Characterization of the cellular participants in tissue immune responses is crucial to understanding infection, cancer, autoimmunity, allergy, graft rejection and other immunological processes. previous reports indicate that leukocytes in lung vasculature fail to be completely removed by perfusion. several studies suggest that intravascular staining may discriminate between tissue-localized and blood-borne cells in the mouse lung. Here we outline a protocol for the validation and use of intravascular staining to define innate and adaptive immune cells in mice. We demonstrate application of this protocol to leukocyte analyses in many tissues and we describe its use in the contexts of lymphocytic choriomeningitis virus and Mycobacterium tuberculosis infections or solid tumors. Intravascular staining and organ isolation usually takes 5–30 min per mouse, with additional time required for any subsequent leukocyte isolation, staining and analysis. In summary, this simple protocol should help enable interpretable analyses of tissue immune responses.

Although the role of radiation therapy and chemotherapy in primary central nervous system lymphoma (PCNSL) has evolved considerably over the past decade, the application of treatment modalities in the community has not been evaluated. We analyzed the use of chemotherapy, radiation therapy, and associated overall survival, among 9165 HIV-negative PCSNL cases reported to the US National Cancer Database in 2004-2013. During this time, the proportion of patients receiving chemotherapy significantly increased from 65.6% to 78.8% ( for trend <.0001), whereas the proportion receiving radiation therapy decreased from 37.6% to 18.8% ( < .0001). Adjusting for the varying distribution of clinical and sociodemographic characteristics by type of treating facility, the risk of not receiving chemotherapy was significantly lower in academic/research cancer programs compared with community programs (adjusted relative risk, 0.69; 95% confidence interval [CI], 0.62-0.76; < .0001). Furthermore, omission of chemotherapy was associated with increasing age, comorbidities, black race, and indicators of poor socioeconomic status. Overall survival at 3 years was 37.7% (95% CI, 36.6-38.8) and ranged from 14.1% for patients treated with radiation therapy alone to 51.8% for those who received multiagent chemotherapy. There was evidence of improved survival over time ( for trend =.0002). The disparities in application of chemotherapy for PCNSL underscore the need to provide access to expert management for this rare disease and improve safe delivery of systemic treatment in the community setting, where most older patients receive their care.

Even for the most pressing study findings, median publication delays approach 1 year. As publication delays hinder research progress and advancements in clinical care, policies that enable early preprint release or public posting of completed data analysis should be pursued.