in induction of CD40L. Although supra-physiologic doses of hormones mildly suppressed CD40L expression on activated T cells, in vitro exposure to typical pregnancy-related physiologic doses of estriol or progesterone showed very little or no suppression of CD40L. On monocytes, neither estriol nor progesterone significantly altered the expression of CD80/CD86. These results suggest that physiologic doses of estriol or progesterone cannot alter CD40L on T cells or CD80/CD86 on monocytes sufficiently to explain the improvement observed in MS during pregnancy. Thus, although amelioration of MS and other autoimmune diseases during pregnancy is thought to be due to increased sex hormones, the present results do not support a role for suppression of costimulation via estriol or progesterone.

Epitope mapping in multiple sclerosis using the ELISPOT assay.

Multiple sclerosis (MS) is thought to be an autoimmune disease in which an unknown trigger initiates an immune response against brain proteins. This autoaggressive response causes the breakdown of the myelin sheaths that protect nerve axons, leading to impaired nerve conduction and subsequent neurodegeneration that are characteristic of MS. Many studies have attempted to determine the exact target within the brain. However, there appear to be multiple targets, which may change over time. No single study has examined all targets nor looked at how they can change over the course of the disease and whether these changes are related to the course of disease. We have approached this by using the single-cell resolution capability of the enzyme-linked immunospot assay to examine cytokine reactivity in MS patients in response to a very large set of overlapping peptides that span the two major proteins of myelin: myelin basic protein and proteolipid protein. Our goal was to use the enzyme-linked immunospot assay to perform comprehensive epitope mapping in relapsing-remitting MS patients in a longitudinal study to help define the role of myelin responses in disease progression.