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.

Using records from the National Cancer Data Base, we studied overall survival of CD20-negative variants of diffuse large B-cell lymphoma (DLBCL): primary effusion (PEL, N = 228), plasmablastic (PBL, N = 481), ALK-positive large B-cell (ALK + LBLC, N = 15), and human herpesvirus-8-positive DLBCL (HHV8 + DLBCL, N = 77). Three-year survival was 27% for PEL, 40% for PBL, 34% for ALK + LBCL, and 63% for HHV8 + DLBCL. Compared with unspecified DLBCL, and adjusting for clinical characteristics (including the HIV status), survival was significantly worse for PEL (hazard ratio [HR], 1.58; 95% confidence interval [CI], 1.31-1.90), PBL (HR 1.66; 95% CI, 1.41-1.95), and ALK + LBCL (HR, 2.70; 95% CI, 1.27-5.75), but not for HHV8 + DLBCL (HR, 0.89; 95% CI, 0.54-1.45). The HIV status was not an independent prognostic factor in PEL, PBL, or HHV8 + DLBCL. Advanced stage was prognostic for PBL (p = .0002), but not for ALK + LBCL (p = .96), or HHV8 + DLBCL (p = .28). In PEL and PBL survival significantly differed according to primary site. Novel therapeutic approaches are urgently needed for these rare diseases.

Introduction: CD20-negative subtypes of diffuse large B-cell lymphoma (DLBCL) are rare, aggressive malignancies. Recently, cancer registries in the United States (US) have distinguished specific subtypes of CD20-negative DLBCL: since 2004-primary effusion lymphoma (PEL), and since 2010-plasmablastic lymphoma (PBL), ALK+ large B-cell lymphoma (ALK+LBCL), and large B-cell lymphoma arising in HHV8+ multicentric Castleman disease (HHV8+MCD). Our objective was to provide the first large-scale analysis of those lymphomas, focusing on their epidemiology, treatment, and outcomes relative to unspecified DLBCL (DLBCL-NOS). Methods: Using data from the National Cancer Data Base (NCDB), we selected patients with PEL (2004-2013) and with PBL, ALK+LBCL and HHV8+MCD (2010-2013), and compared their characteristics and management with contemporaneous DLBCL-NOS. We analyzed receipt of chemotherapy in subsets defined by histology and HIV status. We then compared overall survival (OS) from diagnosis in a multivariate Cox model stratified by age, sex, race, stage, and HIV status, reporting adjusted hazard ratios (HR) with 95% confidence intervals (CI). Predicted survival of DLBCL-NOS matched by the same variables was calculated from flexible parametric models. Results: Of the 801 identified cases, 228 were PEL, 481 PBL, 77 HHV8+MCD, and 15 ALK+LBCL. We compared them with 68,402 contemporary cases of DLBCL-NOS. Patients with CD20-negative lymphomas were significantly younger on average (Table), more often male, HIV+ (except for ALK+LBCL), and less frequently non-Hispanic whites (NHW). Among patients with PEL, PBL, and HHV8+MCD, those who were HIV+ were significantly younger than HIV-negative (median age, 45 versus 70 years), more often male (88% versus 70%), less often NHW (49% versus 80%), and had more B symptoms (44% versus 21%, all P<.0001), but stage distribution did not significantly differ according to HIV+ status (P=.09). PEL cases with specified primary site were pleural/thoracic in 82%, and peritoneal/abdominal in 18%. Among PBL cases, 21% arose from the head/neck area, 21% from gastrointestinal tract, and 58% from other (or unspecified) areas. Almost all ALK+ cases, and 79% of HHV8+MCD, were nodal, whereas 53% of PBL cases were extranodal. Compared with DLBCL-NOS, the use of chemotherapy was significantly less frequent among patients with PEL, PBL, HHV8+MCD (Table, all P<.0001), but not among those with ALK+LBCL (P=.96). In contrast to DLBCL-NOS, HIV+ status was associated with a higher likelihood of receiving chemotherapy in PEL (60%, versus 46% for HIV-negative, P=.035) and PBL (75% versus 59%, respectively, P=.0003), without a significant difference in HHV8+MCD (59% versus 61%, respectively, P=.82). Compared with matched DLBCL-NOS cases, OS was significantly worse for patients with PEL, PBL or ALK+LBCL, but not for those with HHV8+MCD (Table, Figure). HIV+ status was not associated with worse OS in PEL (P=.22), PBL (P=.39) or HHV8+MCD (P=.56) after adjusting for age difference. Advanced stage was associated with worse OS in PBL (P=.0002), but not in ALK+LBCL (P=.98) or HHV8+MCD (P=.27). Conclusions: This large analysis of CD20-negative DLBCL subtypes using nationwide registry data reveals new information about these rare disorders. While PEL, PBL and HHV8+MCD are strongly associated with HIV infection, ALK+LBCL is not, although it still occurs in younger patients with male predominance. HIV+ patients with PEL, PBL, or HHV8+MCD do not have worse survival outcomes than those who are HIV-negative. Furthermore, despite the association with HIV, survival of HHV8+MCD is overall not worse than that of matched DLBCL-NOS cases. In contrast, the other subtypes have a significantly worse survival compared with DLBCL-NOS, highlighting the unmet need for improved therapeutic approaches. ALK+LBCL is either exceptionally rare, or underdiagnosed in the current practice, and has poor OS, warranting studies of ALK-targeted therapy. Disclosures Castillo: Otsuka: Consultancy; Abbvie: Research Funding; Pharmacyclics: Honoraria; Millennium: Research Funding; Janssen: Honoraria; Biogen: Consultancy. Olszewski:TG Therapeutics: Research Funding; Genentech: Research Funding; Bristol-Myers Squibb: Consultancy.

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Introduction: Adult T cell leukemia lymphoma (ATLL) is a rare T cell neoplasm caused by the human T-lymphotropic virus (HTLV-1) virus. Although there are indolent subtypes it is often a highly aggressive and chemotherapy refractory malignancy. We follow one of the largest cohorts in the United States and in this study, we sought to elucidate the prognostic factors associated with inferior survival. Methods: A retrospective analysis of patients diagnosed with ATLL at Montefiore Medical Center was conducted. Subjects included were censored at last point of contact. Variables collected included age, gender, race, ethnicity, ATLL subtype, white blood cell count (WBC), absolute lymphocyte count (ALC), corrected calcium level, lymphadenopathy (LAD) (two or more non-contiguous sites). Associations between WBC, ALC, corrected calcium level, LAD and median overall survival (mOS) were assessed using the Kaplan-Meier method with log-rank test. A four-point prognostic system was designed assigning one point to each: WBC &gt; 11,000; ALC&gt;4000; Corrected Ca≥10.5 and presence of LAD. Three risk groups were assigned based on the number of risk factors as follows: low (0-1 points), intermediate (2 points) and high (3-4 points) (Table 2). Association between these groups and OS was investigated using the Kaplan-Meier method with log-rank test. Results: A total of 61 ATLL subjects were included in this study (table 1). Hypercalcemia (Ca ≥10.5) was observed in 60.6% of subjects at diagnosis and was associated with inferior mOS (234 days) when compared to calcium &lt; 10.5 (747days) (p=0.046), Figure 1A. WBC &gt;11,000 had a strong association with inferior survival (175 days) compared to patients with a WBC ≤11,000 (666 days) (p= 0.0067) (Figure 1B). ALC &gt; 4000 was also associated with inferior mOS (222 days) compared to ALC ≤4000 (666 days) (p=0.015) (Figure 1C). LAD was associated with mOS (188 days) compared with no LAD (847 days) (p=0.022) (Figure 1D). Based on these observations, we designed a prognostic system (0-4 points) (see above) to risk stratify newly diagnosed ATLL patients into: low (0-1 points), intermediate (2 points) and high (3-4 points) risk (Table 2). We divided our cohort into the above-mentioned risk groups and calculated their mOS. Kaplan Meier analysis (Figure 2) revealed a distinct mOS difference between the groups based on their risk score: Low: 419 days, Intermediate: 234 days and High: 181.5 days (p= 0.0042). Conclusions: We identify hypercalcemia (Ca≥10.5), leukocytosis (WBC&gt; 11,000), lymphocytosis (ALC&gt; 4000) and generalized LAD as poor prognostic factors in newly diagnosed ATLL. Using readily available information from basic laboratory and clinical parameters we propose a prognostic system to identify high risk individuals. Further validation will be needed using larger cohorts of this very rare disease. Disclosures Steidl: Aileron Therapeutics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Stelexis Therapeutics: Consultancy, Current equity holder in private company, Membership on an entity's Board of Directors or advisory committees; Pieris Pharmaceuticals: Consultancy; Bayer Healthcare: Research Funding. Verma:stelexis: Current equity holder in private company; BMS: Consultancy, Research Funding; acceleron: Consultancy, Honoraria; Janssen: Research Funding; Medpacto: Research Funding. Janakiram:Takeda, Fate, Nektar: Research Funding. Shah:Celgene/BMS: Research Funding; Physicians Education Resource: Honoraria.

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